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Site-directed mutagenesis of GH10 xylanase A from Penicillium canescens for determining factors affecting the enzyme thermostabilityстатья
Статья опубликована в высокорейтинговом журнале
Информация о цитировании статьи получена из
Web of Science,
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 23 ноября 2017 г.
- Авторы: Denisenko Y.A., Gusakov A.V., Rozhkova A.M., Osipov D.O., Zorov I.N., Matys V.Y., Uporov I.V., Sinitsyn A.P.
- Журнал: International Journal of Biological Macromolecules
- Том: 104
- Год издания: 2017
- Издательство: Elsevier BV
- Местоположение издательства: Netherlands
- Первая страница: 665
- Последняя страница: 671
- DOI: 10.1016/j.ijbiomac.2017.06.079
- Аннотация: In order to investigate factors affecting the thermostability of GH10 xylanase A from Penicillium canescens(PcXylA) and to obtain its more stable variant, the wild-type (wt) enzyme and its mutant forms, carrying single amino acid substitutions, were cloned and expressed in Penicillium verruculosum B1-537 (niaD-) auxotrophic strain under the control of the cbh1 gene promoter. The recombinant PcXylA-wt and I6V,I6L, L18F, N77D, Y125R, H191R, S246P, A293P mutants were successfully expressed and purified for characterization. The mutations did not affect the enzyme specific activity against xylan from wheat as well as its pH-optimum of activity. One mutant (L18F) displayed a higher thermostability relative to the wild-type enzyme; its half-life time at 50–60◦C was 2–2.5–fold longer than that for the PcXylA-wt, and the melting temperature was 60.0 and 56.1◦C, respectively. Most of other mutations led to decrease in the enzyme thermostability. This study, together with data of other researchers, suggests that multiple mutations should be introduced into GH10 xylanases in order to dramatically improve their stability.
- Добавил в систему: Гусаков Александр Васильевич