125971-93-9Relevant articles and documents
Enzymatic preparation of t-butyl-6-cyano-(3R, 5R)-dihydroxyhexanoate by a whole-cell biocatalyst co-expressing carbonyl reductase and glucose dehydrogenase
Wu, Xuri,Gou, Xudong,Chen, Yijun
, p. 104 - 110 (2015)
Statins are the most effective drugs for hyperlipidemia-related diseases by competitively inhibiting 3-hydroxy-3-methylglutaryl coenzyme A reductase. Because of the difficulty and environmental concerns associated with chemical preparation of the chiral diols of statin side chains, different biocatalytic approaches have been explored and the two-step bio-reduction process for the introduction of two chiral hydroxyl groups has been industrialized. However, the high costs and poor stability of nicotinamide cofactors in the process was a major limiting factor. In the present study, a whole-cell biocatalyst simultaneously expressing carbonyl reductase and glucose dehydrogenase was constructed. This biocatalyst was then used to synthesize t-butyl-6-cyano-(3R, 5R)-dihydroxyhexanoate via enzymatic reduction of t-butyl-6-cyano-(5R)-hydroxy-3-carboxylhexanoate, which involves in the self-recycling of endogenous cofactors. After systematic optimization, the bioconversion was complete with a productivity of 120 g l-1 day-1 without exogenous addition of cofactors after 7 h at 35 g/L substrate concentration. Thus, the present system has simplified the process and improved the overall efficiency for the preparation of statin side chains.
Co-evolution of activity and thermostability of an aldo-keto reductase KmAKR for asymmetric synthesis of statin precursor dichiral diols
Chen, Yi,Cheng, Feng,Jin, Ling-Jun,Li, Shu-Fang,Qiu, Shuai,Wang, Ya-Jun,Zheng, Yu-Guo
, (2020)
Aldo-keto reductase KmAKR-catalyzed asymmetric reduction offers a green approach to produce dichiral diol tert-butyl 6-substituted-(3R,5R/S)-dihydroxyhexanoates, which are important building blocks of statins. In our previous work, we cloned a novel gene of NADPH-specific aldo-keto reductase KmAKR (WT) from a thermotolerant yeast Kluyveromyces marxianus ZJB14056 and a mutant KmAKR-W297H/Y296W/K29H (Variant III) has been constructed and displayed strict diastereoselectivity towards tert-butyl 6-cyano-(5R)-hydroxy-3-oxohexanoate ((5R)-1) but moderate activity and stability. Herein, to further co-evolve its activity and thermostability, we performed semi-rational engineering of Variant III by using a combinational screening strategy, consisting of tertiary structure analysis, loop engineering, and alanine scanning. As results, the “best” variant KmAKR-W297H/Y296W/K29H/Y28A/T63M (Variant VI) was acquired, whose Km, kcat/Km towards (5R)-1 was 0.66 mM and 210.77 s?1 mM?1, respectively, with improved thermostability (half-life of 14.13 h at 40 °C). Combined with 1.5 g dry cell weight (DCW) L-1 Exiguobacterium sibiricum glucose dehydrogenase (EsGDH) for NADPH regeneration, 4.5 g DCW L-1 Variant VI completely reduced (5R)-1 of up to 450 g L?1 within 7.0 h at 40 °C, yielding the corresponding optically pure tert-butyl 6-cyano-(3R,5R)-dihydroxyhexanoate ((3R,5R)-3, >99.5% d.e.p) with a space–time yield (STY) of 1.24 kg L?1 day?1, and this was the highest level documented in literatures so far on substrate loading and STY of producing (3R,5R)-3. Besides (5R)-1, Variant VI displayed strong activity on tert-butyl 6-chloro-(5S)-hydroxy-3-oxohexanoate ((5S)-2). 4.5 g DCW L-1 Variant VI completely reduced 400 g L?1 (5S)-2, within 5.0 h at 40 °C, yielding optically pure tert-butyl 6-chloro-(3R,5S)-dihydroxyhexanoate ((3R,5S)-4, >99.5% d.e.p) with a STY of 1.34 kg L?1 day?1. In summary, Variant VI displayed industrial application potential in statins biomanufacturing.
Atorvastatin calcium intermediate as well as preparation method and application thereof
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, (2019/04/04)
The invention discloses an atorvastatin calcium intermediate as well as a preparation method and application thereof. A synthesis process of the intermediate is environmentally-friendly, simple to operate and low in EHS risk; raw materials are easy to obtain; a used chemical reagent is small in toxicity and low in cost; and the synthesis process is a green synthesis process suitable for the industrial production. Moreover, the intermediate provided by the invention is applied to the synthesis of atorvastatin calcium and a key intermediate thereof, the route is relatively short, the yield is high, the industrial production cost of the atorvastatin calcium is effectively reduced, and the atorvastatin calcium intermediate has a relatively high industrial application prospect.