A new member of the short-chain dehydrogenases/reductases superfamily: Purification, characterization and substrate specificity of a recombinant carbonyl reductase from Pichia stipitis

Ye, Qi; Yan, Ming; Yao, Zhong; Lin, Xu; Cao, Hou; Li, Zhengjiang; Chen, Yong; Li, Shuya; Bai, Jianxin; Xiong, Jing; Ying, Hanjie; Ouyang, Pingkai

doi:10.1016/j.biortech.2009.06.014

Cited by 43 publications

(21 citation statements)

References 34 publications

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“…The activity was strongly inhibited by SDS. This inhibition may be the result of the combined effect of factors such as reduction in hydrophobic interactions that normally play a crucial role in maintaining tertiary structure and direct interactions with the protein molecule [22]. Partial inhibition of enzyme activity was observed with some organic solvents, such as dimethylformamide (DMF), and it was insensitive to dimethyl sulphoxide (DMSO) and b-mercaptoethanol.…”

Section: Effect Of Inhibitors and Detergents On Siase Activitymentioning

confidence: 98%

Purification and characterization of a highly selective sucrose isomerase from Erwinia rhapontici NX-5

Ren

et al. 2011

Bioprocess Biosyst Eng

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A highly selective sucrose isomerase (SIase) was purified to homogeneity from the cell-free extract of Erwinia rhapontici NX-5 with a recovery of 27.7% and a fold purification of 213.6. The purified SIase showed a high specific activity of 427.1 U mg(-1) with molecular weight of 65.6 kDa. The K (m) for sucrose was 222 mM while V (max) was 546 U mg(-1). The optimum pH and temperature for SIase activity were 6.0 and 30 °C, respectively. The purified SIase was stable in the temperature range of 10-40 °C and retained 65% of the enzyme activity after 2 weeks' storage at 30 °C. The SIase activity was enhanced by Mg(2+) and Mn(2+), inhibited by Ca(2+), Cu(2+), Zn(2+), and Co(2+), completely inhibited by Hg(2+) and Ag(2+). The purified SIase was strongly inhibited by SDS, while partially inhibited by dimethylformamide, tetrahydrofuran, and PMSF. Additionally, glucose and fructose acted as competitive inhibitors for purified SIase.

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Section: Effect Of Inhibitors and Detergents On Siase Activitymentioning

confidence: 98%

Purification and characterization of a highly selective sucrose isomerase from Erwinia rhapontici NX-5

Ren

et al. 2011

Bioprocess Biosyst Eng

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“…As has been previously reported, carbonyl reductases originated from yeasts preferably utilize NADPH as hydride-donating cofactor. For example, carbonyl reductases from C. parapsilosis CCTCC M203011 (Zhang et al 2009b), K. lactis NRIC 1329 (Yamamoto et al 2002a), Saccharomyces cerevisiae (Katzberg et al 2010;Choi et al 2010), Pichia stipitis CBS 6054 (Ye et al 2009), Candida magnoliae (Wada et al 1998), Sporobolomyces salmonicolor (Kataota et al 1992), and Trichosporon fermentans AJ-5152 (Kira and Onishi 2009) were all NADPH dependent. It is particularly interesting to note that carbonyl reductase from C. krusei SW2026 showed exact specificity to NADPH as cofactor ).…”

Section: Discussionmentioning

confidence: 98%

Asymmetric synthesis of (R)-1,3-butanediol from 4-hydroxy-2-butanone by a newly isolated strain Candida krusei ZJB-09162

Zheng

Qiu

et al. 2012

Appl Microbiol Biotechnol

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Biocatalytic asymmetric preparation of (R)-1,3-butanediol has been attracting much attention in pharmaceuticals industry. A new ideal strain, ZJB-09162, which exhibited high reduction activity and excellent (R)-stereospecificity towards 4-hydroxy-2-butanone, has been successfully isolated from soil samples. Based on morphology, physiological tests (API 20 C AUX), and 5.8S-ITS sequence, the isolate was identified as Candida krusei. Kinetic characterization demonstrated that carbonyl reductase from C. krusei ZJB-09162 preferred NADH to NADPH as cofactor, indicating it might be a new carbonyl reductase. (R)-1,3-Butanediol was produced in 19.8 g/L, 96.6% conversion, and 99.0% ee at optimal pH 8.5, 35 °C with a 2:1 molar ratio of glucose to 4H2B. In order to achieve higher product titer, the substrate loading was optimized in fixed catalysts and fixed substrate/catalysts ratio mode. The bioreduction of 4-hydroxy-2-butanone at a concentration of 45.0 g/L gave (R)-1,3-butanediol in 38.7 g/L and 83.9% conversion. Therefore, C. krusei ZJB-09162 was, for the first time, proven to be a promising biocatalyst for enzymatic preparation of (R)-1,3-butanediol.

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“…Residual activities were normalized as percentages of the maximum residual activity. condition was not commonly seen for SDRs of mesophilic origins, which typically display stability at pH lower than 8.0 [30][31][32][33][34][35][36][37][38].…”

Section: Effects Of Ph and Temperaturementioning

confidence: 99%

“…The results indicated excellent adaptability of ChKRED20 under varied reaction conditions. The high tolerance against thermal treatment and pH variation compared well with the majority of SDRs from mesophilic microorganisms [30][31][32][33][34][35][36][37][38][39][40], which would be beneficial for large-scale application of this enzyme.…”

Section: Effects Of Ph and Temperaturementioning

confidence: 99%

Characterization of a robust anti-Prelog short-chain dehydrogenase/reductase ChKRED20 from Chryseobacterium sp. CA49

Tang

Liu

2014

Journal of Molecular Catalysis B: Enzymatic

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A new member of the short-chain dehydrogenases/reductases superfamily: Purification, characterization and substrate specificity of a recombinant carbonyl reductase from Pichia stipitis

Cited by 43 publications

References 34 publications

Purification and characterization of a highly selective sucrose isomerase from Erwinia rhapontici NX-5

Purification and characterization of a highly selective sucrose isomerase from Erwinia rhapontici NX-5

Asymmetric synthesis of (R)-1,3-butanediol from 4-hydroxy-2-butanone by a newly isolated strain Candida krusei ZJB-09162

Characterization of a robust anti-Prelog short-chain dehydrogenase/reductase ChKRED20 from Chryseobacterium sp. CA49

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