sco1135基因对天蓝色链霉菌M145孢子形成及次级代谢产物合成的调控研究

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

生物技术通报 ›› 2017, Vol. 33 ›› Issue (1): 141-147.doi: 10.13560/j.cnki.biotech.bull.1985.2017.01.016

sco1135基因对天蓝色链霉菌M145孢子形成及次级代谢产物合成的调控研究

马军霞^{1， 2}，张佩佩³，王世立^{1， 2}，曹广祥^{1， 2}

1. 济南大学山东省医学科学院医学与生命科学学院，济南 250022；
2. 山东省医药生物技术研究中心，济南 250062；
3. 山东大学微生物技术国家重点实验室，济南 250100

收稿日期:2016-09-20 出版日期:2017-01-25 发布日期:2017-01-19
作者简介:马军霞，女，硕士，研究方向：微生物次级代谢产物合成调控;E-mail：shuipin_gzuo@163.com
基金资助:
山东省医学科学院医药卫生科技创新工程

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

MA Jun-xia^{1, 2}, ZHANG Pei-pei³, WANG Shi-li^{1, 2}, CAO Guang-xiang¹, ²

1. School of Medicine and Life Sciences,University of Jinan-Shandong Academy of Medical Sciences,Ji’nan 250022;
2. Shandong Medicinal Biotechnology Center,Ji’nan 250062;
3. The State Key Laboratory of Microbial Technology,Shandong University,Ji’nan 250100;

Received:2016-09-20 Published:2017-01-25 Online:2017-01-19

摘要/Abstract

摘要： 旨在研究sco1135基因缺失突变对天蓝色链霉菌M145菌株形态及次级代谢的影响。通过PCR-targeting方法获得重组质粒pSJ1135，通过接合转移将其导入天蓝色链霉菌M145，获得sco1135基因缺失突变菌株△sco1135，并以pMS82为载体构建回补菌株△sco1135com，同时以pMS82为空载对照;随后对野生型菌株、突变菌株和回补菌株进行表型分析和抗生素定量观察。结果显示，表型分析及抗生素定量测定发现，在YBP培养基上△sco1135产孢明显延迟于野生型M145，放线紫红素（ACT）产量明显增加，突变株培养基中ACT产量是野生菌株培养基中的2-3倍;转录分析结果表明，48 h时突变株部分与产孢相关基因的转录水平较野生型降低了50%-75%，72 h时突变株部分与产ACT相关基因的转录水平较野生型提高13-20倍。sco1135基因参与调控M145的孢子形成及次级代谢产物ACT的产生。

关键词: 天蓝色链霉菌M145, sco1135, 表型变化, 孢子形成, 次级代谢产物

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

马军霞，张佩佩，王世立，曹广祥. sco1135基因对天蓝色链霉菌M145孢子形成及次级代谢产物合成的调控研究[J]. 生物技术通报, 2017, 33(1): 141-147.

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.

参考文献

[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.

sco1135基因对天蓝色链霉菌M145孢子形成及次级代谢产物合成的调控研究

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

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	和梦颖, 刘文彬, 林震鸣, 黎尔彤, 汪洁, 金小宝. 一株抗革兰阳性菌的戈登氏菌WA4-43全基因组测序与分析[J]. 生物技术通报, 2023, 39(2): 232-242.
[2]	王楠, 苏誉, 刘文杰, 封明, 毛瑜, 张新国. 植物内生菌中抗耐药微生物活性成分的研究进展[J]. 生物技术通报, 2021, 37(8): 263-274.
[3]	赵江华, 房欢, 张大伟. 微生物次级代谢产物生物合成的研究进展[J]. 生物技术通报, 2020, 36(11): 141-147.
[4]	吴琴琴, 孙敏, 陈雨, 付雅琴, 曾斌, 贺斌. 米曲霉功能基因组研究策略和进展[J]. 生物技术通报, 2019, 35(8): 186-192.
[5]	徐杰, 黄建忠, 李力. 基因组挖掘技术及其在真菌中的研究进展[J]. 生物技术通报, 2019, 35(11): 201-207.
[6]	郝泽婷, 郝晓慧, 阎婧, 谢丙炎, 梁丽琴. 淡紫拟青霉次级代谢产物的研究进展[J]. 生物技术通报, 2018, 34(10): 11-17.
[7]	林海州, 陈洲琴王燕郭俊朱红惠邓名荣. 利用“移树养苗”策略挖掘粤蓝链霉菌隐性活性次级代谢产物[J]. 生物技术通报, 2017, 33(9): 145-152.
[8]	魏志文,孙勇,王菲. 激发子对桦褐孔菌多酚合成的诱导及其生化机制初探[J]. 生物技术通报, 2014, 0(9): 136-141.
[9]	宋恺, 胡洁, 林文翰, 季宇彬. 海绵共附生真菌多样性及其次级代谢产物的研究进展[J]. 生物技术通报, 2014, 0(4): 36-42.
[10]	赵婷峰 ,龚国利. 黏细菌：天然的制药厂[J]. 生物技术通报, 2014, 0(12): 40-46.
[11]	蔡成平;王远山;郑裕国;. 核糖体工程与微生物次级代谢产物合成[J]. , 2012, 0(09): 51-58.
[12]	袁丽杰;刘爱华;张玉琴;余利岩;张月琴;. 一株根际放线菌的分离鉴定及其生物活性初探[J]. , 2012, 0(03): 117-122.
[13]	孙盈盈. 离体培养生产次级代谢产物[J]. , 1997, 0(02): 36-37.
[14]	孙盈盈;. 离体培养生产次级代谢产物[J]. , 1997, 0(01): 26-27.
[15]	. 医药其它[J]. , 1994, 0(05): 75-82.