生物技术通报 ›› 2019, Vol. 35 ›› Issue (3): 87-92.doi: 10.13560/j.cnki.biotech.bull.1985.2018-0784

• 研究报告 • 上一篇    下一篇

过表达INO1基因提高酿酒酵母乙醇耐受性

黄贞杰1, 陈由强2, 薛婷2   

  1. 1. 泉州医学高等专科学校,泉州 362010;
    2. 福建师范大学生命科学学院,福州 350108
  • 收稿日期:2018-09-09 出版日期:2019-03-26 发布日期:2019-04-03
  • 作者简介:黄贞杰,男,讲师,硕士研究生,研究方向:应用微生物与发酵技术;E-mail:hzj887@126.com
  • 基金资助:
    国家现代农业产业技术体系建设专项资金资助项目(CARS-20-4-4)

Improving the Tolerance of Saccharomyces cerevisiae to Ethanol by the Over-expression of Inositol-3-phosphate Synthase Gene INO1

HUANG Zhen-jie1, CHEN You-qiang2, XUE Ting2   

  1. 1. Quanzhou Medical College,Quanzhou 362010;
    2. College of Life Sciences,Fujian Normal University,Fuzhou 350108
  • Received:2018-09-09 Published:2019-03-26 Online:2019-04-03

摘要: 为获得燃料乙醇生产菌株,通过基因工程改造,构建能够利用能源甘蔗汁发酵、乙醇产率高的酿酒酵母工程菌株。即过表达肌醇-3-磷酸合成酶基因ino1,敲除kanMX抗性基因,获得重组菌。对过表达菌株的乙醇耐受性进行分析。利用甘蔗汁进行发酵培养,采用气相色谱(GC)对发酵产物乙醇进行检测。结果显示过表达菌株YI2-1能够耐19%(V/V)的乙醇,利用20oBx甘蔗汁厌氧发酵乙醇积累量为13.10%(V/V),较出发菌提高了8.55%。而过表达菌株YI2-1△KP的最大乙醇积累量为13.17%(V/V),较出发菌提高了9.16%。研究表明通过过表达酿酒酵母ino1基因能够有效提高菌株细胞活力、乙醇耐受性。构建的工程菌可利用甘蔗汁发酵,具有较高的乙醇产量。

关键词: 酿酒酵母, ino1基因过表达, 燃料乙醇, 乙醇耐受性

Abstract: In order to obtain the strains producing fuel ethanol,the engineering strain Saccharomyces cerevisiae,which may ferment using sugarcane juice to yield high ethanol,were constructed by genetic engineering. In other words,the inositol-3-phosphate synthase gene(ino1)was over-expressed in S. cerevisiae;the antibiotic resistance gene(kanMX)were knocked out,thus recombinant S. cerevisiae was acquired. Further,the tolerances(survivability)of the over-expressed strains to ethanol were analyzed. The sugarcane juice was used as a carbon source in the fermentation,and the ethanol content in the fermentation broth of strains was detected by gas chromatography. The experimental results showed that over-expressed strains YI2-1tolerated 19%(V/V)ethanol. The 20oBx of sugarcane juice was employed in the anaerobic fermentation,which yielded 13.10%(V/V)of the ethanol,and this was 8.55% more than that by the initial strain. The biggest output of ethanol by the over-expressed strain YI2-1△KP was accumulated to 13.17%(V/V),and this was 9.16% more than that by the original one. This study indicates that over-expression of ino1 gene efficiently enhances the cell viability and tolerance of S. cerevisiae to ethanol and the constructed strain S. cerevisiae is proved to highly yield ethanol by the fermentation of sugarcane juice.

Key words: Saccharomyces cerevisiae, over-expression of ino1 gene, fuel ethanol, tolerance to ethanol