Abstract
D-phenyllactic acid is a value added chemical with potential uses in wide areas of industry such as antibiotics, biopolymers, and pharmaceutical syntheses. It can be reduced from phenylpyruvic acid by various 2-hydroxy acid dehydrogenases. In this work, the 2-hydroxy acid dehydrogenase from Oenococcus oeni has been expressed in Escherichia coli whole cell along with formate dehydrogenases from two difference sources, Candida boidinii and Pseudomonas species, for regeneration of NADH cofactor. This could enhance the conversion of the product up to 78%, 3.4-fold increase from the one without cofactor regeneration, demonstrating a possibility of an efficient D-phenyllactic acid production system. Structural analysis by molecular dynamics simulation indicated the flexibility of the enzyme was lowered when the bound substrate was phenylpyruvic acid, compared to the natural substrate, pyruvate. This can be exploited to design 2-hydroxy acid dehydrogenase to increase the flexibility for phenylpyruvic acid, in order to further improve the production of D-phenyllactic acid.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- cbFDH:
-
Formate dehydrogenase from Candida boidinii
- psFDH:
-
Formate dehydrogenase from Pseudomonas species
- ooHADH:
-
2-hydroxy acid dehydrogenase from Oenococcus oeni
- D-PLA:
-
D-phenyllactic acid
- PPA:
-
Phenylpyruvic acid
- NADH:
-
Nicotinamide Adenine Dinucleotide, reduced form
- k cat :
-
Catalytic turnover number (unit: s−1)
- K M :
-
Michaelis-Menten constant (unit: mM)
- ΔGbind :
-
Gibbs free energy of binding between the enzyme and the substrate (unit: kcal mol−1)
References
Miltenberger, K. (2000) Hydroxycarboxylic acids, aliphatic. Ullmann’s encyclopedia of industrial chemistry. Wiley-VCH Verlag GmbH & Co. KGaA: Gersthofen, Germany.
Chen, L., Y. Bai, T. P. Fan, X. Zheng, and Y. Cai (2017) Characterization of a D-lactate dehydrogenase from Lactobacillus fermentum JN248 with high phenylpyruvate reductive activity. J. Food Sci. 82: 2269–2275.
Lavermicocca, P., F. Valerio, A. Evidente, S. Lazzaroni, A. Corsetti, and M. Gobbetti (2000) Purification and characterization of novel antifungal compounds from the sourdough Lactobacillus plantarum strain 21B. Appl. Environ. Microbiol. 66: 4084–4090.
Valerio, F., M. Favilla, P. De Bellis, A. Sisto, S. De Candia, and P. Lavermicocca (2009) Antifungal activity of strains of lactic acid bacteria isolated from a semolina ecosystem against Penicillium roqueforti, Aspergillus niger and Endomyces fibuliger contaminating bakery products. Syst. Appl. Microbiol. 32: 438–448.
Xu, G. C., L. L. Zhang, and Y. Ni (2016) Enzymatic preparation of D-phenyllactic acid at high space-time yield with a novel phenylpyruvate reductase identified from Lactobacillus sp. Cgmcc 9967. J. Biotechnol. 222: 29–37.
Zhou, L., Z. Zuo, and M. S. S. Chow (2005) Danshen: An overview of its chemistry, pharmacology, pharmacokinetics, and clinical use. J. Clin. Pharmacol. 45: 1345–1359.
Nguyen, H. D., X. Jin, D. Kaneko, and T. Kaneko (2011) Syntheses of high molecular weight poly(L-phenyllactic acid)s by a direct polycondensation in the presence of stable lewis acids. Chem. Lett. 40: 584–585.
Sohn, Y. J., H. T. Kim, S. Y. Jo, H. M. Song, K. A. Baritugo, J. Pyo, J. Choi, J. C. Joo, and S. J. Park (2020) Recent advances in systems metabolic engineering strategies for the production of biopolymers. Biotechnol. Bioprocess Eng. 25: 848–861.
Yu, S., H. Jiang, B. Jiang, and W. Mu (2012) Characterization of D-lactate dehydrogenase producing D-3-phenyllactic acid from Pediococcus pentosaceus. Biosci. Biotechnol. Biochem. 76: 853–855.
Jia, J., W. Mu, T. Zhang, and B. Jiang (2010) Bioconversion of phenylpyruvate to phenyllactate: Gene cloning, expression, and enzymatic characterization of D- and L1-lactate dehydrogenases from Lactobacillus plantarum SK002. Appl. Biochem. Biotechnol. 162: 242–251.
Mu, W., S. Yu, L. Zhu, T. Zhang, and B. Jiang (2012) Recent research on 3-phenyllactic acid, a broad-spectrum antimicrobial compound. Appl. Microbiol. Biotechnol. 95: 1155–1163.
Zheng, Z., B. Sheng, C. Gao, H. Zhang, T. Qin, C. Ma, and P. Xu (2013) Highly stereoselective biosynthesis of (R)-alpha-hydroxy carboxylic acids through rationally re-designed mutation of dlactate dehydrogenase. Sci. Rep. 3: 3401.
Wang, M., L. Zhu, X. Xu, L. Wang, R. Yin, and B. Yu (2016) Efficient production of enantiomerically pure D-phenyllactate from phenylpyruvate by structure-guided design of an engineered D-lactate dehydrogenase. Appl. Microbiol. Biotechnol. 100: 7471–7478.
Zheng, Z., C. Ma, C. Gao, F. Li, J. Qin, H. Zhang, K. Wang, and P. Xu (2011) Efficient conversion of phenylpyruvic acid to phenyllactic acid by using whole cells of Bacillus coagulans sdm. PLoS One. 6: e19030.
Razeto, A., S. Kochhar, H. Hottinger, M. Dauter, K. S. Wilson, and V. S. Lamzin (2002) Domain closure, substrate specificity and catalysis of D-lactate dehydrogenase from Lactobacillus bulgaricus. J. Mol. Biol. 318: 109–119.
Lee, H. S., J. Park, Y. J. Yoo, and Y. J. Yeon (2019) A novel D-2-hydroxy acid dehydrogenase with high substrate preference for phenylpyruvate originating from lactic acid bacteria: Structural analysis on the substrate specificity. Enzyme Microb. Technol. 125: 37–44.
Berrios-Rivera, S. J., G. N. Bennett, and K. Y. San (2002) Metabolic engineering of Escherichia coli: Increase of nadh availability by overexpressing an nad(+)-dependent formate dehydrogenase. Metab. Eng. 4: 217–229.
Kaup, B., S. Bringer-Meyer, and H. Sahm (2004) Metabolic engineering of Escherichia coli: Construction of an efficient biocatalyst for d-mannitol formation in a whole-cell biotransformation. Appl. Microbiol. Biotechnol. 64: 333–339.
Sathesh-Prabu, C., K. S. Shin, G. H. Kwak, S. K. Jung, and S. K. Lee (2019) Microbial production of fatty acid via metabolic engineering and synthetic biology. Biotechnol. Bioprocess Eng. 24: 23–40.
Schrödinger Release 2018-4 Prime, Schrödinger, LLC, New York, NY, USA.
Schrödinger Release 2018-4 Ligprep, Schrödinger, LLC, New York, NY, USA.
Schrödinger Release 2018-4 Glide, Schrödinger, LLC, New York, NY, USA.
Mukherjee, J. and M. N. Gupta (2015) Increasing importance of protein flexibility in designing biocatalytic processes. Biotechnol. Rep. (Amst). 6: 119–123.
Acknowledgements
This research was supported by the Technology Development Program to Solve Climate Changes of the National Research Foundation (NRF) funded by the Ministry of Science and ICT (Grant Number: 2017M1A2A2087630).
This study was supported by 2017 Academic Research Support Program in Gangneung-Wonju National University. The authors declare no conflict of interest.
No ethical approval and no informed consent required.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Lee, W., Park, YT., Lim, S. et al. Efficient Production of Phenyllactic Acid by Whole-cell Biocatalysis with Cofactor Regeneration System. Biotechnol Bioproc E 26, 402–407 (2021). https://doi.org/10.1007/s12257-020-0270-8
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12257-020-0270-8