【摘要】 介绍高产菊粉酶的克鲁维酵母 (K luyveromyces sp.) Y85菌株与产山梨醇的酿酒酵母(Saccharomyces cerevisiae) E15菌株 ,通过属间原生质体融合构建直接利用菊粉产山梨醇的酵母融合菌株 .结果认为 ,以 0 .1% EDTA-巯基乙醇为脱壁预处理剂 ,蜗牛酶浓度 2 % (W/V) ,酶解温度 30℃ ,E15和 Y85两菌株细胞酶解时间分别为 4 0 min和 30 min,原生质体形成率和再生率分别达90 %和 2 5 %以上 .融合的适宜条件为 :添加 30 % (W/V) PEG和 10 mmol/L Ca Cl2 ,融合时间为 30min,融合率可达 2 .64× 10 -6 以上 .利用两亲株自然性状差异检出融合菌株 ,并经 DNA含量、菊粉酶活性、细胞形态与大小、菌落形态等测定加以确证 .其中 F2 7融合菌株在适宜条件下 ,山梨醇摇瓶发酵产量可达 4 .87g/10 0 m L,并且具有遗传稳定性能更多还原

【Abstract】 The construction of inulin utilizing and sorbitol producing fusants by interge neric protoplast fusion between Kluyveromyces sp. Y 85 and Saccharomy ces cerevisiae E 15 is discussed. The strains protoplast were prepared with 0.1 % EDTA and 2.0%(w/v)helicase at 30 ℃ for 30~40 min, The formation rate was above 9 0% and regeneration rate reached 25%. The suitable conditions for the protoplas t fusion was added with 10mmol/L each and 30%PEG(MW6,000)for 30 min. The fusio n frequency was about 2.64×10 -6 . The fusants were sorted by using the d ifferent character between strans Y 85 and E 15. DNA estimation, inulinase ac tivity examination, and comparison of cell size and morphology indicated that the fusants were fused from strans E 15 and Y 85. Under optimal fermentation c onditions, a maximum sorbitol production reached 4.87g/100mL, and the fermenta tion ability of F27 fusant was quite stable.更多还原