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http://dx.doi.org/10.7845/kjm.2015.5030

Effect of SeaR gene on virginiamycins production in Streptomyces virginiae  

Ryu, Jae-Ki (Department of Biomedical Laboratory Science, Gimcheon University)
Kim, Hyun-Kyung (Department of Biomedical Laboratory Science, Gimcheon University)
Kim, Byung-Won (Department of Biomedical Laboratory Science, Gimcheon University)
Kim, Dong-Chan (Department of Biomedical Laboratory Science, Gimcheon University)
Lee, Hyeong-Seon (Department of Biomedical Laboratory Science, Jungwon University)
Publication Information
Korean Journal of Microbiology / v.51, no.3, 2015 , pp. 256-262 More about this Journal
Abstract
In order to study the effect of the receptor protein (SeaR), which is isolated from Saccharopolyspora erythraea, we introduced the SeaR gene to Streptomyces virginiae as host strains. An effective transformation procedure for S. virginiae was established based on transconjugation by Escherichia coli ET12567/pUZ8002 with a ${\varphi}C31$-derived integration vector, pSET152, which contained int, oriT, attP, and $ermEp^{\ast}$ (erythromycin promotor). Therefore, the pEV615 was introduced into S. virginiae by conjugation and integrated at the attB locus in the chromosome of the recipients by the ${\varphi}C31$ integrase (int) function. Transformants of S. virginiae containing the SeaR gene were confirmed by PCR and transcriptional expression of the SeaR gene in the transformants was analyzed by RT-PCR, respectively. And, we examined the production time of virginiamycins in the culture media of both the transformants and the wild type. The production time of virginiamycins in the wild type and transformants was the same. When 100 ng/ml of synthetic $VB-C_6$ was added to the state of 6 or 8 hour cultivation of wild type and transformants, respectively, the virginiamycins production was induced, meaning that the virginiamycins production in the wild type was detected 2 h early than transformants. From these results, SeaR expression was also affected to virginiamycins production in transformants derived from S. virginiae. In this study, we showed that the SeaR protein worked as a repressor in transformants.
Keywords
Saccharopolyspora erythraea; SeaR gene; Streptomyces virginiae; transformant; virginiamycin;
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1 Alderson, G., Ritchie, D.A., Cappellano, C., Gool, R.H., Ivanova, N.M., Huddleston, A.S., Flaxman, C.S., Kristufek, V., and Lounes, A. 1993. Physiology and genetics of antibiotic production and resistance. Res. Microbiol. 144, 665-672.   DOI
2 Bibb, M.J. 2005. Regulation of secondary metabolism in Streptomycetes. Curr. Opin. Microbiol. 8, 208-215.   DOI
3 Bierman, M., Logan, R., O'Brien, K., Seno, E.T., Rao, R.N., and Schoner, B.E. 1992. Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp. Gene 116, 43-49.   DOI
4 Butler, M.J., Takano, E., Bruheim, P., Jovatic, S., Marinell, F., and Bibb, M.J. 2003. Deletion of scbA enhances antibiotic production in Streptomyces lividans. Appl. Microbiol. Biotechnol. 61, 512-516.   DOI
5 Chater, K.F. 1993. Genetics of differentiation in Streptomyces. Annu. Rev. Microbiol. 47, 685-713.   DOI
6 Choi, S.U., Lee, C.K., Hwang, Y.I., Kinishita, H., and Nihira, T. 2004. Intergeneric conjugal tansfer of plasmid DNA from Escherichia coli to Kitasatospora setae, a bafilomycin $B_1$ producer. Arch. Microbiol. 181, 294-298.   DOI
7 Horinouchi, S., Kito, M., Nishiyama, M., Furuya, K., Hong, S.K., Miyake, K., and Beppu, T. 1990. Primary structure of AfsR, a global regulatory protein for secondary metabolite formation in Streptomyces coelicolor A3(2). Gene 95, 49-56.   DOI
8 Hwang, J.H., Lee, C.K., Lee, K.M., Jo, B.K., Park, H.R., and Hwang, Y.I. 2005. Development of a recombinant Streptomyces griseus with sprA and sprB genes for proteolytic enzyme production. Kor. J. Microbiol. 41, 87-92.
9 Kieser, T., Bibb, M.J., Buttner, M.J., Chater, K.F., and Hopwood, D.A. 2000. Practical Streptomyces Genetics. The John Innes Foundation, Norwich, UK.
10 Kim, H.S., Nihira, T., Tada, H., Yanagimoto, M., and Yamada, Y. 1989. Identification of binding protein of virginiae butanolide C, an autoregulator in virginiamycin production from Streptomyces virginiae. J. Antibiot. 42, 769-778.   DOI
11 Lee, Y.J., Yeo, S.H., Lee, I.S., Lee, S.P., Kitani, S., Nihira, T., and Kim, H.S. 2006. Cloning and characterization of gene encoding $\gamma$-butyrolactone autoregulator receptor from Saccharopolyspora erythraea. J. Microbiol. Biotechnol. 16, 77-83.
12 Nakano, H., Takehara, E., Nihira, T., and Yamada, Y. 1998. Gene replacement analysis of the Streptomyces virginiae barA gene encoding the butyrolactone autoregulator receptor reveals that BarA acts as a repressor in virginiamycin biosynthesis. J. Bacteriol. 180, 3317-3322.
13 Nihira, T., Shimizu, Y., Kim, H.S., and Yamada, Y. 1988. Structureactivity relationships of virginiae butanolide C, an inducer of virginiamycin production in Streptomyces virginiae. J. Antibiot. 41, 1828-1837.   DOI
14 Okamoto, S., Nakamura, K., Nihira, T., and Yamada, Y. 1995. Virginiae butanolide binding protein from Streptomyces virginiae. J. Biol. Chem. 270, 12319-12326.   DOI
15 Onaka, H., Ando, N., Nihira, T., Yamada, Y., Beppu, T., and Horinouchi, S. 1995. Cloning and characterization of the A-factor receptor gene from Streptomyces griseus. J. Bacteriol. 177, 6083-6092.   DOI
16 Ryu, J.K., Kwon, P.S., and Lee, H.S. 2015. Functional analysis of SeaR protein identified from Saccharopolyspora erythraea. Kor. J. Microbiol. 51, 39-47.   DOI
17 Sambrook, J., Fritsch, E.F., and Maniatis, T. 1989. Molecular Cloning:A Laboratory Manual 2nd ed. New York, Cold Spring Harbor Laboratory, USA.
18 Takano, E. 2006. $\gamma$-Butyrolactones: Streptomyces signaling molecules regulating antibiotic production and differentiation. Curr. Opin. Microbiol. 9, 287-294.   DOI
19 Takano, E., Chakraburtty, R., Nihira, T., Yamada, Y., and Bibb, M.J. 2001. A complex role for the $\gamma$-butyrolactone SCB1 in regulating antibiotic production in Streptomyces coelicolor A3(2). Mol. Microbiol. 41, 1015-1028.
20 Umezawa, H. 1988. In Actinomycetes in Biotechnology. London: Academic Press, USA.
21 Yanagimoto, M. and Terui, G. 1971. Physiological studies on staphylomycin production: Formation of a substance effective in inducing staphylomycin production. J. Ferment. Technol. 49, 611-618.