Xu Guo-Chao, Zhang Ling-Ling, Ni Ye
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, China.
J Biotechnol. 2016 Mar 20;222:29-37. doi: 10.1016/j.jbiotec.2015.12.011. Epub 2015 Dec 19.
An NADH-dependent phenylpyruvate reductase (LaPPR) was identified through screening the shotgun library of Lactobacillus sp. CGMCC 9967. It belongs to D-3-phosphoglycerate dehydrogenase (PGDH) subfamily of 2-hydroxy acid dehydrogenase superfamily. LaPPR was stable at pH 6.5 and 30 °C, with a half-life of 152 h. LaPPR has a substrate preference towards aromatic to aliphatic keto acids, and various keto acids could be reduced into D-hydroxy acids with excellent enantioselectivity (>99%). By construction the coexpression system with glucose dehydrogenase, as much as 100 g L(-1) phenylpyruvic acid was asymmetrically reduced into D-phenyllactic acid with 91.3% isolation yield and 243 g L(-1) d(-1) productivity. The results suggest that LaPPR is a promising biocatalyst for the efficient synthesis of optically pure D-phenyllactic acid.
通过筛选嗜酸乳杆菌CGMCC 9967的鸟枪法文库，鉴定出一种NADH依赖的苯丙酮酸还原酶（LaPPR）。它属于2-羟基酸脱氢酶超家族的D-3-磷酸甘油酸脱氢酶（PGDH）亚家族。LaPPR在pH 6.5和30℃下稳定，半衰期为152小时。LaPPR对芳香族酮酸比对脂肪族酮酸具有底物偏好性，各种酮酸可以以优异的对映选择性（>99%）还原为D-羟基酸。通过构建与葡萄糖脱氢酶的共表达系统，高达100 g L(-1)的苯丙酮酸被不对称还原为D-苯乳酸，分离产率为91.3%，生产效率为243 g L(-1) d(-1)。结果表明，LaPPR是高效合成光学纯D-苯乳酸的一种有前景的生物催化剂。
