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| Enhancement of substrate specificity towards unsaturated long-chain fatty acids of Rhizopus chinensis lipase through rational efficient hydrolysis of soybean oil design for |
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| KeyWord:lipase fatty acid rational design substrate specificity hydrolysis of soybean oil |
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| Abstract: |
| Fatty acid specificity of Rhizopus chinensis lipase was altered by rational design for efficient hydrolysis of soybean oil. The enzymatic properties of these variants were studied. The reaction conditions of hydrolysis of soybean oil by the lipase were optimized by single factor experiment. The results showed that a series of variants were obtained by site-directed mutagenesis. Compared with the wild-type lipase, the variant HQL showed enhanced specificity towards unsaturated fatty acids and increased hydrolytic activity for p-NPP(C16) by 2.72 fold. The optimal temperature and optimal pH of the variants were 40?℃ and 8.0, which were consistent with the wild-type lipase. The optimal reaction conditions of hydrolysis of soybean oil by the lipase were obtained as follows: reaction time 24 h, mass ratio of water to oil 1∶?1, dosage of lipase 500 U/g (based on oil mass),pH 8.0 and temperature 40?℃. Under the optimal reaction conditions, the hydrolysis rate of soybean oil by the variant HQL increased to 98% due to the increasing specificity towards unsaturated long-chain fatty acids, while it was only 80% for the wild-type lipase. |
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