福林酚法测定酿酒白曲酸性蛋白酶活力的条件试验  

Conditional Test of the Folinphenol Reagent Method for the Determination of Brewing Bran Koji Acid Protease Activity

作　　者：郑东影 陈玮 张卫卫 司冠儒 张温清 周萍 ZHENG Dongying;CHEN Wei;ZHANG Weiwei;SI Guanru;ZHANG Wenqing;ZHOU Ping(Xuanjiu Group Co.Ltd.,Xuancheng 242000,Anhui,China)

机构地区：[1]安徽宣酒集团股份有限公司,安徽宣城242000

出　　处：《酿酒》2020年第2期63-66,共4页Liquor Making

摘　　要：在国标GBT23527-2009福林酚试剂法测蛋白酶活力的基础上,以自产白曲为材料,研究底物浓度,反应时间,酶液浸取时间以及三氯乙酸沉淀时间对酶活测定结果的影响,并对方法进行部分改进。实验结果显示:国标法测白曲酸性蛋白酶活力时,存在底物不足的问题,利用Hanes作图法求得该酶米氏常数Km约为2.67g/L,确定底物酪蛋白浓度为30g/L;由反应进程曲线得到,反应时间5min更适合酶活测定;酶液浸取时间长会增加比色测定时背景值的干扰,在不影响测定结果的情况下,缩短了酶液的浸取时间;充分摇匀的前提下,三氯乙酸沉淀时间对测定结果无显著影响。Based on the determination of protease activity by GB/T23527-2009 Folin reagent method,this article used self-production Bran Koji as the material,studied the effect of substrate concentration,reaction time,enzymatic liquid leaching time and precipitation time of TCA on the results of enzyme activity determination,and made some improvement to the method.The results showed that there was a problem of insufficient substrate concentration,when used the national standard method to measure the activity of Acid protease,the Km of the enzyme was approximately 2.67g/L by the Hanes method,and determined the concentration of the base casein to be 30g/L;Long leaching time of enzyme solution willed increase the interference of background value during colorimetric determination,and shorten the leaching time of enzyme solution without affecting the determination results;The precipitation time of TCA had no significant effect on the determination results under full shaking condition.

关 键 词：福林酚法 酶活力 底物浓度 反应时间 

分 类 号：TS262.3[轻工技术与工程—发酵工程] TS261.11[轻工技术与工程—食品科学与工程]