Citation: SHI Hongling, WANG Wenjie, LI Xiang, JIAO Zhujin, LIU Yue, TANG Cunduo, KAN Yunchao, YAO Lunguang. Enantioselective Biosynthesis of L-Phenylglycine via Cascade Biocatalysis of D-Mandelate Dehydrogenase and L-Leucine Dehydrogenase[J]. Chinese Journal of Applied Chemistry, ;2020, 37(2): 168-174. doi: 10.11944/j.issn.1000-0518.2020.02.190189 shu

Enantioselective Biosynthesis of L-Phenylglycine via Cascade Biocatalysis of D-Mandelate Dehydrogenase and L-Leucine Dehydrogenase

SHI Hongling ^a,‡ ,
WANG Wenjie ^a ,
LI Xiang ^a ,
JIAO Zhujin ^a ,
LIU Yue ^b ,
TANG Cunduo ^a,b,, ,
KAN Yunchao ^a ,
YAO Lunguang ^a

a.
Henan Provincial Engineering Laboratory of Insect Bio-reactor and Henan Key Laboratory of Ecological Security for Water Source Region of Mid-line of South-to-North, Jiangnan University, Wuxi, Jiangsu 214122, China
b.
State Key Laboratory of Automotive Biofuel Technology, Nanyang, Henan 473061, China

Corresponding author: TANG Cunduo, tcd530@126.com

^‡

Received Date: 8 July 2019
Revised Date: 22 October 2019
Accepted Date: 20 November 2019

Fund Project: the State Key Laboratory of Motor Vehicle Biofuel Technology KFKT2018003Supported by the National Natural Science Foundation of China(No.3190110303, No.31870917), the State Key Laboratory of Motor Vehicle Biofuel Technology(No.KFKT2018003), and the Special Funded Projects of Nanyang Normal University(No.2018QN004)the National Natural Science Foundation of China 31870917the Special Funded Projects of Nanyang Normal University 2018QN004the National Natural Science Foundation of China 3190110303

Figures(7)

L-Phenylglycine is an important class of chiral non-natural amino acids, and can be used to synthesize a variety of important pharmaceutical intermediate. Exploiting the green synthesis process of phenylacetone acid has significant economic value. In this study, novel and highly active D-mandelate dehydrogenase (LhDMDH) and L-leucine dehydrogenase (EsLeuDH) are coupled to achieve bio-transformation of D-mandelic acid into L-phenylglycine on the premise of coenzyme circulation, and this reaction only requires a lower concentration of coenzyme. By optimizing the transformation conditions including added amount of enzyme, β-nicotinamide adenine dinucleotide (NAD⁺) concentration, NH₄⁺ concentration and substrate concentration, we obtain the most economical condition:200 mmol/L D-mandelic acid, 6.5 kU/L enzyme, 0.1 mmol/L NAD ⁺, 0.5 mol/L NH₄⁺ and 30℃ for 12 h. The product yield and enantionmeric excess (e.e.) value can reach more than 98% and 99% under the most economical condition, respectively. This transformation has large industrialization potential, and lays a solid foundation for achieving large-scale biosynthesis of L-phenylglycine.
- L-Phenylglycine,
- D-mandelate dehydrogenase,
- L-leucine dehydrogenase,
- biocatalysis,
- cascade reaction
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