甘肃河西走廊葡萄酒产区高产糖苷酶非酿酒酵母菌株筛选及其酿酒适应性分析  

Screening of non-Saccharomyces yeasts endowed with high glucosidase activity and their adaptability to wine biotope in Hexi Corridor of Gansu Province

作　　者：姚红红 任学梅 严幻汝 祝霞[1,2] 杨学山 YAO Honghong;REN Xuemei;YAN Huanru;ZHU Xia;YANG Xueshan(College of Food Science and Engineering,Gansu Agricultural University,Lanzhou 730070,China;Gansu Key Lab of Viticulture and Enology,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学食品科学与工程学院,甘肃兰州730070 [2]甘肃省葡萄与葡萄酒工程学重点实验室,甘肃兰州730070

出　　处：《食品与发酵工业》2023年第16期49-58,共10页Food and Fermentation Industries

基　　金：国家自然科学基金项目(32260637,32060581);甘肃省葡萄酒产业发展基金项目(20180820-07,20180820-08,GCJ-2019-125-1);甘肃农业大学校列科研项目(GSAU-ZL-2018-8)。

摘　　要：为筛选高产糖苷酶的本土非酿酒酵母菌株,以甘肃河西走廊葡萄酒各子产区主栽酿酒葡萄为原料,在自然酒精发酵过程中分离得到912株酵母菌;采用七叶苷培养基进行产β-葡萄糖苷酶(β-glucosidase,β-Glu)菌株初步筛选,通过对硝基苯酚(p-nitrophenol,p-NP)法分别定量测定初筛菌株的β-Glu、α-L-阿拉伯糖苷酶(α-L-arabinofuranosidase,α-L-Ara)、α-L-鼠李糖苷酶(α-L-rhamnosidase,α-L-Rha)、β-D-木糖苷酶(β-D-xylosidase,β-Xyl)活力,并对优选菌株进行葡萄酒酿造条件适应性分析。结果表明,HX-1、HX-2、HX-3和HX-4具有较高的糖苷酶活力,经WL培养基形态特征及26S rDNA D1/D2区域序列分析鉴定,HX-1和HX-3为葡萄汁有孢汉逊酵母(Hanseniaspora uvarum),HX-2和HX-4为美极梅奇酵母(Metschnikowia pulcherrima)。各糖苷酶的最适pH值为5.0~6.0,最适温度为40~50℃,高乙醇、高SO_(2)和葡萄糖对酶活力均有抑制作用。在葡萄酒酿造条件下,HX-2和HX-4菌株的β-Glu活力、HX-1菌株的α-L-Rha活力、HX-4菌株的α-L-Ara活力、HX-1和HX-3菌株的β-Xyl活力较高。研究结果可为依据葡萄原料中糖苷类型定向选择增香酿造菌株提供技术支持。In order to screen non-Saccharomyces yeasts autochthonous strains endowed with high glycosidase activity,912 yeasts were isolated from spontaneous alcoholic fermentations of the major wine grape cultivars planted in Hexi Corridor of Gansu Province.The β-glucosidase-producing non-Saccharomyces yeast strains were preliminarily screened by esculin medium,and the activities of β-glucosidase(β-Glu),α-L-arabinofuranosidase(α-L-Ara),α-L-rhamnosidase(α-L-Rha) and β-D-xylosidase(β-Xyl) originated from these screened strains were determined quantitatively by p-NP method.Comparing the glucosidase producing ability of non-Saccharomyces yeasts,the adaptability of the selected strains to wine fermentation conditions was analyzed.The results showed that non-Saccharomyces yeast autochthonous strains HX-1,HX-2,HX-3,and HX-4 had high glycosidase activity.According to the morphological characteristics of these strains grown on WL medium and the sequence analysis of 26S rDNA D1/D2 domain,HX-1 and HX-3 were identified as Hanseniaspora uvarum,while HX-2 and HX-4 were Metschnikowia pulcherrima.The individual glycosidase produced by these strains exhibited high activity at pH 5.0-6.0 and 40-50 ℃.In addition,high concentration of alcohol,SO_(2),and glucose can inhibit the glucosidase activity.The activity of β-Glu produced by HX-2 and HX-4 strains,α-L-Rha by HX-1 strain,α-L-Ara by HX-4 strain,and β-Xyl by HX-1 and HX-3 strains was high under the conditions of wine fermentation.Overall,our study provides technical support for the directional selection of the yeast strain with good aroma enhancement performance according to the type of glucosides in wine grapes.

关 键 词：非酿酒酵母 α-L-阿拉伯糖苷酶 α-L-鼠李糖苷酶 β-D-木糖苷酶 酿造因子 

分 类 号：TS262.6[轻工技术与工程—发酵工程]