微生物酮还原酶的生产：新型高通量筛选方法的开发。
Microbial production of ketoreductases: Development of a novel high-throughput screening method.
机构信息
Microbial Processes and Technology Division, CSIR-National Institute for Interdisciplinary Science and Technology, Thiruvananthapuram 695 019, Kerala, India.
Microbial Processes and Technology Division, CSIR-National Institute for Interdisciplinary Science and Technology, Thiruvananthapuram 695 019, Kerala, India; Center of Innovative and Applied Bioprocessing, C-127, II Floor, Phase 8, Industrial Area, SAS Nagar, Mohali 160 071, Punjab, India.
出版信息
Bioresour Technol. 2017 Oct;242:319-323. doi: 10.1016/j.biortech.2017.03.096. Epub 2017 Mar 18.
An assay method for detection of enantiospecific chiral alcohol was developed based on ketoreductase, enantio-selective alcohol oxidase and 2,4-dinitrophenyl hydrazine (DNPH) reagent. The assay method was developed to check the conversion of 1-Acetonapthone to either (S) or (R) specific 1-(1-napthyl) ethanol or its racemic mixture using ketoreductases. Further, estimation was done with the help of 2,4-DNPH method. The resulting orange coloured chromogen showed a maximum absorbance at 560nm. The assay was performed in 96 well microtiter plates and had a linear detection range from 0.05mM to 4mM. The method is found to be suitable for the detection of large numbers of crude samples and screening of ketoreductase producing strains in high-throughput manner.
建立了一种基于酮还原酶、对映选择性醇氧化酶和 2,4-二硝基苯肼（DNPH）试剂检测对映体特异性手性醇的分析方法。该方法旨在检查 1-乙酰萘通过酮还原酶转化为（S）或（R）特异性 1-（1-萘基）乙醇或其外消旋混合物。进一步，借助 2,4-DNPH 法进行了估算。生成的橙色显色剂在 560nm 处显示最大吸收。该测定在 96 孔微量滴定板中进行，线性检测范围为 0.05mM 至 4mM。该方法适用于大量粗样品的检测和酮还原酶产生菌株的高通量筛选。
Zotero 插件