期刊文献+

采用全基因组改组技术选育蛋白酶高产菌株 被引量:2

Whole Genome Shuffling of Bacillus subtilis for Enhanced Protease Production
在线阅读 下载PDF
导出
摘要 为了提高枯草芽孢杆菌B4的蛋白酶产量,采用全基因组改组技术进行菌株选育,先通过紫外诱变构建了B4菌的突变体库,在优化其原生质体制备和再生条件的基础上,以其中4株诱变菌株(BRS1、BRS2、BRS5、BRS6)作为亲本,采用PEG介导的方法进行两次多亲本的全基因组改组,同时,结合双亲灭活的筛选方法,共筛选出3株酶活力大幅度提高并能稳定遗传的优良菌株,为BRS1、BRS5、BRS6,酶活力最高达2175.81U/mL,达到原始菌株的3.01倍。 Whole genome shuffling technique was used to enhance protease production by Bacillus subtilis B4. A candidate mutant library was constructed by UV irradiation of Bacillus subtilis B4. Based on the optimum conditions for protoplast preparation and regeneration, multi-parental whole genome shuffling was carded out with 4 mutant strains (BRS1, BRS2, BRS5 and BRS6) by PEG mediated protoplasts fusion. Then BRS 1, BRS5 and BRS6 were identified by biparental inactivation method as 3 excellent strains with greatly improved protease activity and good genetic stability. The maximum activity was as high as 2175.81 U/mL, which was increased 3.01 fold compared with the initial strain B4.
出处 《食品科学》 EI CAS CSCD 北大核心 2012年第19期211-215,共5页 Food Science
基金 国家"863"计划项目(2008AA100802) 吉林省自然科学基金项目(201215196)
关键词 枯草芽孢杆菌 蛋白酶 基因组改组 Bacillus subtilis protease genome shuffling
  • 相关文献

参考文献24

  • 1赵文婷,邹懿,胡昌华.微生物菌种改良的新方法新策略[J].生物工程学报,2009,25(6):801-805. 被引量:14
  • 2DEMAIN A L, ADRIO J L. Contributions of microorganisms to industrial biology[J]. Mol Biotechnol, 2008, 38(1): 41-55.
  • 3SHIK H A, ADHYAYAN S, NAND A N S, et al. Improved production of alkaline protease from a mutant of alkalophilic Bacillus pan totheneticus using molasses as a substrate[J]. Bioresource Technology, 2007, 15(4): 881-885.
  • 4RAVICHANDR A, POTUMARTH I, ANNAPURNA J. Alkaline protease production by submerged fermentation in stirred tank reactor using Bacillus licheniformis NCIM-2042: effect of aerationand agitationregimes[J]. Biochemical Engineering Journal, 2006, 16(5): 56-65.
  • 5CHI Z, MA C, WANG P, et al. Optimization of medium and cultivation conditions for alkaline protease production by the marine yeast Aureobasidium pullulans[J]. Bioresource Technology, 2007, 15(3): 534-538.
  • 6陈晓明,张良,张建国,周莉薇.枯草芽孢杆菌淀粉酶高产菌株的辐射诱变研究[J].辐射研究与辐射工艺学报,2008,26(3):177-182. 被引量:12
  • 7马明,杜金华.枯草芽孢杆菌酶在工业生产中的应用[J].山东科学,2006,19(3):35-38. 被引量:27
  • 8SCHALLMEY M, SINGH A, WARD O P. Developments in the use of Bacillus species for industrial production[J]. Canadian Journal of Microbiology, 2004, 50(1): 1-17.
  • 9STEMMER W P. Molecular breeding of genes, pathways and genomes by DNA shuffling[J]. Journal of Molecular Catalysis B: Enzymatic, 2003, 20(1): 3-12.
  • 10POWELL K A, RAMER S W, CARDAYRE S B. Directed evolution and biocatalysis[J]. Angew Chem, 2001, 40(10): 3948-3959.

二级参考文献126

共引文献177

同被引文献54

引证文献2

二级引证文献10

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部