利用原生质体融合技术构建梨酒酵母工程菌

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

摘要: 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。

Abstract: 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.