CAJ | 학술논문

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5.8S rDNA-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。
이대초기분분-β-D-포도당감(pNPG)법사선산매균주，병진행매활력정량분석。대산매균적충속정성채용5.8S rDNA-ITS구역RFLP분석적분자감정법。통과세포파벽검측매활력측정균주적산매정위。이용수양감화칠협감위유도물순화균주，제고기산매능력。결과표명，종236주자선효모균중사선출210주산매균주，구분위4충분자류형，분별위：유포한손효모속（Hanseniaspora vineae）、양주효모（Saccharomyces cerevisiae）、포도즙유포한손효모（Hanseniaspora uvarum）화잡리비극필적효모（Pichia caribbica）。산매활성교고적8주양주효모매활성재0.0075～0.0099 U/mL지간，산매정위균위：포외매＞포내매＞포벽매。수양감지대S4균주산매구유유도작용，매활력제고14%。이칠협감가대S1화S4균주산매구유유도작용，매활력분별제고13%화19%。본연구사선출8주고산β-D-포도당감매적양주효모，가용우생산포도주。
In the experiments, p-nitrophenyl-β-D-glucoside(pNPG) was used to select yeast strains withβ-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. Theβ-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 improveβ-D-glucosidase-pro-ducing 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 producingβ-D-glucosidase with high acitivity, and the activity ranged from 0.0075 U/mL to 0.0099 U/mL (enzyme-producing loca-tion positioning, extracellular enzyme>intracellular enzyme>cell wall enzyme);salicin could only induce S4 strain and its enzyme activity in-creased 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.