Identification of catalysis, substrate, and coenzyme binding sites and improvement catalytic efficiency of formate dehydrogenase from Candida boidinii
Abstract Formate dehydrogenases (FDHs) are continually used for the cofactor regeneration in biocatalysis and biotransformation with hiring NAD(P)H-dependent oxidoreductases. Major weaknesses of most native FDHs are their low activity and operational stability in the catalytic reaction. In this work...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
Jiang, Wei [verfasserIn] |
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| Format: |
Artikel |
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| Sprache: |
Englisch |
| Erschienen: |
2016 |
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| Schlagwörter: |
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| Anmerkung: |
© Springer-Verlag Berlin Heidelberg 2016 |
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| Übergeordnetes Werk: |
Enthalten in: Applied microbiology and biotechnology - Springer Berlin Heidelberg, 1984, 100(2016), 19 vom: 20. Mai, Seite 8425-8437 |
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| Übergeordnetes Werk: |
volume:100 ; year:2016 ; number:19 ; day:20 ; month:05 ; pages:8425-8437 |
| Links: |
|---|
| DOI / URN: |
10.1007/s00253-016-7613-6 |
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OLC2050780184 |
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| 520 | |a Abstract Formate dehydrogenases (FDHs) are continually used for the cofactor regeneration in biocatalysis and biotransformation with hiring NAD(P)H-dependent oxidoreductases. Major weaknesses of most native FDHs are their low activity and operational stability in the catalytic reaction. In this work, the FDH from Candida boidinii (CboFDH) was engineered in order to gain an enzyme with high activity and better operational stability. Through comparing and analyzing its spatial structure with other FDHs, the catalysis, substrate, and coenzyme binding sites of the CboFDH were identified. To improve its performance, amino acids, which concentrated on the enzyme active site or in the conserved $ NAD^{+} $ and substrate binding motif, were mutated. The mutant V120S had the highest catalytic efficiency (kcat/Km) with $ COONH_{4} $ as it enhanced the catalytic velocity (kcat) and kcat/Km 3.48-fold and 1.60-fold, respectively, than that of the wild type. And, the double-mutant V120S-N187D had the highest kcat/Km with $ NAD^{+} $ as it displayed an approximately 1.50-fold increase in kcat/Km. The mutants showed higher catalytic efficiency than other reported FDHs, suggesting that the mutation has achieved good results. The single and double mutants exhibited higher thermostability than the wild type. The structure-function relationship of single and double mutants was analyzed by homology models and site parsing. Asymmetric synthesis of L-tert-leucine was executed to evaluate the ability of cofactor regeneration of the mutants with about 100 % conversion rates. This work provides a helpful theoretical reference for the evolution of an enzyme in vitro and promotion of the industrial production of chiral compounds, e.g., amino acid and chiral amine. | ||
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10.1007/s00253-016-7613-6 doi (DE-627)OLC2050780184 (DE-He213)s00253-016-7613-6-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Jiang, Wei verfasserin aut Identification of catalysis, substrate, and coenzyme binding sites and improvement catalytic efficiency of formate dehydrogenase from Candida boidinii 2016 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2016 Abstract Formate dehydrogenases (FDHs) are continually used for the cofactor regeneration in biocatalysis and biotransformation with hiring NAD(P)H-dependent oxidoreductases. Major weaknesses of most native FDHs are their low activity and operational stability in the catalytic reaction. In this work, the FDH from Candida boidinii (CboFDH) was engineered in order to gain an enzyme with high activity and better operational stability. Through comparing and analyzing its spatial structure with other FDHs, the catalysis, substrate, and coenzyme binding sites of the CboFDH were identified. To improve its performance, amino acids, which concentrated on the enzyme active site or in the conserved $ NAD^{+} $ and substrate binding motif, were mutated. The mutant V120S had the highest catalytic efficiency (kcat/Km) with $ COONH_{4} $ as it enhanced the catalytic velocity (kcat) and kcat/Km 3.48-fold and 1.60-fold, respectively, than that of the wild type. And, the double-mutant V120S-N187D had the highest kcat/Km with $ NAD^{+} $ as it displayed an approximately 1.50-fold increase in kcat/Km. The mutants showed higher catalytic efficiency than other reported FDHs, suggesting that the mutation has achieved good results. The single and double mutants exhibited higher thermostability than the wild type. The structure-function relationship of single and double mutants was analyzed by homology models and site parsing. Asymmetric synthesis of L-tert-leucine was executed to evaluate the ability of cofactor regeneration of the mutants with about 100 % conversion rates. This work provides a helpful theoretical reference for the evolution of an enzyme in vitro and promotion of the industrial production of chiral compounds, e.g., amino acid and chiral amine. Biocatalyst Molecular modeling Biocatalysis Chiral compounds Cofactor regeneration Lin, Peng aut Yang, Ruonan aut Fang, Baishan aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 100(2016), 19 vom: 20. Mai, Seite 8425-8437 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:100 year:2016 number:19 day:20 month:05 pages:8425-8437 https://doi.org/10.1007/s00253-016-7613-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4277 GBV_ILN_4305 AR 100 2016 19 20 05 8425-8437 |
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10.1007/s00253-016-7613-6 doi (DE-627)OLC2050780184 (DE-He213)s00253-016-7613-6-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Jiang, Wei verfasserin aut Identification of catalysis, substrate, and coenzyme binding sites and improvement catalytic efficiency of formate dehydrogenase from Candida boidinii 2016 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2016 Abstract Formate dehydrogenases (FDHs) are continually used for the cofactor regeneration in biocatalysis and biotransformation with hiring NAD(P)H-dependent oxidoreductases. Major weaknesses of most native FDHs are their low activity and operational stability in the catalytic reaction. In this work, the FDH from Candida boidinii (CboFDH) was engineered in order to gain an enzyme with high activity and better operational stability. Through comparing and analyzing its spatial structure with other FDHs, the catalysis, substrate, and coenzyme binding sites of the CboFDH were identified. To improve its performance, amino acids, which concentrated on the enzyme active site or in the conserved $ NAD^{+} $ and substrate binding motif, were mutated. The mutant V120S had the highest catalytic efficiency (kcat/Km) with $ COONH_{4} $ as it enhanced the catalytic velocity (kcat) and kcat/Km 3.48-fold and 1.60-fold, respectively, than that of the wild type. And, the double-mutant V120S-N187D had the highest kcat/Km with $ NAD^{+} $ as it displayed an approximately 1.50-fold increase in kcat/Km. The mutants showed higher catalytic efficiency than other reported FDHs, suggesting that the mutation has achieved good results. The single and double mutants exhibited higher thermostability than the wild type. The structure-function relationship of single and double mutants was analyzed by homology models and site parsing. Asymmetric synthesis of L-tert-leucine was executed to evaluate the ability of cofactor regeneration of the mutants with about 100 % conversion rates. This work provides a helpful theoretical reference for the evolution of an enzyme in vitro and promotion of the industrial production of chiral compounds, e.g., amino acid and chiral amine. Biocatalyst Molecular modeling Biocatalysis Chiral compounds Cofactor regeneration Lin, Peng aut Yang, Ruonan aut Fang, Baishan aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 100(2016), 19 vom: 20. Mai, Seite 8425-8437 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:100 year:2016 number:19 day:20 month:05 pages:8425-8437 https://doi.org/10.1007/s00253-016-7613-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4277 GBV_ILN_4305 AR 100 2016 19 20 05 8425-8437 |
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10.1007/s00253-016-7613-6 doi (DE-627)OLC2050780184 (DE-He213)s00253-016-7613-6-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Jiang, Wei verfasserin aut Identification of catalysis, substrate, and coenzyme binding sites and improvement catalytic efficiency of formate dehydrogenase from Candida boidinii 2016 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2016 Abstract Formate dehydrogenases (FDHs) are continually used for the cofactor regeneration in biocatalysis and biotransformation with hiring NAD(P)H-dependent oxidoreductases. Major weaknesses of most native FDHs are their low activity and operational stability in the catalytic reaction. In this work, the FDH from Candida boidinii (CboFDH) was engineered in order to gain an enzyme with high activity and better operational stability. Through comparing and analyzing its spatial structure with other FDHs, the catalysis, substrate, and coenzyme binding sites of the CboFDH were identified. To improve its performance, amino acids, which concentrated on the enzyme active site or in the conserved $ NAD^{+} $ and substrate binding motif, were mutated. The mutant V120S had the highest catalytic efficiency (kcat/Km) with $ COONH_{4} $ as it enhanced the catalytic velocity (kcat) and kcat/Km 3.48-fold and 1.60-fold, respectively, than that of the wild type. And, the double-mutant V120S-N187D had the highest kcat/Km with $ NAD^{+} $ as it displayed an approximately 1.50-fold increase in kcat/Km. The mutants showed higher catalytic efficiency than other reported FDHs, suggesting that the mutation has achieved good results. The single and double mutants exhibited higher thermostability than the wild type. The structure-function relationship of single and double mutants was analyzed by homology models and site parsing. Asymmetric synthesis of L-tert-leucine was executed to evaluate the ability of cofactor regeneration of the mutants with about 100 % conversion rates. This work provides a helpful theoretical reference for the evolution of an enzyme in vitro and promotion of the industrial production of chiral compounds, e.g., amino acid and chiral amine. Biocatalyst Molecular modeling Biocatalysis Chiral compounds Cofactor regeneration Lin, Peng aut Yang, Ruonan aut Fang, Baishan aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 100(2016), 19 vom: 20. Mai, Seite 8425-8437 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:100 year:2016 number:19 day:20 month:05 pages:8425-8437 https://doi.org/10.1007/s00253-016-7613-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4277 GBV_ILN_4305 AR 100 2016 19 20 05 8425-8437 |
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10.1007/s00253-016-7613-6 doi (DE-627)OLC2050780184 (DE-He213)s00253-016-7613-6-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Jiang, Wei verfasserin aut Identification of catalysis, substrate, and coenzyme binding sites and improvement catalytic efficiency of formate dehydrogenase from Candida boidinii 2016 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2016 Abstract Formate dehydrogenases (FDHs) are continually used for the cofactor regeneration in biocatalysis and biotransformation with hiring NAD(P)H-dependent oxidoreductases. Major weaknesses of most native FDHs are their low activity and operational stability in the catalytic reaction. In this work, the FDH from Candida boidinii (CboFDH) was engineered in order to gain an enzyme with high activity and better operational stability. Through comparing and analyzing its spatial structure with other FDHs, the catalysis, substrate, and coenzyme binding sites of the CboFDH were identified. To improve its performance, amino acids, which concentrated on the enzyme active site or in the conserved $ NAD^{+} $ and substrate binding motif, were mutated. The mutant V120S had the highest catalytic efficiency (kcat/Km) with $ COONH_{4} $ as it enhanced the catalytic velocity (kcat) and kcat/Km 3.48-fold and 1.60-fold, respectively, than that of the wild type. And, the double-mutant V120S-N187D had the highest kcat/Km with $ NAD^{+} $ as it displayed an approximately 1.50-fold increase in kcat/Km. The mutants showed higher catalytic efficiency than other reported FDHs, suggesting that the mutation has achieved good results. The single and double mutants exhibited higher thermostability than the wild type. The structure-function relationship of single and double mutants was analyzed by homology models and site parsing. Asymmetric synthesis of L-tert-leucine was executed to evaluate the ability of cofactor regeneration of the mutants with about 100 % conversion rates. This work provides a helpful theoretical reference for the evolution of an enzyme in vitro and promotion of the industrial production of chiral compounds, e.g., amino acid and chiral amine. Biocatalyst Molecular modeling Biocatalysis Chiral compounds Cofactor regeneration Lin, Peng aut Yang, Ruonan aut Fang, Baishan aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 100(2016), 19 vom: 20. Mai, Seite 8425-8437 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:100 year:2016 number:19 day:20 month:05 pages:8425-8437 https://doi.org/10.1007/s00253-016-7613-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4277 GBV_ILN_4305 AR 100 2016 19 20 05 8425-8437 |
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10.1007/s00253-016-7613-6 doi (DE-627)OLC2050780184 (DE-He213)s00253-016-7613-6-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Jiang, Wei verfasserin aut Identification of catalysis, substrate, and coenzyme binding sites and improvement catalytic efficiency of formate dehydrogenase from Candida boidinii 2016 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag Berlin Heidelberg 2016 Abstract Formate dehydrogenases (FDHs) are continually used for the cofactor regeneration in biocatalysis and biotransformation with hiring NAD(P)H-dependent oxidoreductases. Major weaknesses of most native FDHs are their low activity and operational stability in the catalytic reaction. In this work, the FDH from Candida boidinii (CboFDH) was engineered in order to gain an enzyme with high activity and better operational stability. Through comparing and analyzing its spatial structure with other FDHs, the catalysis, substrate, and coenzyme binding sites of the CboFDH were identified. To improve its performance, amino acids, which concentrated on the enzyme active site or in the conserved $ NAD^{+} $ and substrate binding motif, were mutated. The mutant V120S had the highest catalytic efficiency (kcat/Km) with $ COONH_{4} $ as it enhanced the catalytic velocity (kcat) and kcat/Km 3.48-fold and 1.60-fold, respectively, than that of the wild type. And, the double-mutant V120S-N187D had the highest kcat/Km with $ NAD^{+} $ as it displayed an approximately 1.50-fold increase in kcat/Km. The mutants showed higher catalytic efficiency than other reported FDHs, suggesting that the mutation has achieved good results. The single and double mutants exhibited higher thermostability than the wild type. The structure-function relationship of single and double mutants was analyzed by homology models and site parsing. Asymmetric synthesis of L-tert-leucine was executed to evaluate the ability of cofactor regeneration of the mutants with about 100 % conversion rates. This work provides a helpful theoretical reference for the evolution of an enzyme in vitro and promotion of the industrial production of chiral compounds, e.g., amino acid and chiral amine. Biocatalyst Molecular modeling Biocatalysis Chiral compounds Cofactor regeneration Lin, Peng aut Yang, Ruonan aut Fang, Baishan aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 100(2016), 19 vom: 20. Mai, Seite 8425-8437 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:100 year:2016 number:19 day:20 month:05 pages:8425-8437 https://doi.org/10.1007/s00253-016-7613-6 lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_70 GBV_ILN_130 GBV_ILN_267 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4277 GBV_ILN_4305 AR 100 2016 19 20 05 8425-8437 |
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identification of catalysis, substrate, and coenzyme binding sites and improvement catalytic efficiency of formate dehydrogenase from candida boidinii |
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Identification of catalysis, substrate, and coenzyme binding sites and improvement catalytic efficiency of formate dehydrogenase from Candida boidinii |
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Abstract Formate dehydrogenases (FDHs) are continually used for the cofactor regeneration in biocatalysis and biotransformation with hiring NAD(P)H-dependent oxidoreductases. Major weaknesses of most native FDHs are their low activity and operational stability in the catalytic reaction. In this work, the FDH from Candida boidinii (CboFDH) was engineered in order to gain an enzyme with high activity and better operational stability. Through comparing and analyzing its spatial structure with other FDHs, the catalysis, substrate, and coenzyme binding sites of the CboFDH were identified. To improve its performance, amino acids, which concentrated on the enzyme active site or in the conserved $ NAD^{+} $ and substrate binding motif, were mutated. The mutant V120S had the highest catalytic efficiency (kcat/Km) with $ COONH_{4} $ as it enhanced the catalytic velocity (kcat) and kcat/Km 3.48-fold and 1.60-fold, respectively, than that of the wild type. And, the double-mutant V120S-N187D had the highest kcat/Km with $ NAD^{+} $ as it displayed an approximately 1.50-fold increase in kcat/Km. The mutants showed higher catalytic efficiency than other reported FDHs, suggesting that the mutation has achieved good results. The single and double mutants exhibited higher thermostability than the wild type. The structure-function relationship of single and double mutants was analyzed by homology models and site parsing. Asymmetric synthesis of L-tert-leucine was executed to evaluate the ability of cofactor regeneration of the mutants with about 100 % conversion rates. This work provides a helpful theoretical reference for the evolution of an enzyme in vitro and promotion of the industrial production of chiral compounds, e.g., amino acid and chiral amine. © Springer-Verlag Berlin Heidelberg 2016 |
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Abstract Formate dehydrogenases (FDHs) are continually used for the cofactor regeneration in biocatalysis and biotransformation with hiring NAD(P)H-dependent oxidoreductases. Major weaknesses of most native FDHs are their low activity and operational stability in the catalytic reaction. In this work, the FDH from Candida boidinii (CboFDH) was engineered in order to gain an enzyme with high activity and better operational stability. Through comparing and analyzing its spatial structure with other FDHs, the catalysis, substrate, and coenzyme binding sites of the CboFDH were identified. To improve its performance, amino acids, which concentrated on the enzyme active site or in the conserved $ NAD^{+} $ and substrate binding motif, were mutated. The mutant V120S had the highest catalytic efficiency (kcat/Km) with $ COONH_{4} $ as it enhanced the catalytic velocity (kcat) and kcat/Km 3.48-fold and 1.60-fold, respectively, than that of the wild type. And, the double-mutant V120S-N187D had the highest kcat/Km with $ NAD^{+} $ as it displayed an approximately 1.50-fold increase in kcat/Km. The mutants showed higher catalytic efficiency than other reported FDHs, suggesting that the mutation has achieved good results. The single and double mutants exhibited higher thermostability than the wild type. The structure-function relationship of single and double mutants was analyzed by homology models and site parsing. Asymmetric synthesis of L-tert-leucine was executed to evaluate the ability of cofactor regeneration of the mutants with about 100 % conversion rates. This work provides a helpful theoretical reference for the evolution of an enzyme in vitro and promotion of the industrial production of chiral compounds, e.g., amino acid and chiral amine. © Springer-Verlag Berlin Heidelberg 2016 |
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Abstract Formate dehydrogenases (FDHs) are continually used for the cofactor regeneration in biocatalysis and biotransformation with hiring NAD(P)H-dependent oxidoreductases. Major weaknesses of most native FDHs are their low activity and operational stability in the catalytic reaction. In this work, the FDH from Candida boidinii (CboFDH) was engineered in order to gain an enzyme with high activity and better operational stability. Through comparing and analyzing its spatial structure with other FDHs, the catalysis, substrate, and coenzyme binding sites of the CboFDH were identified. To improve its performance, amino acids, which concentrated on the enzyme active site or in the conserved $ NAD^{+} $ and substrate binding motif, were mutated. The mutant V120S had the highest catalytic efficiency (kcat/Km) with $ COONH_{4} $ as it enhanced the catalytic velocity (kcat) and kcat/Km 3.48-fold and 1.60-fold, respectively, than that of the wild type. And, the double-mutant V120S-N187D had the highest kcat/Km with $ NAD^{+} $ as it displayed an approximately 1.50-fold increase in kcat/Km. The mutants showed higher catalytic efficiency than other reported FDHs, suggesting that the mutation has achieved good results. The single and double mutants exhibited higher thermostability than the wild type. The structure-function relationship of single and double mutants was analyzed by homology models and site parsing. Asymmetric synthesis of L-tert-leucine was executed to evaluate the ability of cofactor regeneration of the mutants with about 100 % conversion rates. This work provides a helpful theoretical reference for the evolution of an enzyme in vitro and promotion of the industrial production of chiral compounds, e.g., amino acid and chiral amine. © Springer-Verlag Berlin Heidelberg 2016 |
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