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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
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- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제30권 제1호
- 발행연도
- 2020.1
- 수록면
- 146 - 154 (9page)
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With the advantages of biocatalytic method, enzymes have been excavated for the synthesis of chiral amino acids by the reductive amination of ketones, offering a promising way of producing pharmaceutical intermediates. In this work, a robust phenylalanine dehydrogenase (PheDH) with wide substrate spectrum and high catalytic efficiency was constructed through rational design and active-site-targeted, site-specific mutagenesis by using the parent enzyme from Bacillus halodurans. Active sites with bonding substrate and amino acid residues surrounding the substrate binding pocket, 49L-50G-51G, 74M,77K, 122G-123T-124D-125M, 275N, 305L and 308V of the PheDH, were identified. Noticeably, the new mutant PheDH (E113D-N276L) showed approximately 6.06-fold increment of kcat/Km in the oxidative deamination and more than 1.58-fold in the reductive amination compared to that of the wide type. Meanwhile, the PheDHs exhibit high capacity of accepting benzylic and aliphatic ketone substrates. The broad specificity, high catalytic efficiency and selectivity, along with excellent thermal stability, render these broad-spectrum enzymes ideal targets for further development with potential diagnostic reagent and pharmaceutical compounds applications.
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참고문헌 (46)
참고문헌 신청
Abrahamson MJ / 2012 / Development of an amine dehydrogenase for synthesis of chiral amines / Angew. Chem. Int. Ed Engl 51:3969~3972
Ye LJ / 2015 / Engineering of amine dehydrogenase for asymmetric reductive amination of ketone by evolving Rhodococcus phenylalanine dehydrogenase / ACS Catal 5:1119~1122
Helmchen G / 2000 / Phosphinooxazolines a new class of versatile, modular P, N-ligands for asymmetric catalysis / Acc. Chem. Res 33:336~345
Sheldon RA / 2017 / Biocatalysis engineering : the big picture / Chem. Soc. Rev 46:2678~2691
Sun H / 2017 / Biocatalysis for the synthesis of pharmaceuticals and pharmaceutical intermediates / Bioorg Med Chem 26:1275~1284
Ye LJ / 2015 / Engineering of amine dehydrogenase for asymmetric reductive amination of ketone by evolving Rhodococcus phenylalanine dehydrogenase / ACS Catal 5:1119~1122
Helmchen G / 2000 / Phosphinooxazolines a new class of versatile, modular P, N-ligands for asymmetric catalysis / Acc. Chem. Res 33:336~345
Sheldon RA / 2017 / Biocatalysis engineering : the big picture / Chem. Soc. Rev 46:2678~2691
Sun H / 2017 / Biocatalysis for the synthesis of pharmaceuticals and pharmaceutical intermediates / Bioorg Med Chem 26:1275~1284
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