Identification of efficient amine transaminase and applicability in dual transaminases cascade for synthesis of L-phosphinothricin

IF 3.4 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Enzyme and Microbial Technology Pub Date : 2024-08-22 DOI:10.1016/j.enzmictec.2024.110501

Puhong Yi , Mengdan Liu , Yuhua Hao , Ziwen Wang , Hanlin Liu , Xue Cai , Feng Cheng , Zhiqiang Liu , Yaping Xue , Liqun Jin , Yuguo Zheng

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Abstract

L-phosphinothricin (L-PPT) is the most popular broad-spectrum and highly effective herbicide. Transaminases (TAs) play a pivotal role in asymmetric synthesis of L-PPT, yet encounter the challenge of unfavorable reaction equilibrium. In this study, the novel dual transaminases cascade system (DTCS) was introduced to facilitate the synthesis of L-PPT. The specific amine transaminase BdATA, originating from Bradyrhizobium diazoefficiens ZJY088, was screened and identified. It exhibited remarkable activity, good stability, and required only 2.5 equivalents of isopropylamine to transform pyruvate effectively. By coupling BdATA with previously reported SeTA to construct the DTCS for pyruvate removal in situ, the L-PPT yield escalated from 37.37 % to 85.35 %. Three advantages of the DTCS were presented: the removal of pyruvate alleviated by-product inhibition, the use of isopropylamine reduced reliance on excess L-alanine, and no demand for expensive cofactors like NAD(P)H. It demonstrated an innovative idea for addressing the challenges associated with transaminase-mediated synthesis of L-PPT.

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鉴定高效胺转氨酶并将其应用于合成 L-膦丝菌素的双转氨酶级联中

左旋膦菊酯（L-PPT）是最常用的广谱高效除草剂。转氨酶（TAs）在 L-PPT 的不对称合成中起着关键作用，但却面临着不利反应平衡的挑战。本研究引入了新型双转氨酶级联系统（DTCS）来促进 L-PPT 的合成。筛选并鉴定了来源于 Bradyrhizobium diazoefficiens ZJY088 的特异性胺转氨酶 BdATA。它表现出极高的活性和良好的稳定性，只需要 2.5 等量的异丙基胺就能有效转化丙酮酸。通过将 BdATA 与之前报道的 SeTA 联用，构建了用于原位去除丙酮酸的 DTCS，L-PPT 产率从 37.37% 上升到 85.35%。DTCS 有三个优点：丙酮酸的去除减轻了副产物抑制；异丙胺的使用减少了对过量 L-丙氨酸的依赖；不需要 NAD(P)H 等昂贵的辅助因子。它为解决转氨酶介导的 L-PPT 合成所面临的挑战提供了一种创新思路。

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来源期刊

Enzyme and Microbial Technology 生物-生物工程与应用微生物

CiteScore

7.60

自引率

5.90%

发文量

142

审稿时长

38 days

期刊介绍： Enzyme and Microbial Technology is an international, peer-reviewed journal publishing original research and reviews, of biotechnological significance and novelty, on basic and applied aspects of the science and technology of processes involving the use of enzymes, micro-organisms, animal cells and plant cells. We especially encourage submissions on: Biocatalysis and the use of Directed Evolution in Synthetic Biology and Biotechnology Biotechnological Production of New Bioactive Molecules, Biomaterials, Biopharmaceuticals, and Biofuels New Imaging Techniques and Biosensors, especially as applicable to Healthcare and Systems Biology New Biotechnological Approaches in Genomics, Proteomics and Metabolomics Metabolic Engineering, Biomolecular Engineering and Nanobiotechnology Manuscripts which report isolation, purification, immobilization or utilization of organisms or enzymes which are already well-described in the literature are not suitable for publication in EMT, unless their primary purpose is to report significant new findings or approaches which are of broad biotechnological importance. Similarly, manuscripts which report optimization studies on well-established processes are inappropriate. EMT does not accept papers dealing with mathematical modeling unless they report significant, new experimental data.