Structural-based screening of L-phenylglycine aminotransferase using L-phenylalanine as the optimal amino donor: Recycling of L-phenylalanine to produce L-phenylglycine
| Autor: | Rui Xia Liu, Liang Zhang, Gui Yang Shi, Zheng Hua Gu, Jian Mao, Zhong Yang Ding, Shuang Ping Liu |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification 010405 organic chemistry Stereochemistry Operon Biomedical Engineering Substrate (chemistry) Bioengineering Phenylalanine 01 natural sciences Applied Microbiology and Biotechnology 0104 chemical sciences Amino acid 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology Biochemistry chemistry Biosynthesis Biocatalysis Organic synthesis Binding site Biotechnology |
| Zdroj: | Biotechnology and Bioprocess Engineering. 21:153-159 |
| ISSN: | 1976-3816 1226-8372 |
| Popis: | The aproteinogenic amino acid, L-phenylglycine, is an important side chain building block for some drugs. It would be of great commercial and environmental value to biocatalyse L-phenylalanine to L-phenylglycine, and thus replace the organic synthesis method. To produce L-phenylglycine from L-phenylalanine, an L-phenylglycine aminotransferase was screened and characterized. HpgTAO showed high homology to α-aminoadipate aminotransferase. The L-phenylalanine binding site was near the residues S26, R401, N201, and G46 in HpgTAO, and L-phenylalanine formed a hydrogen bond with Asn20, which was similar to the substrate binding mechanism of α-aminoadipate aminotransferase. HpgTAO showed increased activity in alkalescent environment below 40°C. The kinetic analysis showed that L-phenylalanine had the highest affinity to HpgTAO, which ensured the recycle biosynthesis of Lphenylglycine from L-phenylalanine. To date, it was the only aminotransferase using L-phenylalanine as an optimal amino donor. The L-phenylglycine biocatalysis operon was also constructed by co-expressing the hmaS, hmo and hpgT by a single plasmid. The first in vitro conversion of L-phenylalanine to L-phenylglycine was achieved by directly using the L-phenylalanine fermentation broth as the raw material. |
| Databáze: | OpenAIRE |
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