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KMID : 1094720160210010153
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Biotechnology and Bioprocess Engineering
2016 Volume.21 No. 1 p.153 ~ p.159
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Structural-based screening of L-phenylglycine aminotransferase using L-phenylalanine as the optimal amino donor: Recycling of L-phenylalanine to produce L-phenylglycine
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Liu Shuang-Ping
Liu Rui Xia
Mao Jian
Zhang Liang
Ding Zhong Yang
Gu Zheng Hua
Shi Gui Yang
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Abstract
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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.
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KEYWORD
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L-phenylglycine aminotransferase, L-phenylalanine, structural-based screening, biocatalysis
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