DNS法检测灰略红链霉菌JSD-1产纤维素酶的CMC酶活条件的优化

Optimization of conditions for CMCase activity of cellulase detected with DNS method for Streptomyces griseorubens( JSD-1)

摘要: 为研究3,5-二硝基水扬酸法（DNS）检测纤维素酶的CMC酶活条件,采用单因素实验探讨了波长、酶促反应温度与时间、p H、粗酶液和底物添加量、底物浓度、显色时间及空白实验对测定结果的影响,并通过响应面（BoxBehnken）实验设计及方差分析确定了一种较优的CMC酶活检测条件组合。结果表明,CMC酶活检测最佳条件为:波长520nm、酶促反应温度和时间分别为45℃和30min、p H7.0、底物浓度12g/L、显色15min。

Abstract: In order to study the experimental conditions of 3,5- Dinitrosalicylic acid( DNS) to measure the activity of carboxymethyl cellulase( CMCase),factors including wavelength,enzymatic reaction temperature and time,p H,solvent amount of crude enzyme and substrate,substrate concentration,coloration time and blank experiment were used by a single factor test to explore the influence on the results of CMCase activity.A relatively optimal solution which was applicable to CMCase activity was established by using the method of response surface analysis( RSA)and variance analysis.Results showed that the best conditions for detecting the activity of CMCase was at 520 nm wavelength,45℃ and 30 min for enzymatic reaction,p H7.0,substrate concentration 12 g / L,coloration 15 min.