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浙江大学学报(工学版)
J4  2010, Vol. 44 Issue (2): 344-348    DOI: 10.3785/j.issn.1008-973X.2010.02.024
    
Enhanced production of pristinamycin fermentation by genome shuffling
XU Bo1,2, JIN Zhi-hua1,2, JIN Qing-chao1, CEN Pei-lin2
(1. Department of Biotechnology and Pharmaceutical Engineering, Ningbo Institute of Technology, Zhejiang University Ningbo 315100, China;
2. College of Materials Science and Chemical Engineering, Zhejiang University, Hangzhou 310027, China)
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Abstract  

A series of high pristinamycin-producing mutants were screened after spores and protoplasts of Streptomyces pristinaespiralis ND-23, a producer of pristinamycin, were treated by UV mutagenesis. The pristinamycin production of the highest producer mutant reached 356 mg/L, which was increased by 31% compared with that of the original strain. Genome shuffling was applied to improve pristinamycin production of Streptomyces pristinaespiralis by utilizing the four highest strains derived from the above UV mutants as the starting population. A recombinant Streptomyces pristinaespiralis G-211 with improved phenotype was obtained by four rounds of genome shuffling, and its production reached 832 mg/L, which was increased by 134% and 206% compared with that of the highest parent strain and the original strain, respectively. Fermentation experiments by Streptomyces pristinaespiralis G-211 and ND-23 were conducted in 5 L fermentor. Results show that the biosynthesis of pristinamycins is not coupled to cell growth of Streptomyces pristinaespiralis.The consumption rate of glucose and amino nitrogen of the strain G-211 is more rapid than that of the initial strain ND-23. The results can be useful for medium optimization and scale-up of pristinamycin fermentation.

Published: 09 March 2010
CLC:  Q 939.9  
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Cite this article:

XU Bei, JIN Zhi-Hua, JIN Qiang-Chao, et al. Enhanced production of pristinamycin fermentation by genome shuffling. J4, 2010, 44(2): 344-348.

URL:

http://www.zjujournals.com/eng/10.3785/j.issn.1008-973X.2010.02.024     OR     http://www.zjujournals.com/eng/Y2010/V44/I2/344


应用基因组重排技术提高普那霉素产量

将普那霉素产生菌——始旋链霉菌ND-23的孢子和原生质体经紫外线诱变后获得高产突变菌株,其中高产突变株SP-S73的产量达到356 mg/L,比出发菌株提高了31%.在上述增产突变株中选取4株作为基因组重排的出发亲本,对它们进行了4轮基因组重排育种,筛选得到了高产重排菌株,其中1株重排菌株G-211的普那霉素产量为832 mg/L,比产量最高的亲本菌株提高了134%,比原始出发菌株ND-23(272 mg/L)提高了206%.通过研究高产菌株和出发菌株ND-23在5 L罐上的发酵过程,发现普那霉素产生菌的生物合成属于非生长耦合型；其高产菌株G-211在合成产物的时期对还原糖和氨基氮的消耗量均大于原始菌株ND-23,这些结果将为高产菌株发酵条件优化和发酵放大研究提供有价值的参考.

［1］  PAQUET V, GOMA G, SOUCAILLE P. Induction of pristinamycins production in Streptomyces pristinaespiralis［J］. Biotechnology Letters, 1992, 14(11): 10651070.
［2］ CORVINI P F X, DELAUNAY S, MAUJEAN F, et al. Intracellular pH determination of pristinamycin-producing Streptomyces pristinaespiralis by image analysis［J］. Microbiology, 2000, 146(10): 26712678.
［3］ CORVINI P F X, DELAUNAY S, MAUJEAN F, et al. Intracellular pH of Streptomyces pristinaespiralis is correlated to the sequential use of carbon sources during the pristinamycins-producing process［J］. Enzyme and Microbial Technology, 2004, 34(2): 101107.
［4］ VOELKER F, ALTABA S. Nitrogen source governs the patterns of growth and pristinamycin production in Streptomyces pristinaespiralis［J］. Microbiology, 2001, 147(9): 24472459.
［5］ SUERMONDT G, DENAMUR E, FLEURET A, et al. In vitro activity of miocamycin compared with pristinamycin and four macrolides on Branhamella catarrhalis［J］. Pathologie Biologie, 1989, 37(5): 386389.
［6］ VON EIFF C, PETERS G. In-vitro activity of RP 59500, a new semisynthetic injectable pristinamycin against staphylococci［J］. Zentralblatt fuer Bakteriologie, 1996, 283(4): 497501.
［7］ WEBER P, DUBOIS S, BOUSSOUGANT Y. In vitro activity of pristinamycin and its components against gram-negative anaerobic bacilli and Gardnerella vaginalis［J］. Journal of Antimicrobial Chemotherapy, 1989, 23(6): 825830.
［8］ ZHANG Y X, PERRY K, VINCI V A, et al. Genome shuffling leads to rapid phenotypic improvement in bacteria［J］. Nature, 2002, 415(6872): 644646.
［9］ GONG G L, SUN X, LIU X L, et al. Mutation and a high-throughput screening method for improving the production of epothilones of Sorangium［J］. Journal of Industrial Microbiology and Biotechnology, 2007, 34(9): 615623.
［10］ JIA Bo, JIN Zhi-hua, LEI Yin-lin, et al. Improved production of pristinamycin coupled with an adsorbent resin in fermentation by Streptomyces pristinaespiralis［J］. Biotechnology Letters, 2006, 28(22): 18111815.
［11］ 朱林东,金志华.原生质体诱变育种选育普那霉素高产菌［J］.中国抗生素杂志,2006,31(10): 591594.
ZHU Lin-dong, JIN Zhi-hua. Selection of high pristinamycin-producing strain by protoplast mutation with UV［J］. Chinese Journal of Antibiotics, 2006, 31(10): 591594.
［12］ 陈钧鸣,徐玲娣.抗生素工业分析［M］.北京:中国医药科技出版社,1991: 109139.
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