基因组改组选育产酯化酶地衣芽孢杆菌

中国生物工程杂志

2013, Vol. 33

Issue (8): 45-50

研究报告

基因组改组选育产酯化酶地衣芽孢杆菌

薛正莲, 刘阳, 王洲, 马琦亚, 赵世光, 苏燕南

安徽工程大学生物与化学工程学院微生物发酵安徽省工程技术研究中心芜湖 241000

Breeding of Esterifying Enzyme-producing Bacillus licheniformis by Genome Shuffling

XUE Zheng-lian, LIU Yang, WANG Zhou, MA Qi-ya, ZHAO Shi-guang, SU Yan-nan

Institute of Biologic & Chemical Engineering of Anhui Polytechnic University, Microorganism Fermentation Engineering and Technology Research Center of Anhui Province, Wuhu 241000, China

全文: PDF(1035 KB) HTML

摘要：

从高温大曲中筛选到一株产酯化酶的地衣芽胞杆菌G₂。采用低温等离子体对G₂进行诱变,以筛选酶活提高最多的3株菌株作为亲本。将亲本原生质体灭活后对其进行基因组改组。将融合后的菌体进行筛选纯化之后,对融合子进行发酵测定其酶活并挑选酶活较高菌株进行传代测定其酶活稳定性。结果表明6^#融合子酶活最高且稳定为10.7U/ml,相对于初始菌株的5.3U/ml提高了102 %。对其发酵液进行气相色谱分析表明,6^#融合子代谢产一定量的酸类、酯类及醇类等物质,其中乙偶姻含量达 2565mg/L。

关键词： 酯化酶; 基因组改组; 地衣芽孢杆菌; 等离子诱变

Abstract:

A Bacillus licheniformis strain named G2 with esterifying enzyme activity screened from high temperature Daqu was mutagenized by using low temperature plasma and three strains with high esterifying enzyme activity was screened as parents .The parent protoplast was inactivated for genome shuffling. After the fused strains were purified by selection, the enzyme activity of fusants was determinated by fermentation, the strains of higher enzyme activity were screened to passage and their stability of enzyme activity were detected.The highest and stable enzyme activity of 6^# fusant can reach 10.7U/ml ,which has been improved by 102% compared with the 5.3U/ml of original strain G2.The fermentation broth of the 6^# fusant was analyzed by GC ,the results show that the strain can produce a certain amount of acids ,alcohols and esters, and the acetoin of the fermentation broth can reach 2565mg/L.

Key words: Esterifying enzyme Genome shuffling Bacillus licheniformis Plasma mutagenesis

收稿日期: 2013-01-21 出版日期: 2013-08-25

ZTFLH:

Q819

通讯作者: 薛正莲xuezhen0851@sina.com E-mail: xuezhen0851@sina.com

	服务
	把本文推荐给朋友
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	薛正莲
	刘阳
	王洲
	马琦亚
	赵世光
	苏燕南

引用本文:

薛正莲, 刘阳, 王洲, 马琦亚, 赵世光, 苏燕南. 基因组改组选育产酯化酶地衣芽孢杆菌[J]. 中国生物工程杂志, 2013, 33(8): 45-50.

XUE Zheng-lian, LIU Yang, WANG Zhou, MA Qi-ya, ZHAO Shi-guang, SU Yan-nan. Breeding of Esterifying Enzyme-producing Bacillus licheniformis by Genome Shuffling. China Biotechnology, 2013, 33(8): 45-50.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I8/45

[1] 周恒刚.漫谈己酸乙酯的酯化.酿酒科技1998(3):18-22. Zhou H G. Discussion of esterification of ethyl caproate. Liquor-Making Science ＆ Technology, 1998(3):18-22.
[2] 沈才洪,应鸿, 许德富,等.大曲"酯化力"的探讨.酿酒科技,2005(3):17-20. Shen C H, Ying H, Xu D F, et al. Research on Daqu esterifying power. Liquor-Making Science ＆ Technology, 2005(3):17-20.
[3] 李光辉,程铁辕,黄治国,等.浓香型白酒酒醅微生物群落代谢分析.酿酒科技,2009(3):29-32. Li G H, Cheng T Y, Huang Z G, et al. Metabolic analysis of the microbial community in the fermented grains of luzhou-flavour liquor. Liquor-Making Science ＆ Technology, 2009,3:29-32.
[4] 李大和,刘念,李国红.浓香型大曲酒酿造中酯化菌研究的现状与展望.酿酒科技,2008(2):92-98. Li D H, Liu N, Li G H, et al. Present research situation & prospect of the application of esterifying bacteria in the production of Luzhou-flavor Daqu.Liquor-Making Science ＆ Technology, 2008(2):92-98.
[5] 纪晓俊,黄和,杜军,等. 3-羟基丁酮的合成及应用进展.现代化工,2008,28(4):18-22. Ji X J, Huang H, Du J, et al. Advances in production of 3-hydroxybutanone and its application. Modern Chemical Industry, 2008, 28(4):18-22.
[6] 徐岩,吴群,范文来,等. 中国白酒中四甲基吡嗪的微生物产生途径的发现与证实.酿酒科技,2011,7(205):37-40. Xu Y, Wu Q, Fan W L, et al. The discovery & verification of the production pathway of tetramethylpyrazine (TTMP) in Chiese liquor. Liquor-Making Science ＆ Technology,2011,7(205):37-40.
[7] 吴根福,陈佩华,彭晓国. 己酸乙酯酯化酶产生菌的自然选育及其酶学特性的初步研究.科技通报,1996,12(6):333-337. Wu G F, Chen P H, Peng X G. Sereening of strains producing esterifying enzyme and studies on enzymatie charaeteristics.Bulletin of Science and Technology, 1996, 12(6):333-337.
[8] 董晓宇,李爽,侯英敏,等. 大气压冷等离子体诱变产1,3-丙二醇菌株Klebsiella pneumoniae. 过程工程学报, 2008, 8(3): 82-87. Dong X Y, Li S, Hou Y M, et al. Mutagensis of strain Klebsiella pneumoniae producing 1,3-propanediol by cold plasma at atmospheric pressure. The Chinese Journal of Process Engineering, 2008, 8(3): 82-87.
[9] Wang L Y, Huang Z L, Li G, et al.Novel mutation breeding method for Streptomyces avermitilis using an atmospheric pressure glow discharge plasma. Journal of Applied Microbiology,2010,108:851-858.
[10] Feng Xu, Haojie Jin, Huamin Li, et al. Genome shuffling of Trichoderma viride for enhanced cellulase production. Annals of Microbiology, 2012,62:509-515.
[11] Xia Shu-Qin,Liu Long,Zhang Dong-Xu,et al. Mutation and selection of transglutaminase producing strain by atmospheric pressure glow discharge plasma. Microbiology China,2010,37(11):1642-1649.
[12] 沈怡方. 白酒中四大乙酯在酿造发酵中形成的探讨. 酿酒科技,2003(5):28-31. Shen Y F. Study on the formation mechanism of four main kinds of ethyl ester in the fermentation of liquors. Liquor-Making Science ＆ Technology, 2003(5):28-31.
[13] 鄢洪德,汪钊,李光伟.产酯酶微生物菌种的筛选研究. 工业微物,2007,37(6):44-48. Yan H D, Wang Z, Li G W. Screening of microbial esterase-producing strains. Industrial Microbiology, 2007, 37(6):44-48.
[14] Patnaik R, Louie S, Gavrilovic V, et al .Genome shuffling of Lactobacillus for improved acid tolerance. Nat Biotechnol, 2002, 20: 707-712.
[15] Stephanopoulos G. Metabolic engineering by genome shuffling.Nat Biotechnology, 2002, 20: 666-668.
[16] MingHua Dai, Shelley D Copley. Genome shuffling improves degradation of the anthropogenic pesticide pentachlorophenol by Sphingobium chlorophenolicum ATCC 39723. Applied and Environmental Microbiology,2004,70(4):2391-2397.
[17] Pingying Wei,Zilong Li , Peng He , et al. Genome shuffling in the ethanologenic yeast Candida krusei to improve acetic acid tolerance. Biotechnology and Applied Biochemistry, 2008, 49: 113-120.
[18] Jya-Li Tsau, Arthiur A Guffanti, Thomas J Montville. Conversion of pyruvate to acetoin helps to maintain pH homeostasis in Lactobacillus plantarum. Appl Environ Microbiol, 1992,58:891-894.

[1]	王晓洁,孟凡强,周立邦,吕凤霞,别小妹,赵海珍,陆兆新. 利用基因组改组技术提高短杆菌素产量及其培养条件优化^*[J]. 中国生物工程杂志, 2021, 41(8): 42-51.
[2]	周惠颖,周翠霞,张婷,王雪雨,张会图,冀颐之,路福平. 强化底物利用酶系表达,提升地衣芽孢杆菌生产碱性蛋白酶性能[J]. 中国生物工程杂志, 2021, 41(2/3): 53-62.
[3]	韩海红, 汪俊卿, 王腾飞, 肖静, 韩登兰, 王瑞明. 一种基于单交换原理的地衣芽孢杆菌基因敲除方法及应用[J]. 中国生物工程杂志, 2016, 36(11): 63-69.
[4]	温赛, 杨建国. 地衣芽孢杆菌原生质体电转化方法的研究[J]. 中国生物工程杂志, 2015, 35(7): 76-82.
[5]	朱蓓霖, 周杰, 汪正华, 赵云, 黄静, 吴自荣. 地衣芽孢杆菌谷氨酰内切酶的克隆表达与性质研究[J]. 中国生物工程杂志, 2013, 33(3): 105-110.
[6]	王洲, 薛正莲, 马琦亚, 苏燕南, 赵世光. 基因组改组选育磷脂酶A1生产菌[J]. 中国生物工程杂志, 2013, 33(10): 59-66.

Viewed

Full text

Abstract

Cited

Shared

Discussed