文献类型： 外文期刊

作者： Jia, Yuan-Yuan ¹ ; Xie, Yu-Li ¹ ; Yang, Lu-Lu ¹ ; Shi, Hong-Ling ¹ ; Lu, Yun-Feng ¹ ; Zhang, Si-Pu ³ ; Tang, Cun-Duo ¹ ;

作者机构： 1.Nanyang Normal Univ, Henan Prov Engn Lab Insect Bioreactor, Henan Key Lab Ecol Secur Water Source Reg Midline, Nanyang, Peoples R China

2.Nanyang Normal Univ, Sch Life Sci & Agr Engn, Nanyang, Peoples R China

3.Henan Acad Agr Sci, Zhengzhou, Peoples R China

关键词： leucine dehydrogenase; feed additive; expression; biocatalysis; formate dehydrogenase

期刊名称：FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY （ 影响因子：3.644； ）

ISSN： 2296-4185

年卷期： 2021 年 9 卷

页码：

收录情况： SCI

摘要： Leucine dehydrogenase (LDH) is a NAD(+)-dependent oxidoreductase, which can selectively catalyze alpha-keto acids to obtain alpha-amino acids and their derivatives. It plays a key role in the biosynthesis of L-tert-leucine (L-Tle). As a non-naturally chiral amino acid, L-Tle can be used as an animal feed additive, nutrition fortifier, which is a perspective and important building block in the pharmaceutical, cosmetic, and food additive industry. In this study, four hypothetical leucine dehydrogenases were discovered by using genome mining technology, using the highly active leucine dehydrogenase LsLeuDH as a probe. These four leucine dehydrogenases were expressed in Escherichia coli BL21(DE3), respectively, and purified to homogeneity and characterized. Compared with the other enzymes, the specific activity of PfLeuDH also shows stronger advantage. In addition, the highly selective biosynthesis of L-Tle from trimethylpyruvic acid (TMP) was successfully carried out by whole-cell catalysis using engineered E. coli cells as biocatalyst, which can efficiently coexpress leucine dehydrogenase and formate dehydrogenase. One hundred-millimolar TMP was catalyzed for 25 h, and the yield and space-time yield of L-Tle reached 87.38% (e.e. >99.99%) and 10.90 g L-1 day(-1). In short, this research has initially achieved the biosynthesis of L-Tle, laying a solid foundation for the realization of low-cost and large-scale biosynthesis of L-Tle.