产β-D-葡萄糖苷酶酵母菌的筛选及产酶性质研究  

Screening of Yeast Strains with High-yield of β-D-glucosidase and Study on Their β-D-glucosidase-producing Properties

作　　者：王彩肖 武伟伟[1] 李艳[1,2] 

机构地区：[1]河北科技大学生物科学与工程学院,河北石家庄050018 [2]河北省发酵工程技术研究中心,河北石家庄050018

出　　处：《酿酒科技》2014年第10期14-18,22,共6页Liquor-Making Science & Technology

基　　金：国家农业科技成果转化资金项目(2013GB2A200037);河北省科技支撑计划课题(13397107D);河北科技大学2012年度大学生科技创新基金立项项目(生工14)

摘　　要：以对硝基苯酚-β-D-葡萄糖苷（pNPG）法筛选产酶菌株,并进行酶活力定量分析。对产酶菌的种属定性采用5.8SrDNA-ITS区域RFLP分析的分子鉴定法。通过细胞破壁检测酶活力测定菌株的产酶定位。利用水杨苷和七叶苷为诱导物驯化菌株,提高其产酶能力。结果表明,从236株自选酵母菌中筛选出210株产酶菌株,区分为4种分子类型,分别为：有孢汉逊酵母属（Hanseniaspora vineae）、酿酒酵母（Saccharomyces cerevisiae）、葡萄汁有孢汉逊酵母（Hanseniaspora uvarum）和卡利比克毕赤酵母（Pichia caribbica）。产酶活性较高的8株酿酒酵母酶活性在0.0075-0.0099 U/mL之间,产酶定位均为：胞外酶〉胞内酶〉胞壁酶。水杨苷只对S4菌株产酶具有诱导作用,酶活力提高14%。而七叶苷可对S1和S4菌株产酶具有诱导作用,酶活力分别提高13%和19%。本研究筛选出8株高产β-D-葡萄糖苷酶的酿酒酵母,可用于生产葡萄酒。In the experiments, p-nitrophenyl-β-D-glucoside（pNPG） was used to select yeast strains with β-D-glucosidase-producing capacity, thenβ-D-glucosidase activity was analyzed quantitatively. Those β-D-glucosidase-producing yeast strains were identified by RFLP analysis of 5.8S rDNA-ITS region. The β-D-glucosidase-producing location of the yeast strains was determined through breaking the cell wall and detecting the enzyme activity. Furthermore, salicin and esculin were used as inductors to domesticate yeast strains and further to improve β-D-glucosidase-producing capacity. The results suggested that, 210 β-D-glucosidase-producing strains selected among 236 native yeast strains belonged to four molecular types including Hanseniaspora vineae, Saccharomyces cerevisiae, Hanseniaspora uvarum and Pichia caribbica; among them, there were eight strains producing β-D-glucosidase with high acitivity, and the activity ranged from 0.0075 U/mL to 0.0099 U/mL（enzyme-producing location positioning, extracellular enzyme〉intracellular enzyme〉cell wall enzyme）; salicin could only induce S4 strain and its enzyme activity increased by 14 %, however, esculin could induce S1 strain and S4 strain and their enzyme activity increased by 13 % and 19 %, respectively. The eight strains screened in this study could be used in the production of grape wine.

关 键 词：微生物 酵母菌 β-D-葡萄糖苷酶 筛选 葡萄酒 

分 类 号：Q939.9[生物学—微生物学]