A metagenomics approach for new biocatalyst discovery: application to transaminases and the synthesis of allylic amines
A metagenomics approach for new biocatalyst discovery: application to transaminases and the synthesis of allylic amines
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DOI：
10.1039/c6gc02769e
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发表时间：
2017-02-21
期刊：
影响因子：
9.8
通讯作者：
Hailes, Helen C.
中科院分区：
文献类型：
--
作者：
Baud, Damien;Jeffries, Jack W. E.;Hailes, Helen C.
转氨酶在温和的水相反应条件下可持续合成胺方面具有巨大的潜力。在这里，宏基因组学挖掘策略已用于新的转氨酶发现。从口腔微生物组样品开始，进行DNA测序和生物信息学分析。随后，对从测序数据构建的连续读段文库进行计算机挖掘，鉴定出11种推定的III类转氨酶，这些转氨酶被克隆并过表达。使用了几种筛选方案，并选择了三种感兴趣的酶，这是由于对底物的活性覆盖了广泛的结构多样性。然后研究官能化肉桂醛的氨基转移用于生产有价值的胺结构单元。
Transaminase enzymes have significant potential for the sustainable synthesis of amines using mild aqueous reaction conditions. Here a metagenomics mining strategy has been used for new transaminase enzyme discovery. Starting from oral cavity microbiome samples, DNA sequencing and bioinformatics analyses were performed. Subsequent in silico mining of a library of contiguous reads built from the sequencing data identified 11 putative Class III transaminases which were cloned and overexpressed. Several screening protocols were used and three enzymes selected of interest due to activities towards substrates covering a wide structural diversity. Transamination of functionalized cinnamaldehydes was then investigated for the production of valuable amine building blocks.