食品科学 ›› 2018, Vol. 39 ›› Issue (8): 57-63.doi: 10.7506/spkx1002-6630-201808010

• 生物工程 • 上一篇    下一篇

酿酒酵母W5单倍体的制备及其代谢水平分析

王长丽1,2,佟天奇1,2,刘文娟1,2,张云野1,2,宋刚1,2,葛菁萍1,2,*   

  1. (1.黑龙江大学生命科学学院,微生物学黑龙江省高校重点实验室,黑龙江?哈尔滨 150080;2.黑龙江大学 农业微生物技术教育部工程研究中心,黑龙江?哈尔滨 150500)
  • 出版日期:2018-04-25 发布日期:2018-04-17
  • 基金资助:
    国家自然科学基金面上项目(31570492;31590492;31470537); 黑龙江省高等学校科技创新团队(农业微生物发酵技术)项目(2012td009)

Preparation and Metabolic Profiling of a Haploid Strain of Saccharomyces cerevisiae W5

WANG Changli1,2, TONG Tianqi1,2, LIU Wenjuan1,2, ZHANG Yunye1,2, SONG Gang1,2, GE Jingping1,2,*   

  1. (1. College of Life Sciences, Key Laboratory of Microbiology, Heilongjiang University, Harbin 150080, China; 2. Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education, Heilongjiang University, Harbin 150500, China)
  • Online:2018-04-25 Published:2018-04-17

摘要: 为获得遗传稳定的单倍体酵母菌株,本实验以二倍体酿酒酵母Saccharomyces cerevisiae W5为出发菌株,优化单倍体制备和分离的条件,获得单倍体细胞;利用随机孢子分析法和MAT-聚合酶链式反应法对单倍体菌株进行分离和鉴定,通过连续传代培养的方式验证单倍体菌株的稳定性,并从代谢水平分析单倍体菌株的发酵性能。研究表明:S.?cerevisiae?W5在改良SPM培养基中30?℃条件下培养8?d,产孢率最高为(35.57±0.82)%。经30?mg/mL蜗牛酶液处理2?h后,菌悬液的孢子释放率达到(47.06±0.23)%。本实验共分离出7?株单倍体菌株,其中菌株S.?cerevisiae?H14(MAT-a)遗传性能最稳定。当以葡萄糖为底物,30 ℃、150?r/min发酵30?h,乙醇质量浓度达到最大,为(40.46±1.06)g/L,较S.?cerevisiae?W5发酵24?h乙醇最大质量浓度(51.65±2.39)g/L下降了21.67%,乙酸、乙偶姻的产量分别提高了12.7%和57.14%,获得理想的单倍体菌株。本实验为S. cerevisiae单倍体的制备提供了一定研究经验,也丰富了酵母遗传学研究的理论基础。

关键词: 酿酒酵母, 单倍体制备, 验证, 代谢分析

Abstract: The experimental conditions to obtain a genetically stable haploid strain from the Saccharomyces cerevisiae diploid strain W5 were optimized in this study. Seven haploid strains were isolated and identified by random spore analysis and MAT-PCR. Then, the stability of the haploid strains was evaluated by continuous passage culture. Ultimately, the fermentation performance of two selected haploid strains was analyzed. The results showed that the highest sporulation rate of the strain W5 of (35.57 ± 0.82)% was acquired when it was cultured on a modified SPM medium at 30 ℃ for 8 days. Spore release rate was (47.06 ± 0.23)% after 2 h treatment of the suspension with 3% snailase. S. cerevisiae H14 (MAT-a) was the most genetically stable among seven haploid strains. Strain H14 gave the highest ethanol production of (40.46 ± 1.06) g/L when cultured at 30 ℃ with glucose as a substrate for 30 h with agitation at 150 r/min, which was 21.67% lower than that of W5 at 24 h, whereas the yields of acetic acid and acetoin were increased by 12.7% and 57.14%, respectively. This research can provide useful information for researchers to prepare haploid S. cerevisiae and also enrich the fundamental theory of yeast genetics.

Key words: Saccharomyces cerevisiae, haploid, preparation, identification, metabolism analysis

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