排序方式: 共有110条查询结果,搜索用时 15 毫秒
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Zhang CS Zhang ZJ Li CX Yu HL Zheng GW Xu JH 《Applied microbiology and biotechnology》2012,95(1):91-99
(R)-o-Chloromandelic acid is the key precursor for the synthesis of Clopidogrel?, a best-selling cardiovascular drug. Although nitrilases are often used as an efficient tool in the production of α-hydroxy acids, there is no practical nitrilase specifically developed for (R)-o-chloromandelic acid. In this work, a new nitrilase from Labrenzia aggregata (LaN) was discovered for the first time by genomic data mining, which hydrolyzed o-chloromandelonitrile with high enantioselectivity, yielding (R)-o-chloromandelic acid in 96.5% ee. The LaN was overexpressed in Escherichia coli BL21 (DE3), purified, and its catalytic properties were studied. When o-chloromandelonitrile was used as the substrate, the V(max) and K(m) of LaN were 2.53 μmol min?1 mg?1 protein and 0.39 mM, respectively, indicating its high catalytic efficiency. In addition, a study of substrate spectrum showed that LaN prefers to hydrolyze arylacetonitriles. To relieve the substrate inhibition and to improve the productivity of LaN, a biphasic system of toluene-water (1:9, v/v) was adopted, in which o-chloromandelonitrile of 300 mM (apparent concentration, based on total volume) could be transformed by LaN in 8 h, giving an isolated yield of 94.5%. The development of LaN makes it possible to produce (R)-o-chloromandelic acid by deracemizing o-chloromandelonitrile with good ee value and high substrate concentration. 相似文献
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A new resource of biocatalyst for asymmetric reduction of aromatic ketones has been discovered for the first time from a common plant seed, adzuki bean, i.e. Phaseolus angularis (Willd.) W.F. Wight. The study investigated the best methods to prepare the biocatalyst and its ability to reduce ketones. Our results indicated that the biocatalyst from adzuki bean could reduce various aromatic ketones at relatively high concentrations (e.g. 100 mM), exhibiting excellent stereoselectivity (>98% e.e.). In addition, it was found that NADPH acts as the reducing cofactor, which can be regenerated by the crude enzyme system itself using glucose as an auxiliary substrate. 相似文献
75.
Yi Xu Jian-He Xu Jiang Pan Liang Zhao Si-Liang Zhang 《Journal of Molecular Catalysis .B, Enzymatic》2004,27(4-6):155-159
Biocatalytic resolution of 3-(2′-nitrophenoxy)propylene oxide (1a), 3-(3′-nitrophenoxy)propylene oxide (1b) and 3-(4′-nitrophenoxy)propylene oxide (1c) were exploited by using lyophilized cells of yeast Trichosporon loubierii ECU1040 with epoxide hydrolase (EH) activity, which preferentially hydrolyzes (S)-enantiomers of the epoxides (1a–c), yielding (S)-diols and (R)-epoxides. The activity increased as the nitro group in the phenyl ring was shifted from 4′-position (1c) to 2′-position (1a). When the substrate concentration of 1a was increased from 10 to 80 mM, the E-value increased at first, until reaching a peak at 40 mM, and then decreased at higher concentrations (>40 mM). The optically active epoxide (R)-1a was prepared at gram-scale (97% ee, 41% yield). Furthermore, a simple method was developed to predict the enantiomeric excess of substrate (ees) at any time of the whole reaction course based on the ees value determined at a certain reaction time at a relatively lower substrate concentration. This will be helpful for terminating the reaction at a proper time to get both higher optical purity and higher yield of the remaining epoxides. 相似文献
76.
The lipase from Serratia marcescence ECU1010 (Sml) was capable of enantioselectively catalyzing the synthesis of many chiral drug precursors. This paper investigated
the immobilization of Sml on appropriate supporting materials and its performance in bioreactor. Chitosan, Celite 545, and
DEAE-cellulose were found to be the ideal supports among 8 carriers tested with respect to enzyme load and activity recovery
of lipase. When Sml was immobilized, significant improvements of stability against pH, thermal, and operational deactivation
were observed with all the 3 better supports, and the best stability was observed when the lipase was immobilized on glutaraldehyde
activated chitosan. As for the effect of organic solvent in the biphasic reaction system, the hydrolytic activity of the immobilized
lipase on trans-3-(4′-methoxyphenyl)glycidic acid methyl ester ((±)-MPGM) observed in isopropyl ether was 6 and 3 times higher than those
in toluene and methyl tert-butyl ether. The lipasecatalyzed production of (−)-MPGM by enzymatic resolution of (±)-MPGM with chitosan-Sml in isopropyl
etherwater biphasic system was carried out in a 2 L stirred-tank reactor. The batch operation was more efficient operation
mode for the enantioselective hydrolysis of (±)-MPGM, affording enantiopure (−)-MPGM in 44.3% overall yield, in contrast to
29.3% in a continuous reactor. 相似文献
77.
北柴胡不定根培养及茉莉酸甲酯处理对柴胡皂苷含量的影响 总被引:2,自引:0,他引:2
目的:建立北柴胡不定根培养体系,明确茉莉酸甲酯(MeJA)处理对北柴胡不定根中柴胡皂苷含量积累的影响。方法:利用固体和液体相结合的组织培养技术培养北柴胡不定根;分别以不同浓度的MeJA处理不定根不同时间,利用HPLC测定处理后不定根中柴胡皂苷含量的积累变化。结果:培养了北柴胡不定根;MeJA处理对北柴胡不定根中柴胡皂苷含量的积累有明显促进作用,当MeJA浓度为200μmol/L时,柴胡皂苷含量最高,为0.45%;以200μmol/L MeJA处理北柴胡不定根26 d时,柴胡皂苷含量最高,为0.51%。结论:北柴胡不定根培养结合MeJA诱导,可做为柴胡皂苷次生代谢合成途径及其积累规律研究的有效技术体系。 相似文献
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Xiao-Yan Zhang Xiang Fan Yong-Jun Qiu Cheng-Yuan Li Shuai Xing Yi-Tao Zheng Jian-He Xu 《Applied and environmental microbiology》2014,80(22):6870-6878
EstS1, a newly identified thermostable esterase from Sulfobacillus acidophilus DSM10332, was heterologously expressed in Escherichia coli and shown to enzymatically degrade phthalate esters (PAEs) to their corresponding monoalkyl PAEs. The optimal pH and temperature of the esterase were found to be 8.0 and 70°C, respectively. The half-life of EstS1 at 60°C was 15 h, indicating that the enzyme had good thermostability. The specificity constant (kcat/Km) of the enzyme for p-nitrophenyl butyrate was as high as 6,770 mM−1 s−1. The potential value of EstS1 was demonstrated by its ability to effectively hydrolyze 35 to 82% of PAEs (10 mM) within 2 min at 37°C, with all substrates being completely degraded within 24 h. At 60°C, the time required for complete hydrolysis of most PAEs was reduced by half. To our knowledge, this enzyme is a new esterase identified from thermophiles that is able to degrade various PAEs at high temperatures. 相似文献
80.
Jun-Feng Liu Zhi-Jun Zhang Ai-Tao Li Jiang Pan Jian-He Xu 《Applied microbiology and biotechnology》2011,89(3):665-672
The production of a recombinant nitrilase expressed in Escherichia coli JM109/pNLE was optimized in the present work. Various culture conditions and process parameters, including medium composition,
inducer, induction condition, pH and temperature, were systematically examined. The results showed that nitrilase production
in E. coli JM109/pNLE was greatly affected by the pH condition and the temperature in batch culture, and the highest nitrilase production
was obtained when the fermentation was carried out at 37°C, initial pH 7.0 without control and E. coli was induced with 0.2 mM isopropyl-β-d-thiogalactoside at 4.0 h. Furthermore, enzyme production could be significantly enhanced by adopting the glycerol feeding
strategy with lower flow rate. The enzyme expression was also authenticated by sodium dodecyl phosphate polyacrylamide gel
electrophoresis analysis. Finally, under the optimized conditions for fed-batch culture, cell growth, specific activity and
nitrilase production of the recombinant E. coli were increased by 9.0-, 5.5-, and 50-fold, respectively. 相似文献