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Engineering of Candida glabrata Ketoreductase 1 for Asymmetric Reduction of α-Halo Ketones

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논문

Engineering of Candida glabrata Ketoreductase 1 for Asymmetric Reduction of α-Halo Ketones

학술지

ACS catalysis

저자명

Qin, Fengyu; Qin, Bin; Mori, Takahiro; Wang, Yan; Meng, Lingxin; Zhang, Xin; Jia, Xian; Abe, Ikuro; You, Song

초록

<P>Enantiopure halohydrins, which are important building blocks for pharmaceutical agents, could be synthesized by biocatalytic reduction of &alpha;-halo ketones using ketoreductases. In this study, <I>Candida glabrata</I> ketoreductase 1 (CgKR1) variants with >99% stereoselectivity toward &alpha;-halo ketones, such as 2-chloroacetophenone, 2-chloro-4&prime;-fluoroacetophenone, and 2-bromoacetophenone, were obtained through engineering of CgKR1 at residues Phe92 and Tyr208. Interestingly, asymmetric reduction of these &alpha;-halo ketones by all the variants of CgKR1 followed anti-Prelog&rsquo;s rule, which is rarely found in natural ketoreductases. Moreover, the biocatalytic processes for reduction of these aromatic &alpha;-halo ketones with high substrate loading were achieved by coexpression of glucose dehydrogenase (GDH) for NADPH regeneration, indicating the potential of practical applications of these variants.</P><P><B>Graphic Abstract</B><BR><IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/accacs/2016/accacs.2016.6.issue-9/acscatal.6b01552/production/images/medium/cs-2016-01552q_0005.gif'></P><P><A href='http://pubs.acs.org/doi/suppl/10.1021/cs6b01552'>ACS Electronic Supporting Info</A></P>

발행연도

2016

발행기관

American Chemical Society

ISSN

2155-5435

6

9

페이지

pp.6135-6140

주제어

ketoreductases; protein engineering; asymmetric reduction; α-halo ketones; halohydrins

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논문; 2016-12-31

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