Biotechnology Advances 27 (2009) 4052

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Biotechnology Advances
j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / b i o t e c h a d v

Research review paper

A holistic approach to managing palm oil mill efuent (POME): Biotechnological advances in the sustainable reuse of POME
Ta Yeong Wu, Abdul Wahab Mohammad , Jamaliah Md. Jahim, Nurina Anuar
Scale-up and Downstream Processing Research Group, Department of Chemical and Process Engineering, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor Darul Ehsan, Malaysia

a r t i c l e

i n f o

a b s t r a c t
During the last century, a great deal of research and development as well as applications has been devoted to waste. These include waste minimization and treatment, the environmental assessment of waste, minimization of environmental impact, life cycle assessment and others. The major reason for such huge efforts is that waste generation constitutes one of the major environmental problems where production industries are concerned. Until now, an increasing pressure has been put on nding methods of reusing waste, for instance through cleaner production, thus mirroring rapid changes in environmental policies. The palm oil industry is one of the leading industries in Malaysia with a yearly production of more than 13 million tons of crude palm oil and plantations covering 11% of the Malaysian land area. However, the production of such amounts of crude palm oil result in even larger amounts of palm oil mill efuent (POME), estimated at nearly three times the quantity of crude palm oil. Normally, POME is treated using end-of-pipe processes, but it is worth considering the potential value of POME prior to its treatment through introduction of a cleaner production. It is envisaged that POME can be sustainably reused as a fermentation substrate in the production of various metabolites, fertilizers and animal feeds through biotechnological advances. The present paper thus discusses various technically feasible and economically benecial means of transforming the POME into low or preferably high value added products. 2008 Elsevier Inc. All rights reserved.

Article history: Received 24 December 2007 Received in revised form 19 August 2008 Accepted 21 August 2008 Available online 27 August 2008 Keywords: Cleaner production Palm oil mill efuent (POME) Waste reusability Fermentation substrate Fertilizer Animal feeds

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1.1. A brief glance at palm oil mill efuent (POME) . . . . . . . . . . . 1.2. Cleaner production as a sustainable strategy for POME management 2. POME as a reusable product . . . . . . . . . . . . . . . . . . . . . . . 3. Biotechnological advances in the sustainable reuse of POME. . . . . . . . 3.1. Sustainable reuse of POME as fermentation media . . . . . . . . . 3.1.1. Antibiotics . . . . . . . . . . . . . . . . . . . . . . . . 3.1.2. Bioinsecticides . . . . . . . . . . . . . . . . . . . . . . 3.1.3. Solvents (acetonebutanolethanol: ABE) . . . . . . . . . 3.1.4. Polyhydroxyalkanoates (PHA) . . . . . . . . . . . . . . . 3.1.5. Organic acids . . . . . . . . . . . . . . . . . . . . . . 3.1.6. Enzymes . . . . . . . . . . . . . . . . . . . . . . . . 3.1.7. Hydrogen . . . . . . . . . . . . . . . . . . . . . . . . 3.2. Sustainable reuse of POME as fertilizer . . . . . . . . . . . . . . 3.3. Sustainable reuse of POME as live food for animals and aquacultural 4. Conclusions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . organisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 41 41 42 45 45 45 45 45 46 46 47 47 48 49 50 50

Corresponding author. E-mail addresses: tayeong@hotmail.com (T.Y. Wu), wahabm@vlsi.eng.ukm.my, wahabm@yahoo.com (A.W. Mohammad), jamal@eng.ukm.my (J.M. Jahim), drnurina@eng.ukm.my (N. Anuar). 0734-9750/$ see front matter 2008 Elsevier Inc. All rights reserved. doi:10.1016/j.biotechadv.2008.08.005

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1. Introduction 1.1. A brief glance at palm oil mill efuent (POME) The Malaysian palm oil industry has grown rapidly over the years and Malaysia has become the world's largest producer and exporter of palm oil and its products. In 2003, more than 3.79 million hectares of land were under oil palm cultivation, occupying more than one-third of the total cultivated area in Malaysia and 11% of the total land area (Yusoff and Hansen, 2007). In total, the palm oil industry contributes signicantly towards the country's foreign exchange earnings and the increased standard living among Malaysians. In general, the palm oil milling process can be categorized into a dry and a wet (standard) process. The wet process of palm oil milling is the most common and typical way of extracting palm oil, especially in Malaysia. It is estimated that for each ton of crude palm oil that is produced, 57.5 t of water are required, and more than 50% of this water ends up as palm oil mill efuent (POME) (Ahmad et al., 2003). Raw POME is a colloidal suspension containing 9596% water, 0.60.7% oil and 45% total solids. Included in the total solids are 24% suspended solids, which are mainly constituted of debris from palm fruit mesocarp generated from three main sources, i.e. sterilizer condensate, separator sludge and hydrocyclone wastewater (Borja and Banks, 1994; Khalid and Wan Mustafa, 1992; Ma, 2000). If the untreated efuent is discharged into watercourses, it is certain to cause considerable environmental problems (Davis and Reilly, 1980) due to its high biochemical oxygen demand (25,000 mg/l), chemical oxygen demand (53,630 mg/l), oil and grease (8370 mg/l), total solids (43,635 mg/l) and suspended solids (19,020 mg/l) (Ma, 1995). The palm oil mill industry in Malaysia has thus been identied as the one discharging the largest pollution load into the rivers throughout the country (Hwang et al., 1978). Ponding system is the most conventional method for treating POME (Ma and Ong, 1985; Khalid and Wan Mustafa, 1992) but other processes such as aerobic and anaerobic digestions, physicochemical treatments and membrane ltration may also provide the palm oil industries with a possible insight into the improvement of current POME treatment process. However, the treatment that is based mainly on biological treatments of anaerobic and aerobic systems, is quite inefcient to treat POME, which unfortunately leads to environmental pollution issues (Ahmad et al., 2005). This is because the high BOD loading and low pH of POME, together with the colloidal nature of the suspended solids, render

Fig. 1. The 5 R policy (Olgun et al., 2004).

treatments by conventional methods difcult (Olie and Tjeng, 1972; Stanton, 1974). A detailed cost calculation for Indonesia has also shown that the conventional system of POME treatment, such as the ponding system, is not only the system with the highest environmental pollution and the lowest utilization of renewable resources, but also the system giving rise to the lowest prot (Schuchardt et al., 2005). 1.2. Cleaner production as a sustainable strategy for POME management Currently, end-of-pipe standards imposed through command and control regulations are the basis of environmental legislation (Olgun et al., 2004). However, an international trend promoting pollution prevention through cleaner production, which is based on the 5 R policy (Fig. 1); namely reduction, replacement, reuse, recovery and recycling, is emerging. Within this context, it is proposed herewith that a wastewater management based on the promotion of cleaner production and environmentally sound biotechnologies could be

Table 1 The approximate composition (%) of major constituents, amino acids, fatty acids and minerals in raw POME (adapted from Habib et al., 1997) Major constituents Moisture Crude protein Crude lipid Ash Carbohydrate Nitrogen-free extract Total carotene Total Composition (%) 6.99 12.75 10.21 14.88 29.55 26.39 0.019 100.789 Amino acids Aspartic acid Glutamic acid Serine Glycine Histidine Arginine Threonine Alanine Proline Tyrosine Phenylalanine Valine Methionine Cystine Isoleucine Leucine Lysine Tryptophan Total Composition (%) 9.66 10.88 6.86 9.43 1.43 4.25 2.58 7.70 4.57 3.16 3.20 3.56 6.88 3.37 4.53 4.86 2.66 1.26 90.84 Fatty acids Caprylic acid (8:0) Capric acid (10:0) Lauric acid (12:0) Myristic acid (14:0) Pentadecanoic acid (15:0) Palmitic acid (16:0) Heptadecanoic acid (17:0) 10-Heptadecanoic acid (17:1) Stearic acid (18:0) Oleic acid (18:1n 9) Linoleic acid (18:2n6) Linolenic acid (18:3n3) -linolenic acid (18:3n6) Arachidic acid (20:0) Eicosatrienoic acid (20:3n6) Eicosatetraenoic acid (20:4n6) Eicosapentaenoic acid (20:5n3) Total Composition (%) 2.37 4.29 3.22 12.66 2.21 22.45 1.39 1.12 10.41 14.54 9.53 4.72 0 3.56 2.04 1.12 0.36 95.99 Minerals Fe Zn P Na Mg Mn K Ca Co Cr Cu Ni S Se Si Sn Al B Mo As V Pb Cd Composition (g/g dry weight) 11.08 17.58 14377.38 94.57 911.95 38.81 8951.55 1650.09 2.40 4.02 10.76 1.31 13.32 12.32 10.50 2.30 16.60 7.60 6.45 9.09 0.12 5.15 0.44

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T.Y. Wu et al. / Biotechnology Advances 27 (2009) 4052

included as a part of the POME management in Malaysia in order to attain a sustainable development. Such a strategy could take advantage of the current international interest in promoting cleaner production as the driving force of a new and sustainable industrial development style. This review paper thus describes various technically feasible means of transforming the POME into different addedvalue products through cleaner production and environmentally sound biotechnologies for enabling the balance between environmental protection and a sustainable reuse of bioresources found in the POME. 2. POME as a reusable product The high compositions and concentrations of carbohydrate, protein, nitrogenous compounds, lipids and minerals in POME (Hwang et al., 1978; Phang, 1990; Habib et al., 1997) render it possible to reuse the efuent for biotechnological means (Table 1). Chemical analysis indicates that the rod-like particle fraction (Fig. 2) available in the POME corresponds to carbohydrate in nature. After treatment with phenol/sulfuric acid, the fraction shows a maximum absorption at approximately 480 nm (Fig. 3), indicating the possible presence of pentose, a building unit for insoluble carbohydrates (Ho and Tan, 1983). The presence of pentose in POME has been reported previously (Hwang et al., 1978) and its most likely sources are the cell walls. Water-soluble carbohydrates, in terms of glucose, reducing sugars and pectin, are also found to be present in the soluble fraction of POME. However, the low concentrations of total soluble carbohydrate (0.390 g/100 ml POME) may restrict the usefulness of the soluble fraction of POME as a possible feedstock for substrate conversion via direct single-cell protein production (Ho et al., 1984). Preliminary investigations on enzymatically hydrolyzed substrates from POME have indeed demonstrated the possibility of such substrates supporting the growth of Candida tropicalis (Wang et al., 1981). On the other hand, Barker and Worgan (1981) noted that unhydrolyzed POME

Fig. 3. Absorption curves for the rod-like particles after treatment with phenol/ sulphuric acid and for a number of other simple sugars (Ho and Tan, 1983).

could support good growth of Aspergillus oryzae in the presence of an added inorganic nitrogen source. Their results also revealed that celluloses, polyphenols and nitrogenous compounds were the least biodegradable of the substrate constituents. This lends further support to the view that a proper hydrolysis step is essential in obtaining an optimal level of readily biodegradable sugars from the plant cell materials for a meaningful microbial bioconversion.

Fig. 2. Centrifugal fractionation of POME (Ho and Tan, 1983).

Table 2 Various products or metabolites produced in bioprocesses during the reuse of POME or its derivatives as substrates Product Penicillin Microorganism Fermentation medium based on POME 50% (v/v) concentrated POME + KH2PO4 + (NH4)2SO4 50% (v/v) concentrated POME + KH2PO4 + (NH4)2SO4 + NH4 lactate Raw POME Fermentation conditions 300 rpm, 30 C, 2% (v/v) inoculum, ask fermentation 300 rpm, 30 C, 2% (v/v) inoculum, ask fermentation 150 rpm, 30 C, initial pH = 6.9, ask fermentation 150 rpm, 30 C, initial pH = 6.9, ask fermentation 30 C, 10% (v/v) inoculum, initial pH = 5.8, ask fermentation 30 C, 10% (v/v) inoculum, initial pH = 5.8, ask fermentation 200 rpm, 37 C, 10% (v/v) inoculum, pH was switched from 6.2 to 5.5 at 9 h, 2 g/l/h glucose feeding started at 10 h, bioreactor fermentation 35 C, 10% (v/v) inoculum, initial pH = 5.8, ask fermentation Not clearly stated Fermentation timea (h) 72 72 72 72 Not clearly stated Not clearly stated 35 (for A) 45 (for B) 48 (for ABE) 36 (for ABE) Maximum production 602 U/ml 715 U/ml 7.0 10 spores/ml 2.0 10 spores/ml A = 0.45 g/l, B = 2.47 g/l, E = 0.49 g/l A = 0.33 g/l, B = 2.30 g/l, E = 0.43 g/l A = 5.78 g/l, B = 6.78 g/l
8 3

Reference Suwandi (1991) Suwandi (1991) Suwandi (1991) Suwandi (1991) Mun et al. (1995) Mun et al. (1995) Somrutai et al. (1996) Kalil et al. (2003), Pang et al. (2004) Kalil et al. (2003)

Penicillium chrysogenum FR2284 Penicillin Penicillium chrysogenum FR2284 Bioinsecticide Bacillus thuringiensis H-14

Bioinsecticide Bacillus thuringiensis H-14 ABE ABE ABE

1% (w/v) concentrated POME in powder form Clostridium saccharoperbutylacetonicum Separator sludge N1-4 (ATCC 13564) Clostridium saccharoperbutylacetonicum Sterilized condensate N1-4 (ATCC 13564) Model medium for raw POME Clostridium aurantibutyricum ATCC 17777 90% (v/v) particulate fraction of raw POME Raw POME

ABE ABE

ABE ABE

Clostridium acetobutylicum NCIMB 13357 Immobilized Clostridium saccharoperbutylacetonicum N1-4 Clostridium acetobutylicum NCIMB 13357 Clostridium acetobutylicum NCIMB 13357 Clostridium acetobutylicum NCIMB 13357 Clostridium acetobutylicum NCIMB 13357 Rhodobacter sphaeroides IFO 12203 Rhodobacter sphaeroides IFO 12203 Alcaligenes eutrophus H16 (ATCC 17699) Ralstonia eutropha ATCC 17699 Mixed cultures

A = 1.97 g/l, B = 1.74 g/l, E = 0.3 g/l ABE = 3.8 g/l

T.Y. Wu et al. / Biotechnology Advances 27 (2009) 4052

Particulate fraction of raw POME Particulate fraction of raw POME

ABE

Particulate fraction of raw POME

ABE

Particulate fraction of raw POME

PHA

PHA PHA

PHA

PHA

Organic acids Organic acids Citric acid Citric acid

Mixed cultures Mixed cultures Aspergillus (A103) Aspergillus niger (A103)

Itaconic acid

Aspergillus terreus IMI 282743

30 C, pH was controlled at 7, photobioreactor fermentation 400 rpm with an aeration rate of 0.75 l/min, Standard medium with feeding of 30 C, pH was controlled at 7, stirred tank acetic acid obtained from bioreactor fermentation anaerobically digested POME Concentrated organic acids from the anaerobically digested 400 rpm with an aeration rate of 0.75 l/min, POME (100 g/l of total acids with acetic:propionic = 3:1) 30 C, pH was controlled at 7, bioreactor fermentation 1,000 rpm with an aeration rate of 1.5 l/min, High concentration of POME with 30 C, pH was controlled at 7, sequencing batch 490 COD/N ratio (g COD/g N) and reactor 160 COD/P ratio (g COD/g P) fermentation POME + palm oil sludge 30 C, pH was controlled at 7, bioreactor fermentation POME + palm oil sludge in the ratio of 1:1 300 rpm, 30 C, pH was controlled at 5, stirred tank bioreactor fermentation 1% (w/w) POME + 2% (w/w) wheat our 150 rpm, 2730 C, initial pH = 3, inoculum size of 2% (106 spores/ml), ask fermentation 150 rpm, 32 C, initial pH = 5, inoculum size of 2% (w/w) POME + 4% (w/w) 2% (106 spores/ml), wheat our + 4% (w/w) glucose with no ask fermentation added ammonium nitrate (optimized medium) Retentate of POME 300 rpm, 35 C, 5% (v/v) inoculum, ask fermentation

Synthetic waste based on organic acids proles obtained during POME treatment Anaerobically digested POME

35 C, 10% (v/v) inoculum, initial pH = 6, ask fermentation Oscillated at 0.45 Hz, 35 C, 10% (v/v) inoculum, initial pH = 6, oscillatory ow bioreactor fermentation Oscillated at 0.78 Hz, 35 C, 10% (v/v) inoculum, initial pH = 5.8, oscillatory ow bioreactor fermentation 250 rpm, 35 C, 10% (v/v) inoculum, initial pH = 5.8, stirred tank bioreactor fermentation 30 C, pH was controlled at 7, photobioreactor fermentation

30 (for A) 24 (for E) 42 (for A) 30 (for E) 48 (for ABE)

A = 1.2 g/l, B = 0 g/l, E = 0.5 g/l A = 0.7 g/l, B = 0 g/l, E = 0.6 g/l A = 0.05 g/l, B = 1.54 g/l, E = 0 g/l A = 0.13 g/l, B = 0.50 g/l, E = 0.24 g/l 4 g/l

Takriff et al. (2005) Takriff et al. (2005)

Masngut et al. (2006, 2007) Masngut et al. (2007)

60 (for ABE)

200

Hassan et al. (1996)

Dilution rate = 0.024 d 1 17

N2 g/l 1.8 g/l

Hassan et al. (1997b) Hassan et al. (1997c)

65

6.25 g/l

Hassan et al. (2002)

Not clearly stated

24.24 g/l

Md Din et al. (2006b)

24 84 48 168

7.8 g/l 1014 g/l 0.28 g/l 5.2 g/l

Hassan et al. (1996) Yee et al. (2003) Jamal et al. (2005) Alam et al. (2008)

120

0.079 g/l

Wu et al. (2005) 43 (continued on next page)

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Table 2 (continued) Product Cellulase (CMCase) Cellulase (CMCase) Cellulase (CMCase) Cellulase (CMCase) Cellulase (CMCase) Microorganism Aspergillus niger ATCC 6275 Fermentation medium based on POME POME with added nutrients + 0.6 g/l NH4NO3 50% (v/v) raw POME 50% (v/v) raw POME 50% (v/v) raw POME 50% (v/v) raw POME Fermentation conditions 200 rpm, room temperature, inoculum size was 7.5 105 spores/ml, ask fermentation 300 rpm with an aeration rate of 4 l/min, 30 C, 10% (v/v) inoculum, bioreactor fermentation 300 rpm with an aeration rate of 4 l/min, 30 C, 10% (v/v) inoculum, bioreactor fermentation 300 rpm with an aeration rate of 4 l/min, 30 C, 10% (v/v) inoculum, bioreactor fermentation 300 rpm with an aeration rate of 4 l/min, 45 C, 10% (v/v) inoculum, initial pH 5.5 was uncontrolled until 6.0, bioreactor fermentation 250 rpm, 30 C, 3% (v/v) inoculum, initial pH = 4, ask fermentation Not clearly stated 200 rpm, 55 C, 5% (v/v) inoculum, initial pH = 7, ask fermentation 250 rpm, 40 C, 1% (v/v) inoculum, initial pH = 6, ask fermentation Not clearly stated 200 rpm, 37 C, 10% (v/v) inoculum, constant pH at 6.8, bioreactor fermentation 200 rpm, room temperature, inoculum size was 7.5 x 105 spores/ml, ask fermentation Aeration rate of 2.5 l/min, 30 C, 10% (v/v) inoculum, bioreactor fermentation 200 rpm, 55 C, 5% (v/v) inoculum, initial pH = 7, ask fermentation 250 rpm, 37 C, 5% (v/v) inoculum, ask fermentation 60 C, pH was uncontrolled, bioreactor fermentation 200 rpm, 60 C, pH was controlled at 5.5, bioreactor fermentation pH was controlled at 5, anaerobic contact lter fermentation Fermentation timea (h) 72 Maximum production 1.09 U/ml Reference Prasertsan et al. (1997) Mashitah (2002) Mashitah (2002) Mashitah (2002) Mashitah (2002) et al. et al. et al. et al.

Aspergillus niger Trichoderma harzianum Mixed culture (1:1) of Aspergillus niger and Trichoderma harzianum Myceliophthora thermophila

51 45 45 24

1.040 U/ml 1.227 U/ml 0.656 U/ml 3.495 U/ml

Cellulase (FPase) Cellulase (FPase) Cellulase Lignin peroxidase Lignin peroxidase Lipase

Trichoderma harzianum Penicillium (P1-EFB) Isolate SO1 Phanerochaete chrysosporium Penicillium (P1-EFB) Clostridium aurantibutyricum ATCC 17777 Aspergillus niger ATCC 6275

2% (w/v) particulate fraction of POME + 3% (w/v) wheat our 1% (w/w) POME sludge 10% (v/v) supernatant of POME + another nine different types of supporting nutrients 2% particulate fraction of POME + 1% wheat our 1% (w/w) POME sludge Model medium for raw POME

96 144 48 96 144 20

13.44 U/ml 33 U/ml 12.11 U/ml 3.373 U/ml 5.038 U/ml 0.4 U/ml

Alam et al. (2006a) Chowdhury et al. (2006) Laohaprapanon et al. (2007) Alam et al. (2006b) Chowdhury et al. (2006) Somrutai et al. (1996) Prasertsan et al. (1997) Cheng (2006) Laohaprapanon et al. (2007) Wu et al. (2006a) Morimoto et al. (2004) Atif et al. (2005) Vijayaraghavan and Ahmad (2006)

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Xylanase

POME with added nutrients + 0.6 g/l NH4NO3 Raw POME 10% (v/v) supernatant of POME + another nine different types of supporting nutrients 75% (v/v) retentate of POME 1% glucose + 0.2% yeast extract + 0.018% magnesium chloride hexahydrate + 0.5% (w/v) POME sludge Raw POME + 2.5% (w/v) POME sludge Raw POME

96

22.77 U/ml

Xylanase Xylanase

Aspergillus terreus SUK-1 Isolate SO1

96 96

0.448 U/ml 50.98 U/ml

Protease Hydrogen

Aspergillus terreus IMI 282743 Mixed culture from POME sludge

96 Not clearly stated

129 U/ml 23.82 mmol H2/ (1-medium) 4,708 ml H2/(l-medium) 0.42 l biogas/g CODdestroyed with 57% hydrogen content 4.4 l H2/(1-medium) per day 6.1 l H2/(1-medium) per day 6.33 H2/(1-medium)

Hydrogen Hydrogen

Mixed culture from POME sludge Mixed culture (isolated from cow dung)

38 168

Hydrogen

Thermophilic microora

Raw POME

Hydrogen

Thermophilic microora

Raw POME + Fe2+ + peptone + Na2 HPO42H2O Raw POME + 257 mg Fe2+/l + a C/N ratio of 74 + a C/P ratio of 559 + 0.3 g NaHCO3 (optimized medium)

Hydrogen

Thermoanaerobacterium-rich sludge

200 rpm, 60 C, pH was controlled at 5.5, anaerobic sequencing batch reactor fermentation 200 rpm, 60 C, pH was controlled at 5.5, anaerobic sequencing batch reactor fermentation 60 C, initial pH = 5.5, 10 ml seed sludge inoculum, serum bottle fermentation

At steady state (during day 2228) At steady state (during day 2228) 48

O-Thong et al. (2007)

O-Thong et al. (2007)

O-Thong et al. (2008a)

A = Acetone, B = Butanol, E = Ethanol. PHA = Polyhydroxyalkanoates. a The fermentation time is the time required to reach a maximum product concentration.

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Nitrogen is originally present in POME in the form of organic (protein) nitrogen and as time progresses the organic nitrogen is gradually converted to ammoniacal nitrogen with a molecular weight of 1735 kg/ kmol (Chow, 1991). According to Ho and Tan (1988), the nutrient balance in terms of the average ratio of BOD:N:P for raw POME is 100:4:0.3. Muhrizal et al. (2006) reported that POME is characterized by a low C:N ratio (C:NPOME = 6.54) as compared to sawdust (C:Nsawdust = 185.74), purun (C:Npurun =88.32) and peat (C:Npeat = 50.31). The amino acids of POME and their approximate compositions (%) are shown in Table 1. The major portion of proteins in POME is tightly associated with its insoluble part (Ho et al.,1984). This may perhaps account for the low digestibility of proteins in POME found by Devendra et al. (1981). Only N, P, K, Mg and Ca are consistently present in relatively large amounts in the POME (Ho et al., 1984; Habib et al., 1997; Muhrizal et al., 2006) (Table 1). Muhrizal et al. (2006) also reported that POME has a high content of Al as compared to chicken manure and composted sawdust. It would thus seem that the probable usefulness of POME as fertilizer or animal feed substitute, in terms of providing sufcient mineral requirements, depends mainly on the soluble fraction of POME. Toxic metals, such as Pb, can also be found in POME (Habib et al., 1997) but their concentrations are usually below sublethal levels (N17.5 g/g) (James et al., 1996). POME is thus not toxic for plants and animals. Pb is found in POME due to contamination from plastic and metal pipes, tanks and containers where Pb is widely employed in paints and glazing materials (James et al., 1996). The reusability of POME especially as animal feed is arguable because another important factor that needs to be taken into consideration is its lignin content. Lignin is a recalcitrant, waterinsoluble organic chemical (Palm and Rowland, 1997). A high content of lignin in organic materials is known to slow down their decomposition (Klepper et al., 1990; Tian et al., 1992). High lignin contents (0.412 g/100 ml POME) also present major barriers with regard to digestion of roughage by-product feeds from eld and tree crops by both ruminants and non-ruminants (Ho et al., 1984). Lignin is known to be highly resistant to chemical degradation thus giving rise to its low digestibility. This has been clearly illustrated by the work of Devendra et al. (1981) on the use of POME, in both raw and dehydrated forms, as animal feed. Therefore, to allow the POME to be reused effectively as animal feed, Webb et al. (1975) have suggested that efuent products can be nutritionally improved by reducing the ash and ber content as well as increasing the protein content on par with imported feed meal. The composition of raw POME with regard to saturated and unsaturated fatty acids other than polyunsaturated fatty acids is shown in Table 1. Here, myristic acid, palmitic acid, stearic acid and oleic acid are present in compositions higher than other saturated fatty acids. The 20 carbon chained polyunsaturated fatty acids such as eicosatrienoic acid (20:3n6), eicosatetraenoic acid (20:4n6) and eicosapentaenoic acid (20:5n3), which are available in raw POME, are essential for the proper development of marine sh, shrimp larvae and fry (Oka et al., 1982). According to Habib et al. (1997), these substances have been accumulated and synthesized in higher amounts for chironomid larvae grown in POME than those grown in algal culture. 3. Biotechnological advances in the sustainable reuse of POME Chemical analyses of POME with respect to its proximate composition have been carried out (Wood, 1977; Hwang et al., 1978; Ho et al., 1984; Habib et al.,1997), and these analyses are of vital importance in the understanding of the properties of POME in relation to formulating waste-utilization programs and efcient wastewater management processes. This is particularly true in view of the increasing emphasis placed on the zero discharge concept and innovative technology for sustainable development. An important case is the production of biogas and other metabolites by fermentation processes. Of no less importance is the possibility of recovering bioresources from POME, or its conversion into useful substitutes for animal feed and fertilizer.

3.1. Sustainable reuse of POME as fermentation media The possibility of reusing POME as fermentation media is largely due to the fact that POME contains high concentrations of carbohydrate, protein, nitrogenous compounds, lipids and minerals (Hwang et al., 1978; Phang, 1990; Habib et al., 1997). Suwandi, (1991) and Wu et al., (2006b) pointed out the possibility of recovering and concentrating the available bioresources in POME by an ultraltration process in order for the concentrated bioresources to be reused more effectively as fermentation media. According to Wu et al. (2007), POME and its derivatives have been exploited as fermentation media to produce various products/metabolites such as antibiotics, bioinsecticides, solvents, polyhydroxyalkanoates, organic acids as well as enzymes to varying degrees of success. The hydrogen production from POME during anaerobic treatment has also been intensively studied (Atif et al., 2005; Vijayaraghavan and Ahmad, 2006) since the generated hydrogen and its combustion products do not count as green house gases (Koroneos et al., 2004). However, it has been reported that POME also contains certain powerful water-soluble antioxidants, phenolic acids and avonoids (Wattanapenpaiboon and Wahlqvist, 2003) that may inhibit the growth development in microorganisms (Lin et al., 2005; Uzel et al., 2005). Table 2 displays the various products or metabolites produced in bioprocesses by reusing POME or its derivatives as substrates. 3.1.1. Antibiotics Studies on ultraltered POME concentrates or retentates as growth media for Penicillium chrysogenum in the production of antibiotics have been conducted some time ago (Suwandi and Mohammad, 1984; Suwandi, 1991). It was found that a supplementary nitrogen source was required in the ratio C:N = 20:1 to produce a maximum mass of P. chrysogenum (Suwandi and Mohammad, 1984). However, no attempts were made to assay the penicillin produced during the process of incubation. Suwandi (1991) later determined the effect of the POME concentration and the addition of chemicals on the production of penicillin in cultured broths for periods of 4 days. He found that smaller amounts of penicillin were produced in POME as compared to in Deo and Gaucher's (1984) standard medium. Suwandi (1991) argued that the low production of penicillin was due to the lower concentration of carbohydrates (23 g/l) in POME as compared to in the standard medium, which contained 35 g/l carbohydrates. The addition of ammonium lactate to POME stimulated the production of penicillin. 3.1.2. Bioinsecticides Nor and Mahadi (1986) as well as Suwandi (1991) embarked on a research topic related to the use of ultraltered POME concentrate or retentate as a medium for Bacillus thuringiensis to produce bioinsecticide for mosquito control. Suwandi (1991) observed that a medium containing 1% (w/v) retentate in powder form was as good as the standard medium of glucose yeast extract salts in terms of spore production by B. thuringiensis. The ability of the retentate to support and stimulate the growth of B. thuringiensis could be attributed to the proper ratio of carbon and nitrogen as well as to sufcient levels of ions (such as Mg, Ca, Mn, etc.) in the POME. 3.1.3. Solvents (acetonebutanolethanol: ABE) Separator sludge and sterilized condensate from POME have been tested for their suitability to be reused as fermentation media for ABE production by Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564) (Mun et al., 1995). Separator sludge was found to be the better medium between the two for supporting the production of ABE as no mineral supplements were required. The enzymes produced by C. saccharoperbutylacetonicum N1-4 (ATCC 13564) were sufcient to hydrolyze the mixed carbohydrates and celluloses found in the separator sludge. Hipolito et al. (2007) later reconrmed that enzymatic hydrolysates of separator sludge could be used as media for inoculum

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development since the cultures inoculated with C. saccharoperbutylacetonicum N1-4 (ATCC 13564) using the sludge hydrolysate produced the same concentration of butanol as compared to in a potato glucose medium, whereas the corresponding ethanol production was increased by over 100%. The authors concluded that the enzymatic hydrolysates of separator sludge could serve as growth and ABE fermentation media as well as a source of nitrogen and trace elements. Somrutai et al. (1996) investigated the possibility of acetonebutanol fermentation by C. aurantibutyricum ATCC 17777 in a model medium for raw POME. They found that by decreasing the pH from 6.2 to 5.5 at 9 h and starting an hourly glucose feeding (2 g/l) at 10 h, it was possible to obtain 30% of the oil hydrolysis as well as a production of 5.78 g/l acetone and 6.78 g/l butanol. Kalil et al. (2003) studied the direct use of raw POME as a fermentation medium for ABE production by C. acetobutylicum NCIMB 13357 and immobilized C. saccharoperbutylacetonicum N1-4 in a batch culture system. It was found that C. acetobutylicum NCIMB 13357 produced the highest total ABE in 90% (v/v) particulate fraction of raw POME after 48 h of fermentation at an initial pH of 5.8 while immobilized cells of C. saccharoperbutylacetonicum N1-4 could be reused for at least 5 times in 100% (v/v) particulate fraction of raw POME without losing their performance (Kalil et al., 2003). Similar results were also obtained by Pang et al. (2004) with the addition of hydrogen production up to 28.5 ml. An oscillatory ow bioreactor was used to enhance the production of ABE in the raw POME (Takriff et al., 2005; Masngut et al., 2006, 2007), and initial results showed that by using a particulate fraction of raw POME as the fermentation medium, C. acetobutylicum NCIMB 13357 could produce 31% higher concentrations of ABE, especially acetone, in ask as compared to an oscillatory ow bioreactor (Takriff et al., 2005). On the other hand, Masngut et al. (2006, 2007) found that C. acetobutylicum NCIMB 13357 could only produce higher amounts of butanol in shorter periods of time when particulate fractions of raw POME were used in an oscillatory ow bioreactor as opposed to with a reinforced clostridial medium in a stirred tank bioreactor. Pang et al. (2004) also claimed that the concentration of ABE produced by C. acetobutylicum NCIMB 13357 in a particulate fraction of raw POME was 20-fold that obtained in the reinforced clostridial medium. 3.1.4. Polyhydroxyalkanoates (PHA) Over 40% of the total polyhydroxyalkanoates (PHA) production cost is estimated to account for the raw materials of the overall process and more than 70% of this cost is attributed to the carbon source (Lee et al., 1999). POME can be considered as an alternative, nocost reusable substrate for PHA production. According to Hassan et al., (1997a), with a content of 50% PHA in the dried cells and 2% dissolved in the chloroform, the calculated minimum cost for obtaining PHA from POME is below 2 US$/kg. By increasing the PHA content in the cell from 50% to 80%, the unit cost of PHA could be slightly reduced; whereas an increase in the amount of PHA dissolved in chloroform from 2% to 5% would result in a remarkable reduction of the PHA cost to less than 1 US$/kg (Hassan et al., 1997a). Nevertheless, POME is usually presented in complicated forms that cannot be directly reused by PHA-producing species such as Ralstonia eutropha, a representative bacterium for PHA synthesis (Salim et al., 2006). It was proposed that an anaerobic treatment of POME could be coupled with PHA production using photosynthetic bacteria to reduce PHA production costs (Hassan et al., 1996, 1997b). According to Hassan et al. (1996), it was critical to maintain the pH at 7 in the anaerobic treatment of POME by sludge in the rst stage of the process, in order for only acetic and propionic acid to be produced and not formic acid and biogas. With increasing concentrations of formic acid (for a pH maintained below 4), the PHA yield and content in Rhodobacter sphaeroides IFO 12203 dropped from 0.50 g/g and 67% to 0.21 g/g and 18%, respectively. Hassan et al. (1997b) later found that the presence of sludge in the anaerobically treated POME inhibited PHA accumulation by R. sphaeroides IFO 12203. This was attributed to the PHA being

produced in a POME without sludge as opposed to a treated POME with sludge. A low concentration of ammonium would accelerate the PHA production in a synthetic waste with an organic acid prole, which was observed during POME treatment (Hassan et al., 1996). However, Hassan et al. (1997b) found that addition of ammonium and phosphate to anaerobically treated POME was required to maintain the cell activity and production of PHA since neither ammonium nor phosphate was present in the anaerobically treated POME. In total, the organic acid concentrations obtained from anaerobically treated POME were too low (Hassan et al., 1996) for it to be reused as raw material in the production of PHA on an industrial scale. The underlying reason was that this would require a production reactor with a much larger size than that of a reactor for normal bioplastic production. Therefore, Hassan et al. (1997c) used an anion exchange resin to separate and concentrate the acetic acid from the anaerobically treated POME so that the concentrated acetic acid could be reused as a substrate in the fed-batch production of PHA by Alcaligenes eutrophus H16 (ATCC 17699). They found that the PHA content was comparable to that of the batch and fed-batch PHA production by Alcaligenes (around 18% to 76%), but the overall PHA productivity obtained was less than the 0.53 g PHAs/l h obtained by other researchers (Suzuki et al., 1986; Ishizaki and Tanaka, 1991; Lee et al., 1993; Yamane et al., 1996). This might be due to the low cell concentration when concentrated acetic acid separated from POME was incorporated into the standard medium. Hassan et al. (2002) showed that organic acids in the anaerobically digested POME could be concentrated by evaporation for use as substrates in the fed-batch non-sterile PHA fermentation system using R. eutropha ATCC 17699. Although the proposed overall zero emission system appeared to be practical, major drawbacks were found, including the rather low yield and productivity of PHA by R. eutropha when the concentrated organic acids from POME were used as compared to synthetic organic acids. This could be due to the high presence of ammonium (1.5 g/l) or other compounds in the anaerobically digested POME concentrate (Hassan et al., 2002). According to Md Din et al. (2006a,b), it was possible to use mixed cultures to produce PHA in POME since most prokaryotes are capable of PHA production (Chua et al., 2003). Md Din et al. (2006a) noted that by using mixed cultures and POME, different types of PHA-constituents could be obtained. The harvesting of these PHA-constituents was more reliable for use as biodegradable plastics materials as opposed to a single PHA-constituent. Md Din et al. (2006b) maintained a type of mixed culture in a sequencing batch reactor, and a high concentration of POME was proposed for this system in order to generate autotrophic rather than heterotrophic bacteria in the production of PHA. However, the average PHA production by using POME could only reach 44% of the cells' dry weight, indicating that an optimization of the PHA sludge content must be carried out by varying the oxygen rate, feeding regime or transient conditions. 3.1.5. Organic acids It is a well-known fact that a variety of organic acids are produced as intermediates during the anaerobic treatment of biological wastes (Kotz et al., 1969; Zeikus, 1980; Archer, 1983). As mentioned earlier, POME could be put to sustainable use for organic acid production, whereby the latter could be utilized as raw material for PHA production (Hassan et al., 1996, 1997b,c, 2002). According to Hassan et al. (1996), the conversion to organic acids from the BOD sources in POME by R. sphaeroides IFO 12203 was more than 70%, and acetic acid and formic acid were the predominant substances at higher and lower pH, respectively. Yee et al. (2003) showed that by incorporating a sludge recycling system with the freezingthawing method in the anaerobic treatment of POME, the retention time could be lowered to 3.5 days without affecting the organic acid production. Moreover, Yee et al. (2003) found that the effect of freezing and thawing produced concentrated viable and ruptured cells in recycled sludge with a total

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nitrogen source of 45 g/l as a result of the ruptured cells releasing the nitrogen sources that were required to support the growth of higher cell densities. Some organic acids, such as citric acid (Jamal et al., 2005; Alam et al., 2008) and itaconic acid (Wu et al., 2005), could be produced under aerobic conditions using POME as the substrate. Jamal et al. (2005) screened potential microorganisms for citric acid production and found that Aspergillus (A103) produced the highest concentration of citric acid after 2 days of fermentation by using POME and wheat our as medium. Alam et al. (2008) later found that higher production of citric acid could be obtained with longer time of fermentation (up to 7 days of fermentation) and addition of co-substrates (glucose and wheat our) into POME. Wu et al. (2005) employed raw POME and its derivatives to produce itaconic acid by using Aspergillus terreus IMI 282743. However, only little itaconic acid could be obtained. It was postulated that this low production was mainly due to the wild and unsuitable strain of A. terreus IMI 282743. 3.1.6. Enzymes The particulates in the POME, comprised mainly of plant cell debris and fragments, are entirely organic in nature as indicated by the very low ash contents (Ho et al., 1984). The availability of such particulates may provide a potential substrate for production of cellulase (Prasertsan et al., 1997; Mashitah et al., 2002; Alam et al., 2006a; Chowdhury et al., 2006; Laohaprapanon et al., 2007), xylanase (Prasertsan et al., 1997; Cheng, 2006; Laohaprapanon et al., 2007) and lignin peroxidase (Alam et al., 2006b; Chowdhury et al., 2006) in POME. The optimization of cellulase (CMCase) and xylanase productions from Aspergillus niger ATCC 6275 was investigated under both submerged and solid state fermentation (Prasertsan et al., 1997). Prasertsan et al. (1997) found that the addition of 0.6 g/l NH4NO3 into the POME during submerged fermentation increased the maximum production of xylanase and CMCase with up to 156% and 43%, respectively. Prasertsan et al. (1997) also revealed that the enzyme production was lower in POME when the fermentation process was conducted in a fermenter, which might be due to the destruction of mycelium by shearing forces, thus causing the cessation of enzyme synthesis and the induction of enzyme inhibition (Wase et al., 1985). Contrarily, Cheng (2006) reported on 107% higher activity of xylanase obtained by A. terreus SUK-1 in the fermenter as compared to the shake ask when raw POME was used as the substrate. However, Cheng (2006) also found a 31% lower activity of xylanase with the raw POME as opposed to the Mandels medium (Mandels, 1974). Mashitah et al. (2002) used axenic and mixed cultures of mesophilic and thermophilic fungi to produce cellulase in the POME, in which case they found that the axenic culture of Trichoderma harzianum was superior to the mixed culture of A. niger and T. harzianum in terms of CMCase and exoglucanase production. This low production of cellulase in the mixed culture was believed to be due to the competition of nutrient consumptions between the fungi in the POME. Moreover, the genus Aspergillus is known to release other metabolites such as endotoxins (Debeaupuis and Lafont, 1978) and proteases (Aunstrup, 1974), which may inhibit the activity of cellulase. Mashitah et al. (2002) also noted that the thermophilic fungi, namely Myceliophthora thermophila grew well and produced a higher amount of cellulase in POME as compared to the mesophilic fungus. Alam et al. (2006a) utilized a combination of POME and wheat our as the substrate in the optimization process for maximizing cellulase production by T. harzianum. They found that the linear effect of agitation was not highly signicant for cellulase production but this parameter should not be totally overruled because of its interactive effect with wheat our. Six out of twenty strains of Penicillium were isolated from four different sources of POME sludge and selected for the production of cellulase and lignin peroxidase in the POME (Chowdhury et al., 2006).

The results revealed Penicillium (P1-EFB), which was isolated from empty fruit bunches, displayed the best potential strain for biodegradation in liquid state bioconversion of POME at pH 7.3. The whiterot fungus Phanerochaete chrysosporium was used for lignin peroxidase production with a combination of POME and wheat our as the substrate (Alam et al., 2006b). It was observed that, although wheat our could be used as an additional carbon source to enhance the initial growth of P. chrysosporium, more than 2% wheat our was expected to further decrease the enzyme activity of the lignin peroxidase. Laohaprapanon et al. (2007) found that the isolate SO1, which was isolated from a soil near to the rst anaerobic pond of palm oil mill, produced the highest activity of cellulase and xylanase in comparison with other isolate of microorganisms. Isolate SO1 was an aerobic, Gram-positive, rod-shaped and thermo-tolerant microorganism, which was able to reduce the oil content in the sediment of POME up to 85.32%. In view of the fact that POME contains considerable amounts of oil and grease, it is possible to reuse POME as a substrate in lipase production and isolate oil-degrading microorganisms from efuent treatment ponds of POME. Somrutai et al. (1996) found that Clostridium aurantibutyricum ATCC 17777 was able to produce lipase in the model medium for POME in which high rate of oil hydrolysis (46.0%) was observed at pH of 6.8. Razak et al. (1997) were able to isolate thermophilic fungi, namely Rhizopus oryzae and Rhizopus rhizopodiformis, from POME. They found that the fungi could produce remarkable amounts of extracellular lipases in the dened medium. Furthermore, the isolated R. oryzae from POME could produce membrane-bound lipases that were active in both acidic and alkaline conditions as opposed to extracellular lipases from the same fungi (Razak et al., 1999). Apart from thermophilic fungi, thermophilic bacteria such as Geobacillus sp. strain T1 (Leow et al., 2004; Rahman et al., 2007) and Bacillus sp. strain 42 (Eltaweel et al., 2005), which were isolated from POME, were found to be lipase producers too. The concentrated bioresources from POME or its retentate (Wu et al., 2006b, 2007) could be reused as effective substrates to produce protease (Wu et al., 2006a,b). According to Wu et al. (2006a), a wildtype strain of A. terreus IMI 282743 produced a maximum protease activity in the medium containing 75% (v/v) POME retentate as the sole carbon and nitrogen source without addition of extra nutrients or adjustment of the initial pH. However, using pure retentate, i.e. without slightly diluting it, was not recommended since it was presumed that an increase in the retentate concentration would bring about a decrease in the free water level in the medium and consequently reduce the solubility and availability of nutrients to the culture (Wu et al., 2006a). 3.1.7. Hydrogen According to Morimoto et al. (2004), it was possible to use natural anaerobic microora from POME sludge, instead of pure culture of isolated strain, to produce signicant amounts of hydrogen under anaerobic fermentation in a batch culture. The anaerobic microora in the POME sludge was found to produce hydrogen whereas no methane gas was observed in the evolved gas (Morimoto et al., 2004; Atif et al., 2005). Thus, anaerobic microora found in the POME sludge might be useful for the production of hydrogen from POME biomass resources without sterilization. Vijayaraghavan and Ahmad (2006) tested a new source of microora from cow dung for its hydrogen-generating capability in POME. They pointed out that the highest biogas generation and hydrogen content occurred at pH 5. OThong et al. (2007) highlighted that a raw POME supplemented with nutrients (N, P and Fe) gave a 20% increase in hydrogen production yields as well as 5861% higher hydrogen contents as compared to raw POME. The hydrogen production rate could be increased by 60% if raw POME was adjusted to an optimized iron concentration of 257 mg/l, a C/N ratio of 74 and a C/P ratio of 559 (O-Thong et al., 2008a). The nutrient supplementation strategy was found to increase the bacterial

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T.Y. Wu et al. / Biotechnology Advances 27 (2009) 4052 Table 4 The application of POME (m3/acre/year) as fertilizer for palm oil plantations (Onyia et al., 2001) Crops Young palms Adults palms Old palms N 2570 90128 162 P 27.532 52.5 52 K 5.110 1018.5 18 Mg 1.210 15 20

diversity in the anaerobic sequencing batch reactor and promote the growth of hydrogen-producing bacteria, namely Thermoanaerobacterium thermosaccharolyticum (O-Thong et al., 2007). Later, O-Thong et al. (2008b) were able to isolate T. thermosaccharolyticum PSU-2 from a sequencing batch reactor, which was used to digest POME for continuous hydrogen production. They found that the isolated strain of PSU-2 produced higher amounts of hydrogen, up to 68%, in organic nitrogen amended medium as compared to inorganic nitrogen amended medium. 3.2. Sustainable reuse of POME as fertilizer The application of raw or digested POME as fertilizer on land was initially thought to be impractical because of the efuent killing vegetation and leading to the blocking of percolation and waterlogging, thus resulting in anaerobic conditions. However, Wood et al. (1979) found that although raw POME would readily cause clogging and waterlogging of the soil, these problems could be overcome by the controlled application of small quantities of POME at a time. The testing of ground waters after 6 to 12 months of trial applications of raw POME as fertilizer showed no substantial percolation of oxygendemanding or other polluting elements without excessive run-off over the surface during wet weather (Wood et al., 1979). It was thus established that the water quality in the applied areas was unaffected (Dolmat et al., 1987). Oviasogie and Aghimien (2003) later reconrmed that a proper use and safe disposal of POME in the land environment would lead to improved soil fertility and contribute to environmental sustainability. Their results showed an enrichment of the soils with regard to phosphorus, nitrogen, calcium, magnesium, sodium and potassium following the application of the POME. Copper, iron and lead were predominant in their organic forms, while zinc was particularly present in its exchangeable form. The potential for using POME as a cheap organic fertilizer may offer an alternative to the excessive application of chemical fertilizers, especially phosphorus, for which cost is a severe economic constraint. For example, biologically treated POME has been widely used in the oil palm plantations for irrigation purposes and can be employed as a liquid fertilizer. It is estimated that each 15 million tonnes of POME would have a fertilizer value of RM 95.41 million (Table 3). According to Wood et al. (1979), an application of POME at 4.5 106 l per applied hectare was estimated to represent a fertilizer application of about 30 kg ammonium sulfate, 7 kg rock phosphate, 52 kg potash and 18 kg kieserite per palm per year. The nutrient composition of the fertilizers is shown in Table 3. An incorporation of POME may help to increase the organic matter in the soil, which may turn into humus after decomposition and become an active soil component. Thus, POME application would result in changes in the chemical properties of the soil. According to Ferreira and Araujo (2002), average contents of calcium, magnesium, potassium and phosphorus were found to increase in the soil with an increase in POME dosage, especially at a depth of 020 cm but an application of 120 m3/ha of POME to the soil reduced the aluminum content to zero at a depth of 20 cm after 12 months. Such a reduction

Table 3 Estimated fertilizer values from POME, which is based on 15 million tonnes of POME Fertilizer Ammonium sulphate Rock phosphate Muriate of potash Kieserite Total Tonnes (1000) 75.5 19.5 68.6 59.6 December 2002 price (RM/ton) 580 545 250 400 Fertilizer value (RM million) 43.79 10.63 17.15 23.84 95.41

of aluminum in the soil would eventually help prevent toxicity and growth hindrance for plants in acid soils (Matsumoto, 2000; Guo et al., 2007). Nevertheless, variations in POME quality among the mills and the rate of application as well as other details need to be determined in relation to local situations (Agamuthu et al., 1992). Moreover, the use of POME as fertilizer must be carried out with caution because of imbalances in the nutrient composition. A prolonged improper utilization may cause an accumulation of magnesium and thereby inhibit the availability of potassium (Onyia et al., 2001). Table 4 shows the nutrient requirements for the various growth stages of plants as well as the suitable amount of POME for reuse as fertilizer in order to avoid soil damage. According to Chan et al. (1980), the use of POME has been shown to improve soil productivity and increase the yield of crops as well as contribute to better root health by improving the soil structure. An increase in crop yield on the order of 10 to 24% has been reported (Tam et al., 1982; Lim et al., 1984). Teoh and Chew (1983) have further shown that mixtures of soil and POME in a ratio of 1:5 resulted in more vigorous growth of cocoa seedlings and decreased nursery rotation without the addition of supplementary fertilizers. With the help of organic matter consisting of peat and the sludge from POME, Shamshuddin et al. (2004) conrmed that aluminum toxicity towards the growth of cocoa seedlings on acid sulfate soil could be reduced to a certain extent. Agamuthu (1994) stated that the application of POME alone as fertilizer provided the highest yield of Napier grass (Pennisetum purpureum), up to 3276 kg/ha, as a result of POME containing almost all the major and minor elements required for its growth (Agamuthu et al., 1992). Although the application of fresh dung also gave rise to high yields of Napier grass, up to 2574 kg/ha, POME was preferred since fresh dung releases an unwanted odor that might attract ies (Agamuthu, 1994). Shamshuddin et al. (1998) indicated that the application of POME together with ground magnesian limestone, which might last for 3 years, was a sound agronomic option to alleviate the soil acidity and improve the fertility in Ultisol for maize production. They also revealed that a POME application up to 40 t/ha did not signicantly change the topsoil pH and exchangeable calcium, magnesium and aluminum, in which case the calcium and magnesium from the POME were held by the negative charge present on the exchange complex. Saltes et al. (2004) conducted a trial on a composting platform in windrows comprised of shredded empty fruit bunches that were watered weekly with POME. They found that the resulting compost had a good agronomic value but that the mineral balance was considerably affected due to the nutrients provided by POME being poorly retained by the substrate and partially lost in percolation following the weekly watering operations. According to Saltes et al. (2004), a better distribution of POME applications together with a system for recovering the leaching might substantially reduce the nutrient losses while maintaining a suitable humidity for microbial degradation. In a similar case to that of Saltes et al. (2004), Aisueni and Omoti (2002) also used shredded empty fruit bunches together with POME in the composting process but with the addition of poultry droppings as nutrient supplements. They noted that the use of POME in the composting process was particularly benecial in signicantly reducing the amount of poultry droppings, which were required to produce the same amount of nal compost.

T.Y. Wu et al. / Biotechnology Advances 27 (2009) 4052 Table 5 Chemical attributes of humic substances derived from POME (adapted from Siva et al, 2000) Chemical attribute Percent yield Percent loss on ignition Elemental make-up (%) C H N O O:H C:N C:H Functional groups (meq/g) Quinonoid (C=O) Carboxylic (COOH) Phenolic (OH) Total acidity (carboxylic + phenolic) Optical density (E4:E6 ratio) Humic acid Claried POME 3.47 97.5 57.87 8.26 2.91 30.96 3.75 19.89 7.01 2.52 2.22 3.34 5.56 4.22 Decomposed POME 1.82 99.5 48.94 5.76 8.05 37.25 6.47 6.08 8.50 2.85 2.08 3.27 5.35 6.09

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POME did not signicantly affect the pH and redox potential in the iron-poor acid sulfate soil during submergence. They also claimed that POME contains high concentrations of lignin that presumably decomposes slowly under anaerobic condition. Thus, these materials could not become active electron donors in the reduction process. 3.3. Sustainable reuse of POME as live food for animals and aquacultural organisms The reuse of POME as a dietary substitute for pigs, poultry and small ruminants as well as aquacultural organisms is gaining importance. Apart from oil palm fronds, palm press ber and palm kernel cake, Devendra (2004) pointed out that POME was especially important for feeding ruminants. Using POME as animal feeds, however, could only be considered as a co-management of the efuent because, according to Agamuthu (1995), a 40-ton-per-hour mill would require around 44,000 pigs or 43,000 cattle for the entire efuent to be utilized. Hutagalung et al. (1977) investigated the use of POME as animal feed for growingnishing pigs, in which case two types of meals known as censor tk8 (35% palm oil sludge, 32.5% cassava root meal, 32.5% palm kernel cake) and tkg (32% palm oil sludge, 34% cassava root meal, 17% palm kernel cake, 17% grass meal) were used. They found that it was economical to replace 50% maize (the regular diet constituent) with a POME-based animal feed, thus saving up to RM 0.02 per pig per day. In Colombia, POME has been fed with good results directly to pig (1012 l/head/day) together with palm oil and other ingredients (Devendra, 2004). POME could also be used as supplementary food in poultry farming. According to Ho (1976), animal feed production from palm oil wastes can replace at least half of the amount of imported maize for poultry diets and up to 100% for pig diets. Yeong et al. (1980) investigated the nutritive values of a POME product known as Prolima (Table 6) as the protein source in broiler chicken diets. It was observed that the amino acid content of palm kernel cake and palm oil sludge were somewhat close to cereal by-products and that of Prolima was between soybean meal and peanut meal, in which case the overall percentage of amino acid availability for palm kernel cake, palm oil sludge and Prolima were 74.4%, 24.8% and 71.0%, respectively. Therefore, the concentrations of Prolima up to 30% could be included in broiler diets as a replacement for soybean meal without causing any adverse effect on the growth performance of the chickens (Yeong et al., 1980). Later, Yeong and Azizah (1987) reported the optimum levels of using 1015% of dried POME in chicken feed for the growth and egg production. Pasha (2007) also reported that the optimum levels of POME in the diet for broilers and layers are 15% and 10%, respectively. The Malaysian Agricultural Research Development (MARDI) proved that wastes from the palm oil industry (such as oil palm sludge and palm press ber) alone or in combination, dried to moisture contents of 7%,

The slow release of the total N and P after the rst 12 weeks could also become a limiting factor if these nutrients were to be made available for plant growth (Palaniappan et al., 1983). Therefore, Azizah Chulan (1991) suggested that if POME were to be reused as fertilizer, the soil should be inoculated with vesiculararbuscular mycorrhizal (VAM) fungus Scutellospora calospora because the combination between POME and VAM will form mycorrhizae that may enhance the breakdown of certain soluble phosphates and insoluble organic phosphate such as phytate by roots (Gianinazzi-Pearson, 1985). Onyia et al. (2001) stated that the application of organic nitrogen from raw POME has been associated with lower yields due to ammonia being liberated during the mineralization of organic matter. Onyia et al. (2001) therefore suggested that nitrication of POME was necessary since a nitried POME would be more easily absorbed by most plants than a raw POME with a high organics content, especially in the tropics where nitrate leaching does not present a major problem. Numerous studies have identied ammonia volatilization as the major cause of low N efciencies in urea (Mikkelsen et al., 1978; Fillery et al., 1984), in which case up to 80% of the applied urea-N may be lost within 23 weeks of application (Hargrove and Kissel, 1979; Torello et al., 1983). Siva et al. (2000) reported that POME is rich in organic matter and varying amounts of humic substances across their respective organic matrices (Table 5). Seeing as humic substances have been reported to interact with ammonia compounds (Banerjee and Basak, 1978; Thorn and Mikita, 1992) and urea (Patti et al., 1992), Siva et al. (1999, 2000) investigated the effects of POME-derived humic substances on ammonia volatilization from urea. Initial studies by Aminuddin (1994) showed that POME could introduce a preferred environment within the ureasoil reaction zone (microsite) and successfully reduce ammonia volatilization to 8% of the applied N. Siva et al. (1999) displayed that this reduction in ammonia volatilization was accompanied by a corresponding increase in ammonium recovery and a decrease in pH, particularly at the microsite. The performance of humic fractions from POME also indicated an interplay of several mechanisms that could possibly include urease inhibition, urea absorption and ammonia xation (Siva et al., 2000). These results have implications to the reduction of N loss by ammonia volatilization from urea applied to the soil during crop production. According to Muhrizal et al (2006), the incorporation of organic material into iron-poor acid sulfate soil might enhance the benecial effects of reducible Fe(III) oxides or S in the soil and eventually promote an increase in pH under ooded conditions. However, not all organic materials are able to alleviate acid sulfate soil infertility with equal efcacy (Muhrizal et al., 2003). Although POME contains considerable amounts of organic materials, Muhrizal et al. (2006) revealed that

Table 6 The chemical composition of Prolima as compared to palm oil sludge (Agamuthu, 1995) Composition Moisture, % Crude protein (N 6.25), % Crude ber, % Ether extract, % Ash, % Nitrogen-free extract, % Calcium, % Phosphorus, % Magnesium, % Iron, mg/l Copper, mg/l Manganese, mg/l Zinc, mg/l Gross energy, MJ/kg Prolima 5.1 43.3 7.6 12.0 4.1 27.9 0.19 0.52 0.17 365 42 56 145 18.5 Palm oil sludge 6.9 12.4 15.2 24.1 11.2 46.7 0.28 0.18 0.25 1757 36 62 1075 19.6

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could be used as supplementary food for sheep (Devendra and Muthurajah, 1976). Vadiveloo (1988) fed Katjang and Katjang German Fawn crossbred goats with Napier grass and Leucaena leucocephala, which were supplemented with dried POME. He found that supplementing with POME at all levels did not signicantly depress forage intake but rather increased the total intake. Vadiveloo (1989b) further conrmed that L. leucocephala, which was supplemented with dehydrated POME, allowed mature regrowths of the crop (increased with regard to both dry matter and neutral detergent ber digestibility) to be fed to goats. Agamuthu et al. (1996) also found that goats and sheep digested POME-treated Napier grass signicantly faster than Napier grass alone. Feeding studies with goats have also shown that rice straw, which was properly supplemented with dehydrated POME, could promote satisfactory performance levels (Phang and Vadiveloo, 1991). According to Vadiveloo (1989a), NaOH treatment of rice straw together with dehydrated POME and Leucaena promoted an even higher straw intake among the goats. The increased intake and dry matter digestibility with the NaOH treatment might be due to an enhanced edibility and digestible energy of the roughage (Kellaway and Leibholz, 1983). It was concluded by Vadiveloo (1989a) that diets comprising 25% NaOH-treated straw, 50% dehydrated POME and 25% Leucaena permitted dietary nutrients to be reused efciently and maximized the inclusion of agro by-products. According to Devendra (2004), 10% of POME in diets for sheep gave the best result in terms of digestibility because crude ber digestibility dropped signicantly from 80.6% in a 10% POME diet to 27.0% in a 60% POME diet. Ether extract digestibility decreased progressively with increasing dietary POME. It should be stressed that the utilization of POME as animal feeds could be enhanced further by addition of molasses and palm press ber or other oil palm by-products. According to Devendra (2004), the maximum suitable level of inclusion of molasses appeared to be one part molasses for every 1.2 parts of palm press ber + POME. It was found that combining palm kernel cake and oil palm fronds with POME could create a low-cost and excellent feeding system. According to Pasha (2007), 50% palm kernel cake, 30% oil palm fronds and 20% POME can produce a reasonably good diet for moderate growth rate and acceptable meat quality in beef cattle. Two agricultural waste products, namely YM20 (a mixture of pea and corn) and POME, were evaluated in a closed recirculation system for their suitability to replace a costly diet of live algae in the culture of the Sudanese fairy shrimp, Streptocephalus proboscideus (Jawahar Ali and Brendonck, 1995). They found that the results in terms of growth (increase in length), cyst production and mortality were more successful when S. proboscideus were supplied with high densities of YM20 as compared to POME and algae. Habib et al. (1997) pointed out that POME could also be reused as a food source by aquatic organisms such as chironomid larvae known as bloodworms. They reported that the production of chironomid larvae was signicantly higher in POME (580 g/20 l POME) than in algal cultures (35 g/20 l algal culture). These chironomid larvae, in turn, present valuable live food for sh or cultured invertebrates (Shaw and Mark, 1980; Yusoff et al., 1996). Babu et al. (2001) studied the use of POME for the culturing of four species of sh, i.e. silver carp, catla, rohu and mrigal. The sh were harvested after 9 months and the maximum individual growths of silver carp, rohu and mrigal were 700, 550 and 600 g, respectively whereas the average growth of catla was 35.3 g. Vairappan and Yen (in press) showed that Isochrysis sp., which was grown in a modied medium of aerobically digested POME, was suitable to be reused as a supplement to further enrich and improve the rotifer cultures. These rotifer cultures, in turn, play their role as food organisms for sh larvae (Lubzens et al., 2001). 4. Conclusions On the whole, it is an undeniable fact that POME has its own potential for sustainable reuse through biotechnological advances. Moreover, it is understood that a cleaner production is, in the long run, a better option

for managing POME as opposed to end-of-pipe processes. However, although the emphasis on a cleaner production for POME management is well-intentioned, it may sometimes raise false expectations. The limits imposed by the economic and social frameworks present obvious limitations to sustainable practices that could be applied in the industries (Fricker, 2003), especially in most developing countries such as Malaysia. Consequently, there is usually no economic incentive to develop wastefree processes. A cleaner production is therefore limited unless it is subsidized, externalities are factored in, products are successfully designed for commercial reuse and, most importantly, the government takes the initiative in legislating for a sustainable industrial development. Since the economic framework depends on growth, production and consumption, the initiatives to promote a cleaner production for POME management can only come from the palm oil industries themselves since their subtle actions could accelerate the research and development for an enhanced POME management. In short, we need to become responsible citizens rather than mere consumers. References
Agamuthu P. Composting of goat dung with various additives for improved fertilizer capacity. World J Microbiol Biotechnol 1994;10:1948. Agamuthu P. Palm oil mill efuent treatment and utilization. In: Sastry CA, Hashim MA, Agamuthu P, editors. Waste treatment plant. New Delhi: Narosa Publishing House; 1995. p. 33860. Agamuthu P, Sivaraj S, Mukherjee TK. Agronomic and nutrition studies using Napier grass (Pennisetum purpureum) as fodder for goat and sheep grown with palm oil mill efuent (POME) fertilizer. In: Lokeshwar RR, Kumar AT, editors. Recent advances in goat production, 5th International Conference on Goats. New Delhi, India; 1992. p. 68593. Agamuthu P, Sivaraj S, Mukherjee TK. Nutrition and in vitro digestion aspects of Napier grass (Pennisetum purpureum) grown with palm oil mill efuent (POME) as fertiliser for goat and sheep. Indian J Anim Sci 1996;66:9357. Ahmad AL, Ismail S, Bhatia S. Water recycling from palm oil mill efuent (POME) using membrane technology. Desalination 2003;157:8795. Ahmad AL, Ismail S, Bhatia S. Ultraltration behavior in the treatment of agro-industry efuent: pilot scale studies. Chem Eng Sci 2005;60:538594. Aisueni NO, Omoti U. Nutrient status and economic potentials of composted oil palm bunch refuse as organic fertilizer. Moor J Agri Res 2002;3:1923. Alam MZ, Karim MIA, Hamisan AF, Jamal P, Jalal KCA. Liquid state bioconversion of palm oil mill efuent for cellulase production: statistical optimization of process conditions. Proceedings of the 1st International Conference on Natural Resources Engineering & Technology 2006. Putrajaya, Malaysia; 2006a. p. 22632. Alam MZ, Karim MIA, Mespuan MT. Statistical optimization of physicochemical factors for lignin peroxidase production using POME as substrate. Proceedings of the 20th Symposium of Malaysian Chemical Engineers (SOMChE 2006). Shah Alam, Malaysia; 2006b. p. 1516. Alam MZ, Jamal P, Nadzir MM. Bioconversion of palm oil mill efuent for citric acid production: statistical optimization of fermentation media and time by central composite design. World J Microbiol Biotechnol 2008;24:117785. Aminuddin H. Ammonia volatilization loss from surface placed urea-treated POME pellets. Malaysian Science and Technology Congress. Kuala Lumpur, Malaysia; 1994. Archer DB. The microbiological basis of process control in methanogenic fermentation of soluble wastes. Enzyme Microbiol Technol 1983;5:16270. Atif AAY, Fakhru'l-Razi A, Ngan MA, Morimoto M, Iyuke SE, Veziroglu NT. Fed batch production of hydrogen from palm oil mill efuent using anaerobic microora. Int J Hydrogen Energy 2005;30:13937. Aunstrup K. Industrial production of proteolytic enzymes. In: Spencer B, editor. Industrial aspects of biochemistry, Part A. North Holland-American Amsterdam: Elsevier; 1974. p. 2346. Azizah Chulan H. Effect of fertilizer and endomycorrhizal inoculum on growth and nutrient uptake of cocoa (Theobroma cacao L.) seedlings. Biol Fertil Soils 1991;11:2504. Babu MK, Sukumaran N, Kumar MGV, Murugesan AG, Jose GS, Hussain MIZ. Fish culture in palm oil mill efuent. Int J Oil Palm Res 2001;2:479. Banerjee SK, Basak RK. Studies on ammonia and urea treated humic substances and their interaction with some heavy metal ions. J Indian Soc Soil Sci 1978;26:34751. Barker TW, Worgan JT. The utilization of palm oil processing efuents as substrates for microbial protein production by the fungus Aspergillus oryzae. European J Appl Microbiol Biotechnol 1981;11:23440. Borja R, Banks CJ. Anaerobic digestion of palm oil mill efuent using an up-ow anaerobic sludge blanket (UASB) reactor. Biomass Bioenergy 1994;6:3819. Chan KW, Watson I, Lim KC. Use of oil palm waste material for increase production. In: Pushparajah E, Chin SL, editors. Soil science and agricultural development. Kuala Lumpur: Malaysian Society of Soil Science; 1980. p. 21342. Cheng JW. Penghasilan dan penulenan separa enzim xilanase daripada Aspergillus terreus SUK-1. Bachelor thesis, Universiti Kebangsaan Malaysia, Bangi, Malaysia; 2006. Chow MC. Palm oil mill efuent analysis. Kuala Lumpur: Palm Oil Research Institute of Malaysia; 1991. p. 118.

T.Y. Wu et al. / Biotechnology Advances 27 (2009) 4052 Chowdhury AJK, Alam MZ, Shahlizah SH. solation, purication and screening of fungal strain fro effective bioconversion of palm oil mill efuent. Proceedings of the 1st International Conference on Natural Resources Engineering & Technology 2006. Putrajaya, Malaysia; 2006. p. 16775. Chua ASM, Takabatake H, Satoh H, Mino T. Production of polyhydroxyalkanoates (PHA) by activated sludge treating municipal wastewater: effect of pH, sludge retention time (SRT) and acetate concentration in inuent. Water Res 2003;37:360211. Davis JB, Reilly PJA. Palm oil mill efuent a summary of treatment methods. Oleagineux 1980;35:32330. Debeaupuis JP, Lafont P. Fumitoxins, new mycotoxins from Aspergillus fumigatus Fres. Appl Environ Microbiol 1978;36:8-10. Deo YM, Gaucher GM. Semicontinuous and continuous production of penicillin-G by Penicillium chrysogenum cells immobilized in -carrageenan beads. Biotechnol Bioeng 1984;26:28595. Devendra C. Integrated tree crops-ruminants systems potential importance of the oil palm. Outlook Agri 2004;33:15766. Devendra C, Muthurajah RN. The utilization of oil palm by-products by sheep. Proceedings of the Malaysian International Symposium of Palm Oil Processing and Marketing. Kuala Lumpur; 1976. Devendra C, Yeong SW, Ong HK. The potential value of palm oil mill efuent (POME) as a feed source for farm animals in Malaysia. Palm Oil Research Institute Malaysia Workshop on Oil Palm By-products Utilization. Kuala Lumpur; 1981. p. 145. Dolmat MT, Lim KH, Zakaria ZZ, Hassan H. Recent studies on the effects of land application of palm oil mill efuent on oil palm and the environment. Proceedings of the 1987 International Oil Palm/Palm Oil Conference Agriculture; 1987. p. 596604. Eltaweel MA, Rahman RNZRA, Salleh AB, Basri M. An organic solvent-stable lipase from Bacillus sp. strain 42. Ann Microbiol 2005;55:18792. Ferreira WdeA, Araujo SMB. Effect of application of agroindustrial oil palm efuent on chemical characteristics of a yellow alic latosol with medium texture. Bol Pesqui Desenvolv - Embrapa Amazon Orient 2002;11:26. Fillery IRP, Simpson JR, De Datta SK. Inuence of eld environment and fertilizer management on ammonia loss from ooded rice. Soil Sci Soc Am J 1984;48:91420. Fricker A. Waste reduction in focus. Futures 2003;35:50919. Gianinazzi-Pearson V. Mycorrhizal effectiveness in phosphate nutrition: how, when and where? In: Molina R, editor. Proc 6th North Am Conf on Mycorrhizae. Bend. Oregon: Forest Research Laboratory; 1985. p. 1504. Guo TR, Zhang GP, Zhang YH. Physiological changes in barley plants under combined toxicity of aluminum, copper and cadmium. Colloids Surf B 2007;57:1828. Habib MAB, Yusoff FM, Phang SM, Ang KJ, Mohamed S. Nutritional values of chironomid larvae grown in palm oil mill efuent and algal culture. Aquaculture 1997;158: 95-105. Hargrove WL, Kissel DE. Ammonia volatilization from surface application of urea in the eld and laboratory. Soil Sci Soc Am J 1979;43:35963. Hassan MA, Shirai Y, Kusubayashi N, Karim MIA, Nakanishi K, Hashimoto K. Effect of organic acid proles during anaerobic treatment of palm oil mill efuent on the production of polyhydroxyalkanoates by Rhodobacter sphaeroides. J Ferment Bioeng 1996;82:1516. Hassan MA, Karim MIA, Shirai Y, Inagaki M, Nakanishi K, Hashimoto K. Economic analysis on production of bacterial polyhydroxyalkanoates from palm oil mill efuent. J Chem Eng Jpn 1997a;30:7515. Hassan MA, Shirai Y, Kusubayashi N, Karim MIA, Nakanishi K, Hashimoto K. The production of polyhydroxyalkanoate from anaerobically treated palm oil mill efuent by Rhodobacter sphaeroides. J Ferment Bioeng 1997b;83:4858. Hassan MA, Shirai Y, Umeki H, Yamazumi H, Jin S, Yamamoto S, et al. Acetic acid separation from anaerobically treated palm oil mill efuent by ion exchange resins for the production of polyhydroxyalkanoate by Alcaligenes eutrophus. Biosci Biotechnol Biochem 1997c;61:14658. Hassan MA, Nawata O, Shirai Y, Rahman NAA, Yee PL, Ariff A, et al. A proposal for zero emission from palm oil industry incorporating the production of polyhydroxyalkanoates from palm oil mill efuent. J Chem Eng Jpn 2002;35:9-14. Hipolito CN, Crabbe E, Badillo CM, Zarrabal OC, Mora MAM, Flores GP, et al. Bioconversion of industrial wastewater from palm oil processing to butanol by Clostridium saccharoperbutylacetonicum N1-4 (ATCC 13564). J Clean Prod 2007;16:6328. Ho KP. Malaysia: sup the wastes. Planter 1976;52:2967. Ho CC, Tan YK. Centrifugal fractionation studies on the particulates of palm oil mill efuent. Water Res 1983;17:6138. Ho CC, Tan YK. The treatment of anaerobically digested palm oil mill efuent by pressurized activated sludge. J Chem Technol Biotechnol 1988;41:7584. Ho CC, Tan YK, Wang CW. The distribution of chemical constituents between the soluble and the particulate fractions of palm oil mill efuent and its signicance on its utilization/treatment. Agric Wastes 1984;11:6171. Hutagalung RI, Chang CC, Toh KM, Chan HC. Potential of palm oil mill efuent as feed for growingnishing pigs. Planter 1977;53:29. Hwang TK, Ong SM, Seow CC, Tan HK. Chemical composition of palm oil mill efuents. Planter 1978;54:74956. Ishizaki A, Tanaka K. Production of poly--hydroxybutyric acid from carbon dioxide by Alcaligenes eutrophus ATCC 17697T. J Ferment Bioeng 1991;71:2547. Jamal P, Alam MZ, Ramlan M, Salleh M, Nadzir MM. Screening of Aspergillus for citric acid production from palm oil mill efuent. Biotechnology 2005;4:2758. James R, Sampath K, Alagurathinam S. Effect of lead on respiratory enzyme activity, glycogen and blood sugar levels of the teleost Oreochromis mossambicus (Peters) during accumulation and depuration. Asian Fish Sci 1996;9:87-100. Jawahar Ali A, Brendonck L. Evaluation of agro-industrial wastes as diets for culture of the fairy shrimp Streptocephalus proboscideus. Hydrobiologia 1995;298:16773. Kalil MS, Pang WK, Wan Yusoff WM, Sadazo Y, Abdul Rahman R. Direct fermentation of palm oil mill efuent to acetonebutanolethanol by solvent producing Clostridia. Pak J Biol Sci 2003;6:12735.

51

Kellaway RC, Leibholz J. Effects of nitrogen supplements on intake and utilization of low quality roughages. World Anim Rev 1983;48:337. Khalid AR, Wan Mustafa WA. External benets of environmental regulation: resource recovery and the utilization of efuents. Environmentalist 1992;12:27785. Klepper O, Hatta GT, Sunardi M, Rijksen HD. Organic matter input and decomposition before and after development of acid sulfate soils at Pulau Petak, Kalimantan, Indonesia. Workshop on Acid Sulfate Soils in the Humid Tropics. International Institute for Land Reclamation and Improvement, Wageningen, Netherlands; 1990. p. 34773. Koroneos C, Dompros A, Roumbas G, Moussiopoulos N. Life cycle assessment of hydrogen fuel production processes. Int J Hydrogen Energy 2004;29:144350. Kotz JP, Thiel PG, Hattingh WHJ. Anaerobic digestion II. The characterization and control of anaerobic digestion. Water Res 1969;3:45994. Laohaprapanon T, Prasertsan P, H-Kittikun A. Screening of thermotolerant microorganisms and application for oil separation from palm oil wastewater. Songklanakarin J Sci Technol 2007;29:8018. Lee IY, Nam SW, Choi ES, Chang HN, Park YH. Production of poly--hydroxybutyrate and measurement of related enzyme activities in Alcaligenes eutrophus. J Ferment Bioeng 1993;76:4168. Lee SY, Choi J, Wong H. Recent advances in polyhydroxyalkanoate production by bacterial fermentation: mini review. Int J Biol Macromol 1999;25:316. Leow TC, Rahman RNZRA, Basri M, Salleh AB. High level expression of thermostable lipase from Geobacillus sp strain T1. Biosci Biotechnol Biochem 2004;68:96-103. Lim KH, Quah SK, Gillies D, Wood BJ. Palm oil mill efuent treatment and utilization in Sime Darby Plantations the current position. Workshop Proceedings of the Palm Oil Research Institute Malaysia, vol. 9. ; 1984. p. 4252. Lin YT, Labbe RG, Shetty K. Inhibition of Vibrio parahaemolyticus in seafood systems using oregano and cranberry phytochemical synergies and lactic acid. Innovative Food Sci Emerging Technol 2005;6:4538. Lubzens E, Zmora O, Barr Y. Biotechnology and aquaculture of rotifers. Hydrobiologia 2001;446447:33753. Ma AN. A novel treatment for palm oil mill efuent. Palm Oil Res Instit Malaysia (PORIM) 1995;29:20112. Ma AN. Environmental management for the palm oil industry. Palm Oil Dev 2000;30:19. Ma AN, Ong ASH. Pollution control in palm oil mills in Malaysia. J Am Oil Chem Soc 1985;62:2616. Mandels M. Production and applications of cellulase. Laboratory procedures handbook. U.S: Army Materials Laboratories; 1974. December. Mashitah MD, Masitah H, Kamaruddin MY. Palm oil mill efuent: a potential substrate for cellulase production. The Proceedings of RSCE and 16th SOMChE. Malaysia; 2002. p. 20916. Masngut N, Takriff MS, Kalil MS, Kadhum AAH, Mohammad AW. Production of acetonebutanolethanol in oscillatory ow bioreactor. Proceedings of the 20th Symposium of Malaysian Chemical Engineers (SOMChE 2006). Shah Alam, Malaysia; 2006. p. 14550. Masngut N, Takriff MS, Mohammad AW, Kalil MS, Kadhum AAH. Performance of oscillatory ow reactor and stirred tank reactor in solvent fermentation from palm oil mill efuent. J Teknol 2007;47(F):4554. Matsumoto H. Cell biology of aluminum toxicity and tolerance in higher plants. Int Rev Cytol 2000;200:1-46. Md Din MF, Ujang Z, Muhd Yunus S, van Loosdrecht MCM. Storage of polyhydroxyalkanoates (PHA) in fed-batch mixed culture using palm oil mill efuent (POME). 4th Seminar on Water Management (JSPS-VCC). Johor; 2006a. p. 11927. Md Din MF, Ujang Z, van Loosdrecht MCM, Ahmad A, Sairan MF. Optimization of nitrogen and phosphorus limitation for better biodegradable plastic production and organic removal using single fed-batch mixed cultures and renewable resources. Water Sci Technol 2006b;53:1520. Mikkelsen DS, De Datta SK, Obcemea WN. Ammonia volatilization losses from ooded rice soils. Soil Sci Soc Am J 1978;42:72530. Morimoto M, Atsuko M, Atif AAY, Ngan MA, Fakhru'l-Razi A, Iyuke SE, et al. Biological production of hydrogen from glucose by natural anaerobic microora. Int J Hydrogen Energy 2004;29:70913. Muhrizal S, Shamshuddin J, Husni MHA, Fauziah I. Alleviation of aluminum toxicity in an acid sulfate soil using organic materials. Commun Soil Sci Plant Anal 2003;34:29933011. Muhrizal S, Shamshuddin J, Fauziah I, Husni MAH. Changes in iron-poor acid sulfate soil upon submergence. Geoderma 2006;131:11022. Mun LT, Ishizaki A, Yoshino S, Furukawa K. Production of acetone, butanol and ethanol from palm oil waste by Clostridium saccharoperbutylacetonicum N1-4. Biotechnol Lett 1995;17:64954. Nor MT, Mahadi NM. Ultraltration makes POME suitable medium for Bacillus thuringiensis H-14. Proc. of 3rd ASEAN Workshop on Membrane Technology. Singapore; 1986. p. 11824. O-Thong S, Prasertsan P, Intrasungkha N, Dhamwichukorn S, Birkeland NK. Improvement of biohydrogen production and treatment efciency on palm oil mill efuent with nutrient supplementation at thermophilic condition using an anaerobic sequencing batch reactor. Enzyme Microb Technol 2007;41:58390. O-Thong S, Prasertsan P, Intrasungkha N, Dhamwichukorn S, Birkeland NK. Optimization of simultaneous thermophilic fermentative hydrogen production and COD reduction from palm oil mill efuent by Thermoanaerobacterium-rich sludge. Int J Hydrogen Energy 2008a;33:122131. O-Thong S, Prasertsan P, Karakashev D, Angelidaki I. Thermophilic fermentative hydrogen production by the newly isolated Thermoanaerobacterium thermosaccharolyticum PSU-2. Int J Hydrogen Energy 2008b;33:120414. Oka A, Suzuki N, Watanabe T. Effect of fatty acids in Moina on the fatty acid composition of larvae Ayu Plecoglossus altivelis. Bull Jpn Sci Soc Fish 1982;48:115962.

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T.Y. Wu et al. / Biotechnology Advances 27 (2009) 4052 International Conference on Chemical and Bioprocess Engineering / 19th Symposium of Malaysian Chemical Engineers: ICCBPE/SOMChE 2005. Sabah, Malaysia; 2005. p. 1435. Tam TK, Yeow KH, Poon YC. Land application of POME, H & C experience. Proceedings of the Regional Workshop on Palm Oil Mill Technology and Efuent Treatment. Kuala Lumpur, Malaysia; 1982. p. 21624. Teoh KC, Chew PS. Use of palm oil sludge cake in oil palm and cocoa polybag nurseries. In: Lim KH, Bachik AT, Chin PY, editors. Proc seminar on land application of palm oil mill and rubber factory efuent. Kuala LumpurMalaysian Soil Sci Soc; 1983. p. 14262. Thorn KA, Mikita MA. Ammonia xation by humic substances: a nitrogen-15 and carbon-13 NMR study. Sci Total Environ 1992;113:6787. Tian G, Kang BT, Brussaard L. Biological effects of plant residues with contrasting chemical compositions under humid tropical conditions decompositions and nutrient release. Soil Biol Biochem 1992;24:105160. Torello WA, Wehner DJ, Turgeon AJ. Ammonia volatilization from fertilized turfgrass stands. Agron J 1983;75:4546. Uzel A, Sorkun K, na , oulu D, Genay , Salih B. Chemical compositions and antimicrobial activities of four different Anatolian propolis samples. Microbiol Res 2005;160:18995. Vadiveloo J. Performance of young indigenous and crossbred goats fed forages supplemented with palm oil mill efuent. Small Rumin Res 1988;1:36979. Vadiveloo J. Supplementation of alkali-treated rice straw. Biol Wastes 1989a;27:1537. Vadiveloo J. The intake and digestibility in goats of Leucaena leucocephala supplemented with dehydrated palm oil mill efuent. Anim Feed Sci Technol 1989b;24:4555. Vairappan CS, Yen AM. Palm oil mill efuent (POME) cultured marine microalgae as supplementary diet for rotifer culture. J Appl Phycol (in press); doi:10.1007/s10811007-9305-1 Vijayaraghavan K, Ahmad D. Biohydrogen generation from palm oil mill efuent using anaerobic contact lter. Int J Hydrogen Energy 2006;31:128491. Wang CW, Chong CN, Rahim Bidin. Growth of SCP on hydrolysate of palm oil sludge. Proceedings of the First ASEAN Workshop on the Technology of Animal Feed Production Utilizing Food Waste Materials. Bandung; 1981. p. 1-13. Wase DAJ, Raymahasay S, Wang WC. Production of -D-glucosidase, endo-1,4--Dglucanase and D-xylanase from straw by Aspergillus fumigatus IMI 255091. Enzyme Microb Technol 1985;7:2259. Wattanapenpaiboon N, Wahlqvist ML. Phytonutrient deciency: the place of palm fruit. Asia Pacic J Clin Nutr 2003;12:3638. Webb BH, Rajagopalan K, Cheam ST, Dhiaddin MNJ. Palm oil mill waste recovery as a byproduct industry, Part I mechanical aspects. Planter 1975;51:85-101. Wood BJ. A review of current methods for dealing with palm oil mill efuent. Planter 1977;53:47795. Wood BJ, Pillai KR, Rajaratnam JA. Palm oil mill efuent disposal on land. Agric Wastes 1979;1:10327. Wu TY, Mohammad AW, Md Jahim J, Anuar N. Penghasilan asid itakonik oleh Aspergillus terreus IMI 282743 dalam pelbagai jenis medium penghasilan. Pascasidang Kolokium Jabatan Kejuruteraan Kimia dan Proses. Malaysia; 2005. p. 14751. Wu TY, Mohammad AW, Md Jahim J, Anuar N. Investigations on protease production by a wild-type Aspergillus terreus strain using diluted retentate of pre-ltered palm oil mill efuent (POME) as substrate. Enzyme Microb Technol 2006a;39:12239. Wu TY, Mohammad AW, Md Jahim J, Anuar N. Treatment of palm oil mill efuent (POME) using ultraltration membrane and sustainable reuse of recovered products as fermentation substrate. 4th Seminar on Water Management (JSPSVCC). Johor, Malaysia; 2006b. p. 12835. Wu TY, Mohammad AW, Md Jahim J, Anuar N. Palm oil mill efuent (POME) treatment and bioresources recovery using ultraltration membrane: effect of pressure on membrane fouling. Biochem Eng J 2007;35:30917. Yamane T, Fukunaga M, Lee YW. Increased PHB productivity by high-cell-density fedbatch culture of Alcaligenes latus, a growth-associated PHB producer. Biotechnol Bioeng 1996;50:197202. Yee PL, Hassan MA, Shirai Y, Wakisaka M, Karim MIA. Continuous production of organic acids from palm oil mill efuent with sludge recycle by the freezingthawing method. J Chem Eng Jpn 2003;36:70710. Yeong SW, Azizah A. Effect of processing on feeding values of palm oil mill efuent (POME) in non-ruminants. Proceedings of 10th Annual Conference, Malaysian Society of Animal Production. Genting Highlands, Malaysia; 1987. p. 3026. Yeong SW, Syed Ali AB, Faizah M. The nutritive value of a palm oil mill efuent product (PROLIMA) as a protein source in broiler diets. MARDI Res Bull 1980;8:24759. Yusoff S, Hansen SB. Feasibility study of performing an life cycle assessment on crude palm oil production in Malaysia. Int J Life Cycle Assess 2007;12:508. Yusoff MF, Om AD, Cheah SH. Use of agro-industrial efuent in augmenting microalgae production and sh fry growth in hatchery tanks. J Aqua Trop 1996;11:11926. Zeikus JG. Chemical and fuel production by anaerobic bacteria. Annu Rev Microbiol 1980;34:42364.

Olgun EJ, Snchez G, Mercado G. Cleaner production and environmentally sound biotechnology for the prevention of upstream nutrient pollution in the Mexican coast of the Gulf of Mxico. Ocean Coast Manag 2004;47:6170. Olie JJ, Tjeng TD. Traitment et evacuation des eaux rsiduaires d'une huilerie de palm. Oleagineux 1972;27:2158. Onyia CO, Uyub AM, Akunna JC, Norulaini NA, Omar AKM. Increasing the fertilizer value of palm oil mill sludge: bioaugmentation in nitrication. Water Sci Technol 2001;44:15762. Oviasogie PO, Aghimien AE. Macronutrient status and speciation of Cu, Fe, Zn and Pb in soil containing palm oil mill efuent. Global J Pure Appl Sci 2003;9:7180. Palaniappan S, Syed So, Palaniappan VM. Land application of raw palm oil mill efuent (POME). Studies on leaching and decomposition and nutrient release of the residues. In: Lim KH, Bachik AT, Chin PY, editors. Proc seminar on land application of palm oil mill and rubber factory efuents. Kuala LumpurMalaysian Soil Sci Soc; 1983. p. 180217. Palm CA, Rowland AP. A minimum data set for characterization of plant quality for decomposition. In: Cadisch G, Gillier KE, editors. Driven by nature: plant litter quality and decompositon. Oxford, UK: CAB International, Wallingford; 1997. p. 37992. Pang WK, Kalil MS, Hassan O, Wan Yusoff WM. Penghasilan asetonbutanoletanol dan gas hidrogen daripada euen kilang kelapa sawit secara fermentasi anaerobik dengan Clostridium acetobutylicum NCIMB 13357. Sains Malaysiana 2004;33:7381. Pasha TN. Boosting animal feed. Global Oil & Fats Business Magazine 2007;4:4344. Patti AF, Verheyen TV, Douglas L, Wang X. Nitrohumic acids from Victorian brown coal. Sci Total Environ 1992;113:4965. Phang SM. Algal production from agro-industrial and agricultural waste in Malaysia. Ambio 1990;19:4158. Phang OC, Vadiveloo J. Effects of varieties, botanical fractions and supplements of palm oil byproducts on the feeding value of rice straw in goats. Small Rumin Res 1991;6:295301. Prasertsan P, H-Kittikul A, Kunghae A, Maneesri J, Oi S. Optimization for xylanase and cellulase production from Aspergillus niger ATTC 6275 in palm oil mill wastes and its application. World J Microbiol Biotechnol 1997;13:5559. Rahman RNZRA, Leow TC, Salleh AB, Basri M. Geobacillus zalihae sp. nov., a thermophilic lipolytic bacterium isolated from palm oil mill efuent in Malaysia. BMC Microbiol 2007. doi:10.1186/1471-2180-7-77 Razak CNA, Salleh AB, Musani R, Samad MY, Basri M. Some characteristics of lipases from thermophilic fungi isolated from palm oil mill efuent. J Mol Catal B: Enzym 1997;3:1539. Razak CNA, Musani R, Basri M, Salleh AB. Characterization of membrane-bound lipase from a thermophilic Rhizopus oryzae isolated from palm oil mill efuent. J Am Oil Chem Soc (JAOCS) 1999;76:1714. Saltes S, Siregar FA, Caliman JP, Liwang T. Ligno-cellulose composting: case study on monitoring oil palm residuals. Compost Sci Util 2004;12:37282. Salim MR, Ujang Z, Muhd Yunus S, Md Din MF, Ahmad MA. Biodegradable polymer production from POME using activated enriched sludge by coupled aerobic and anaerobic system. 4th Seminar on Water Management (JSPS-VCC). Johor, Malaysia; 2006. p. 1539. Schuchardt F, Wulfert K, Damoko D. New process for combined treatment of waste (EFB) and waste water (POME) from palm oil mills technical, economical and ecological aspects. Landbauforsch Volkenrode 2005;55:4760. Shamshuddin J, Sharifuddin HAH, Bell LC. Changes in chemical properties of an ultisol as affected by palm oil mill efuent application. Commun Soil Sci Plant Anal 1998;29:2395406. Shamshuddin J, Muhrizal S, Fauziah I, Husni MHA. Effects of adding organic materials to an acid sulfate soil on the growth of cocoa (Theobroma cacao L.) seedlings. Sci Total Environ 2004;323:3345. Shaw PC, Mark KK. Chironomid farming a means of recycling farm manure and potentially reducing water pollution in Hong Kong. Aquaculture 1980;21:15563. Siva KB, Aminuddin H, Husni MHA, Manas AR. Ammonia volatilization from urea as affected by tropical-based palm oil mill efuent (POME) and peat. Commun Soil Sci Plant Anal 1999;30:785804. Siva KB, Aminuddin H, Husni MHA, Manas AR. Ammonia volatilization from urea as affected by humic substances derived from palm oil mill efuent (POME) and tropical peat. Trop Agric (Trinidad) 2000;77:1320. Somrutai W, Takagi M, Yoshida T. Acetonebutanol fermentation by Clostridium aurantibutyricum ATCC 17777 from a model medium for palm oil mill efuent. J Ferment Bioeng 1996;81:5437. Stanton WR. Treatment of efuent from palm oil factories. The Planter 1974;50:3827. Suwandi MS. POME, from waste to antibiotic and bioinsecticide. Jurutera Kim Malays 1991;1:7999. Suwandi MS, Mohammad AA. Growth of Penicillium chrysogenum in ultraltered POME concentrate. Regional Workshop in Biotechnology in Industrial Development. Universiti Pertanian Malaysia; 1984. Suzuki T, Yamane T, Shimizu S. Mass production of poly--hydroxybutyric acid by fedbatch culture with controlled carbon/nitrogen feeding. Appl Microbiol Biotechnol 1986;24:3704. Takriff MS, Masngut N, Kalil MS, Kadhum AAH, Mohammad AW. Production of solvents from palm oil mill efuent in oscillatory ow bioreactor. Proceedings of the 2nd