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

doi:10.11944/j.issn.1000-0518.2020.02.190189

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

• Full Papers • Previous Articles Next Articles

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^*(),KAN Yunchao^a,YAO Lunguang^a

^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

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

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.

References

[1]	Yasukawa K,Asano Y.Enzymatic Synthesis of Chiral Phenylalanine Derivatives by a Dynamic Kinetic Resolution of Corresponding Amide and Nitrile Substrates with a Multi-enzyme System[J]. Adv Synth Catal,2012,354(17):3327-3332.
[2]	Fan C W,Xu G C,Ma B D,et al. A Novel D-Mandelate Dehydrogenase Used in Three Enzyme Cascade Reaction for Highly Efficient Synthesis of Non-natural Chiral Amino Acids[J]. J Biotechnol,2015,195:67-71.
[3]	Wang Z M,Kolb H C,Sharpless K B.Large-Scale and Highly Enantioselective Synthesis of the Taxol C-13 Side Chain Through Asymmetric Dihydroxylation[J]. J Org Chem,1994,59(17):5104-5105.
[4]	Han S W,Shin J S.One-Pot Preparation of d-Amino Acids Through Biocatalytic Deracemization Using Alanine Dehydrogenase and ω-Transaminase[J]. Catal Lett,2018,148(12):3678-3684.
[5]	Resch V,Fabian W M F,Kroutil W. Deracemisation of Mandelic Acid to Optically Pure Non-natural L-Phenylglycine via a Redox-Neutral Biocatalytic Cascade[J]. Adv Synth Catal,2010,352(6):993-997.
[6]	FAN Changwei.Gene Mining of D-Mandelate Dehydrogenase and Its Application in the Synthesis of L-Phenylglycine by Multi-enzyme Conjugation[D]. East China University of Science and Technology,2014(in Chinese). 范长伟. D-扁桃酸脱氢酶的基因挖掘及在多酶偶联合成L-苯甘氨酸的应用[D].华东理工大学,2014(in Chinese).
[7]	Bornscheuer U T,Huisman G W,Kazlauskas R J,et al. Engineering the Third Wave of Biocatalysis[J]. Nature,2012,485(7397):185-194.
[8]	TANG Cunduo,SHI Hongling,HE Zihan,et al. Green Biosynthesis of Phenylglyoxylic Acid by Biotransformation Using Recombinant Escherichia Coli Whole Cells[J]. CIESC J,2018,69(6):2627-2631(in Chinese). 唐存多,史红玲,和子涵,等. 重组大肠杆菌全细胞用于苯乙酮酸的绿色生物合成[J]. 化工学报,2018,69(6):2627-2631.
[9]	Tang C D,Shi H L,Xu J H,et al. Biosynthesis of Phenylglyoxylic Acid by LhDMDH, a Novel D-Mandelate Dehydrogenase with High Catalytic Activity[J]. J Agric Food Chem,2018,66(11):2805-2811.
[10]	Gokhale D V,Bastawde K B,Patil S G,et al. Chemoenzymatic Synthesis of D-Phenylglycine Using Hydantoinase of Pseudomonas Desmolyticum Resting Cells[J]. Enzyme Microb Technol,1996,18(5):353-357.
[11]	Wiyakrutta S,Meevootisom V.A Stereo-Inverting D-Phenylglycine Aminotransferase from Pseudomonas Stutzeri ST-201:Purification, Characterization and Application for D-Phenylglycine Synthesis[J]. J Biotechnol,1997,55(3):193-203.
[12]	Mast Y J,Wohlleben W,Schinko E.Identification and Functional Characterization of Phenylglycine Biosynthetic Genes Involved in Pristinamycin Biosynthesis in Streptomyces Pristinaespiralis[J]. J Biotechnol,2011,155(1):63-67.
[13]	Li D,Zeng Z,Yang J,et al. Mandelate Racemase and Mandelate Dehydrogenase Coexpressed Recombinant Escherichia Coli in the Synthesis of Benzoylformate[J]. Biosci Biotechnol Biochem,2013,77(6):1236-9.
[14]	Chen F F,Liu Y Y,Zheng G W,et al. Asymmetric Amination of Secondary Alcohols by Using a Redox-Neutral Two-Enzyme Cascade[J]. ChemCatChem,2015,7(23):3838-3841.
[15]	TANG Cunduo,SHI Hongling,MA Yue,et al. Gene Mining, Expression and Characterization of Novel R-Mandelate Dehydrogenases[J]. China Biotechnol,2018,38(2):30-37(in Chinese). 唐存多,史红玲,马越,等. 新型R-扁桃酸脱氢酶的基因挖掘及表达鉴定[J]. 中国生物工程杂志,2018,38(2):30-37.
[16]	Gong X M,Qin Z,Li F L,et al. Development of an Engineered Ketoreductase with Simultaneously Improved Thermostability and Activity for Making a Bulky Atorvastatin Precursor[J]. ACS Catal,2019,9(1):147-153.
[17]	Zhou J,Wang Y,Xu G,et al. Structural Insight into Enantioselective Inversion of an Alcohol Dehydrogenase Reveals a “Polar Gate” in Stereorecognition of Diaryl Ketones[J]. J Am Chem Soc,2018,140(39):12645-12654.
[18]	Tang C D,Shi H L,Jiao Z J,et al. Exploitation of Cold-Active Cephalosporin C Acylase by Computer-Aided Directed Evolution and Its Potential Application in Low-Temperature Biosynthesis of 7-Aminocephalosporanic Acid[J]. J Chem Technol Biotechnol,2018,93(10):2925-2930.

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

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 1

Recommended Articles

Metrics

Comments