利用麻风树油驯化筛选高活性脂肪酶产生菌及其催化酯化反应

以1株分解麻风树油的脂肪酶产生菌Pseudomonas sp. LP-1为出发菌株, 通过麻疯树油定向驯化筛选获得1株酶活较高且产酶稳定的菌株P. sp. X-2-45, 其水解酶活为29.79 U/mL, 比原始菌株提高了288%。对P. sp. X-2-45生长与产酶特征、对植物油脂水解能力及在有机相中催化脂肪酸和有机醇间的酯化反应研究发现, 该菌株生长速率和产酶速率明显加快, 培养30 h时生物量和酶活达到最大, 稳定期延长, 培养过程中脂肪酶在培养基中的稳定性提高。以麻疯树油诱导合成的P. sp. X-2-45脂肪酶对麻疯树油的水解能力比原始菌株提高了378%, 说明采用麻风树油定向驯化可提高脂肪酶对相应底物的水解能力。X-2-45脂肪酶可以催化月桂酸与正丁醇、正辛醇、月桂醇和丙三醇之间, 棕榈酸、硬脂酸与甲醇、正辛醇、月桂醇和丙三醇之间, 油酸与甲醇、正丁醇、正辛醇、月桂醇和丙三醇之间发生酯化反应。

Breeding of High Lipolytical Bacteria by Acclimation to Jatropha Oil and Resultant Strain-catalyzed Esterification Reaction

A strain, Pseudomonas sp. X-2-45, with high and stable lipolytical activity was screened by continuously subculturing a lipase-producing bacterium P. sp. LP-1 in culture medium containing Jatropha oil as a sole carbon source. Its hydrolytic activity was 29.79 U/mL, which was increased by 288% as compared to that of parent strain. Furthermore, the growth and lipase synthesis of X-2-45, its catalytic ability to hydrolyze vegetable oils, as well as ester synthesis between fatty acids and organic alcohols were studied. Results showed that rates of bacterial growth and lipase synthesis were significantly raised. Bacterial biomass and lipase activity reached the highest level after 30 h of incubation. Moreover, growth stationary period was prolonged and lipase produced exhibited good stability in culture media during incubation period. Hydrolytic activity of P. sp. X-2-45 lipase toward Jatropha oil was increased by 378% as compared to parent strain, suggesting that acclimation to Jatropha oil was an effective approach for improving substrate selectivity of lipase. Finally, results of ester synthesis catalyzed by P. sp. X-2-45 lipase indicated that this lipase could catalyze esterification reactions between lauric acid and n-butanol, n-octanol, 1-dodecanol or glycerol, palmitic acid or stearic acid and methanol, n-octanol, 1-dodecanol or glycerol, oleic acid and methanol, n-butanol, n-octanol, 1-dodecanol or glycerol.