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DOI 10.1002/ejlt.200401051
809
Rudolf Brenneis
Burkhard Baeck
Gerd Kley
Commercial native lipase A from Candida antarctica was used to produce alkyl esters
through the alcoholysis of (waste) fats with 2-ethyl-1-hexanol. The process was carried
out in batch stirred tank reactors (from 100 mL up to 3000 L).
The content of alkyl esters in reaction mixtures was determined by gradient HPLC
using an evaporative light scattering detector and the reaction progress was controlled
by determining the ratio of the palmitic acid ester peak area to the oleic acid ester peak
area in HPLC chromatograms.
The results show that alcoholysis is the favoured reaction in presence of excess water
and water-insoluble alcohols in comparison with hydrolysis (fatty acid content ,5%).
The optimum amount of water for the alcoholysis was found to be 80100% of the
amount of fat. In the presence of low quantities of water both alcoholysis and hydrolysis are slow.
Conversion rate increases with increasing temperature to 6570 7C.
Based on these results a large-scale test to produce 3000 L of alkyl ester (to be used as
lubricant coolant) was carried out. The experiments have proved that alcoholysis is
completed after about 710 h depending on temperature.
1 Introduction
The direct conversion of triacylglycerols (TAG) by lipasecatalysed alcoholysis in alkyl esters (AE) and glycerol is of
increasing practical interest. This process can be used to
synthesise value-added products, for instance biodiesel
or biodegradable lubricants (ester oils).
Ester oils have many technological and ecological advantages over petroleum oils in certain fields of (total loss)
lubrication. But the conventional production of these
esters is expensive which makes an extensive use prohibitive. Lower-cost feedstock, such as restaurant grease
or tallow, would be more advantageous to be used.
The goal of the work performed was to develop a biotechnological process to produce ester oils from waste fat
[1]. In [2] we reported the lipase-catalysed alcoholysis of
various model fats with different linear, branched-chain
and cyclic water-insoluble monohydric alcohols by lipases from Candida antarctica, A-component. We also ex-
Correspondence: Rudolf Brenneis, Bundesanstalt fr Materialforschung und -prfung (BAM), Richard-Willsttter-Str. 11,
12489 Berlin, Germany. Phone: 149-30-6392-5849, Fax: 14930-6392-5917; e-mail: rudolf.brenneis@bam.de
Research Paper
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R. Brenneis et al.
Candida antarctica (A-component). The lipase is nonposition specific, the activity level can vary from batch
to bach, but it will typically range around 5 KLU/g [12].
2.3 Apparatus
Most of the lipase-catalysed alcoholysis experiments
were carried out in 100-mL cylindric glass tubes of an
Apparatus for determination of demulsibility characteristics of petroleum, oils or synthetic fluids (Herschel Stirring Method based on ASTM D 401-DIN
51599-FTM 7913201-ISO 6614) by Petrotest Instruments (Dahlewitz, Germany). In this misused tank
reactor the components were stirred with a blade mixer
(1600 rpm) and their temperature was kept constant by
a water bath.
2.1 Materials
2-Ethyl-1-hexanol was obtained from Merck-Schuchardt
(Hohenbrunn, Germany) and European Oxo GmbH
(Oberhausen, Germany).
Recycled restaurant grease was supplied by Kanzler &
Co. (Groalmerode, Germany) = F1 and by B. Vierhouten
BV (Ermelo, Netherlands) = F2, and model fat (mixture of
vegetable fats on the basis of palm and peanut oils) was
supplied by Gerlicher (Berlin, Germany) = F3. The fatty
acid compositions of these input fats are shown in Tab. 1.
TAG is favourable to use with a low proportion of polyunsaturated chains as starting material, because the
processes of autoxidation and photo-oxidation as well as
polymerisation must be prevented or inhibited since they
lead to the deterioration of ester oil products.
Glycerol was purchased from Merck (Darmstadt, Germany).
Tab. 1. Fatty acid compositions of the input fats.
Fatty acids
F1
F2
F3
[%]
C16:0
C18:0
C18:1
C18:2
C18:3
48.1
1.2
44.2
5.1
0.4
48.4
11.8
35.1
3.5
0.8
54.0
8.7
33.7
2.1
0
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2.2 Enzymes
811
The content of AE and TAG as well as di- and monoacylglycerols (DAG, MAG) and free fatty acids (FFA) in
reaction mixtures was determined by gradient HPLC
(degasser DG-1310, liquid chromatograph LC-10AD,
diode array detector SPD-M10A, column oven CTO-10A,
system controller SCL-10A) from Shimadzu (Kyoto,
Japan), dynamic mixing chamber from Knauer (Berlin,
Germany) and a standard column (C18) EC 250/4 Nucleosil 1203 from Macherey-Nagel (Dren, Germany) with an
evaporative light scattering detector Sedex 55 from
S.E.D.E.R.E. (Alfortville, France). Purified air was used as
carrier gas at constant pressure (200 kPa); the column was
held at 35 7C.
Acetonitrile#
[%]
Dichloromethane##
[%]
Methanol##
[%]
0
30
45
60
100
30
10
0
0
35
45
55
0
35
45
45
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##
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4 Conclusion
Commercial native lipase (Candida antarctica lipase A) was
used for the synthesis of alkyl esters using alcoholysis of
recycled restaurant grease with 2-ethyl-1-hexanol. Water
has been found to play an important role in this process.
Our studies have shown that the determination of palmitic/oleic acid value by means of HPLC is a suitable method
to get information about the conversion rate of enzymatic
alcoholysis using input fats with dominant contents of
palmitic and oleic acids in the fatty acid composition.
Acknowledgements
This work was financially supported within the programme Innovations Centrum Biocatalysis (ICBio) by the
Deutsche Bundesstiftung Umwelt (DBU).
References
[1] R. Brenneis, P. Kcher, G. Kley, B. Baeck, T. Schwilling:
Untersuchungen zur Herstellung von biologisch abbaubaren
Verlustschmierstoffen aus Altfetten. Mll und Abfall 34 (2002)
244249.
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