葡萄酒相关酵母β-葡萄糖苷酶活性及影响因素研究

Activity and influence factors of β-glucosidase in wine-making yeasts

以内蒙古西部地区分离到的6个属7个种共340株葡萄酒相关酵母菌株为材料,对产β-葡萄糖苷酶菌株进行初筛,进而采用对硝基苯基-β-D-吡喃葡萄糖苷(p NPG)法检测菌株胞外、胞壁结合及胞内β-葡萄糖苷酶活力,并对高酶活性菌株中β-葡萄糖苷酶的理化性质进行了研究。结果表明,分属5个属5个种(葡萄汁有孢汉逊酵母(Hanseniaspora uvarum)、酿酒酵母(Saccharomyces cerevisiae)、浅黄隐球酵母(Cryptococcus flavescens)、异常毕赤酵母(Pichia anomala)及星形假丝酵母(Candida stellata))的66株酵母菌株产β-葡萄糖苷酶,且胞内酶活性均高于胞壁结合酶活性;不同菌株中β-葡萄糖苷酶具有相同的理化性质,4%以上葡萄糖可抑制该酶的活性,5%~15%(V/V)的乙醇添加量对该酶活性无明显影响,其最适p H值为5.0~6.0,最适温度为40℃。

Using 340 wine-making related yeasts strains as raw material which were classified into 7 species of 6 genera and isolated from the western district of Inner Mongolia of China, β-glucosidase producing strains were screened firstly. Then the activity of β-glucosidase in extracellular, wall-bound and intracellular was assayed by p-nitrophenyl-β-D-glucoside （pNPG） methodology. Furthermore, physical and chemical properties of β-glucosidase originated from strain with high enzyme activity were also studied. The results showed that 66 yeast strains, classified into 5 species of 5 genera （Hanseniaspora uvarum, Saccharomyces cerevisiae, Cryptococcus flavescens, Pichia anomala and Candida stellata）, had the ability to produce β-glucosidase, and the activity of intracellular β-glucosidase was higher than that of wall-bound β-glucosidase. Physical and chemical properties of β-glucosidase in different strains were same, glucose with concentration more than 4% could inhibit β-glucosidase activity, 5%-15% （V/V） of alcohol addition had no significant influence on the enzyme activity, its optimal pH value and temperature were 5.0-6.0 and 40 ℃, respectively.