重组Kex2蛋白酶在毕赤酵母BG10中的胞外表达

Q3 Biochemistry, Genetics and Molecular Biology Biointerface Research in Applied Chemistry Pub Date : 2022-09-24 DOI:10.33263/briac134.360

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引用次数: 0

摘要

Kex2是酵母和真菌中一种约67kD的蛋白酶，专门消化-Lys-Arg-或-Arg-Arg-。由于具有相同的切割位点，Kex2有可能应用于蛋白质工程，例如蛋白质纯化。在最近的研究中，使用pD902设计并构建了酿酒酵母KEX2基因，然后将其与毕赤酵母BG10的基因组整合。在AOX1启动子控制下，用甲醇诱导表达Kex2，并在细胞外部分中发现。使用单克隆抗His-tag抗体和LC/MS/MS通过蛋白质印迹成功地证实了重组Kex2。

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Extracellular Expression of Recombinant Kex2 Protease in Pichia pastoris BG10

Kex2 is a ~67 kD protease in yeast and fungi that specifically digest at -Lys-Arg- or -Arg-Arg-. Because of the same cleaving site, Kex2 is the potential to be applied in protein engineering, for instance, for protein purification. In the recent study, the Saccharomyces cerevisiae KEX2 gene was designed and constructed using pD902 and then integrated with the genome of Pichia pastoris BG10. Under AOX1 promoter control, Kex2 was expressed with methanol induction and found in extracellular fractions. The recombinant Kex2 was successfully confirmed with a western blot using a monoclonal anti-His-tag antibody and LC/MS/MS.

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来源期刊

Biointerface Research in Applied Chemistry CHEMISTRY, APPLIED-

CiteScore

4.80

自引率

0.00%

发文量

256

期刊介绍： Biointerface Research in Applied Chemistry is an international and interdisciplinary research journal that focuses on all aspects of nanoscience, bioscience and applied chemistry. Submissions are solicited in all topical areas, ranging from basic aspects of the science materials to practical applications of such materials. With 6 issues per year, the first one published on the 15th of February of 2011, Biointerface Research in Applied Chemistry is an open-access journal, making all research results freely available online. The aim is to publish original papers, short communications as well as review papers highlighting interdisciplinary research, the potential applications of the molecules and materials in the bio-field. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible.