Microbial Transformation of Steroids Essay Example for Free

Microbial Transformation of Steroids EssayAbstractHydroxylations are possibly the most far-flung type of steroid bioconversion. Hydroxylations can be used to buildintermediates for further chemical implication to provide the steroid molecule with the adequate structure for therapeutic applications. Microorganisms adequate to hydroxylate steroids in positions C1 to C21 and in position C26 have been reported. 11 hydroxylation is regarded as essential for anti-inflammatory action. Present study was promoted to ext complete the investigations to evaluate potentiality of this local vegetable oil colour contaminated soil isolates for transformation of progesterone. Enrichment is carried out in the inorganic medium providing progesterone as an only carbon source. Metabolites are place by GC/MS and isolates by 16 s rRNA. 11 hydroxyprogesterone is the metabolite identified having 60 percent of yield. and Bacillus circulans Tex 01-S3c has been found most efficient strain.Key words Hydroxylation of Progesterone, oil contaminated, Bacillus circulans Tex 01-S3c,INTRODUCTIONApplication of molecular biology techniques and genetic engineering of microorganisms for their improvement as steroid transforming agents are the areas of drug development. Site selection oxygenation, oddly hydroxylation of exogenous steroids is frequent in bacteria genera Bacillus. Several microbial bioconversion of steroids and sterols have been reported, focusing mainly on steroid hydroxylations, 1-dehydrogenation and sterol side-chain cleavage.These biotransformations, mostly associated to chemical synthetic thinking steps, have provided adequate tools for the large scale production of natural or modified steroid analogues. The complex structure of the steroid molecule requires complicated, multi-step schemes for the chemical synthesis of steroid compounds. Microbial steroid conversions are performed in mild temperature and pressure conditions and can provide an efficient alternative to chemical synthesis.Screening and isolation of busy microbial strains for steroid bioconversion is presently an important part of the research and development effort in the steroid drug industry. Oil contaminated soil is a rich source of variety of bacteria capable of biotransformations and degradations of complex chemicals.MATERIAL AND METHODS1 gm of oil contaminated soil from different local places was added in the sterile 9 ml saline and 1 ml from this is added in the sterile progesterone liquid enrichment medium (KH2PO4, Na2HPO4. 12 H2O, (NH4)2SO4, MgSO4. 7 H2O, CaCl2. 6H2O, FeSO4. 7 H2O, pH 7) providing progesterone as a only carbon source. As progesterone is not soluble in water, it was dissolved in acetone (50 mg / ml). The flasks are incubated at 300 C for 7 days. Regular microscopic observation and turbidity checking carried out. By serial dilution techniques 0.1 ml was spread on the nutrient agar-agar plates and well isolated colonies are studied for biotransformation po tential.The medium of composition (g/liter) glucose, 40.0 peptone, 1.0 KH2PO4 0.74 MgSO4. 7H2O, 1.0 Yeast extract, 1.0 and asparagine, 0.7, pH was adjusted to 7 used for transformation. The cultivation was performed in 250-ml Erlenmeyer flasks each containing 50 ml medium. The flasks were sterilized and inoculated with 2 ml inoculum of 24 hr cultures of the pure organism. The culture flasks were incubated on incubator shaker at 34 C for 48 hr. Thereafter, 50 mg of progesterone, dissolved in 1 ml acetone, was added to each flask and fermentation was continued for another 72 hr.At the end of fermentation, metabolites are analyzed by GC/MS. The efficient bacterium identified by 16S rRNA. Standardization of parameters like temperature, pH, incubation period, carried out spectrophotometrically.RESULTS AND DISCUSSION11 hydroxyprogesterone is the metabolite identified having 60 percent of yield. Bacillus circulans Tex 01-S3c is the efficient bacterium identified. Temperature 340 C, pH 6.5 , incubation period 60 hours are the optimum parameters for maximum yield.REFRENCES1. Al-Awadi, S., Afzal, M. and Oommen, S. (2003) Studies on Bacillus stearothermophilus. founder III. Transformation of testosterone. Applied Microbiology and Biotechnology 62 , pp. 48-52. 2. Garai S, Banerjee S, Mahato SB (1995). Selective 1-dehydrogenation of progesterone by Aspergillus fumigatus. J Chem Res (Suppl) 408-409. 3. Fernandes, P., Cruz, A., Angelova, B., Pinheiro, H. and Cabral, J. (2003) Microbial conversion of steroid compounds youthful developments. Enzyme and Microbial Technology 32 , pp. 688-705 4. Mahato, S.B. and S. Garai, 1997. Advances in microbial steroid biotransformation. Steroids, 62332-345. 5. Sameera Al-Awadi, M.A. Fzal and S. Oommen, 2001. Studies on Bacillus stearithermophilus part Transformation of progesterone to a new metabolites 9,10-seco-4-pregnene-3,9,20-trione, diary of steroid biochemistry and Molecular biology 78 493-498. 6. Smith, KF and Kirk, DN (1989) Micr obial transformation of steroids-II. Transformations of progesterone, testosterone and androstenedione by Phycomyces blakesleeanus. Journal ofSteroid Biochemistry 32 , pp. 445-451