Abstract
A bacterial artificial chromosomal library of Nonomuraea sp. ATCC39727 was constructed using Escherichia coli–Streptomyces artificial chromosome (ESAC) and screened for the presence of dbv genes known to be involved in the biosynthesis of the glycopeptide A40926. dbv genes were cloned as two large, partially overlapping, fragments and transferred into the host Streptomyces lividans, thus generating strains S. lividans∷NmESAC50 and S. lividans∷NmESAC57. The heterologous expression of Nonomuraea genes in S. lividans was successfully demonstrated by using combined RT–PCR and proteomic approaches. MALDI-TOF analysis revealed that a Nonomuraea ABC transporter is expressed as two isoforms in S. lividans. Moreover, its expression may not require a Nonomuraea positive regulator at all, as it is present at similar levels in both clones even though S. lividans∷NmESAC57 lacks regulatory genes. Considered together, these results show that S. lividans expresses Nonomuraea genes from their own promoters and support the idea that S. lividans can be a good host for genetic analysis of Nonomuraea.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ahmad S, Selvapandiyan A, Bhatnagar RK (2000) Phylogenetic analysis of Gram-positive bacteria based on grpE, encoded by the dnaK operon. Int J Syst Evol Microbiol 50:1761–1766
Alduina R, De Grazia S, Dolce L, Salerno P, Sosio M, Donadio S, Puglia AM (2003) Artificial chromosome libraries of Streptomyces coelicolor A3(2) and Planobispora rosea. FEMS Microbiol Lett 218:181–186 DOI 10.1016/S0378-1097(02)01125-4
Beyazova ML, Brodsky BC, Shearer MC, Horan AC (1995) Preparation of actinomycete DNA for pulse-field gel electrophoresis. Int J Syst Bacteriol 45:852–854
Candiani G, Abbondi M, Borgonovi M, Romano G, Parenti F (1999) In-vitro and in-vivo antibacterial activity of BI 397, a new semi-synthetic glycopeptide antibiotic. J Antimicrob Chemother 44:179–192
Chiu ML, Folcher M, Katoh T, Puglia AM, Vohradsky J, Yun BS, Seto H, Thompson CJ (1999) Broad spectrum thiopeptide recognition specificity of the Streptomyces lividans TipAL protein and its role in regulating gene expression. J Biol Chem 274:20578–20586
Clauser KR, Baker PR, Burlingame AL (1999) Role of accurate mass measurement (+/−10 ppm) in protein identification strategies employing MS or MS/MS and database searching. Anal Chem 71:2871–2882
Doekel S, Eppelmann K, Marahiel MA (2002) Heterologous expression of nonribosomal peptide synthetases in B. subtilis: construction of a bi-functional B. subtilis/E. coli shuttle vector system. FEMS Microbiol Lett 216:185–191 DOI 10.1016/S0378-1097(02)00993-X
Donadio S, Monciardini P, Alduina R, Mazza P, Chiocchini C, Cavaletti L, Sosio M, Puglia AM (2002) Microbial technologies for the discovery of novel bioactive metabolites. J Biotechnol 99:187–198 DOI 10.1016/S0168-1656(02)00209-2
Evans M, Dyson P (1993) Pulsed-field gel electrophoresis of Streptomyces lividans DNA. Trends Genet 9:72
Hesketh AR, Chandra G, Shaw AD, Rowland JJ, Kell DB, Bibb MJ, Chater KF (2002) Primary and secondary metabolism, and post-translational protein modifications, as portrayed by proteomic analysis of Streptomyces coelicolor. Mol Microbiol 46:917–932 DOI 10.1046/j.1365-2958.2002.03219.x
Kang JG, Hahn MY, Ishihama A, Roe JH (1997) Identification of sigma factors for growth phase-related promoter selectivity of RNA polymerases from Streptomyces coelicolor A3(2). Nucleic Acids Res 25:2566–2573
Keller U, Schauwecker F (2003) Combinatorial biosynthesis of non-ribosomal peptides. Comb Chem High Throughput Screen 6:527–540
Kieser T, Bibb MJ, Buttner MJ, Chater KF, Hopwood DA (2000) Practical Streptomyces genetics. John Innes Centre, Norwich
Novotna J, Vohradsky J, Berndt P, Gramajo H, Langen H, Li XM, Minas W, Orsaria L, Roeder D, Thompson CJ (2003) Proteomic studies of diauxic lag in the differentiating prokaryote Streptomyces coelicolor reveal a regulatory network of stress-induced proteins and central metabolic enzymes. Mol Microbiol 48:1289–1303 DOI 10.1046/j.1365-2958.2003.03529.x
Puglia AM, Vohradsky J, Thompson CJ (1995) Developmental control of the heat-shock stress regulon in Streptomyces coelicolor. Mol Microbiol 17:737–746
Redenbach M, Scheel J, Schmidt U (2000) Chromosome topology and genome size of selected actinomycetes species. Antonie van Leeuwenhoek 78:227–235
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y.
Shevchenko A, Jensen ON, Podtelejnikov AV, Sagliocco F, Wilm M, Vorm O, Mortensen P, Shevchenko A, Boucherie H, Mann M (1996) Linking genome and proteome by mass spectrometry: large-scale identification of yeast proteins from two dimensional gels. Proc Natl Acad Sci USA 93:14440–14445
Sosio M, Giusino F, Cappellano C, Bossi E, Puglia AM, Donadio S (2000) Artificial chromosomes for antibiotic-producing actinomycetes. Nat Biotechnol 18:343–345 DOI 10.1038/73810
Sosio M, Stinchi S, Beltrametti F, Lazzarini A, Donadio S (2003) The gene cluster for the biosynthesis of the glycopeptide antibiotic A40926 by Nonomuraea species. Chem Biol 10:541–549 DOI 10.1016/S1074-5521(03)00120-0
Steiert M, Schmitz FJ (2002) Dalbavancin (Biosearch Italia/Versicor). Curr Opin Invest Drugs 3:229–233
Stinchi S, Azimonti S, Donadio S, Sosio M (2003) A gene transfer system for the glycopeptide producer Nonomuraea sp ATCC39727. FEMS Microbiol Lett 225:53–57 DOI 10.1016/S0378-1097(03)00490-7
Talamo F, D’Ambrosio C, Arena S, Del Vecchio P, Ledda L, Zehender G, Ferrara L, Scaloni A (2003) Proteins from bovine tissues and biological fluids: defining a reference electrophoresis map for liver, kidney, muscle, plasma and red blood cells. Proteomics 3:440–460 DOI 10.1002/pmic.200390059
Wilkinson CJ, Hughes-Thomas ZA, Martin CJ, Bohm I, Mironenko T, Deacon M, Wheatcroft M, Wirtz G, Staunton J, Leadlay PF (2002) Increasing the efficiency of heterologous promoters in actinomycetes. J Mol Microbiol Biotechnol 4:417–426
Zhang WZ, Chair BT (2000) Profound. An expert system for protein identification using mass spectrometric peptide mapping information. Anal Chem 72:2482–2489
Zirkle R, Ligon JM, Molnar I (2004) Heterologous production of the antifungal polyketide antibiotic soraphen A of Sorangium cellulosum So ce26 in Streptomyces lividans. Microbiology 150:2761–2774 DOI 10.1099/mic.0.27138-0
Acknowledgements
We thank Margherita Sosio for helpful suggestions and Maurizio Noto for technical assistance. This work was supported by the European Union via the MEGA-TOP project QLK3-1999-00650 and the COMBIG-TOP project LSHB-CT-2003-503491. We acknowledge our colleagues participating in these projects for stimulating discussions.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Alduina, R., Giardina, A., Gallo, G. et al. Expression in Streptomyces lividans of Nonomuraea genes cloned in an artificial chromosome. Appl Microbiol Biotechnol 68, 656–662 (2005). https://doi.org/10.1007/s00253-005-1929-y
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-005-1929-y