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Effects of Asn-33 glycosylation on the thermostability of Thermomyces lanuginosus lipase.

J Appl Microbiol.. 2014-04;

Zhu J, Liu H, Zhang J, Wang P, Liu S, Liu G, Wu L. CAS Key Laboratory of Genome Sciences and Information, Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing, China.

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摘要

AIMS: The study is to examine whether glycosylation could improve the thermostability of recombinant Thermomyces lanuginosus lipase (Tll) expressed in Pichia pastoris.METHODS AND RESULTS: The Tll gene was synthesized and transformed into Pichia pastoris GS115.The recombinant Tll protein was expressed and purified, and its glycosylation site was identified by LCMS/ MS as Asn-33. Two non-glycosylated mutants were constructed and the variant proteins were also expressed and purified. Effects of temperature on activities of the wild type Tll and variants were analyzed. The glycosylated Tll exhibited better thermostability than non-glycosylated variants.CONCLUSIONS: Our experiments have demonstrated the improvement ... More