Esterase and lipase activity in Jatropha curcas L. seeds.

Two new esterases (JEA and JEB) and a lipase (JL) were extracted from the seeds of Jatropha curas L. Lipase activity was only found during germination of the seeds and increased to a maximum after 4 days of germination. All enzymes were found to be most active in the alkaline range at around pH 8 and the purified (fractionated precipitation with ethanol and gel filtration) esterases were very stable at high temperatures. The molecular weight (SDS-PAGE) of both esterases was determined to be 21.6-23.5 kDa (JEA) and 30.2 kDa (JEB) and the isoelectric point was 5.7-6.1 for esterase JEA and 9.0 for esterase JEB. Most ions caused a negative influence on the activity of both esterases. Using p-nitrophenyl butyrate as a substrate JEA showed a K(m) of 0.02 mM and a v(max) of 0.26 micromol mg(-1) min(-1). Under the same conditions JEB showed a K(m) of 0.07 mM and a v(max) of 0.24 micromol mg(-1) min(-1). Both esterases hydrolyzed tributyrin, nitrophenyl esters up to a chain length of =C4 and naphtylesters up to a chain length =C6. In transesterification reactions, JL was found to be most active at very low water activities (0.2) and in high water activities, the lipase hydrolysed triglycerides into conversions above 80%. The lipase hydrolysed both short chain and long chain triglycerides at about the same rate but was inactive on alpha-methylbenzyl acetate. JL is a potentially useful biocatalyst in the hydrolysis of triglycerides in organic solvents.

Enzyme-supported oil extraction from Jatropha curcas seeds.

Jatropha curcas is a tropical plant widely distributed in arid areas. The seeds contain about 55% of oil, which is mainly used for the production of soap as a fuel and after transesterification as biodiesel. Various methods for recovering of oil from the seeds, including extraction with organic solvents and water, have been investigated. Compared to hexane extraction (98%) the oil extraction using water only yielded 38% of the total oil content of the seeds. Using several cell wall degrading enzymes during aqueous extraction a maximum yield of 86% was obtained. The influence of cellulolytic, hemicellulolytic enzymes, as well as proteases was studied. The experiments were carried out at different pH-values and temperatures to find out the optimum for oil recovering using enzymes. , Surprisingly, the best results (86%) were obtained using an alkaline protease. Combinations of proteases with hemicellulases and/or cellulases did not further increase the extraction yield. The enzyme-supported aqueous extraction offers a nontoxic alternative to common extraction methods using organic solvents with reasonable yields.

Biogas production from Jatropha curcas press-cake.

Seeds of the tropical plant Jatropha curcas (purge nut, physic nut) are used for the production of oil. Several methods for oil extraction have been developed. In all processes, about 50% of the weight of the seeds remain as a press cake containing mainly protein and carbohydrates. Investigations have shown that this residue contains toxic compounds and cannot be used as animal feed without further processing. Preliminary experiments have shown that the residue is a good substrate for biogas production. Biogas formation was studied using a semicontinous upflow anaerobic sludge blanket (UASB) reactor; a contact-process and an anaerobic filter each reactor having a total volume of 110 L. A maximum production rate of 3.5 m3 m"3 d"1 was obtained in the anaerobic filter with a loading rate of 13 kg COD m~3 d"1. However, the UASB reactor and the contact-process were not suitable for using this substrate. When using an anaerobic filter with Jatropha curcas seed cake as a substrate, 76% of the COD was degraded and 1 kg degraded COD yielded 355 L of biogas containing 70% methane.