产β-D-葡萄糖苷酶酵母菌的筛选及产酶性质研究.pdf

产β-D-葡萄糖苷酶酵母菌的筛选及产酶性质研究

王彩肖;武伟伟;李艳

【摘要】Intheexperiments,p-nitrophenyl-β-D-glucoside(pNPG)wasused

toselectyeaststrainswithβ-D-glucosidase-producingcapacity,thenβ-D-

glucosidaseactivitywasanalyzedquantitatively.Thoseβ-D-glucosidase-

producingyeaststrainswereidentifiedbyRFLPanalysisof5.8SrDNA-ITS

region.Theβ-D-glucosidase-producinglocationoftheyeaststrainswas

determinedthroughbreakingthecellwallanddetectingtheenzyme

activity.Furthermore,salicinandesculinwereusedasinductorsto

domesticateyeaststrainsandfurthertoimproveβ-D-glucosidase-pro-

ducingcapacity.Theresultssuggestedthat,210β-D-glucosidase-

producingstrainsselectedamong236nativeyeaststrainsbelongedto

fourmoleculartypesincludingHanseniasporavineae,Saccharomyces

cerevisiae,HanseniasporauvarumandPichiacaribbica;amongthem,there

wereeightstrainsproducingβ-D-glucosidasewithhighacitivity,andthe

activityrangedfrom0.0075U/mLto0.0099U/mL(enzyme-producing

loca-tionpositioning,extracellularenzymeintracellularenzymecellwall

enzyme);salicincouldonlyinduceS4strainanditsenzymeactivityin-

creasedby14%,however,esculincouldinduceS1strainandS4strainand

theirenzymeactivityincreasedby13%and19%,respectively.Theeight

strainsscreenedinthisstudycouldbeusedintheproductionofgrape

wine.%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（Hanseniasporavineae）、

酿酒酵母（Saccharomycescerevisiae）、葡萄汁有孢汉逊酵母

（Hanseniasporauvarum）和卡利比克毕赤酵母（Pichiacaribbica）。产酶活

性较高的8株酿酒酵母酶活性在0.0075～0.0099U/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。

【期刊名称】《酿酒科技》

【年(卷),期】2014(000)010

【总页数】6页(P14-18,2