响应面法优化酿酒酵母乙醛脱氢酶和乙醇脱氢酶酶活检测方法.pdfVIP

2011 No．7 · · Serial No．232 China Brewing 84 Research Report 响应面法优化酿酒酵母乙醛脱氢酶和乙醇脱氢酶酶活检测方法 1，2 1 2 张彦青 ，贾士儒*，张五九 （1.天津科技大学工业微生物重点实验室，天津 300457 ；2. 中国食品发酵工业研究院，北京 100027） 摘 要：从酿酒酵母前处理方式方面对ALD和ADH酶活测定条件进行了优化，采用单因素试验对前处理方式进行了分析研究，结果 表明破壁时间、超声波功率、超声时间、反应温度和缓冲液pH值对酶活测定值产生一定的影响。通过Box-Behnken 中心组合设计和响 应面分析法，确定了测定这2种酶的最理想酵母前处理条件，即超声功率420W，超声时间为11min，破碎时间为12min。 关 键 词：酿酒酵母；乙醛脱氢酶；乙醇脱氢酶；响应面；优化 中图分类号：TS261.1 文献标识码：A 文章编号：0254-5071 （2011）07-0084-05 Optimization of assay method of aldehyde dehydrogenase and alcohol dehydrogenase activity in Saccharomyces cerevisiae by response surface methodology 1，2 1 2 ZHANG Yanqing , JIA Shiru *, ZHANG Wujiu ( 1.Tianj in University of Science and Technology, Tianj in Key Laboratory of Industry Microbiology, Tianj in 300457, China; 2.China National Research Institute of Food and Fermentation Industries, Beij ing 100027, China) Abstract : Pretreatment conditions of aldehyde dehydrogenase (ALD) and alcohol dehydrogenase (ADH) activity assay method was optimized with single-factor experiment. The results showed that t broken cell wall time, power and treating time of ultrasonic, temperature and buffer pH value had effects on detection of ALD and ADH activity. By Box-Behnken central composite design and response surface analysis method, the optimal pretreat- ment conditions for determination of two enzymes of yeast were obtained as follows: ultrasonic power 420W, ultrasonic treating time 11min, break- ing cell wall for 12m