Regulatory Effects of Gene sco1135 on the Sporulation and Secondary Metabolite Synthesis of Streptomyces coelicolor M145

doi:10.13560/j.cnki.biotech.bull.1985.2017.01.016

Abstract

Abstract: This work aims to study the effects of deletion and mutation of gene sco1135 on the morphogenesis and secondary metabolism in Streptomyces coelicolor strain M145. Recombinant plasmid pSJ1135 was obtained by PCR-targeting. Gene deletion mutant of sco1135（△sco1135）was acquired by introducing the recombinant plasmid into the Streptomyces coelicolor M145 via conjugation and transformation,and the complemented strain △sco1135com was constructed using the vector pMS82. Using pMS82 as a control,the phenotypes of the parental wild strain（M145）,mutant strain（△sco1135）and complemented strain（△sco1135com）were analyzed and observed with quantitative antibiotics. The results of phenotypic analysis and observation with quantitative antibiotics revealed that the sporulation in △sco1135 obviously delayed than that of the wild type M145 on YBP medium;while the actinorhodin（ACT）production in △sco1135 increased to 2-3 folds of that in M145. The transcriptions of some sporulation-related genes decreased by 50%-75% in △sco1135 at 48 h,while the transcriptional expressions of genes related to ACT in △sco1135 increased to 13-20 folds at 72 h,compared to the M145. Conclusively,sco1135 is involved in regulating the sporulation in M145 and also the production of secondary metabolite ACT.

Key words: Streptomyce coelicolor M145, sco1135, phenotypic changes, sporulation, secondary metabolite

MA Jun-xia, ZHANG Pei-pei, WANG Shi-li, CAO Guang-xiang,. Regulatory Effects of Gene sco1135 on the Sporulation and Secondary Metabolite Synthesis of Streptomyces coelicolor M145[J]. Biotechnology Bulletin, 2017, 33(1): 141-147.

References

[1] Challis GL, Hopwood DA. Synergy and contingency as driving forces for the evolution of multiple secondary metabolite production by Str-eptomyces species[J]. Proc Natl Acad Sci USA, 2003, 100（Suppl 2）：14555-14561.
[2] Hopwood DA. Therapeutic treasures from the deep[J]. Nat Chem Biol, 2007, 3（8）：457-458.
[3] Nodwell JR. Novel links between antibiotic resistance and antibiotic production[J]. J Bacteriol, 2007, 189（10）：3683-3685.
[4] Bentley SD, Chater KF, Cerdeño-Tárraga AM, et al. Complete genome sequence of the model actinomycete Streptomyces coelicolor A3（2）[J]. Nature, 2002, 417（6885）：141-147.
[5] Hwang KS, Kim HU, Charusanti P, et al. Systems biology and biotechnology of Streptomyces species for the production of secondary metabolites[J]. Biotechnol Adv, 2014, 32（2）：255-268.
[6] Gomez-Escribano JP, Song L, Fox DJ, et al. Structure and biosynthesis of the unusual polyketide alkaloid coelimycin P1, a metabolic product of the cpk gene cluster of Streptomyces coelicolor M145[J]. Chemical Science, 2012, 3：2716-2720.
[7] Liu G, Chandra G, Tan H, et al. Molecular regulation of antibiotic biosynthesis in Streptomyces[J]. Microbiol Mol Biol Rev, 2013, 77（1）：112-143.
[8] Wang W, Shu D, Lu Y, et al. Cross-talk between an orphan response regulator and a noncognate histidine kinase in Streptomyces coelicolor[J]. FEMS Microbiol Lett, 2009, 294（2）：150-156.
[9] Kieser T, Bibb M, Buttner M, et al. Practical Streptomyces Genetics：a laboratory Manual[M]. Norwich：John Innes Foundation, 2000.
[10] Gust B, Challis GL, Fowler K, et al. PCR-targeted Streptomyces gene replacement identifies a protein domain needed for biosynthesis of the sesquiterpene soil odor geosmin[J]. Proc Natl Acad Sci USA, 2003, 100：1541-1546.
[11] Gregory MA, Till R, Smith MC. Integration site for Streptomyces phage phiBT1 and development of site-specific integrating vectors[J]. Journal of Bacteriol, 2003, 185（17）：5320-5323.
[12] Gust B, Kieser T, Chater KF. Redireccted technology：PCR-targeting sysrem in Streptomyces coelicolor A3（2）：a laboratory manual[M]. Norwich：John Innes Foundation, 2002.
[13] Sambrook J, Russell D. Molecular Cloning：A Laboratory Manual[M]. New York：Cold Spring Harbor Laboratory Press, 2001.
[14] Yu Z, Zhu H, Dang F, et al. Differential regulation of antibiotic biosynthesis by DraR-K, a novel two-component system in Strepto-myces coelicolor[J]. Mol Microbiol, 2012, 85（3）：535-556.
[15] Hillerich B, Westpheling J. A new TetR family transcriptional regulator required for morphogenesis in Streptomyces coelicolor[J]. Journal of Bacteriol, 2008, 190（1）：61-67.
[16] Chater KF. Regulation of sporulation in Streptomyces coelicolor A3（2）：A checkpoint multiplex?[J]. Current Opinion in Microbiology, 2001（4）：667-673.
[17] Keijser BJ, van Wezel GP, Canters GW, et al. The Ram-dependence of Streptomyces lividans difference is by bypassed by copper[J]. J Molecul Microbiol Biotechnol, 2014, 82（6）：1093-1098.
[18] Kim JM, Won HS, Kang SO. The C-terminal domain of the transcriptional regulator BldD from Streptomyces coelicolor A3（2）constitutes a novel fold of winged-helix domains[J]. Proteins, 2014, 82（6）：1093-1098.
[19] Bibb MJ, Domonkos A, Chandra G, et al. Expression of the chaplin and rodlin hydrophobic sheath proteins in Streptomyces venezuelae is controlled by σBldN and a cognate anti-sigma factor, RsbN[J]. Molecular Microbiology, 2012, 84（6）：1033-1049.
[20] Taguchi T, Itou K, Ebizuka Y, et al. Chemical characterisation of disruptants of the Streptomyces coelicolor A3（2）actVI genes involved in actinorhodin biosynthesis[J]. Journal of Antibiotics, 2000, 53（2）：144-152.
[21] Iqbal M, Mast Y, Amin R, et al. Extracting regulator activity profiles by integration of de novo motifs and expression data：characterizing key regulators of nutrient depletion responses in Streptomyces coelicolor[J]. Nucleic Acids Res, 2012, 40（12）：5227-5239.