Asymmetric Synthesis of Enantiomerically Pure Aliphatic and Aromatic D-Amino Acids Catalyzed by Transaminase from <i>Haliscomenobacter hydrossis</i>
D-amino acids are valuable building blocks for the synthesis of biologically active compounds and pharmaceuticals. The asymmetric synthesis of chiral amino acids from prochiral ketones using stereoselective enzymes is a well-known but far from exhausted approach for large-scale production. Herein, w...
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MDPI AG
2022-12-01
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| Online Access: | https://www.mdpi.com/2073-4344/12/12/1551 |
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| author | Alina K. Bakunova Tatiana Y. Isaikina Vladimir O. Popov Ekaterina Yu. Bezsudnova |
| author_facet | Alina K. Bakunova Tatiana Y. Isaikina Vladimir O. Popov Ekaterina Yu. Bezsudnova |
| author_sort | Alina K. Bakunova |
| collection | DOAJ |
| description | D-amino acids are valuable building blocks for the synthesis of biologically active compounds and pharmaceuticals. The asymmetric synthesis of chiral amino acids from prochiral ketones using stereoselective enzymes is a well-known but far from exhausted approach for large-scale production. Herein, we investigated a pyridoxal-5′-phosphate-dependent D-amino acid transaminase from <i>Haliscomenobacter hydrossis</i> as a potential biocatalyst for the enzymatic asymmetric synthesis of optically pure aliphatic and aromatic D-amino acids. We studied the catalytic efficiency and stereoselectivity of transaminase from <i>H. hydrossis</i> in the amination of aliphatic and aromatic α-keto acids, using D-glutamate as a source of the amino group. We constructed a one-pot three-enzyme system, which included transaminase and two auxiliary enzymes, hydroxyglutarate dehydrogenase, and glucose dehydrogenase, to produce D-amino acids with a product yield of 95–99% and an enantiomeric excess of more than 99%. We estimated the stability of the transaminase and the cofactor leakage under reaction conditions. It was found that a high concentration of α-keto acids as well as a low reaction temperature (30 °C) can reduce the cofactor leakage under reaction conditions. The obtained results demonstrated the efficiency of transaminase from <i>H. hydrossis</i> in the asymmetric synthesis of enantiomerically pure D-amino acids. |
| first_indexed | 2024-03-09T17:13:36Z |
| format | Article |
| id | doaj.art-afc58eac9ec840ae892ec77a146f3d2c |
| institution | Directory Open Access Journal |
| issn | 2073-4344 |
| language | English |
| last_indexed | 2024-03-09T17:13:36Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Catalysts |
| spelling | doaj.art-afc58eac9ec840ae892ec77a146f3d2c2023-11-24T13:50:54ZengMDPI AGCatalysts2073-43442022-12-011212155110.3390/catal12121551Asymmetric Synthesis of Enantiomerically Pure Aliphatic and Aromatic D-Amino Acids Catalyzed by Transaminase from <i>Haliscomenobacter hydrossis</i>Alina K. Bakunova0Tatiana Y. Isaikina1Vladimir O. Popov2Ekaterina Yu. Bezsudnova3Bach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, RussiaBach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, RussiaBach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, RussiaBach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, Bld. 2, Moscow 119071, RussiaD-amino acids are valuable building blocks for the synthesis of biologically active compounds and pharmaceuticals. The asymmetric synthesis of chiral amino acids from prochiral ketones using stereoselective enzymes is a well-known but far from exhausted approach for large-scale production. Herein, we investigated a pyridoxal-5′-phosphate-dependent D-amino acid transaminase from <i>Haliscomenobacter hydrossis</i> as a potential biocatalyst for the enzymatic asymmetric synthesis of optically pure aliphatic and aromatic D-amino acids. We studied the catalytic efficiency and stereoselectivity of transaminase from <i>H. hydrossis</i> in the amination of aliphatic and aromatic α-keto acids, using D-glutamate as a source of the amino group. We constructed a one-pot three-enzyme system, which included transaminase and two auxiliary enzymes, hydroxyglutarate dehydrogenase, and glucose dehydrogenase, to produce D-amino acids with a product yield of 95–99% and an enantiomeric excess of more than 99%. We estimated the stability of the transaminase and the cofactor leakage under reaction conditions. It was found that a high concentration of α-keto acids as well as a low reaction temperature (30 °C) can reduce the cofactor leakage under reaction conditions. The obtained results demonstrated the efficiency of transaminase from <i>H. hydrossis</i> in the asymmetric synthesis of enantiomerically pure D-amino acids.https://www.mdpi.com/2073-4344/12/12/1551D-amino acid transaminasesubstrate specificitysynthesis of D-amino acidsstereoselective amination of keto acidsPLP leakage |
| spellingShingle | Alina K. Bakunova Tatiana Y. Isaikina Vladimir O. Popov Ekaterina Yu. Bezsudnova Asymmetric Synthesis of Enantiomerically Pure Aliphatic and Aromatic D-Amino Acids Catalyzed by Transaminase from <i>Haliscomenobacter hydrossis</i> Catalysts D-amino acid transaminase substrate specificity synthesis of D-amino acids stereoselective amination of keto acids PLP leakage |
| title | Asymmetric Synthesis of Enantiomerically Pure Aliphatic and Aromatic D-Amino Acids Catalyzed by Transaminase from <i>Haliscomenobacter hydrossis</i> |
| title_full | Asymmetric Synthesis of Enantiomerically Pure Aliphatic and Aromatic D-Amino Acids Catalyzed by Transaminase from <i>Haliscomenobacter hydrossis</i> |
| title_fullStr | Asymmetric Synthesis of Enantiomerically Pure Aliphatic and Aromatic D-Amino Acids Catalyzed by Transaminase from <i>Haliscomenobacter hydrossis</i> |
| title_full_unstemmed | Asymmetric Synthesis of Enantiomerically Pure Aliphatic and Aromatic D-Amino Acids Catalyzed by Transaminase from <i>Haliscomenobacter hydrossis</i> |
| title_short | Asymmetric Synthesis of Enantiomerically Pure Aliphatic and Aromatic D-Amino Acids Catalyzed by Transaminase from <i>Haliscomenobacter hydrossis</i> |
| title_sort | asymmetric synthesis of enantiomerically pure aliphatic and aromatic d amino acids catalyzed by transaminase from i haliscomenobacter hydrossis i |
| topic | D-amino acid transaminase substrate specificity synthesis of D-amino acids stereoselective amination of keto acids PLP leakage |
| url | https://www.mdpi.com/2073-4344/12/12/1551 |
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