一步降解苹果酸葡萄酒酵母的构建及特性研究——葡萄酒酵母与裂殖酵母属间原生质体融合

Construction of Malic Acid -Reducing Yeast Fusants and Studies On Characterization

以葡萄酒酵母和解苹果酸裂殖酵母为亲本,通过正交实验优化了亲本原生质体的制备,再生以及融合的最佳条件,融合率为1.76×10 -6,经筛选获得了一株降解苹果酸能力强且发酵性能优良的融合菌株。融合株的体积较亲株大,为4 7.5 4 89μm3 ,融合子的DNA含量约为二亲株之和,为7.36×10 -8μg/cell,证明确为融合子;连续15次传代,融合子各项性能稳定。

With Saccharomyces cerevisiae and Schizosaccharomyces malidevorans as primarystrains,through the cross cut tests,the optimism conditions of the preparation,regeneration and fusion of protoplast was studied,main factors influencing the formationand regneration of the protoplast were analyzed,It was found that they are enzyme concentration and. osmotic stabilizer. Several fusants were obtained.and the complementation frequency of fusion was 1.76×10-6～2.31×10-7,The fusions was tested in several aspects including cell morphology, the amount of chromosomal DNA and the fermention ability Through all these experiments,One fusant was selected out which has bigger cell volume,increased amount of chromosomal DNA per cell compared with that of the parental strains.and expresses fine properties of reducing malic acid and fermentation speed , The genetic stability was valued after 15 generations,showed that the degration rate of malic acid and other features came from parental strains was stable.