Biosynthesis of Phenylglyoxylic Acid by LhDMDH, a Novel d -Mandelate Dehydrogenase with High Catalytic Activity
| Autor: | Hong-Ling Shi, Fei Liu, Zhu-Jin Jiao, Hongfei Shi, Tang Cunduo, Kan Yunchao, Jian-He Xu, Lun-Guang Yao, Peng-Ju Ding |
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| Rok vydání: | 2018 |
| Předmět: |
0106 biological sciences
0301 basic medicine Phenylglyoxylic acid Stereochemistry Dehydrogenase 01 natural sciences Chemical synthesis Cofactor 03 medical and health sciences chemistry.chemical_compound Bacterial Proteins 010608 biotechnology Enzyme kinetics biology Glyoxylates General Chemistry Mandelic acid Chiral resolution Alcohol Oxidoreductases Kinetics Lactobacillus 030104 developmental biology chemistry Biocatalysis biology.protein Mandelic Acids NAD+ kinase General Agricultural and Biological Sciences |
| Zdroj: | Journal of Agricultural and Food Chemistry. 66:2805-2811 |
| ISSN: | 1520-5118 0021-8561 |
| DOI: | 10.1021/acs.jafc.7b05835 |
| Popis: | d-Mandelate dehydrogenase (DMDH) has the potential to convert d-mandelic acid to phenylglyoxylic acid (PGA), which is a key building block in the field of chemical synthesis and is widely used to synthesize pharmaceutical intermediates or food additives. A novel NAD+-dependent d-mandelate dehydrogenase was cloned from Lactobacillus harbinensi (LhDMDH) by genome mining and expressed in Escherichia coli BL21. After being purified to homogeneity, the oxidation activity of LhDMDH toward d-mandelic acid was approximately 1200 U·mg–1, which was close to four times the activity of the probe. Meanwhile, the kcat/Km value of LhDMDH was 28.80 S–1·mM–1, which was distinctly higher than the probe. By coculturing two E. coli strains expressing LhDMDH and LcLDH, we developed a system for the efficient synthesis of PGA, achieving a 60% theoretical yield and 99% purity without adding coenzyme or cosubstrate. Our data supports the implementation of a promising strategy for the chiral resolution of racemic mandelic acid an... |
| Databáze: | OpenAIRE |
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