Abstract
Prodigiosin is a natural red pigment with algicidal activity against Cochlodinium polykrikoides, a major harmful red-tide microalga. To increase the yield of prodigiosin, a mutant of Hahella chejuenesis KCTC 2396, assigned M3349, was developed by an antibiotic mutagenesis using chloramphenicol. When cultured in Sucrose-based Marine Broth medium (SMB), M3349 could produce prodigiosin at 1.628±0.06 g/L, while wild type producing at 0.658±0.12 g/L under the same conditions. To increase the yield of prodigiosin production by M3349, significant medium components were determined using a two-level Plackett-Burman statistical design technique. Among fourteen components included in SMB medium, NaCl, Na2SiO3, MgCl2, H3BO3, Na2HPO4, Na2SO4, and CaCl2 were determined to be important for prodigiosin production. The medium formulation was finally optimized using a Box-Behnken design as follows: sucrose 10.0, peptone 8.0, yeast extract 2.0, NaCl 10.0, Na2SO4 12.0, CaCl2 1.8, MgCl2 0.7 g/L; and H3BO3 22.0, Na2HPO4 20.0, Na2SiO3 8.0 mg/L. The predicted maximum yield of prodigiosin in the optimized medium was 2.43 g/L by the Box-Behnken design, while the practical production was 2.60±0.176 g/L, which was 3.9 times higher than wild type with SMB Medium (0.658 g/L).
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Adsul, M.G., K.B. Bastawde, A.J. Varma, and D.V. Gokhale. 2007. Strain improvement of Penicillium janthinellum NCIM 1171 for increased cellulose production. Bioresour. Technol. 98, 1467–1473.
Alonzo, V., M.E. Scoglio, and L. Mangione. 1979. Effect of the carbon/nitrogen ratio on the antibiotics activity of prodigiosin. Bacteriol. Virol. Immunol. 71, 3–15.
Anwar, M.N., M. Suto, and F. Tomita. 1996. Isolation of mutants of Penicillium purpurogenum resistant to catabolite repression. Appl. Microbiol. Biotechnol. 45, 684–687.
Box, G.E.P. and D.W. Behnken. 1960. Some new three level designs for the study of quantitative variables. Technometrics 2, 455–475.
Chand, P., A. Aruna, A.M. Maqsood, and L.V. Rao. 2005. Novel mutation method for increased cellulase production. J. Appl. Microbiol. 98, 318–323.
D’Aoust, J.Y. and N.N. Gerber. 1974. Isolation and purification of prodigiosin from Vibrio psychroreythrus. J. Bacteriol. 118, 756–757.
Fürstner, A. 2003. Chemistry and biology of roseophilin and the prodigiosin alkaloids: a survey of the last 2500 years. Angew. Chem. Int. Ed. Engl. 42, 35820–3603.
Jeong, H., J.H. Yim, C. Lee, S.H. Choi, Y.K. Park, S.H. Yoon, C.G. Hur, H.Y. Kang, D. Kim, H.H. Lee, K.H. Park, S.H. Park, H.K. Lee, T.K. Oh, and J.F. Kim. 2005. Genomic blueprint of Hahella chejuensis, a marine microbe producing an algicidal agent. Nucleic Acids Res. 33, 7066–7073.
Lee, H.K., J. Chun, E.Y. Moon, S.H. Ko, D.S. Lee, H.S. Lee, and K.S. Bae. 2001. Hahella chejuensis gen. nov., sp. nov., an extracellular-polysaccharide-producing marine bacterium. Int. J. Syst. Evol. Microbiol. 51, 661–666.
Lewis, S.M. and W.A. Corpe. 1964. Prodigiosin-producing bacteria from marine sources. Appl. Microbiol. 12, 13–17.
Liming, X. and S. Xueliang. 2004. High-yield cellulase production by Trichoderma reseei ZU-02 on corn cob residue. Bioresour. Technol. 91, 259–262.
Naveena, B.J., M. Altaf, K. Bhadriah, and G. Reddy, 2005. Selection of medium components by Plackett-Burman design for production of l(+) lactic acid by Lactobacillus amylophilus GV6 in SSF using wheat bran. Bioresour. Technol. 96, 485–490.
Plackett, R.L. and J.P. Burman. 1946. The design of optimum multifactorial experiments. Biometrika. 7, 305–325.
Rjazantseva, I.N., I.N. Andreeva, and T.I. Ogorodnikova. 1994. Effect of various growth conditions on pigmentation of Serratia marcescens. Microbes 79, 155–161.
Skomarovsky, A.A., A.V. Gusakov, O.N. Okunev, I.V. Solov’eva, T.V. Bubnova, E.G. Kondrat’eva, and A.P. Sinitsyn. 2005. Studies of hydrolytic activity of enzyme preparations of Penicillium and Trichoderma fungi. Appl. Biochem. Microbiol. 41, 182–184.
Sole, M., N. Rius, A. Francia, and J.G. Loren. 1994. The effect of pH on prodigiosin production by non-proliferating cells of Serratia marcescens. Lett. Appl. Microbiol. 19, 341–344.
Stanbury, P.F., A. Whitaker, and S.J. Hall. 1997. Principles of fermentation technology, p. 93–122. 2th ed. Aditya Books, New Delhi, India.
Strobel, R. and G. Sullivan. 1999. Experimental design for improvement of fermentations. In A.L. Demain and J.E. Davies (eds.). Manual of industrial microbiology and biotechnology, p. 80–93. ASM Press. Washington, D.C., USA.
Szengyel, Z., G. Zacchi, A. Varga, and K. Reczey. 2000. Cellulase production of Trichoderma reseei Rut C-30 using steam-pretreated spruce. Hydrolytic potential of cellulases on different substrate. Appl. Biochem. Biotechnol. 84, 679–691.
Walid, A.L., M.G. Khaled, and R.E.H. Ehab. 2007. Citric acid production by a novel Aspergillus niger isolate: I. Mutagenesis and cost reduction studies. Bioresour. Technol. 98, 3464–3469.
Wei, Y.H. and W.C. Chen. 2005. Enhanced production of prodigiosin-like pigment from Serratia macescens SMΔR by medium improvement and oil-supplementation strategies. J. Biosci. Bioeng. 99, 616–622.
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Kim, S.J., Lee, H.K., Lee, Y.K. et al. Mutant selection of Hahella chejuensis KCTC 2396 and statistical optimization of medium components for prodigiosin yield-up. J Microbiol. 46, 183–188 (2008). https://doi.org/10.1007/s12275-008-0037-y
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DOI: https://doi.org/10.1007/s12275-008-0037-y