4.2 Article

Sequencing, cloning and high-yield expression of a fungal β-N-acetylhexosaminidase in Pichia pastoris

期刊

PROTEIN EXPRESSION AND PURIFICATION
卷 82, 期 1, 页码 212-217

出版社

ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2012.01.004

关键词

Talaromyces flavus; Pichia pastoris; Yeast expression system; beta-N-Acetylhexosaminidase; Gene sequencing; N-Deglycosylation

资金

  1. Czech Science Foundation [203/09/P024, 305/09/H008, P207/11/0629]
  2. EU [MSMT 7E11011]

向作者/读者索取更多资源

The beta-N-acetylhexosaminidase from Talaromyces flavus has a remarkable synthetic ability, processing even carbohydrates with various functionalities. Its broader use is partially hampered by low-yield production in the native fungus. Here, we present an optimized 3-day production of this enzyme in the eukaryotic host of Pichia pastoris, in ca 10-fold higher volume activity (10 U/ml) and close-to-perfect purity (one chromatographic step needed). Importantly, the recombinant enzyme features the same biochemical and catalytic properties, including the syntheses with derivatized carbohydrate substrates. This is the first example of the overexpression of a fungal beta-N-acetylhexosaminidase by a single-cell producer in liquid medium. It represents a promising solution for wider biotechnological applications of this outstanding enzyme.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.2
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据