Steroid C1, 2-site dehydrogenase enzyme activity detection method

• Main Authors: LIU ZHIQIANG, ZHENG YUGUO, ZHANG BO, NI YEWEN, KE XIA, CHEN XINXIN
• Format: Patent
• Language: Chinese; English
• Published: 26.03.2021
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; ENZYMOLOGY; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; SPIRITS; TESTING; VINEGAR; WINE

The invention relates to a steroid C1, 2-site dehydrogenase enzyme activity detection method. The steroid C1, 2-site dehydrogenase can metabolize and reduce WST-1, a water-soluble orange-yellow formazan product is generated under the combined action of PMS, the absorbance is measured at 440 nm, the WST-1 reduction amount is used for representing the dehydrogenase activity, the generation amount oforange-yellow formazan is calculated according to a standard curve, and then the steroid C1, 2-site dehydrogenase activity is obtained. The invention provides an oxidoreductase activity detection method which is strong in specificity, high in sensitivity, convenient to operate, short in detection time, accurate in quantification and suitable for high-flux steroid C1, 2-site dehydrogenase activitydetection. 本发明涉及一种甾体C1,2位脱氢酶酶活检测方法。甾体C1,2位脱氢酶能够代谢还原WST-1，同时在PMS的联合作用下生成还原成水溶性的橙黄色甲臢产物，在440nm下测定吸光度，以WST-1还原量表示脱氢酶活性，根据标准曲线计算橙黄色甲臢的生成量，进而求出甾体C1,2位脱氢酶活性。本发明提供了一种氧化还原酶活力检测方法，特异性强、灵敏度高、操作方便、检测时间短、定量准确，适合高通量的甾体C1,2位脱氢酶活性检测。