期刊
BIOTECHNOLOGY LETTERS
卷 33, 期 12, 页码 2431-2438出版社
SPRINGER
DOI: 10.1007/s10529-011-0705-6
关键词
Esterification activity; Pichia pastoris; Rational design; Rhizomucor miehei lipase; Surface display
资金
- National Natural Science Foundation of China [30900017, 20976062]
To increase the activity of Rhizomucor miehei lipase (RML) in organic solvent, multiple sequence alignments and rational site-directed mutagenesis were used to create RML variants. The obtained proteins were surface-displayed on Pichia pastoris by fusion to Flo1p as an anchor protein. The synthetic activity of four variants showed from 1.1- to 5-fold the activity of native lipase in an esterification reaction in heptane with alcohol and caproic acid as substrates. The increase in esterification activity may be attributed to the four mutations changing the flexibility of RML or facilitating the reaction. In conclusion, this method demonstrated that multiple sequence alignments and rational site-directed mutagenesis combined with yeast display technology is a faster and more effective means of obtaining high-efficiency esterification lipase variants compared with previous similar methods.
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