Please wait a minute...

中国生物工程杂志

China Biotechnology
China Biotechnology  2013, Vol. 33 Issue (8): 45-50    DOI:
    
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
Download: HTML   PDF(1035KB) HTML
Export: BibTeX | EndNote (RIS)      

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 wordsEsterifying enzyme      Genome shuffling      Bacillus licheniformis      Plasma mutagenesis     
Received: 21 January 2013      Published: 25 August 2013
ZTFLH:  Q819  
Cite this article:

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.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/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] WANG Xiao-jie,MENG Fan-qiang,ZHOU Li-bang,LV Feng-xia,BIE Xiao-mei,ZHAO Hai-zhen,LU Zhao-xin. Breeding of Brevibacillin Producing Strain by Genome Shuffling and Optimization of Culture Conditions[J]. China Biotechnology, 2021, 41(8): 42-51.
[2] SONG Jia-wen, TIAN Su, ZHANG Yu-ru, WANG Zhi-zhen, CHANG Zhong-yi, GAO Hong-liang, BU Guo-jian, JIN Ming-fei. Genome Shuffling Enhances Transglutaminase Production of Streptomyces mobaraensis[J]. China Biotechnology, 2017, 37(9): 105-111.
[3] HAN Hai hong, WANG Jun qing, WANG Teng fei, XIAO Jing, HAN Deng lan, WANG Rui ming. Method and Application of Gene Knockout Based Single Cross in Bacillus licheniformis 20085[J]. China Biotechnology, 2016, 36(11): 63-69.
[4] WEN Sai, YANG Jian-guo. Transformation of Undomesticated Strains of Bacillus licheniformis by Protoplast Electroporation[J]. China Biotechnology, 2015, 35(7): 76-82.
[5] HUANG Jun, WU Ren-zhi, CHEN Ying, LU Zhi-long, CHEN Xiao-ling, CHEN Dong, HUANG Ri-bo. Screening and Breeding of high Ethanol-producing Strains by Genome Shuffling[J]. China Biotechnology, 2014, 34(7): 56-62.
[6] WANG Zhou, XUE Zheng-lian, MA Qi-ya, SU Yan-nan, ZHAO Shi-guang. Breeding of Phospholipase A1-producing Strains by Genome Shuffling[J]. China Biotechnology, 2013, 33(10): 59-66.
[7] ZHENG Lian-bao, QIU Juan-ping. The Application of Genome Shuffling in Developing New Metabolites[J]. China Biotechnology, 2012, 32(03): 100-105.
[8] . The applications and progress of genome shuffling[J]. China Biotechnology, 2010, 30(07): 0-0.
[9] MAO Yu, WANG Dan, LI Jiang, GENG Jian-Min, HUANG Tie-Bin. Protoplast Preparation and Regeneration of Actinobacillus succinogenes[J]. China Biotechnology, 2010, 30(06): 103-108.
[10] MAO Yu, WANG Dan, HUANG Tie-Bin, GENG Jian-Min. Application of Microbial Protoplast Fusion Technology in Genetic Breeding[J]. China Biotechnology, 2010, 30(01): 93-97.
[11] . Whole genome shuffling to enhance activity of fibrinolytic enzyme-producing strains[J]. China Biotechnology, 2007, 27(10): 39-43.