Chinese Journal of Applied Chemistry ›› 2020, Vol. 37 ›› Issue (2): 168-174.DOI: 10.11944/j.issn.1000-0518.2020.02.190189

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

SHI Honglinga,WANG Wenjiea,LI Xianga,JIAO Zhujina,LIU Yueb,TANG Cunduoa*(),KAN Yunchaoa,YAO Lunguanga   

  1. aHenan 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,Nanyang,Henan 473061,China
    bState Key Laboratory of Automotive Biofuel Technology,Nanyang,Henan 473061,China
  • Received:2019-07-08 Accepted:2019-11-20 Published:2020-02-01 Online:2020-02-06
  • Contact: TANG Cunduo
  • Supported by:
    Supported 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)

Abstract:

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.

Key words: L-Phenylglycine, D-mandelate dehydrogenase, L-leucine dehydrogenase, biocatalysis, cascade reaction