提高放线菌次级代谢产物产量方法的研究进展

摘要/Abstract

摘要： 放线菌的次级代谢产物一直是抗生素等药物开发的重要来源。通过基因技术来提高放线菌次级代谢产物产量的方法已被广泛接受。结合近年的研究成果，概述了利用基因技术过表达或者敲除代谢途经关键基因及加倍基因簇，诱变核糖体相关蛋白的基因，以及异源表达外源基因簇等提高次级代谢产物产量的方法。旨在为寻找新药及改造高产菌株提供相应的技术参考。

关键词: 次级代谢, 核糖体工程, 异源表达, 放线菌

Abstract: Actinomycetes secondary metabolites have been the important sources of developing antibiotics and other drugs. Gene technology has been widely practiced for improving the yield of Actinomycetes secondary metabolites. This review summarized a series of technical methods used for improving Actinomycetes secondary metabolites，consisting of over-expressing the key structural genes and/or positive regulation genes，knocking out the negative regulatory genes，doubling the clusters，mutating ribosomal protein genes，and expressing the foreign cluster heterologously. These methods could be used for new drug discovery and strains improvement.

Key words: Secondary, Netabolites, Ribosome engineering, Heterologous expression, Actinomycetes

雷秀清, 李力, 黄建忠. 提高放线菌次级代谢产物产量方法的研究进展[J]. 生物技术通报, 2014, 0(5): 45-51.

Lei Xiuqing, Li Li, Huang Jianzhong. Research Progress of Improving the Actinomycetes Secondary Metabolite Production Method[J]. Biotechnology Bulletin, 2014, 0(5): 45-51.

参考文献

[1] Page MG. Antibiotic discovery and development[M]. Springer, 2012：79-117.
[2] 李力, 贺新义, 邓子新. Streptoverticillium rimofaciens ZJU5119提取液中米多霉素及其衍生物的分析[J]. 上海交通大学学报, 2009(1)：1-4.
[3] Park CN, Lee JM, Lee D, et al. Antifungal activity of valinomycin, a peptide antibiotic produced by Streptomyces sp. Strain M10 antagonistic to Botrytis cinerea[J]. J Microbiol Biotechnol, 2008, 18(5)：880-884.
[4] Robbel L, Marahiel MA. Daptomycin, a bacterial lipopeptide synthe-sized by a nonribosomal machinery[J]. Journal of Biological Chemistry, 2010, 285(36)：27501-27508.
[5] 亓芳, 王振东, 刘霆. 纳他霉素及其生产菌育种研究进展[J]. 生物技术通报, 2010(9)：42-47.
[6] Wohlleben W, Mast Y, Muth G, et al. Synthetic Biology of secondary metabolite biosynthesis in actinomycetes：Engineering precursor supply as a way to optimize antibiotic production[J]. FEBS Letters, 2012, 586(15)：2171-2176.
[7] 陈路劼, 赵薇, 连云阳. 聚酮合酶与药物筛选的研究进展[J]. 中国抗生素杂志, 2012, 37(9)：655-661.
[8] 段月娇, 薛超友, 卢文玉. 异源表达聚酮类化合物前体的研究进展[J]. 中国生物工程杂志, 2012, 32(11)：107-114.
[9] Shen B. Polyketide biosynthesis beyond the type I, II and III polyketide synthase paradigms[J]. Curr Opin Chem Biol, 2003, 7(2)：285-295.
[10] Chen Y, Smanski MJ, Shen B. Improvement of secondary metabolite production in Streptomyces by manipulating pathway regulation[J]. Appl Microbiol Biotechnol, 2010, 86(1)：19-25.
[11] Hertweck C, Luzhetskyy A, Rebets Y, et al. Type II polyketide synthases：gaining a deeper insight into enzymatic teamwork[J]. Natural Product Reports, 2007, 24(1)：162-190.
[12] Watanabe K, Praseuth AP, Wang CC. A comprehensive and engag-ing overview of the type III family of polyketide synthases[J]. Curr Opin Chemi Biol, 2007, 11(3)：279-286.
[13] Chiang YM, Chang SL, Oakley BR, et al. Recent advances in awakening silent biosynthetic gene clusters and linking orphan clusters to natural products in microorganisms[J]. Current Opinion in Chemical Biology, 2011, 15(1)：137-143.
[14] Laureti L, Song LJ, Huang S, et al. Identification of a bioactive 51-membered macrolide complex by activation of a silent polyketide synthase in Streptomyces ambofaciens[J]. Proceedings of the National Academy of Sciences, 2011, 108(15)：6258-6263.
[15] Ryu YG, Butler MJ, Chater KF, et al. Engineering of primary carbohydrate metabolism for increased production of actinorhodin in Streptomyces coelicolor[J]. Applied and Environmental Microbiology, 2006, 72(11)：7132-7139.
[16] 白林泉, 邓子新. 微生物次级代谢产物生物合成基因簇与药物创新[J]. 中国抗生素杂志, 2006, 31(2)：80-86.
[17] Antón N, Mendes MV, Martin JF, et al. Identification of PimR as a positive regulator of pimaricin biosynthesis in Streptomyces natalensis[J]. J Bacteriol, 2004, 186(9)：2567-2575.
[18] Antón N, Santos-Aberturas J, Mendes MV, et al. PimM, a PAS domain positive regulator of pimaricin biosynthesis in Streptomyces natalensis[J]. Microbiology, 2007, 153(9)：3174-3183.
[19] Jang BY, Hwang YI, Choi SU. Effects of pimM and pimR on the Increase of Natamycin Production in Streptomyces natalensis[J]. Journal of the Korean Society for Applied Biological Chemistry, 2011, 54(1)：141-144.
[20] Santos-Aberturas J, Vicente CM, Payero TD, et al. Hierarchical control on polyene macrolide biosynthesis：PimR modulates pimaricin production via the PAS-LuxR transcriptional activator pimM[J]. PloS One, 2012, 7(6)：1-10.
[21] Karray F, Darbon E, Oestreicher N, et al. Organization of the biosynthetic gene cluster for the macrolide antibiotic spiramycin in Streptomyces ambofaciens[J]. Microbiology, 2007, 153(12)：4111-4122.
[22] Juguet M, Lautru S, Francou FX, et al. An iterative nonribosomal peptide synthetase assembles the pyrrole-amide antibiotic congocidine in Streptomyces ambofaciens[J]. Chemistry & Biology, 2009, 16(4)：421-431.
[23] Nicolaou KC, Tria GS, Edmonds DJ. Total synthesis of platencin[J]. Angewandte Chemie, 2008, 120(9)：1804-1807.
[24] Wang J, Kodali S, Lee SH, et al. Discovery of platencin, a dual FabF and FabH inhibitor with in vivo antibiotic properties[J]. Proc Nat Acad Sci USA, 2007, 104(18)：7612-7616.
[25] Smanski MJ, Peterson RM, Rajski SR, et al. Engineered Streptomy-ces platensis strains that overproduce antibiotics platensimycin and platencin[J]. Antimicrobial Agents and Chemotherapy, 2009, 53(4)：1299-1304.
[26] Liu W, Christenson SD, Standage S, et al. Biosynthesis of the enediyne antitumor antibiotic C-1027[J]. Science, 2002, 297(5584)：1170-1173.
[27] Wang L, Hu Y, Zhang Y, et al. Role of sgcR3 in positive regulation of enediyne antibiotic C-1027 production of Streptomyces globispo-rus C-1027[J]. BMC Microbiology, 2009, 9(1)：1-14.
[28] Chen YH, Yin M, Horsman GP, et al. Improvement of the enediyne antitumor antibiotic C-1027 production by manipulating its biosynthetic pathway regulation in Streptomyces globisporus[J]. Journal of Natural Products, 2011, 74(3)：420-424.
[29] Yanai K, Murakami T, Bibb M. Amplification of the entire kanamycin biosynthetic gene cluster during empirical strain improvement of Streptomyces kanamyceticus[J]. Proceedings of the National Academy of Sciences, 2006, 103(25)：9661-9666.
[30] Murakami T, Burian J, Yanai K, et al. A system for the targeted amplification of bacterial gene clusters multiplies antibiotic yield in Streptomyces coelicolor[J]. Proceedings of the National Academy of Sciences, 2011, 108(38)：16020-16025.
[31] Liao G, Li J, Li L, et al. Cloning, reassembling and integration of the entire nikkomycin biosynthetic gene cluster into Streptomyces ansochromogenes lead to an improved nikkomycin production[J]. Microbial Cell Factories, 2009, 9：1-6.
[32] Yu L, Cao N, Wang L, et al. Oxytetracycline biosynthesis improvement in Streptomyces rimosus following duplication of minimal PKS genes[J]. Enzyme and Microbial Technology, 2012, 50(6-7)：318-324.
[33] Ochi K, Hosaka T. New strategies for drug discovery：activation of silent or weakly expressed microbial gene clusters[J]. Applied Microbiology and Biotechnology, 2013, 97(1)：87-98.
[34] Hu H, Zhang Q, Ochi K. Activation of antibiotic biosynthesis by specified mutations in the rpoB gene encoding the RNA polymerase beta subunit of Streptomyces lividans[J]. Journal of Bacteriology, 2002, 184(14)：3984-3891.
[35] Shima J, Hesketh A, Okamoto S, et al. Induction of actinorhodin production by rpsL(encoding ribosomal protein S12)mutations that confer streptomycin resistance in Streptomyces lividans and Streptomyces coelicolor A3(2)[J]. Journal of Bacteriology, 1996, 178(24)：7276-7284.
[36] Wang G, Inaoka T, Okamoto S, et al. A novel insertion mutation in Streptomyces coelicolor ribosomal S12 protein results in paromomycin resistance and antibiotic overproduction[J]. Antimicrob Agents Chemothera, 2009, 53(3)：1019-1026.
[37] Hu HF, Ochi K. Novel approach for improving the productivity of antibiotic-producing strains by inducing combined resistant muta-tions[J]. App Environ Microbiol, 2001, 67(4)：1885-1892.
[38] Wang G, Hosaka T, Ochi K. Dramatic activation of antibiotic prod-uction in Streptomyces coelicolor by cumulative drug resistance mut-ations[J]. Appl Environ Microbiol, 2008, 74(9)：2834-2840.
[39] Li L, Wu J, Deng ZX, et al. Streptomyces lividans blasticidin S deaminase and its application in engineering a blasticidin S-producing strain for ease of genetic manipulation[J]. Applied and Environmental Microbiology, 2013, 79(7)：2349-2357.
[40] Gomez-Escribano JP, Bibb MJ. Heterologous expression of natural product biosynthetic gene clusters in Streptomyces coelicolor：from genome mining to manipulation of biosynthetic pathways[J]. J Industrial Microbiol Biotechnol, 2014, 41(2)：425-431.
[41] Komatsu M, Komaetu K, Koiwai H, et al. Engineered Streptomyces avermitilis host for heterologous expression of biosynthetic gene cluster for secondary metabolites[J]. ACS Synthetic Biology, 2013, 2(7)：384-396.
[42] Gomez-Escribano JP, Song LJ, 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(9)：2716-2720.
[43] Gomez-Escribano JP, Bibb MJ. Engineering Streptomyces coelicolor for heterologous expression of secondary metabolite gene clusters[J]. Microbial Biotechnology, 2011, 4(2)：207-215.
[44] Zhou M, Jing XY, Xie PF, et al. Sequential deletion of all the polyketide synthase and nonribosomal peptide synthetase biosynthetic gene clusters and a 900-kb subtelomeric sequence of the linear chromosome of Streptomyces coelicolor[J]. FEMS Microbiology Letters, 2012, 333(2)：169-179.
[45] Komatsu M, Uchiyama T, Omura S, et al. Genome-minimized Streptomyces host for the heterologous expression of secondary metabolism[J]. PNASUSA, 2010, 107(6)：2646-2651.
[46] Komatsu M, Tsuda M, Omura S, et al. Identification and functional analysis of genes controlling biosynthesis of 2-methylisoborneol[J]. PNAS, 2008, 105(21)：7422-7427.
[47] Li L, Xu ZN, Xu XY, et al. The mildiomycin biosynthesis：initial steps for sequential generation of 5-hydroxymethylcytidine 5'-monophosphate and 5-hydroxymethylcytosine in Streptoverticillium rimofaciens ZJU[J]. Chem Bio Chem, 2008, 9(8)：1286-1294.
[48] Wu J, Li L, Deng ZX, et al. Analysis of the mildiomycin biosynthesis gene cluster in Streptoverticillum remofaciens ZJU5119 and characterization of MilC, a hydroxymethyl cytosyl-glucuronic acid synthase[J]. Chem Bio Chem, 2012, 13(11)：1613-1621.
[49] 李力, 贺新义, 邓子新. 米多霉素生物合成机理的研究[D]. 上海：上海交通大学, 2008.
[50] Wong FT, Khosla C. Combinatorial biosynthesis of polyketides—a perspective[J]. Curr Opin Chem Biol, 2012, 16(1)：117-123.