A Rapid Method for the Selection of Amidohydrolases from Metagenomic Libraries by Applying Synthetic Nucleosides and a Uridine Auxotrophic Host
A Rapid Method for the Selection of Amidohydrolases from Metagenomic Libraries by Applying Synthetic Nucleosides and a Uridine Auxotrophic Host
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应用合成核苷和尿苷营养缺陷型宿主快速筛选超基因组文库中的氨基水解酶
DOI：
10.3390/catal10040445
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发表时间：
2020-04-01
期刊：
影响因子：
3.9
通讯作者：
Meskys, Rolandas
中科院分区：
文献类型：
--
作者：
Urbeliene, Nina;Meskiene, Rita;Meskys, Rolandas
在这项研究中，描述了一种快速、高通量地从超基因组中选择酰胺水解酶的方法。该方法是以尿苷营养缺陷型大肠杆菌DH10B菌株为基础，以N-4-苯甲酰基-2‘-脱氧胞苷为唯一尿苷来源，在最小微生物M9培养基上进行的。这里描述的方法允许选择独特的生物催化剂，例如来自激活信号共整合同源(Asch)家族的新的氨基水解酶和聚对苯二甲酸乙二酯水解酶(PETase)相关的酶。
In this study, the development of a rapid, high-throughput method for the selection of amide-hydrolysing enzymes from the metagenome is described. This method is based on uridine auxotrophic Escherichia coli strain DH10B increment pyrFEC and the use of N-4-benzoyl-2'-deoxycytidine as a sole source of uridine in the minimal microbial M9 medium. The approach described here permits the selection of unique biocatalysts, e.g., a novel amidohydrolase from the activating signal cointegrator homology (ASCH) family and a polyethylene terephthalate hydrolase (PETase)-related enzyme.