Genomic data mining reveals the transaminase repertoire of Komagataella phaffii (Pichia pastoris) strain GS115 and supports a systematic nomenclature
Genomic data mining reveals the transaminase repertoire of Komagataella phaffii (Pichia pastoris) strain GS115 and supports a systematic nomenclature
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DOI：
10.1007/s12041-020-01201-1
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发表时间：
2020-06-03
影响因子：
1.5
通讯作者：
Nesbeth, Darren Nicholas
中科院分区：
文献类型：
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作者：
Henriquez, Maria-Jose;Cardos-Elena, Rosalia Paula;Nesbeth, Darren Nicholas
转氨酶是工业上重要的一类酶，因为它们能够催化用于产生手性胺的胺化反应，并且是小分子药物的关键构件。我们分析了菌株GS115的甲基营养型酵母Komagataella phaffii，以前称为毕赤酵母，以确定转氨酶基因，并提出了一个系统的命名的基础上，遗传学和结构功能的特点。K. Phaffii是一种越来越有吸引力的工业宿主细胞，这是由于其能够使用甲醇作为碳源和转基因表达的诱导剂生长到高生物量，高达60%湿细胞重量/体积。基于序列相似性和隐马尔可夫模型，将39个UniProt数据库命中减少到19个。在这19个基因中，3个（KpTam I-II.1b、KpTam I-II.7和KpTam V.2）的开放阅读框与没有特征性蛋白具有很强的同源性，4个（KpTam III.1a、KpTam III.1b、KpTam III.2a和KpTam III.2b）与omega型转氨酶具有相对较高的序列相似性，omega型转氨酶是一种通常接受最广泛底物的亚型。与酿酒酵母S288 C的比较表明KpTam I-II.1b和KpTam I-II.7的功能。K. phaffii GS 115最初是通过诱变K. phaffii CBS 7435的突变体，比较结果表明，在该诱变过程中可能缺失了一个转氨酶基因。这些见解可以推进对酵母生物学的基本理解，并可以为酵母转氨酶的工业筛选和工程提供信息。
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 Komagataella phaffii, formerly known as Pichia pastoris, to identify the transaminase genes and propose a systematic nomenclature based on both phylogeny and structuro-functional features. K. phaffii 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 protein and four (KpTam III.1a, KpTam III.1b, KpTam III.2a and KpTam III.2b) had relatively high sequence similarity to omega-type transaminases, a subtype that typically accepts the broadest range of substrates. Comparison with Saccharomyces cerevisiae S288C suggested functions for KpTam I-II.1b and KpTam I-II.7. K. phaffii GS115 was originally generated by mutagenesis of K. phaffii 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.