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基因组数据挖掘揭示了()GS115 菌株的转氨酶系谱,并支持系统命名法。

Genomic data mining reveals the transaminase repertoire of () strain GS115 and supports a systematic nomenclature.

机构信息

Department of Biochemical Engineering, University College London, Bernard Katz Building, London WC1E 6BT, UK.

出版信息

J Genet. 2020;99.

PMID:32661202
Abstract

Transaminases are an industrially important class of enzyme, due to their ability to catalyse amination reactions for production of chiral amines, and are key building blocks of small molecule pharmaceuticals. We analysed the genome of strain GS115 of the methylotrophic yeast , formerly known as , to identify the transaminase genes and propose a systematic nomenclature based on both phylogeny and structuro-functional features. is an increasingly attractive industrial host cell due to its ability to grow to high biomass, up to 60% wet cell weight by volume, using methanol as carbon source and inducer of transgene expression. Thirty-nine UniProt database hits were reduced to 19 on the basis of sequence similarity and hidden Markov model. Of the 19 genes, the open-reading frames of three (KpTam I-II.1b, KpTam I-II.7 and KpTam V.2) had strong homology with no characterized proteinand four (KpTam III.1a, KpTam III.1b, KpTam III.2a and KpTam III.2b) had relatively high sequence similarity to x-type transaminases, a subtype that typically accepts the broadest range of substrates. Comparison with S288C suggested functions for KpTam I-II.1b and KpTam I-II.7. GS115 was originally generated by mutagenesis of CBS7435 and comparison revealed that one transaminase gene may have been deleted during this mutagenesis. These insights can advance fundamental understanding of yeast biology and can inform industrial screening and engineering of yeast transaminases.

摘要

转氨酶是一类具有工业重要性的酶,因为它们能够催化氨基化反应以生产手性胺,并且是小分子药物的关键构建块。我们分析了甲醇营养型酵母菌株 GS115 的基因组,以鉴定转氨酶基因,并基于系统发育和结构功能特征提出了系统命名法。由于其能够利用甲醇作为碳源和转基因表达的诱导剂生长到高生物量,体积可达 60%湿细胞重量,因此成为越来越有吸引力的工业宿主细胞。根据序列相似性和隐马尔可夫模型,从 UniProt 数据库中减少到 19 个。在 19 个基因中,三个(KpTam I-II.1b、KpTam I-II.7 和 KpTam V.2)的开放阅读框与无特征蛋白具有很强的同源性,四个(KpTam III.1a、KpTam III.1b、KpTam III.2a 和 KpTam III.2b)与 x 型转氨酶具有相对较高的序列相似性,x 型转氨酶通常接受最广泛的底物。与 S288C 的比较表明 KpTam I-II.1b 和 KpTam I-II.7 的功能。GS115 最初是通过 CBS7435 的诱变产生的,比较表明在诱变过程中可能删除了一个转氨酶基因。这些见解可以促进对酵母生物学的基本理解,并为酵母转氨酶的工业筛选和工程提供信息。

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