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Effects of A sn‐33 glycosylation on the thermostability of T hermomyces lanuginosus lipase
Author(s) -
Zhu J.,
Liu H.,
Zhang J.,
Wang P.,
Liu S.,
Liu G.,
Wu L.
Publication year - 2014
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12519
Subject(s) - thermostability , pichia pastoris , glycosylation , lipase , biochemistry , recombinant dna , pichia , enzyme , mutant , chemistry , biology , gene
Aims The study was 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 GS 115 .The recombinant Tll protein was expressed and purified, and its glycosylation site was identified by LCMS / MS as Asn‐33. Two nonglycosylated 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 analysed. The glycosylated Tll exhibited better thermostability than nonglycosylated variants. Conclusions Our experiments have demonstrated the improvement of Tll thermostability by Asn‐33 glycosylation. Significance and Impact of the Study This work has deepened our understanding in the mechanism of Tll thermostability and will guide us to directional improvement of lipases and even other industrial enzymes.