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  • Title: Effects of Asn-33 glycosylation on the thermostability of Thermomyces lanuginosus lipase.
    Author: Zhu J, Liu H, Zhang J, Wang P, Liu S, Liu G, Wu L.
    Journal: J Appl Microbiol; 2014 Jul; 117(1):151-9. PubMed ID: 24724829.
    Abstract:
    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 GS115.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.
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