CN108179120B - 一株酿酒酵母及其应用 - Google Patents
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Abstract
本发明公开了一株酿酒酵母及其应用,属于发酵技术领域。酿酒酵母(Saccharomyces cerevisiae)TC‑JRN已于2017年12月14日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC No.15064。该菌株发酵性能优良,果酒风味柔和,香气馥郁,适合作为橙汁发酵的酵母。
Description
技术领域
本发明涉及一株酿酒酵母及其应用,属于发酵技术领域。
背景技术
酿酒酵母菌(Saccharomyces cerevisiae)在分类学上属于真核生物原界、真菌界、子囊菌门、酵母亚门、半子囊菌纲、酵母目、酵母科、酵母属。酿酒酵母菌落呈圆形,有光泽,平坦,边缘整齐,在麦芽汁中培养细胞呈小圆形或卵圆形。适宜生长温度为28℃~30℃,最适pH值为4~5。酿酒酵母是与人类关系最广泛的一种酵母,不仅因为传统上它用于制作面包和馒头等食品及酿酒,在现代分子和细胞生物学中用作真核模式生物,其作用相当于原核的模式生物大肠杆菌。酿酒酵母是发酵中最常用的生物种类。酿酒酵母的细胞为球形或者卵形,直径5–10μm。其繁殖的方法为出芽生殖。
甜橙是我国广泛种植的水果,属于芸香科柑橘属植物,是柑橘的一种。甜橙含有多种对人体有益的营养物质,多种维生素、氨基酸、柠檬苦素、多种矿物质等。目前甜橙主要以鲜食为主,其副产品加工主要集中在橙汁产品上,使得甜橙产量过剩,造成极大的浪费。把橙汁发酵成橙酒不仅解决了甜橙浪费的问题,还大大的增加了产品的附加值。
目前橙酒在市场上比较罕见,并且市场中大多橙酒都是采用葡萄酒酵母发酵而成或者调配勾兑而成,因此造成市场上并无典型性好、香气浓郁、口感优雅的橙酒。对于果酒企业,企业的命脉就是珍贵的菌种,果酒酵母直接影响着果酒发酵后的口感和风味,因此筛选出一株适用于橙酒发酵的专用果酒酵母就显得非常重要。
发明内容
本发明的第一个目的是提供一株酿酒酵母(Saccharomyces cerevisiae),已于2017年12月14日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC No.15064,保藏地址为北京市朝阳区北辰西路1号院3号。
本发明的第二个目的是提供所述的酿酒酵母CGMCC No.15064的培养方法,是将酿酒酵母CGMCC No.15064接种至YEPD液体培养基中培养。
在本发明的一种实施方式中,所述YEPD培养基为:蛋白胨10~30g/L,酵母粉5~20g/L,葡萄糖10~30g/L,pH自然。
本发明的第三个目的是提供一种应用所述酿酒酵母生产橙酒的方法,是在橙酒主发酵阶段接种酿酒酵母CGMCC No.15064。
在本发明的一种实施方式中,所述接种是以4~6%的接种量进行接种。
本发明的第四个目的是提供所述酿酒酵母CGMCC No.15064制备的橙酒。
本发明的第五个目的是提供所述酿酒酵母CGMCC No.15064在食品中的应用。
在本发明的一种实施方式中,所述应用包括制备以橙为发酵原料的果汁或酒精饮品。
本发明的第六个目的是提供包含所述酿酒酵母CGMCC No.15064的微生物菌剂。
在本发明的一种实施方式中,所述微生物菌剂为固体菌剂或液体菌剂。
本发明有益效果
本发明提供的酿酒酵母CGMCC No.15064用于橙酒的酿造,该菌株在含糖量24oBx的橙汁中,25℃条件下发酵旺盛,发酵后果酒口感柔和,香气浓郁,并有典型橙汁香气。
生物材料保藏
一株酿酒酵母,其分类学命名为酿酒酵母Saccharomyces cerevisiae,已于2017年12月14日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCCNO.15064,保藏地址为北京市朝阳区北辰西路1号院3号中国科学院微生物研究所。
附图说明
图1酵母PCR产物琼脂糖电泳图;其中1:20号菌株;Y:阴性对照;M:Marker;
图2基于18S rDNA测序构建酵母菌株系统发育树。
具体实施方式
下面结合说明书附图和具体实施方式对本发明作进一步说明。
实施例1:酿酒酵母筛选及鉴定
1菌种分离
准备已灭菌的三角瓶,将成熟的鲜橙榨汁后装入无菌的三角瓶内,放入28℃恒温培养箱中静置,使其自然发酵。当橙汁发酵液出现大量的气泡时,取10mL发酵液加入已灭菌的富集培养基中,放置于28℃恒温培养箱中。富集2d后,取100μL菌液稀释5个浓度梯度,每个梯度3个平行,将稀释的菌液均匀涂布在含有氨苄的抗菌YPD平板上。将涂布好的平板放置于28℃恒温培养箱培养2d,并观察菌落生长情况。典型的酵母菌落表面光滑湿润,呈乳白色,有酒香味。将培养后的平板中具有典型的酵母菌的特征的单菌落挑出,进一步在YPD平板中进行二次划线分离,将分离出的酵母单菌落进行进一步镜检。将分离到的酵母菌培养后接入甘油管在-80℃下保存,并编号
2菌种一级筛选:以菌株的发酵能力和发酵液香气为指标,保留发酵能力旺盛,香气宜人的酵母,共44株。
3菌种二级筛选:以酒精度和感官品评为指标,保留酒精度高,品评结果良好的菌株,共9株。
4菌种的三级筛选:以感官品评和GC-MS分析得到的香气组分种类和含量为指标,最终保留一株发酵性能良好,香气馥郁、口感优良的酵母。
5菌种鉴定:TC-JRN基因组DNA的提取采用酵母基因组DNA提取试剂盒,对提取物进行PCR扩增,得到的PCR产物经电泳检测,结果显示良好。将扩增产物测序,测序结果在NCBI数据库中进行BLAST序列比对,发现TC-JRN与一株酿酒酵母Saccharomyces cerevisiae具有100%的18S rDNA基因序列相似度,故TC-JRN为酿酒酵母。该菌已于2017年12月14日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCCNO.15064,保藏地址为北京市朝阳区北辰西路1号院3号中国科学院微生物研究所。
实施例2:酿酒酵母的活化
将甘油管保藏的酿酒酵母接入YEPD液体培养基,在28℃、150rpm摇瓶培养18h,达到菌种活化的目的。用于橙酒的接种发酵。
实施例3:酿酒酵母在橙酒发酵中的应用效果
以橙汁为原料,调节初始糖浓度为240g/L,SO2浓度40mg/L,,接种总体积4%的酿酒酵母TC-JRN,在25℃静置发酵,直至残糖含量低于4g/L发酵结束。
TC-JRN发酵能力旺盛,发酵1d即有大量气泡产生。发酵结束后对橙酒各项理化指标进行检测,专业品评小组从口感和风味协调性等方面进行感官评价,如图1~2,并使用GC-MS半定量的方法分析了发酵后的橙酒的各个香气组分,将筛选所得TC-JRN与葡萄酒酵母D254的发酵的橙酒的挥发性物质的OAV值进一步比较,如表3所示,TC-JRN发酵的橙酒各种香气组分均大于D254,只有β-大马酮的OAV值略小于D254发酵的橙酒。
结合感官评定结果,20号菌株更适合橙酒的酿造,发酵后的橙酒口感清新,香气馥郁。
表1橙酒感官评价表(总分100分)
表2两种橙酒各指标分析结果
表3橙酒中挥发性成分的OAV值(仅列出OAV>1的香气成分)
虽然本发明已以较佳实施例公开如上,但其并非用以限定本发明,任何熟悉此技术的人,在不脱离本发明的精神和范围内,都可做各种的改动与修饰,因此本发明的保护范围应该以权利要求书所界定的为准。
Claims (9)
1.一株酿酒酵母(Saccharomyces cerevisiae)TC-JRN,于2017年12月14日保藏于中国微生物菌种保藏管理委员会普通微生物中心,保藏编号为CGMCC No.15064,保藏地址为北京市朝阳区北辰西路1号院3号。
2.权利要求1所述的酿酒酵母TC-JRN的培养方法,其特征在于,所述方法是将酿酒酵母TC-JRN接种至YEPD液体培养基中培养。
3.根据权利要求2所述的方法,其特征在于,所述YEPD培养基为:蛋白胨10~30g/L,酵母粉2~20g/L,葡萄糖10~30g/L,pH自然。
4.应用权利要求1所述的酿酒酵母生产橙酒的方法,其特征在于,所述方法是在橙酒主发酵阶段接种酿酒酵母TC-JRN。
5.根据权利要求4所述的方法,其特征在于,所述接种是以4~6%的接种量进行接种。
6.权利要求1所述的酿酒酵母TC-JRN在食品中的应用。
7.根据权利要求6所述的应用,其特征在于,所述应用包括制备以橙为发酵原料的果汁或酒精饮品。
8.包含权利要求1所述的酿酒酵母TC-JRN的微生物菌剂。
9.根据权利要求8所述的微生物菌剂,其特征在于,所述微生物菌剂为固体菌剂或液体菌剂。
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