ISSN 0003 6838, Applied Biochemistry and Microbiology, 2012, Vol. 48, No. 4, pp. 377384.

Pleiades Publishing, Inc., 2012. Original Russian Text E.A. Tsavkelova, M.A. Egorova, E.V. Petrova, A.I. Netrusov, 2012, published in Prikladnaya Biokhimiya i Mikrobiologiya, 2012, Vol. 48, No. 4, pp. 417 424.

Biogas Production by Microbial Communities via Decomposition of Cellulose and Food Waste
E. A. Tsavkelova, M. A. Egorova, E. V. Petrova, and A. I. Netrusov
Department of Biology, Moscow State University, Moscow, Russia e mail: tsavkelova@mail.ru
Received May 24, 2011

AbstractSeveral active microbial communities that form biogas via decomposition of cellulose and domes tic food waste (DFW) were identified among 24 samples isolated from different natural and anthropogenic sources. The methane yield was 190260 ml CH4/g from microbial communities grown on cellulose sub strates, office paper, and cardboard at 37C without preprocessing. Under mesophilic conditions, bioconver sion of paper waste yields biogas with a methane content from 47 to 63%; however, the rate of biogas produc tion was 1.52.0 times lower than under thermophilic conditions. When microbial communities were grown on DFW under thermophilic conditions, the most stable and effective of them produced 230353 ml CH4/g, and the methane content in biogas was 5458%. These results demonstrates the significance of our studies for the development of a technology for the biotransformation of paper waste into biogas and for the need of selection of microbial communities to improve the efficiency of the process. DOI: 10.1134/S0003683812040126

INTRODUCTION Methanogenesis is the final step in the process of organic compound destruction under anaerobic con ditions. Different groups of microorganisms are capa ble of down organic compounds into substrates that participate in methanogenesis: acetate, carbon diox ide, and hydrogen, from which methane can be later formed. As a consequence of microorganism activity, about 1 billion tons of CH4/year is formed during the carbon cycle [1]. Methane, which is a greenhouse gas with some 25 times more infrared absorbing capacity per molecule than CO2 [2, 3]. Biogas, in which meth ane is present, is one of the major sources of renewable fuel. The reason for the interest in the search for alter native energy sources is not only the ever increasing demand for energy, but also the safety issues associated with the use of traditional energy sources, the steadily deteriorating environment, and the depletion of primary resources in the world (oil, gas, coal, and wood). Never theless, renewable energy makes up approximately 14% of the primary energy consumed in the world [4]. Biogas mainly consists of methane (5570% 4) and carbon dioxide (3045% 2). It may also con tain trace amounts of hydrogen and hydrogen sulfide, ammonia, nitrogen, aromatic hydrocarbons, and halogenated aromatic hydrocarbons [5]. Biogas has several advantages over other types of alternative fuels; for example, methane contains three times more energy than biohydrogen fuel [6]. Besides, biogas pro duction does not require the cultivation of agricultural plants, which are commonly used for the production

of biodiesel and ethanol [4], since biogas is mainly obtained via the decomposition of livestock waste (mainly cattle manure) and during wastewater treat ment, wherein biogas formation occurs in methane tanks during the last step. However, despite the fact that anaerobic treatment of municipal and agricultural wastewater has been widely used for a long time, the use of the technology for biogas production from solid domestic waste has been less successful [3]. Any biom ass, as well as the organic portion of waste that is capa ble of biodegradation, can be used as a substrate for biogas production. In addition to energy production and reduction in air pollution by greenhouse gases after anaerobic digestion and biogas generation, the remaining mass can be used as a high quality fertilizer. However, for the complete and extensive application of the biogas technology, the issues of full substrate uti lization, increasing the quality and quantity of biogas production, and maintenance of the stability and functional activity of the microbial community remain open [3]. A range of microorganisms belonging to several dif ferent groups is involved in biogas formation. Micro bial communities are responsible for major processes, such as hydrolysis of polymer substrates, fermentation of sugars and amino acids, anaerobic oxidation, aceto genesis, and acetoclastic or hydrogenotrophic metha nogenesis [7]. Also, the composition of microbial communities and, therefore, the efficacy of biogas generation depend on the composition of the growth medium, growth conditions, temperature, pH, and

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other factors [8]. A large number of projects are devoted to the study of biogas generation from live stock manure, sewage sludge, and other organic waste, but information about the use of cellulose as a sub strate is limited. Also, cellulose and hemicellulose are predominant in solid domestic waste (SDW) [9]. In recent years, conversion of paper materials into biogas has generated considerable interest [10, 11]. This is due to separate waste collection, as well as the fact that paper and cardboard are the most suitable parts of SDW for biodegradation [11, 12]. However, the mod ern approach to separate waste processing is not com mon among domestic producers and municipal ser vices; therefore, microbial processes of substrate transformation and the biotechnological potential of such industries remain poorly understood. Recently [13], several active microbial communi ties that can degrade cellulose with the formation of biogas at 55 have been identified among 24 samples isolated from different natural and anthropogenic econiches. The isolated microbial communities showed stable functional activity during five passages (more than six months of growth). The objectives of this study were to select anaerobic microbial communities during their cultivation on cellulose, office paper, and cardboard, as well as on domestic food waste (DFW), and to compare their ability to produce biogas under anaerobic conditions at 37 and 55. MATERIALS AND METHODS Biomass sources. Samples of inoculum were col lected from different natural and anthropogenic eco logical niches: samples nos. 1 and 2 are compost heap (Moscow region); samples nos. 3 and 4 are bagasse of red and white grapes (Dagestan); sample no. 5 is rabbit droppings (Moscow region); sample no. 6 is livestock manure no. 1 (Moscow region); sample no. 7 is live stock manure (Moscow region); samples nos. 813 are samples from freshwater thermophilic reservoirs from the Kamchatka Peninsula; samples nos. 1418 are silt and pond sedimentation from ponds and fresh water reservoirs (Tver suburb); samples nos. 1923 are zebra, pony, wildebeest, black antelope, and elephant manure, respectively (zoo, Moscow); and sample no. 24 is coprolites of earthworms (botanic garden, Mos cow). Growth. Cultivation and selection of active micro bial communities that form biogas were carried out on a medium of the following composition (g/l, distilled water): 1.0 K2HPO4, 1.0 KH2PO4, 2.5 NH4Cl, 0.5 MgSO4 7H2O, 0.1 CaCl2 6H2O, 0.1 NaCl, 1.0 CaCO3, 5.0 NaHCO3, 2.0 yeast extract, 1 peptone, 1 ml microelement solution, 0.5 mg/l rezazurin, and pH 7.07.5. The trace solution contained the follow ing (mg/l, distilled water): 70.0 ZnCl2, 100.0 MnCl2 4H2O, 190.0 CoCl2 6H2O, 6.0 H3BO3, 36.0 Na2MoO4 2H2O, 2.0 CuCl2 2H2O, 24.0 NiCl2 6H2O, 15.0

Na2WO4 2H2O, and 1.0 g/l FeSO4 7H2O. Ferrous sulfate was first dissolved in 10 ml of 25% HCl. The following items were used as cellulose containing sub strates (15 g/l): ash free MRTU 6 06 2411 65 filters, office paper with black and white print, and corru gated cardboard. All items were cut into 0.5 cm2 pieces. When DFW with diverse plant and animal components was used, all ingredients were first dried first, mixed, and ground in a mortar until the largest pieces did not exceed 0.5 cm2. To determine the dry weight, thesample substrates (10 g) were dried at 105. Inoculum (30% of the total medium volume) was introduced into 30 ml of growth medium, which was contained in 100 ml bottles. Each vial was sealed with a rubber stopper and was then sealed with an alumi num cap; air was replaced with argon. Cultures were incubated in the dark at 55 under thermophilic conditions or in an incubator at 37 under meso philic conditions; if needed, pH of the medium was adjusted to 7.0 with 1N HCl. The cultures were stored in 25% glycerol under anaerobic conditions at 20. The activity of microbial communities was deter mined by the increase in the methane concentration in biogas. The stability of the selected communities was examined by repeated passages onto a fresh growth medium after the community reached the maximum level of biogas production. Chromatography. Measurements of the 4, 2, and 2 concentrations were performed on a Crystal 2000 M gas chromatograph (Chromatec, Russia), equipped with an FFIP microcapillary column (15000 0.5 mm), argon carrier gas, 15 ml/min flow rate, FID detector; detector temperature 200C the temperature gradient within the incubator varied from 70 to 160C. The results were analyzed using the Chro matec Analytic 2.5 software (Chromatec, Russia). The rate of gas formation was estimated by measur ing the pressure in the sealed vials with growing com munities. The gas concentrations in the mixture and methane content were determined under standard temperature and pressure conditions. All experiments were repeated 35 times. The data were analyzed using standard statistical methods. The deviation from the mean values was less than 510% for the data pre sented in the tables and figures. Microscopy. A Nikon Eclipse E100 optical light microscope (Japan) was used to monitor the composi tion of microbial communities and changes during the cultivation of microorganisms. Fixed samples were stained with aqueous solution of fuchsin for 3 min and examined with an oil immersion lens at 900 magnifi cation. RESULTS AND DISCUSSION Biogas production by microbial communities from cellulose containing materials. To identify effective microbial communities that can degrade cellulose
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BIOGAS PRODUCTION BY MICROBIAL COMMUNITIES VIA DECOMPOSITION % 35 30 25 20 15 10 5 4 5, 6, 9, 10, 11, 12 0 4 8 12 16 days 0 4 8 8 % 70 7 60 50 40 3 30 1 2 20 10 15 23 13 17, 18 14, 16 12 16 20 24 28 32 36 40 44 days

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(b)

24 22 19 21 20

Fig.1. Dynamics of methane production (%) by communities nos. 112 (a) and 1324 (b), grown at 37C (first passage).

under mesophilic growth conditions (37), we stud ied 24 samples from the same econiches as the ther mophilic communities [13]. However, the majority of these cultures were either unable to produce biogas under these conditions (Fig. 1) or decreased methane production during subsequent passages. It was shown that communities isolated at 55 (mostly from manure of herbivores) were able to convert cellulose into methane with high efficacy: on average, about 15 mmol of 4/g of starting substrate [13]. When the communities were grown under mesophilic condi tions, the methane yield was significantly lower: from 24 to 811 mmol 4/g of cellulose. During the selection process, only communities nos. 1 and 21 that were isolated from compost and wildebeest manure, respectively, retained the ability to produce biogas at a high level (Table 1). Community no. 21 was the most stable one, producing about 11.5 mmol 4/g of cel lulose (250260 ml of 4/g). The high yield of methane during the first passage can be explained by the high concentration of nutrients and other biologi cally active substances in the starting inoculate (wilde beest manure). Previously [13], we reported that no active micro bial community was isolated during the growth of

sample no. 1 under thermophilic conditions (the total methane yield was less than 38.8%; the efficacy of cel lulose decomposition was only 4.9 mmol 4/g of cellulose), while an active community that was able to convert cellulose into methane (with a yield of 15.6 mmol of 4/g of cellulose and a methane con tent of 58%) was isolated from sample no. 21. Active microbial communities that can effectively produce biogas from cellulose under both mesophilic and ther mophilic conditions have been isolated from individ ual samples. At the same time, significantly more sta ble communities that yield up to 55 60% biogas were isolated under thermophilic conditions. Under the following conditions, biogas formation was the most effective: ~16 mmol 4/g of cellulose (358.3 ml of 4/g) formed during 1525 days. For comparison, under mesophilic conditions, cultures nos. 1 and 21 took more than 1.5 months to synthesize 260267 ml of 4/g. Table 2 shows the change in the rate of biogas for mation (RBF) during cellulose decomposition under thermophilic and mesophilic conditions. During the selection process, changes in this value depended on the growing community: it increased for some and did not significantly change for others. Also, the RBF was

Table 1. Methane accumulation during the growth of microbial communities nos.1 and 21 on cellulose at 37C Sample number 1 Methane yield indicators CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g First passage 45.87 36.49 1.63 61.40 395.51 17.67 Second passage 63.98 347.15 15.49 50.48 256.51 11.45
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Third passage 47.07 202.22 9.03 63.33 246.67 11.02

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Fourth passage 58.25 250.20 11.17 60.78 258.17 11.53

Fifth passage 57.96 266.73 11.91 62.46 260.20 11.62

21

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Table 2. Rate of biogas formation (RBF) during the decomposition of office paper and cardboard under thermofilic (T) and mesophilic (M) conditions Maximum RBF, ml of CH4/ day ml of medium No. I passage 3T 4T 6T 7T 17 T 18 T 19 T 20 T 21 T 22 T 1M 21 M 3T 6T 17 T 18 T 19 T 20 T 21 T 22 T 1M 21 M 1.30 0.66 0.45/1.30 0.64/0.50 0.64/0.52 0.59 0.91/0.96 1.00 0.93 1.36 0.40 0.40/0.35 Office paper 0.60 1.00 0.68 0.86 1.00 1.91 0.86 0.84 0.60 0.49 V passage 0.76/0.54 0.72 0.71 0.48/0.47 0.72/0.45 0.64/0.48 0.80 0.82 0.54/0.50 0.90 0.46 0.43 Cardboard 0.66 0.74 0.76 0.70 0.80 0.78 0.82 0.62 0.60 0.46 Maximum RBF, ml of CH4/day g Cellulose I passage 86.7 44.0 30.0/86.7 42.7/33.3 42.7/34.7 39.3 60.6/64.0 66.7 62.0 90.7 26.7 26.7/23.3 Office paper 40.0 66.7 45.3 75.3 66.7 127.3 75.3 56.0 40.0 32.7 V passage 50.7/36.0 48.0 47.3 32.0/31.3 48.0/30.0 42.7/32.0 53.4 54.7 36.0/33.3 60.0 30.7 28.7 Cardboard 44.0 49.3 50.7 46.7 53.3 52.0 54.7 41.3 40.0 30.7 I passage 5 7 5/12 5/14 5/14 5 3/14 21 7 11 7 8/24 Office paper 6 6 3 6 6 3 6 6 7 7 V passage 7/21 7 5 14/21 7/26 7/21 5 5 5/33 12 8 8 Cardboard 7 7 7 7 7 7 7 7 13 13 Maximum RBF, day

at least two times higher under thermophilic condi tions than under mesophilic ones. It was shown that, for some cultures, there are two peaks of biogas forma tion, with the first peak for the majority of cultures occurring on days 57. It is known that at a high tem perature and in the presence of inhibitors, such as ammonium and volatile fatty acids, methane forma tion occurs in two stages: first, acetate is oxidized by syntrophic acetate oxidizing bacteria into 2 and 2; then, hydrogenotrophic methanogens transform these compounds into methane [14]. Acetoclastic methanogens are more sensitive to changes in pH (acidification), while hydrogenotrophic methanogens are able to grow at low pH [15]. Paper and cardboard, which constitute the major part of municipal solid waste, are good materials for biodegradation [11, 12]. Biogas production by micro bial communities isolated from cellulose was investi gated while cultures were growing on office paper and corrugated cardboard. The most active communities were isolated from different substrates: bagasse of red grapes (no. 3), manure of herbivores (nos. 6 and 20

22), and slit pond sediment (nos. 17 and 18). Commu nities were grown under thermophilic and mesophilic conditions; the results are shown in Fig. 2 and in Tables 2 and 3. The communities isolated from cellu lose were effective for transforming paper into biogas. Under thermophilic conditions, microbial communi ties nos. 3 and 1722 grown on office paper were able to convert cellulose into methane with high efficiency (the methane content in biogas was about 50%). Active substrate hydrolysis occurred during growth on corrugated cardboard at 55, followed by biogas for mation, which was less effective (Fig. 2, Table 3). It is known from literature that during the decomposition of different types of paper sources, the methane yield can amount to as much as 217.3 ml of CH4/g for office paper and 152.3183.0 ml of CH4/g for corrugated cardboard [11, 12]. During the growth of microbial communities nos. 1 and 21 under mesophilic conditions, the cultures pro duced methane with 47.3 and 51.7% yield on office paper and with 48.9 and 49.6% yield on cardboard, respectively. Community no. 21 produced more meth
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BIOGAS PRODUCTION BY MICROBIAL COMMUNITIES VIA DECOMPOSITION % 50 () % 50 40 30 20 10 22 20 21 3 19 6 45 40 18 17 35 30 25 20 15 10 5 5 10 15 20 25 days 0 5 10 15 20 25 days 20 6 19

381

(b)

18

22 3 17 21

Fig. 2. Dynamics of methane production (%) by the most active communities (nos. 3, 6, and 1722) from office paper with black and white print (a) and packaging cardboard (b) at 55C.

ane and did it more effectively (Table 3). Under meso philic conditions, cardboard biodegradation was as effective as at 55, but at a lower rate. Nevertheless, under both mesophilic and thermophilic conditions, the selected microbial communities retained their activity during the decomposition and transformation of the paper source into biogas. The biodegradation of the studied substrates occurred without preprocessing, i.e., without acid or alkaline hydrolysis used for the destruction of lignin, which degrades slowly under anaerobic conditions and reduces cellulose availability for hydrolysis [11]. The lignin content in such sub strates usually varies from 2% in office paper to 24% in newsprint [16]. The RBF by microbial communities on office paper under thermophilic conditions (~0.91.0 ml/day ml of medium) was higher than on cellulose, except commu nities nos. 3 and 17. The highest biogas yield (by cul ture no. 20) was almost 2.0 ml/day per ml of medium. The maximum RBF during the office paper and card board decomposition was observed on days 67 for the majority of cultures, except cultures nos. 17 and 21, in which the bioconversion was carried out over the first 3 days. The RBF was slightly lower when cardboard was used as a source (Table 2). Despite the fact that under mesophilic conditions, the bioconversion of paper resulted in biogas formation, the RBF was 1.5 2 times lower than under thermophilic conditions. The study of the dynamics of biogas formation showed that the concentration and hydrogen pressure first increased and then decreased with an increase in hydrogenotrophic organisms. Then, the decrease in pH, caused by the formation of large amounts of vola tile fatty acids (VFAs) during cellulose hydrolysis, inhibited methanogenesis. The increase in pH led to
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the formation of methane from acetic acid and an increase in methane concentration and pressure. Sim ilar processes occurring during biodegradation of cel lulose and office paper under mesophilic conditions were described in [10]. Thus, by selecting microbial communities and adapting them to the substrate, we were able to increase significantly the yield of the methane content in biogas. Moreover, the increase was equally effective under both thermophilic and meso philic growth conditions. Biogas production by microbial communities that can degrade DFW. We compared the process of biogas formation by isolated communities when DFW was used as a substrate. Microbial communities were iso lated at 55, because the most effective bioconver sion of organic matter occurred under thermophilic conditions (the first passage); however, the majority of communities produce more than 50% of methane in gas mixtures (Fig. 3). A distinctive feature of biogas production on this substrate was that a number of communities, for example, nos. 15, 21, and 23, pro duced a significant amount of hydrogen at early growth stages5.95, 4.24, and 4.72%, respectively which then became involved in methanogenesis. We selected several active microbial communities that effectively processed organic waste into biogas (Table 4) and did not reduce the efficiency of bioconversion during passages. The maximum rate of methane for mation (7 days of growth) for communities nos. 2, 3, 19, and 23 was 0.60, 0.88, 0.43, and 0.44 ml/day ml of medium, respectively. The communities, isolated from compost heap (no. 2) and bagasse of red grapes (no. 3), were the most active ones for DFW decomposition: 353 and 313 ml of 4 /g, respectively; the methane content in biogas was 58%. According to R. Zhang et al. [17], up to 440 ml of methane/g were produced
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Table 3. Methane accumulation during the cultivation of microbial communities on office paper and cardboard un der thermophilic (T) and mesophilic (M) conditions Sample number 3T Methane yield indicators CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g Office paper 51.25 247.95 11.07 48.61 241.14 10.76 48.61 246.60 11.00 49.78 259.05 11.57 49.09 270.88 12.09 50.33 281.80 12.58 48.77 255.84 11.42 50.41 265.35 11.85 47.32 211.38 9.43 51.69 245.50 10.96 Cardboard 50.13 223.44 9.97 48.02 231.52 10.33 51.02 252.43 11.27 49.33 244.49 10.92 47.91 234.26 10.43 48.70 241.79 10.79 50.12 240.28 10.72 50.04 246.98 11.02 48.90 193.65 8.64 49.63 240.88 10.75

6T

17 T

18 T

19 T

20 T

21 T

22 T

1M

21 M

during anaerobic microbial degradation (within 28 days at 50) of food waste from different public catering establishments and commercial sources (res taurants, food markets, hotels). Other authors [18],

studying the decomposition of various food waste types under mesophilic and thermophilic conditions during the same time, demonstrated that the methane yield did not depend on the growth conditions and selected species of the substrate. Thus, during the con version of waste from soup processing, cafeteria, and commercial kitchen into biogas, the methane content ranged from 250 to 450 ml/g under mesophilic condi tions and from 240 to 470 ml/g under thermophilic conditions. The greatest amount of methane (more than 500 ml of 4/g) was only formed during the decomposition of fish wastes grease trap wastes. Nev ertheless, because of the heterogeneity of these sub strates, it should be taken into account that the meth ane yield can vary depending on the type of the organic substrate and the composition of the microbial com munity. All selected communities were able to effec tively produce biogas from cellulose and organic waste, and the selection of microbial communities resulted in methane yields that were comparable with or exceeded the reported methane yield on similar substrates [11, 12, 17, 18]. It is known that a methanogenic microbial com munity consists of different types of bacteria and archaea; the close relationship between them is prima rily based on the nutritional needs [19, 20]. During the analysis of both thermophilic [13] and mesophilic communities decomposing cellulose, we have noticed differences in the culture composition, including the initial and final steps (passages) of their selection. Cells of different morphotypes were found in these communities, as well as a large number of spore forms, which may indicate the presence of clostridia. Clostridia are usually well represented in anaerobic communities hydrolyzing cellulose [21]. A greater microbial diversity was observed in thermophilic com munities degrading DFW. The diversity is determined by the presence of different types of substrates (pro teins, fats, carbohydrates, and cellulose components). It was noted that microbial cells bind with substrate particles and fibers of cellulose. It is known that adhe siveness is an important feature of anaerobic hydro lytic (including cellulolytic) organisms. OSullivan

Table 4. Mathane accumulation during the cultivation of microbial communities nos. 2, 3, 19, and 23 on DFW at 55C Sample number 2 3 19 23 Methane yield indicators CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g CH4, % CH4, ml/g CH4, mmol/g First passage 58.88 350.17 15.63 60.88 486.92 21.74 54.50 252.03 11.25 52.71 227.41 10.15 Second passage 45.34 188.08 8.40 52.18 247.02 11.03 47.47 197.12 8.80 47.22 212.12 9.47 Third passage 62.00 303.86 13.57 62.96 323.86 14.46 60.88 223.34 9.97 62.92 224.36 10.02 Fourth passage 61.20 261.85 11.70 63.56 333.38 14.88 56.55 182.31 8.14 56.64 182.57 8.15
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Fifth passage 58.08 352.91 15.76 57.73 313.28 14.00 54.60 230.75 10.30 54.5 250.11 11.16
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BIOGAS PRODUCTION BY MICROBIAL COMMUNITIES VIA DECOMPOSITION % 70 60 50 40 30 20 10 0 5 10 15 20 25 30 6 15 35 40 days 13 7 30 20 10 18 17 % 60 50 40

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4 3 2 1

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24 19

22 23 21

10

15

20

25

30

35

40 days

Fig. 3. Dynamics of methane production (%) from DFW at 55C (first passage) by selected microbial communities: nos. 14, 6, 7, 13, 15 (a) and nos. 1719, 2124 (b).

et al. [22] demonstrated that the ability of a microbial community to degrade cellulose was largely attributed to the colonization density of hydrolitic organisms capable of adhesion onto substrate, rather than intrin sic hydrolitic activity of individual microorganisms of the community. In our study, microbial communities were able to produce biogas with a high methane content under both mesophilic and thermophilic [13] growth condi tions. Despite the fact that food waste is biodegraded faster, cellulose and its derivatives have been no less effective as biogas sources. In addition, the commu nity selected on cellulose showed a greater stability during passages for several months. It should be noted that the growth medium did not contain expensive components, and the substrate was not preprocessed; this permits the use of a large variety of cellulose con taining substrates. It is known that thermophilic anaerobic destruction of organic matter is more effec tive and requires less time [2325]. Thermophilic reactors provide a higher biogas yield and a high rate of its formation, with better conversion coefficient, and can be loaded with a large amount of substrate [24, 26]. In addition, the growth of pathogenic microor ganisms (such as E. coli and Salmonella spp.) and par asites is suppressed at high temperatures [2729]. At the same time, biogas production under mesophilic conditions requires less energy consumption, and the process is to a lesser extent inhibited by the formation of ammonia and VFAs [24]. Therefore, depending on the requirements, it is necessary to select an optimal microbial community, which allows for the processing of organic substrates into biogas with maximum effi ciency. Our data are valuable for understanding and evaluation of different strategies that can be used for processing and more efficient use of solid organic waste, including those containing cellulose.
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ACKNOWLEDGMENTS The work was supported by the federal program Human Capital for Science and Education in Inno vative Russia, 20092013 (state contract no. P2470). REFERENCES
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APPLIED BIOCHEMISTRY AND MICROBIOLOGY

Vol. 48

No. 4

2012