利用原生质体融合技术构建梨酒酵母工程菌

以菌株酿酒酵母Saccharomyces cerevisiae YDJ05及产香酵母Issatchenkia orientalis YS03为融合亲本X和Y,通过原生质体融合技术构建适合梨酒酿造的酵母工程菌,研究结果表明:利用EMS诱变亲本菌株Y,得到了一株精氨酸营养缺陷型菌株,亲本菌株X和Y采用蜗牛酶液在35℃下处理100min,得到菌株X、Y的原生质体融合率分别为93.6%、94.2%,再生率分别为27.8%、31.6%。以加热的方式灭活亲本菌株X,灭活时间为14min。在以PEG为促融剂的条件下对两株菌进行原生质体融合,经过优选得到融合子DJ02,其产酒精率、产香率等指标最优,分别达到了9.87%、0.37g/L。 

Study on construction of yeast engineering strain for pear wine by protoplast fusion method

The yeast engineering strain was structured by the protoplast fusion technology.Saccharomyces cerevisiae YDJ05 and Issatchenkia orientalis YS03 were used as the fusion parents X and Y.A arginine nutrition unfairness strain was selected by EMS mutagenesis from parents strains Y.The strains X and Y were dealt with the snail enzyme liquid in 35℃ for 100min.The best protoplast formation rate and regeneration rate were 93.6%, 94.2% and 27.8%, 31.6%.Inactivate strains of the parents of X with heating them in 14min.In the circumstance with PEG as promotion melting agent DJ01 ~ DJ06 six strains fusant were gotten, one of them DJ02 with the best alcohol producting rate and aroma producting rate were reached to 9.87% and 0.37g/L.