首页 | 本学科首页   官方微博 | 高级检索  
文章检索
  按 检索   检索词:      
出版年份:   被引次数:   他引次数: 提示:输入*表示无穷大
  收费全文   5篇
  免费   0篇
  国内免费   1篇
  2020年   1篇
  2019年   1篇
  2014年   1篇
  2010年   1篇
  2008年   1篇
  2006年   1篇
排序方式: 共有6条查询结果,搜索用时 15 毫秒
1
1.
Previously we have demonstrated the reduction of ethyl and t-butyl diketoesters 1 to the corresponding syn-(3R,5S)-dihydroxy esters 2a by Acinetobacter sp. 13874. The syn-(3R,5S)-dihydroxy ester 2a was obtained with an enantiomeric excess (e.e.) of 99% and a diastereomeric excess (de) of 63%. In this report, we identified a gene encoding desired ketoreductase III which catalyzed the diastereoselective reduction of diketoesters 1 to syn-(3R,5S)-dihydroxy esters 2a and describe cloning and expression of ketoreductase III into Escherichia coli. Cells or extracts of recombinant E. coli efficiently reduced the diketoester 1 to the corresponding syn-(3R,5S)-dihydroxy ester 2a in 99.3% yield, 100% e.e., and 99.8% de.  相似文献   
2.
An efficient route to the pharmaceutically important (6S,9R)-11-oxo-5,6,7,8,9,10-hexahydro-6,9-methanobenzocyclooctene intermediate has been demonstrated via kinetic resolution of 11-oxo-5,6,7,8,9,10-hexahydro-6,9-methanobenzocyclooctene using a commercially available ketoreductase. Biocatalytics KRED 101 has been shown to selectively reduce the (6R,9S) enantiomer leaving behind the desired (6S,9R) enantiomer. This novel reaction is the first demonstration of a high yielding (44% versus 50% maximum theoretical yield) highly stereoselective (>99% ee) resolution of a bicyclic ketone, via enzymatic reduction using a commercially available ketoreductase, where the stereochemistry of the substrate is determined by a bridged ring system. Several challenges were overcome, including enhancing the selectivity of the enzyme by controlling temperature and increasing substrate solubility by employing a combination of cyclodextrin and organic co-solvent in the aqueous reaction system.  相似文献   
3.
《Process Biochemistry》2014,49(9):1527-1532
Herein we describe the kinetic characterization of a fusion protein from the 3-ketoacyl-[acyl-carrier-protein]-reductase (KR) from Synechococcus PCC 7942 and a mutant formate dehydrogenase from Mycobacterium vaccae N10 (MycFDH). Upon purification, a specific proteolytic cleavage of the MycFDH was observed. The cleavage site was elucidated, which is ubiquitously spread among prokaryotic FDHs. After depletion of the cleavage site the correct, full length fusion protein was obtained. In asymmetric reductions of ethylbenzoyl acetate (EBA) this fusion protein performed equal or even better than the free enzymes, yielding up to 39% more of the fluoxetine precursor ethyl-(S)-3-hydroxy-3-phenylpropanoate ((S)-HPPE). The rate acceleration is due to an improved Km,EBA of the KR subunit.  相似文献   
4.
The immobilization conditions of commercial ketoreductase for continuous enantioselective reduction in the gas-phase reaction were investigated with respect to the immobilization efficiency (residual activity and protein loading) and the gas-phase reaction efficiency (initial reaction rate, half-life, and enantioselectivity). For the analyses, ketoreductase was first immobilized by physical deposition on glass supports and the reduction of 2-butanone to (S)-2-butanol with the concomitant regeneration of NADH by 2-propanol was used as a model reaction. The optimal conditions of enzyme immobilization were obtained using an absolute pressure of 100 hPa for drying, a pH between 6.5 and 7.0, and a buffer concentration of 50 mM. The buffer concentration in particular had a strong effect on both the enzyme activity and enantioselectivity. Under optimal immobilization conditions, the thermostability of ketoreductase in the gas-phase system was enhanced compared to the aqueous-phase system, while the enantioselectivity was successfully maintained at a level identical to that of the native enzyme. These results indicate that the gas-phase reaction has a great potential for industrial production of chiral compounds, but requires careful optimization of immobilization conditions for the reaction to progress effectively.  相似文献   
5.
徐徐  郑舰艇 《微生物学通报》2020,47(7):2082-2093
【背景】在模块化聚酮合成酶(polyketidesynthase,PKS)的催化过程中,催化结构域与同源酰基载体蛋白(acyl-carrierprotein,ACP)之间的蛋白质-蛋白质相互作用起重要作用,但这种瞬时可逆的相互作用难以捕捉分析。【目的】获得ACP和酮基还原酶(ketoreductase,KR)相互作用的蛋白复合物。【方法】在KR和ACP之间的Linker上插入烟草蚀纹病毒(tobacco etch virus,TEV)蛋白酶切位点,通过双功能马来酰亚胺试剂BMH将KR和ACP共价交联,随后TEV酶切检测交联结果。调整反应条件,使交联效率最大化。根据KR-ACP交联复合物与体系内其他蛋白标签和分子量的差异,通过亲和层析和凝胶过滤等纯化手段,获得纯度较高的KR-ACP稳定交联复合物。【结果】单独表达的KR和ACP结构域交联不成功,融合表达的KR+ACP双结构域可以有效交联,结合使用亲和层析和凝胶过滤等纯化手段成功获得纯度较高的复合物。该策略可运用于多个KR和ACP的共价交联。【结论】建立了捕获并纯化KR和ACP瞬时相互作用复合物的有效方法,为后期晶体结构的解析、KR与ACP相互作用机理的揭示及参与相互作用关键氨基酸的鉴定提供了实验基础。  相似文献   
6.
Enzymatic production of chemicals typically includes fermentation of engineered bacteria, preparation of enzymes, and bioconversion processes. Here coupled fermentation-bioconversion process for production of chiral atazanavir intermediate α-chlorohydrin was established. In the fermentation step, the ketoreductase was released from recombinant E. coli cells into medium after high-temperature induction, and the enzyme activity reached 5960 U/mL in the fermentation supernatant. In the bioconversion step, the fermentation supernatant was used directly for conversion of 200 g/L α-chloroketone in heterogeneous reaction mixture, and the residual amount of chloroketone reached less than 0.1% after 24 h of reaction. After filtration and drying, powder α-chlorohydrin was obtained with total yield of 95.7%. Finally, the filtrate of the reaction mixture was used for 2nd batch conversion, and powder α-chlorohydrin was obtained with yield of 95.6%. The above coupled process simplified enzyme preparation procedure, allowed the bioproduction of α-chlorohydrin to be carried out smoothly with reduced waste water discharge, and was advantageous for industrial scale-up.  相似文献   
1
设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号