Doxorubicin is a clinically important anticancer polyketide compound that is typically produced by Streptomyces peucetius var. caesius. To improve doxorubicin productivity by S. peucetius, a doxorubicin pathway-specific regulatory gene, dnrI, was cloned into a high-copy-number plasmid containing a catechol promoter system. The S. peucetius containing the recombinant plasmid exhibited approximately 9.5-fold higher doxorubicin productivity compared with the wild-type S. peucetius. The doxorubicin productivity by this recombinant S. peucetius strain was further improved through the optimization of culture media composition. Based on the Fractional Factorial Design (FFD), cornstarch, $K_2HPO_4$, and $MgSO_4$ were identified to be the key factors influencing doxorubicin productivity. The Response Surface Method (RSM) results based on 20 independent culture conditions with varying amounts of key factors predicted the highest theoretical doxorubicin productivity of 11.1 mg/l with corn starch of 46.33 g/l, $K_2HPO_4$ of 4.63 g/l, and $MgSO_4$ of 9.26 g/l. The doxorubicin productivity of the recombinant S. peucetius strain with the RSM-based optimized culture condition was experimentally verified to be 11.46 mg/l, which was approximately 30.8-fold higher productivity compared with the wild-type S. peucetius without culture media optimization.
Berdy, J. 1984. New ways to obtain antibiotics. Chin. J. Antibiot. 7: 272- 290
Hopwood, D. A., K. F. Chater, J. E. Dowding, and A Vvian. 1973. Advances in Streptomyces coelicolor genetics. Bacteriol. Rev. 37: 371-405
Hopwood, D. A., T. Kieser, M. J. Bibb, M. J. Buttner, and K. F. Chater. 2000. Practical Streptomyces Genetics. The John Innes Foundation, Norwich, U.K.
Hwang, Y. S., J. Y. Lee, E. S. Kim, and C. Y. Choi. 2001. Optimization of transformation procedure in avermectin high-producing Streptomyces avermitilis. Biotechnol. Lett. 23: 457-462
Myers, R. H. and D. C. Montgomery. 1995. Response Surface Methodology: Process and Product Optimization using Designed Experiments, 1st Ed. Wiley-Interscience, U.S.A.
Nicholls, G., B. J. Chark, and J. E. Brown. 1992. Solid-phase extraction and optimized separation of doxorubicin, epirubicin and their metabolites using reverse-phase high performance liquid chromatography. J. Pharm. Biomed. Anal. 10: 949-957
Otten, S. L., J. Ferguson, and C. R. Hutchinson. 1995. Regulation of daunorubicin production in Streptomyces peucetius by the $dnrR_2$ locus. J. Bacteriol. 177: 1216-1224
Peggy, A. R., P. A. Mcann, M. A. Pentella, and B. M. Pogell. 1979. Simultaneous loss of multiple differentiated functions in aerial mycelium-negative isolates of Streptomyces. J. Bacteriol. 137: 891- 899
Hamseveni, D. R., S. G. Prapulla, and S. Divakar. 2001. Response surface methodological approach for the synthesis of isobutyl isobutyrate. Proc. Biochem. 36: 1103-1109
Vetrivel, K. S. and K. Dharmalingam. 2001. Isolation and characterization of stable mutants of Streptomyces peucetius defective in daunorubicin biosynthesis. J. Genet. 80: 31-38
Park, H. J. and E. S. Kim. 2003. An inducible Streptomyces gene cluster involved in aromatic compound metabolism. FEMS Microbiol. Lett. 226: 151- 157
Volff, J. N. and J. Altenbuchner. 1998. Genetic instability of the Streptomyces chromosome. Mol. Microbiol. 27: 239-246
Box, G. E. P., W. G. Hunter, and J. S. Hunter. 1978. Statistics for Experimenters, pp. 291- 334. John Wiley and Sons, New York, U.S.A.
Demain, A. L. 2000. Small bugs, big business: The economic power of the microbe. Biotechnol. Adv. 18: 499- 514
Elibol, M. and F. Mavituna. 1998. Effect of sucrose on actinorhodin production by Streptomyces coelicolor A3(2). J. Biochem. 33: 307- 311
Chatterjee, S. and L. C. Vining. 1981. Nutrient utilization in actinomycetcs. Induction of $\alpha-glucosidases$ in Streptomyces venezualae. Can. J. Microbiol. 27: 639- 645
Cochran, W. G. and G. M. Cox. 1957. Experimental Design, 2nd Ed. pp. 346- 354. John Wiley and Sons, New York, U.S.A.
Kim, C.-Y., H.-J. Park, Y. J. Yoon, H.-Y. Kang, and E.-S. Kim. 2004. Stimulation of actinorhodin production by Streptomyces lividans with a chromosomally-integrated antibiotic regulatory gene afsR2. J. Microbiol. Biotechnol. 14: 1089-1092
Stutzman-Engwall, K. J., S. L. Otten, and C. R. Hutchinson. 1992. Regulation of secondary metabolism in Streptomyces spp. and overproduction of daunorubicin in Streptomyces peucetius. J. Bacteriol. 174: 144- 154
Jung, W.-S., E.-S. Kim, H.-Y. Kang, C.-Y. Choi, D. H. Sherman, and Y. J. Yoon. 2003. Site-directed mutagenesis on putative macrolactone ring size determinant in the hybrid pikrornycin-tylosin polyketide synthase. J. Microbiol. Biotechnol. 13: 823- 827
Hounsa, C. G., J. M. Aubry, and H. C. Dubourguier. 1996. Application of factorial and Doehlert design for optimization of pectate lyase production by a recombinant Escherichia coli. Appl. Microbiol. Biotechnol. 45: 764- 770
Kim, C.-Y., H.-J. Park, and E.-S. Kim. 2003. Heterologous expression of hybrid type II polyketide synthase system in Streptomyces species. J. Microbiol. Biotechnol. 13: 819-822
Kalakoutskii, L. V. and N. S. Agre. 1976. Comparative aspects of development and differentiation in actinomycetes. Bacteriol. Rev. 40: 469- 524
Rao, K. J., C. H. Kim, and S. K. Rhee. 2000. Statistical optimization of medium for the production of recombinant hirudin from Saccharomyces cerevisiae using response surface methodology. Proc. Biochem. 35: 639- 647
Park, H. S., H. J. Park, Y. H. Kim, S. M. Lim, D. I. Kim, W. S. Ry, and E. S. Kim. 2003. Development of doxorubicin overproducing Streptomyces strain using protoplast regeneration. K. J. Biotech. Bioeng. 18: 289- 293
Dey, G., A. Mitra, R. Banerjee, and B. R. Maiti. 2001. Enhanced production of amylase by optimization of nutritional constituents using response surface methodology. Biochem. Eng. J. 7: 227- 233
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