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Construction of ivermectin producer by domain swaps of avermectin polyketide synthase in Streptomyces avermitilis

  • Applied Genetics and Molecular Biotechnology
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Abstract

Ivermectin, 22, 23-dihydroavermectin B1, is commercially important in human, veterinary medicine, and pesticides. It is currently synthesized by chemical reduction of the double bond between C22 and C23 of avermectins B1, which are a mixture of B1a (>80%) and B1b (<20%) produced by fermentation of Streptomyces avermitilis. The cost of ivermectin is much higher than that of avermectins B1 owing to the necessity of region-specific hydrogenation at C22–C23 of avermectins B1 with rhodium chloride as the catalyst for producing ivermectin. Here we report that ivermectin can be produced directly by fermentation of recombinant strains constructed through targeted genetic engineering of the avermectin polyketide synthase (PKS) in S. avermitilis Olm73-12, which produces only avermectins B and not avermectins A and oligomycin. The DNA region encoding the dehydratase (DH) and ketoreductase (KR) domains of module 2 from the avermectin PKS in S. avermitilis Olm73-12 was replaced by the DNA fragment encoding the DH, enoylreductase, and KR domains from module 4 of the pikromycin PKS of Streptomyces venezuelae ATCC 15439 using a gene replacement vector pXL211. Twenty-seven of mutants were found to produce a small amount of 22, 23-dihydroavermectin B1a and avermectin B1a and B2a by high performance liquid chromatography and liquid chromatography mass spectrometry analysis. This study might provide a route to the low-cost production of ivermectin by fermentation.

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References

  • Aziz MA, Diallo S, Diop IM, Lariviere M, Parta M (1982) Efficacy and tolerance of ivermectin in human onchocerciasis. Lancet 2:171–173

    Article  CAS  Google Scholar 

  • Bierman M, Logan R, O’Brien K, Seno ET, Rao RN, Schoner BE (1992) Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116:43–49

    Article  CAS  Google Scholar 

  • Burg RW, Miller BM, Baker EE, Birnbaum J, Currie SA, Hartman R, Kong Y-L, Monaghan RL, Olson G, Putter I, Tunac JB, Wallick H, Stapley EO, Oiwa R, Ōmura S (1979) Avermectins, new family of potent anthelmintic agents: producing organism and fermentation. Antimicrob Agents Chemother 15:361–367

    Article  CAS  Google Scholar 

  • Cane DE, Liang T-C, Kaplan LK, Nallin MK, Schulman MD, Hensens OD, Douglas AW, Albers-Schőnberg G (1983) Biosynthetic origin of the carbon skeleton and oxygen atoms of the avermectins. J Am Chem Soc 105:4110–4112

    Article  CAS  Google Scholar 

  • Chabala JC, Mrozik H, Tolman RL, Eskola P, Lusi A, Peterson LH, Woods MF, Fisher MH, Campbell WC, Egerton JR, Ostling DA (1980) Ivermectin, a new broad-spectrum antiparasitic agent. J Med Chem 23:1134–1136

    Article  CAS  Google Scholar 

  • Chen TS, Hensens OD, Schulman MD (1989) Biosynthesis. In: Campbell WC (ed) Ivermectin and abamectin. Springer, Berlin Heidelberg New York, pp 55–72

    Chapter  Google Scholar 

  • Chen Z, Wen Y, Song Y, Li JL (2002) Effect of gene deletion of aveD on avermectins production in Streptomyces avermitilis. Acta Microbiol Sin 42:534–538

    CAS  Google Scholar 

  • Egerton JR, Ostling DA, Blair LS, Eary CH, Suhayda D, Cifelli S, Riek RF, Campbell WC (1979) Avermectins, new family of potent anthelmintic agents: efficacy of the B1a component. Antimicrob Agents Chemother 15:372–378

    Article  CAS  Google Scholar 

  • Hopwood DA, Bibb MJ, Chater KF, Kieser T, Bruton CJ, Kieser HM, Lydiate DJ, Smith CP, Ward JM, Schrempf H (1985) Genetic manipulation of Streptomyces: a laboratory manual. John Innes Foundation, Norwich

    Google Scholar 

  • Hutchinson CR (1998) Combinatorial biosynthesis for new drug discovery. Curr Opin Microbiol 1:319–329

    Article  CAS  Google Scholar 

  • Ikeda H, Ōmura S (1993) Genetic aspects of the selective production of useful components in the avermectin producer Streptomyces avermitilis. Actinomycetologica 7:133–144

    Article  Google Scholar 

  • Ikeda H, Ōmura S (1995) Control of avermectin biosynthesis in Streptomyces avermitilis for the selective production of a useful component. J Antibiot 48:549–562

    Article  CAS  Google Scholar 

  • Ikeda H, Ōmura S (1997) Avermectin biosynthesis. Chem Rev 97:2591–2609

    Article  CAS  Google Scholar 

  • Ikeda H, Kotaki H, Ōmura S (1987) Genetic studies of avermectin biosynthesis in Streptomyces avermitilis. J Bacteriol 169:5615–5621

    Article  CAS  Google Scholar 

  • Ikeda H, Kotaki H, Tanaka H, Ōmura S (1988) Involvement of glucose catabolism in avermectin production by Streptomyces avermitilis. Antimicrob Agents Chemother 32:282–284

    Article  CAS  Google Scholar 

  • Ikeda H, Takada Y, Pang C-H, Tanaka H, Ōmura S (1993) Transposon mutagenesis by Tn4560 and applications with avermectin-producing Streptomyces avermitilis. J Bacteriol 175:2077–2082

    Article  CAS  Google Scholar 

  • Ikeda H, Pang C-H, Endo H, Ohta T, Tanaka H, Ōmura S (1995) Construction of a single component producer from the wild type avermectin producer Streptomyces avermitilis. J Antibiot 48:532–534

    Article  CAS  Google Scholar 

  • Ikeda H, Wang L-R, Ohta T, Inokoshi J, Ōmura S (1998) Cloning of the gene encoding avermectin B 5-O-methyltransferase in avermectin-producing Streptomyces avermitilis. Gene 206:175–180

    Article  CAS  Google Scholar 

  • Ikeda H, Nonomiya T, Usami M, Ohta T, Ōmura S (1999) Organization of the biosynthetic gene cluster for the polyketide anthelmintic macrolide avermectin in Streptomyces avermitilis. Proc Natl Acad Sci U S A 96:9509–9514

    Article  CAS  Google Scholar 

  • Ikeda H, Ishikawa J, Hanamoto A, Shinose M, Kikuchi H, Shiba T, Sakaki Y, Hattori M, Ōmura S (2003) Complete genome sequence and comparative analysis of the industrial microorganism Streptomyces avermitilis. Nat Biotechnol 21:526–531

    Article  Google Scholar 

  • Khosla C (1997) Harnessing the biosynthetic potential of modular polyketide synthase. Chem Rev 97:2577–2590

    Article  CAS  Google Scholar 

  • Kieser T (1984) Factors affecting the isolation of CCC DNA from Streptomyces lividans and Escherichia coli. Plasmid 12:19–36

    Article  CAS  Google Scholar 

  • Lambalot RH, Cane DE (1992) Isolation and characterization of 10-deoxymethynolide produced by Streptomyces venezuelae. J Antibiot 45:1981–1982

    Article  CAS  Google Scholar 

  • MacNeil DJ, Klapko LM (1987) Transformation of Streptomyces avermitilis by plasmid DNA. J Ind Microbiol 2:209–218

    Article  CAS  Google Scholar 

  • MacNeil DJ, Occi JL, Gewain KM, MacNeil T, Gibbons PH, Ruby CL, Danis SJ (1992) Complex organization of the Streptomyces avermitilis genes encoding the avermectin polyketide synthase. Gene 115:119–125

    Article  CAS  Google Scholar 

  • MacNeil T, Gewain KM, MacNeil DJ (1993) Deletion analysis of the avermectin biosynthetic genes of Streptomyces avermitilis by gene cluster displacement. J Bacteriol 175:2552–2563

    Article  CAS  Google Scholar 

  • McDaniel R, Thamchaipenet A, Gustafsson C, Fu H, Betlach M, Betlach M, Ashley G (1999) Multiple genetic modifications of the erythromycin polyketide synthase to produce a library of novel “unnatural” natural products. Proc Natl Acad Sci U S A 96:1846–1851

    Article  CAS  Google Scholar 

  • Ranganathan A, Timoney M, Bycroft M, Cortes J, Thomas LP, Wilkinson B, Kellenberger L, Hanefeld U, Galloway LS, Staunton J, Leadlay PF (1999) Knowledge-based design of bimodular and trimodular polyketide synthases based on domain and module swaps: a route to simple statin analogues. Chem Biol 6:731–741

    Article  CAS  Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Shikiya K, Kinjo N, Uehara T, Uechi H, Ohshiro J, Arakaki T, Kinjo F, Saito A, Iju M, Kobari K (1992) Efficacy of ivermectin against Strongyloidesstercoralis in humans. Intern Med 31:310–312

    Article  Google Scholar 

  • Song Y, Cao GM, Chen Z, Li JL (2000) Selection of high avermectins producing strain and identification of avermectin B1. Chin J Biotechnol 16:31–35

    CAS  Google Scholar 

  • Stutzman-Engwall K, Conlon S, Fedechko R, Kaczmarek F, McArthur H, Krebber A, Chen Y, Minshull J, Raillard SA, Gustafsson C (2003) Engineering the aveC gene to enhance the ratio of doramectin to its CHC-B2 analogue produced in Streptomyces avermitilis. Biotechnol Bioeng 82:359–369

    Article  CAS  Google Scholar 

  • Tang L, Fu H, McDaniel R (2000) Formation of functional heterologous complexes using subunits from the picromycin, erythromycin and oleandomycin polyketide synthases. Chem Biol 7:77–84

    Article  CAS  Google Scholar 

  • Yoon YJ, Beck BJ, Kim BS, Kang H-Y, Reynolds KA, Sherman DH (2002) Generation of multiple bioactive macrolides by hybrid modular polyketide synthases in Streptomyces venezuelae. Chem Biol 9:203–214

    Article  CAS  Google Scholar 

  • Zhang XL, Chen Z, Zhao JL, Song Y, Wen Y, Li JL (2004) Deletion analysis of oligomycin PKS genes (olmA) in Streptomyces avermitilis. Chin Sci Bull 49:350–354

    CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr. Huarong Tan (Institute of Microbiology, Chinese Academy of Sciences) for giving us the plasmid pKC1139 and for valuable comments on the manuscript. This work was supported by grants from National Basic Research Project (Grant No. 2003CH114205) and National High Technology Research and Development Program (Grant No. 2001AA214071).

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Authors and Affiliations

  1. State Key Laboratory for Agro-biotechnology, China Agricultural University, No.2 Yuanmingyuan Xi Lu, Haidian, Beijing, 100094, People’s Republic of China

    Zhi Chen, Meng Li, Ying Wen, Yuan Song & Jilun Li

  2. Department of Microbiology, China Agricultural University, No.2 Yuanmingyuan Xi Lu, Haidian, Beijing, 100094, People’s Republic of China

    Zhi Chen, Meng Li, Ying Wen, Yuan Song & Jilun Li

  3. Academy of State Administration of Grain, No. 11 Baiwanzhuang Avenue, Beijing, 100037, People’s Republic of China

    Xiaolin Zhang

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  1. Xiaolin Zhang
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  2. Zhi Chen
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  3. Meng Li
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  4. Ying Wen
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  5. Yuan Song
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  6. Jilun Li
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Correspondence to Jilun Li.

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Zhang, X., Chen, Z., Li, M. et al. Construction of ivermectin producer by domain swaps of avermectin polyketide synthase in Streptomyces avermitilis . Appl Microbiol Biotechnol 72, 986–994 (2006). https://doi.org/10.1007/s00253-006-0361-2

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  • DOI: https://doi.org/10.1007/s00253-006-0361-2

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