Please wait a minute...

中国生物工程杂志

China Biotechnology
China Biotechnology  2015, Vol. 35 Issue (11): 23-28    DOI: 10.13523/j.cb.20151104
    
The Effects of ste3 and ste4 Genes Double Disruption in Ebosin Biosynthesis
BAI Li-ping, JIANG Rong, GUO Lian-hong, ZHANG Yang, LI Yuan
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China
Download: HTML   PDF(903KB) HTML
Export: BibTeX | EndNote (RIS)      

Abstract  

Objective: The biosynthesis cluster (ste) of a novel exololysaccharide called Ebosin producing by Streptomyces had been identified previously,that ste3 and ste4 maybe encode a membrane spanning protein probably for sugar import on the basis of bioinformatics analysis. This work studied If ste3 and ste4 were involved in Ebosin biosynthesis. Methods: the ste3 and ste4 genes were disrupted together with a double crossover via homologous recombination. The mutant strain was identified by southern blot and gene complementation also performed. The production of Ebosin through isolating it from wild type, mutant and complementation was analyzed. Results: Ebosin isolated from the supernatants of fermentation cultures of the mutant strain was 153mg/L, remarkably lower than that of the wild-type strain (319mg/L). Complementation of the mutant strain resulted in a recovery of the EPS production (299mg/L) comparing with the wild-type strain. Conclusion: This research firstly elucidate that genes ste3 and ste4 encoding a membrane spanning protein involved in Ebosin biosynthesis and lay the foundation for studying the relationship between primary and second metabolism of Streptomyces sp. 139.



Key wordsste3 ste4      Ebosin      Streptomyces      Gene double disruption     
Received: 25 June 2015      Published: 25 November 2015
ZTFLH:  Q78  
Cite this article:

BAI Li-ping, JIANG Rong, GUO Lian-hong, ZHANG Yang, LI Yuan. The Effects of ste3 and ste4 Genes Double Disruption in Ebosin Biosynthesis. China Biotechnology, 2015, 35(11): 23-28.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20151104     OR     https://manu60.magtech.com.cn/biotech/Y2015/V35/I11/23

[1] Jing C, Jianbo W, Yuan L, et al. A new IL-1 receptor inhibitor 139A: fermentation, isolation, physico-chemical properties and structure. J Antibiot (Tokyo), 2003, 56(2):87-90.
[2] Zhang Y, Wang L F, Bai J Y, et al. Anti-inflammatory effect of Ebosin on rat collagen-induced arthritis through suppessing production of interleukin-1β, interleukin-6 and tumor necrosis factor. Eur J Inflamm, 2013, 11(3):677-688.
[3] Wang L Y, Li S T, Li Y. Identification and characterization of a new exopolysaccharide biosynthesis gene cluster from Streptomyces. FEMS Microbiol Lett, 2003, 220(1):21-27.
[4] Bai L, Chang M, Shan J, et al. Identification and characterization of a novel spermidine/spermine acetyltransferase encoded by gene ste26 from Streptomyces sp. 139. Biochimie, 2011, 93(9):1401-1407.
[5] Zhang Y, Zhou J, Chang M, et al. Characterization and functional evidence for Ste27 of Streptomyces sp. 139 as a novel spermine/spermidine acetyltransferase. Biochem J, 2012, 443(3):727-734.
[6] 白利平,李元. 链霉菌调控蛋白DasRABC的研究进展. 微生物学通报, 2010, 37(9):1369-1373. Bai L P, Li Y. Study of DasRABC in Streptomyces. Microbiology China, 2010, 37(9):1369-1373.
[7] Rigali S, Titgemeyer F, Barends S, et al. Feast or famine: the global regulator DasR links nutrient stress to antibiotic production by Streptomyces. EMBO Rep, 2008, 9(7):670-675.
[8] Rigali S, Nothaft H, Noens E E, et al. The sugar phosphotransferase system of Streptomyces coelicolor is regulated by the GntR-family regulator DasR and links N-acetylglucosamine metabolism to the control of development. Mol Microbiol, 2006, 61(5):1237-1251.
[9] Seo J W, Ohnishi Y, Hirata A, et al. ATP-binding cassette transport system involved in regulation of morphological differentiation in response to glucose in Streptomyces griseus. J Bacteriol, 2002, 184(1):91-103.
[10] Bai L, Qi X, Zhang Y, et al. A new GntR family regulator Ste1 in Streptomyces sp. 139. Appl Microbiol Biotechnol, 2013, 97(19):8673-8682.
[11] Kieser T, Bibb M J, Butter M J, et al. Practical Streptomyces Genetics. Norwich:The John Innes Foundation, 2000.
[12] Marchler-Bauer A, Derbyshire M K, Gonzales N R, et al. CDD: NCBI's conserved domain database. Nucleic Acids Res, 2015, 43(Database issue):D222-226.
[13] Herrou J, Crosson S. Myo-inositol and D-ribose ligand discrimination in an ABC periplasmic binding protein. J Bacteriol, 2013, 195(10):2379-2388.
[14] Colson S, van Wezel G P, Craig M, et al. The chitobiose-binding protein, DasA, acts as a link between chitin utilization and morphogenesis in Streptomyces coelicolor. Microbiology, 2008, 154(Pt 2):373-382.
[15] Saito A, Shinya T, Miyamoto K, et al. The dasABC gene cluster, adjacent to dasR, encodes a novel ABC transporter for the uptake of N,N'-diacetylchitobiose in Streptomyces coelicolor A3(2). Appl Environ Microbiol, 2007, 73(9):3000-3008.
[16] Lebeer S, Verhoeven T L, Francius G, et al. Identification of a gene cluster for the biosynthesis of a long, galactose-rich exopolysaccharide in Lactobacillus rhamnosus GG and functional analysis of the priming glycosyltransferase. Appl Environ Microbiol, 2009, 75(11):3554-3563.

[1] LI Xiao-mei, LIN Chun-yan, PANG Ai-ping, LI Xiao-bo, ZHAO Guang-rong. Application of Synthetic Biology in Research and Development of the Secondary Metabolites from Streptomyces[J]. China Biotechnology, 2015, 35(4): 92-97.
[2] TAO Wen-na, XIA Li-qiu, DING Xue-zhi, TANG Ying. Cloning and Function Study of amtS Gene from Saccharopolyspora spinosa[J]. China Biotechnology, 2015, 35(2): 25-30.
[3] HE Jie, SU Ling-qia, WU Jing. Optimization of Fermentation in Shake Flasks for the Xylanase in Recombinant E. coli[J]. China Biotechnology, 2013, 33(2): 41-46.
[4] CHEN Xu-sheng, REN Xi-dong, ZENG Xin, DONG Nan, MAO Zhong-gui. ε-Poly-L-lysine Production from Precursor L-lysine by Streptomyces sp. M-Z18[J]. China Biotechnology, 2013, 33(1): 53-59.
[5] BAI Li-Beng-1, XIE Hong-Guan-1, CHAN Dun-Jie-2, JIANG Rong-1, ZHANG Xiang-1, GUO Lian-Hong-1, LI Yuan-1. The Effects of Glycosyltransferase Genes Double Disruption in Ebosin Biosynthesis[J]. China Biotechnology, 2009, 29(06): 46-51.