Abstract
Streptomyces venezuelae ATCC 10712 produces chloramphenicol in small amounts. To enhance chloramphenicol production, two genes, aroB and aroK, encoding rate-limiting enzymes of the shikimate pathway were overexpressed using the expression vector pIJ86 under the control of the strong constitutive ermE* promoter. The recombinant strains, S. venezuelae/pIJ86-aroB and S. venezuelae/pIJ86-aroK, produced 2.5- and 4.3-fold greater amounts respectively of chloramphenicol than wild type at early stationary phase of growth. High transcriptional levels of aroB and aroK genes were detected at the early exponential growth of both recombinant strains and consistent with the enhanced expression of pabB gene encoding an early enzyme in chloramphenicol biosynthesis. The results suggested that the increment of carbon flux was directed towards intermediates in the shikimate pathway required for the production of chorismic acid, and consequently resulted in the enhancement of chloramphenicol production. This work is the first report of a convenient genetic approach to manipulate primary metabolite genes in S. venezuelae in order to increase chloramphenicol production.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bibb MJ (2005) Regulation of secondary metabolism in streptomycetes. Curr Opin Microbiol 8:208–215. doi:10.1016/j.mib.2005.02.016
Brown MP, Aidoo KA, Vining LC (1996) A role for pabAB, a p-aminobenzoate synthase gene of Streptomyces venezuelae ISP5230, in chloramphenicol biosynthesis. Microbiology 142:1345–1355. doi:10.1099/13500872-142-6-1345
Chatterjee S, Vining LC, Westlake DWS (1983) Nutritional requirements for chloramphenicol biosynthesis in Streptomyces venezuelae. Can J Microbiol 29:247–253. doi:10.1139/m83-041
Chen D, Zhang Q, Zhang Q, Cen P, Xu Z, Liu W (2012) Improvement of FK506 production in Streptomyces tsukubaensis by genetic enhancement of the supply of unusual polyketide extender units via utilization of two distinct site-specific recombination systems. Appl Environ Microbiol 78:5093–5103. doi:10.1128/aem.00450-12
Dell KA, Frost JW (1993) Identification and removal of impediments to biocatalytic synthesis of aromatics from d-glucose: rate-limiting enzymes in the common pathway of aromatic amino acid biosynthesis. J Am Chem Soc 115:11581–11589. doi:10.1021/ja00077a065
Fernández-Martínez LT, Borsetto C, Gomez-Escribano JP, Bibb MJ, Al-Bassam MM, Chandra G, Bibb MJ (2014) New insights into chloramphenicol biosynthesis in Streptomyces venezuelae ATCC 10712. Antimicrob Agents Chemother 58:7441–7450. doi:10.1128/aac.04272-14
Gomez-Escribano JP, Bibb MJ (2011) Engineering Streptomyces coelicolor for heterologous expression of secondary metabolite gene clusters. Microb Biotechnol 4:207–215. doi:10.1111/j.1751-7915.2010.00219.x
He J, Magarvey N, Piraee M, Vining LC (2001) The gene cluster for chloramphenicol biosynthesis in Streptomyces venezuelae ISP5230 includes novel shikimate pathway homologues and a monomodular non-ribosomal peptide synthetase gene. Microbiology 147:2817–2829. doi:10.1099/00221287-147-10-2817
Healy FG, Eaton KP, Limsirichai P, Aldrich JF, Plowman AK, King RR (2009) Characterization of γ-butyrolactone autoregulatory signaling gene homologs in the angucyclinone polyketide WS5995B producer Streptomyces acidiscabies. J Bacteriol 191:4786–4797. doi:10.1128/jb.00437-09
Hobbs G, Frazer C, Gardner DJ, Cullum J, Oliver S (1989) Dispersed growth of Streptomyces in liquid culture. Appl Microbiol Biotechnol 31:272–277
Hodgson DA (2000) Primary metabolism and its control in streptomycetes: a most unusual group of bacteria. Adv Microb Physiol 42:47–238. doi:10.1016/S0065-2911(00)42003-5
Hwang K-S, Kim HU, Charusanti P, Palsson BØ, Lee SY (2014) Systems biology and biotechnology of Streptomyces species for the production of secondary metabolites. Biotechnol Adv 32:255–268. doi:10.1016/j.biotechadv.2013.10.008
Juminaga D, Baidoo EEK, Redding-Johanson AM, Batth TS, Burd H, Mukhopadhyay A, Petzold CJ, Keasling JD (2012) Modular engineering of l-tyrosine production in Escherichia coli. Appl Environ Microbiol 78:89–98. doi:10.1128/aem.06017-11
Kern A, Tilley E, Hunter IS, Legiša M, Glieder A (2007) Engineering primary metabolic pathways of industrial micro-organisms. J Biotechnol 129:6–29. doi:10.1016/j.jbiotec.2006.11.021
Kieser T, Bibb M, Buttner M, Chater K, Hopwood D (2000) Practical Streptomyces Genetics. The John Innes Foundation, Norwich
Laureti L, Song L, Huang S, Corre C, Leblond P, Challis GL, Aigle B (2011) Identification of a bioactive 51-membered macrolide complex by activation of a silent polyketide synthase in Streptomyces ambofaciens. Proc Natl Acad Sci USA 108:6258–6263. doi:10.1073/pnas.1019077108
Li R, Townsend CA (2006) Rational strain improvement for enhanced clavulanic acid production by genetic engineering of the glycolytic pathway in Streptomyces clavuligerus. Metab Eng 8:240–252. doi:10.1016/j.ymben.2006.01.003
Lütke-Eversloh T, Stephanopoulos G (2008) Combinatorial pathway analysis for improved l-tyrosine production in Escherichia coli: identification of enzymatic bottlenecks by systematic gene overexpression. Metab Eng 10:69–77. doi:10.1016/j.ymben.2007.12.001
MacNeil DJ, Gewain KM, Ruby CL, Dezeny G, Gibbons PH, MacNeil T (1992) Analysis of Streptomyces avermitilis genes required for avermectin biosynthesis utilizing a novel integration vector. Gene 111:61–68. doi:10.1016/0378-1119(92)90603-M
Malik V, Vining L (1970) Metabolism of chloramphenicol by the producing organism. Can J Microbiol 16:173–179
Matsumoto A, Ishizuka H, Beppu T, Horinouchi S (1995) Involvement of a small ORF downstream of the afsR gene in the regulation of secondary metabolism in Streptomyces coelicolor A3(2). Actinomycetologica 9:37–43. doi:10.3209/saj.9_37
Ochi K, Hosaka T (2013) New strategies for drug discovery: activation of silent or weakly expressed microbial gene clusters. Appl Microbiol Biotechnol 97:87–98. doi:10.1007/s00253-012-4551-9
Olano C, Lombó F, Méndez C, Salas JA (2008) Improving production of bioactive secondary metabolites in actinomycetes by metabolic engineering. Metab Eng 10:281–292. doi:10.1016/j.ymben.2008.07.001
Oldiges M, Kunze M, Degenring D, Sprenger GA, Takors R (2004) Stimulation, monitoring, and analysis of pathway dynamics by metabolic profiling in the aromatic amino acid pathway. Biotechnol Prog 20:1623–1633. doi:10.1021/bp0498746
Paget MSB, Chamberlin L, Atrih A, Foster SJ, Buttner MJ (1999) Evidence that the extracytoplasmic sigma factor, sigmaE, is required for normal cell wall structure in Streptomyces coelicolor A3(2). J Bacteriol 181:204–211
Phornphisutthimas S, Sudtachat N, Bunyoo C, Chotewutmontri P, Panijpan B, Thamchaipenet A (2010) Development of an intergeneric conjugal transfer system for rimocidin-producing Streptomyces rimosus. Lett Appl Microbiol 50:530–536. doi:10.1111/j.1472-765X.2010.02835.x
Reeves AR, Brikun IA, Cernota WH, Leach BI, Gonzalez MC, Weber JM (2007) Engineering of the methylmalonyl-CoA metabolite node of Saccharopolyspora erythraea for increased erythromycin production. Metab Eng 9:293–303. doi:10.1016/j.ymben.2007.02.001
Rokem JS, Lantz AE, Nielsen J (2007) Systems biology of antibiotic production by microorganisms. Nat Prod Rep 24:1262–1287. doi:10.1039/B617765B
Ryu Y-G, Butler MJ, Chater KF, Lee KJ (2006) Engineering of primary carbohydrate metabolism for increased production of actinorhodin in Streptomyces coelicolor. Appl Environ Microbiol 72:7132–7139. doi:10.1128/aem.01308-06
Sambrook J, Russell D (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York
Tamehiro N, Hosaka T, Xu J, Hu H, Otake N, Ochi K (2003) Innovative approach for improvement of an antibiotic-overproducing industrial strain of Streptomyces albus. Appl Environ Microbiol 69:6412–6417. doi:10.1128/aem.69.11.6412-6417.2003
Tanaka Y, Komatsu M, Okamoto S, Tokuyama S, Kaji A, Ikeda H, Ochi K (2009) Antibiotic overproduction by rpsL and rsmG mutants of various actinomycetes. Appl Environ Microbiol 75:4919–4922. doi:10.1128/aem.00681-09
Tang Z, Xiao C, Zhuang Y, Chu J, Zhang S, Herron PR, Hunter IS, Guo M (2011) Improved oxytetracycline production in Streptomyces rimosus M4018 by metabolic engineering of the G6PDH gene in the pentose phosphate pathway. Enzyme Microb Technol 49:17–24. doi:10.1016/j.enzmictec.2011.04.002
Thanapipatsiri A, Claesen J, Gomez-Escribano J-P, Bibb M, Thamchaipenet A (2015) A Streptomyces coelicolor host for the heterologous expression of Type III polyketide synthase genes. Microb Cell Fact 14:145. doi:10.1186/s12934-015-0335-0
Thykaer J, Nielsen J, Wohlleben W, Weber T, Gutknecht M, Lantz AE, Stegmann E (2010) Increased glycopeptide production after overexpression of shikimate pathway genes being part of the balhimycin biosynthetic gene cluster. Metab Eng 12:455–461. doi:10.1016/j.ymben.2010.05.001
Vining L, Stuttard C (1995) Chloramphenicol. Biotechnology 28:505–530
Xie P, Sheng Y, Ito T, Mahmud T (2012) Transcriptional regulation and increased production of asukamycin in engineered Streptomyces nodosus subsp. asukaensis strains. Appl Environ Microbiol 96:451–460. doi:10.1007/s00253-012-4084-2
Acknowledgments
We thank Mervyn Bibb, John Innes Centre, UK for kindly provide S. venezuelae ATCC 10712 and pIJ86. V. Vitayakritsirikul received Ph.D. scholarship from Huachiew Chalermprakiet University, Thailand. This work was supported financially by BIOTEC, NSTDA under the grant number BT-B-01-XG-11-5001 and the Center of Excellence for Innovation in Chemistry (PERCH-CIC). The Office of the Higher Education Commission, Ministry of Education is also gratefully acknowledged.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Vitayakritsirikul, V., Jaemsaeng, R., Lohmaneeratana, K. et al. Improvement of chloramphenicol production in Streptomyces venezuelae ATCC 10712 by overexpression of the aroB and aroK genes catalysing steps in the shikimate pathway. Antonie van Leeuwenhoek 109, 379–388 (2016). https://doi.org/10.1007/s10482-015-0640-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10482-015-0640-y