Effects of Glycerol and Shikimic Acid on Rapamycin Production in Streptomyces rapamycinicus
2
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
Rapamycin, derived from Streptomyces rapamycinicus, is an important bioactive compound having a therapeutic value in managing Parkinson’s disease, rheumatoid arthritis, cancer, and AIDS. Because of its pharmaceutical activity, studies over the past decade have focused on the biosynthesis of rapamycin to enhance its yield. In this study, the effect of rapG on rapamycin production was investigated. The rapG expression vector was constructed by utilizing the integration vector pSET152 under the control of the erythromycin resistance gene (ermE*), a strong constitutive promoter. The rapamycin yield of wild type (WT) and WT/ rapG overexpression mutant strains, under fermentation conditions, was analyzed by high-performance liquid chromatography (HPLC). Our results revealed that overexpression of rapG increased rapamycin production by approximately 4.9-fold (211.4 mg/l) in MD1 containing 15 g/l of glycerol, compared to that of the WT strain. It was also found that Illicium verum powder (10 g/l), containing shikimic acid, enhanced rapamycin production in both WT and WT/rapG strains. Moreover, the amount of rapamycin produced by the WT/rapG strain was statistically higher than that produced by the WT strain. In conclusion, the addition 15 g/l glycerol and 15 g/l I. verum powder produced the optimal conditions for rapamycin production by WT and WT/rapG strains.
Abstract
Rapamycin, derived from Streptomyces rapamycinicus, is an important bioactive compound having a therapeutic value in managing Parkinson’s disease, rheumatoid arthritis, cancer, and AIDS. Because of its pharmaceutical activity, studies over the past decade have focused on the biosynthesis of rapamycin to enhance its yield. In this study, the effect of rapG on rapamycin production was investigated. The rapG expression vector was constructed by utilizing the integration vector pSET152 under the control of the erythromycin resistance gene (ermE*), a strong constitutive promoter. The rapamycin yield of wild type (WT) and WT/ rapG overexpression mutant strains, under fermentation conditions, was analyzed by high-performance liquid chromatography (HPLC). Our results revealed that overexpression of rapG increased rapamycin production by approximately 4.9-fold (211.4 mg/l) in MD1 containing 15 g/l of glycerol, compared to that of the WT strain. It was also found that Illicium verum powder (10 g/l), containing shikimic acid, enhanced rapamycin production in both WT and WT/rapG strains. Moreover, the amount of rapamycin produced by the WT/rapG strain was statistically higher than that produced by the WT strain. In conclusion, the addition 15 g/l glycerol and 15 g/l I. verum powder produced the optimal conditions for rapamycin production by WT and WT/rapG strains.
참고문헌 (23)
- Vezina C, "Rapamycin (AY-22,989), a new antifungal antibiotic. I. Taxonomy of the producing streptomycete and isolation of the active principle", J. Antibiot, 28, pp.721~726 (1975)
- Park SR, "Biosynthesis of rapamycin and its regulation : past achievements and recent progress", J. Antibiot, 63, pp.434~441 (2010)
- Tain LS, "Rapamycin activation of 4E-BP prevents parkinsonian dopaminergic neuron loss", Nat. Neurosci, 12, pp.1129~1135 (2009)
- Shao P, "Modulation of the immune response in rheumatoid arthritis with strategically released rapamycin", Mol. Med. Rep, 16, pp.5257~5262 (2017)
- Zou CY, "Dual targeting of AKT and mammalian target of rapamycin : a potential therapeutic approach for malignant peripheral nerve sheath tumor", Mol. Cancer Ther, 8, pp.1157~1168 (2009)
- Nicoletti F, "Inhibition of human immunodeficiency virus(HIV-1)infection in human peripheral blood leucocytes-SCID reconstituted mice by rapamycin", Clin. Exp. Immunol, 155, pp.28~34 (2009)
- Molnar I, "Organisation of the biosynthetic gene cluster for rapamycin in Streptomyces hygroscopicus : analysis of genes flanking the polyketide synthase", Gene, 169, pp.1~7 (1996)
- Kuscer E, "Roles of rapH and rapG in positive regulation of rapamycin biosynthesis in Streptomyces hygroscopicus", J. Bacteriol, 189, pp.4756~4763 (2007)
- 김용현, "Production of Rapamycin in Streptomyces hygroscopicus from Glycerol-Based Media Optimized by Systemic Methodology", Journal of Microbiology and Biotechnology, 24(10), pp.1319~1326 (2014), 한국미생물·생명공학회[DOI]
- Zhu X, "Generation of high rapamycin producing strain via rational metabolic pathwaybased mutagenesis and further titer improvement with fedbatch bioprocess optimization", Biotechnol. Bioeng, 107, pp.506~515 (2010)
- Paiva NL, "The cyclohexane moiety of rapamycin is derived from shikimic acid in Streptomyces hygroscopicus", J. Ind. Microbiol. Biotechnol, 12, pp.423~428 (1993)
- Fang A, "Exogenous shikimic acid stimulates rapamycin biosynthesis in Streptomyces hygroscopicus", Folia Microbiol. (Praha), 40, pp.607~610 (1995)
- Lee MS, "Effect of nitrogen source on biosynthesis of rapamycin by Streptomyces hygroscopicus", J. Ind. Microbiol. Biotechnol, 19, pp.83~86 (1997)
- Yen HW, "Effects of dissolved oxygen level on rapamycin production by pellet-form of Streptomyces hygroscopicus", J. Biosci. Bioeng, 116, pp.366~370 (2013)
- Yen H-W, "The enhancement of rapamycin production using Streptomyces hygroscopicus through a simple pHshifted control", J. Taiwan Inst. Chem. E, 44, pp.743~747 (2013)
- Hanahan D, "Studies on transformation of Escherichia coli with plasmids", J. Mol. Biol, 166, pp.557~580 (1983)
- Kurniawan YN, "Regulation of production of the blue pigment indigoidine by the pseudo gamma-butyrolactone receptor FarR2 in Streptomyces lavendulae FRI-5", J. Biosci. Bioeng, 121, pp.372~379 (2016)
- Dai D, "Regulation of dndB gene expression in Streptomyces lividans", Front. Microbiol, 9, pp.2387~ (2018)
- Sallama LAR, "Some physiological factors affecting rapamycin production by Streptomyces hygroscopicus ATCC 29253", Am. J. Sci, 6, pp.188~194 (2010)
- More AS, "Extraction of rapamycin(sirolimus)from Streptomyces rapamycinicus using ultrasound", Prep. Biochem. Biotechnol, 47, pp.627~632 (2017)
- Dutta S, "Kinetics of rapamycin production by Streptomyces hygroscopicus MTCC 4003", 3 Biotech, 4, pp.523~531 (2014)
- Ohira H, "Rapid separation of shikimic acid from Chinese star anise (Illicium verum Hook. f.) with hot water extraction", Sep. Purif. Technol, 69, pp.102~108 (2009)
- Bierman M, "Plasmid cloning vectors for the conjugal transfer of DNA from Escherichia coli to Streptomyces spp", Gene, 116, pp.43~49 (1992)