酵母细胞原生质体拆合技术的研究 Ⅰ.K氏酵母原生质体再生和拆分的研究

STUDIES ON ISOLATION-FUSION TECHNIQUE OF PROTOPLASTS OF THE YEAST SACCHAROMYCES CEREVISIAE Ⅰ. STUDIES ON REGENERATION AND ISOLATION OF PROTOPLASTS IN SACCHAROMYCES CEREVISIAE

微生物细胞原生质体拆合技术是在细胞融合基础上发展起来的一项新兴技术。本课题选择高产酒精的K氏酵母和具有淀粉糖化能力的黑曲霉进行拆合研究,以期形成核体/胞质体一原生质体,核体/胞质体—核体/胞质体等多种类型的融合子,实现远缘杂交。本文深入研究多种因素对K氏酵母原生质体制备的影响,并添加了牛血清白蛋白,聚乙烯吡咯烷酮;酵母细胞壁合成前体物以提高原生质体再生能力,当原生质体形成率>95%时,再生率最高达58.6%。此外,本文初步研究了酵母原生质体的分部拆分。

The isolation-fusion technique of protoplasts of microbial cell is a kind of new and developing technique growing on the basis of the cell fusion. In order to achieve distant hybridization, we choose Saccharomyces cerevisiae which is capable of high alcohol productivity and Aspergillus niger which is capable of starch fermentation to study the isolation-fusion technique. Some types of fusant, such as karyoplasts/cytoplasts-protoplasts or karyoplasts /cytoplasts-karyoplasts/cytoplasts are expected. This paper contains the effects of several factors on formation and regeneration of protoplasts. Moreover, bovine serum albumin and polyvinylpyrrolidone (PVP) and precursor of yeast cell wall syntheses are added to increase the regeneration ability of protoplasts. When the formation rates of protoplasts are more than 95%, the highest regeneration rate comes up to 58.6%. In addition, the paper presents tentative isolation of protoplasts in Saccharomyces cerevisiae.