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›› 2008, Vol. 8 ›› Issue (6): 1185-1189.

• 生化工程专栏 • 上一篇    下一篇

丙酮-丁醇发酵生产菌的快速筛选方法

靳孝庆 周华 吴薛明 吴薛明 张赣道 何冰芳   

  1. 南京工业大学制药与生命科学学院 南京工业大学 南京工业大学制药与生命科学学院 南京工业大学制药与生命科学学院 江苏省生物工业技术重点实验室
  • 收稿日期:2008-05-26 修回日期:2008-07-31 出版日期:2008-12-20 发布日期:2008-12-20
  • 通讯作者: 何冰芳

A Rapid Screening Method of Producing Strain in Acetone-Butanol Fermentation

JIN Xiao-qing ZHOU Hua WU Xue-ming ZHANG Gan-dao HE Bing-fang   

  1. College of Life Science and pharmaceutical engineering, Nanjing University of Technology College of Life Science and pharmaceutical engineering, Nanjing University of Technology College of Life Science and Pharmacy, Nanjing University of Technology, The Provincial Key Laboratory of Industrial Biotechnology
  • Received:2008-05-26 Revised:2008-07-31 Online:2008-12-20 Published:2008-12-20
  • Contact: HE Bing-fang

摘要: 建立了高效丙酮-丁醇生产菌的快速筛选方法,采用氮离子注入诱变技术选育丙酮-丁醇发酵生产菌,以2-脱氧-D-葡萄糖作为抗代谢阻遏物,筛选高淀粉酶活性的突变株. 首次根据丙酮-丁醇发酵代谢途径中先产酸后产溶剂及高活力厌氧发酵细胞具有较强还原力的特点,设计了溴甲酚绿和刃天青2种筛选平板,所筛突变株经发酵验证丁醇和总溶剂产生能力均有显著提高,I4-28突变菌株总溶剂产量和丁醇产量分别较出发菌株提高了27.96%和40.66%,丁醇/总溶剂比由63.39%提高到71.94%,突变菌株具有较好的传代稳定性.

关键词: 丙酮-丁醇发酵, 筛选方法, 离子注入

Abstract: A rapid screening method of producing strains in acetone-butanol fermentation was developed. Nitrogen ion beam implantation was used to breed producing strains in the fermentation. The high amylase activity mutants were obtained by using 2-deoxy-D-glucose as the antimetabolite. Then the high-yielding mutants were further screened by screening plates of bromocresol green and resazurin. It was proved that the butanol and solvent producing ability of selected mutants were significantly improved. The total solvents production and butanol production of mutant I4-28 increased by 27.96% and 40.66%, compared with the parent strain, and the ratio of butanol to total solvents was up to 71.94% from 63.39%. The mutant had good genetic stability after 8 times of subculture. The rapid screening method was simple, efficient and significantly valuable in breeding of acetone-butanol producing strains.

Key words: acetone-butanol fermentation, screening method, ion beam implantation

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