AbstractionHydroxylations are perchance the most widespread type of steroid bioconversion. Hydroxylations can be used to buildintermediates for farther chemical synthesis to supply the steroid molecule with the equal construction for curative applications. Microorganisms able to hydroxylate steroids in places C1 to C21 and in place C26 have been reported. 11 ? hydroxylation is regarded as indispensable for anti-inflammatory action. Present survey was promoted to widen the probes to measure potency of this local oil contaminated dirt isolates for transmutation of Lipo-Lutin.
Enrichment is carried out in the inorganic medium supplying Lipo-Lutin as an lone C beginning. Metabolites are identified by GC/MS and isolates by 16 s rRNA. 11 ? hydroxyprogesterone is the metabolite identified holding 60 per centum of output. and Bacillus circulans Tex 01-S3c has been found most efficient strain.Cardinal words: –Hydroxylation of Progesterone. oil contaminated. Bacillus circulans Tex 01-S3c.
IntroductionApplication of molecular biological science techniques and familial technology of micro-organisms for their betterment as steroid transforming agents are the countries of drug development. Site choice oxygenation. peculiarly hydroxylation of exogenic steroids is frequent in bacteriums genera Bacillus. Several microbic bioconversion of steroids and steroid alcohols have been reported. concentrating chiefly on steroid hydroxylations. ?1-dehydrogenation and sterol side-chain cleavage.These biotransformations.
largely associated to chemical synthesis stairss. have provided equal tools for the big scale production of natural or modified steroid parallels. The complex construction of the steroid molecule requires complicated. multi-step strategies for the chemical synthesis of steroid compounds.
Microbial steroid transitions are performed in mild temperature and force per unit area conditions and can supply an efficient option to chemical synthesis.Screening and isolation of active microbic strains for steroid bioconversion is soon an of import portion of the research and development attempt in the steroid drug industry. Oil contaminated dirt is a rich beginning of assortment of bacteriums capable of biotransformations and debasements of complex chemicals.MATERIAL AND METHODS1 gram of oil contaminated dirt from different local topographic points was added in the unfertile 9 milliliter saline and 1 milliliter from this is added in the unfertile Lipo-Lutin liquid enrichment medium ( KH2PO4. Na2HPO4. 12 H2O. ( NH4 ) 2SO4.
MgSO4. 7 H2O. CaCl2. 6H2O.
FeSO4. 7 H2O. pH 7 ) supplying Lipo-Lutin as a lone C beginning. As Lipo-Lutin is non soluble in H2O. it was dissolved in propanone ( 50 milligram / milliliter ) . The flasks are incubated at 300 C for 7 yearss. Regular microscopic observation and turbidness checking carried out.
By consecutive dilution techniques 0. 1 milliliter was spread on the alimentary agar home bases and good stray settlements are studied for biotransformation potency.The medium of composing ( g/liter ) : glucose. 40. 0 ; peptone. 1.
0 ; KH2PO4 0. 74 ; MgSO4. 7H2O. 1. 0 ; Yeast infusion.
1. 0 and asparagine. 0. 7. pH was adjusted to 7 used for transmutation. The cultivation was performed in 250-ml Erlenmeyer flasks each incorporating 50 ml medium. The flasks were sterilized and inoculated with 2 milliliters inoculant of 24 hr civilizations of the pure being. The civilization flasks were incubated on brooder shaker at 34° C for 48 hour.
Thereafter. 50 milligram of Lipo-Lutin. dissolved in 1 milliliter propanone. was added to each flask and agitation was continued for another 72 hour.At the terminal of agitation. metabolites are analyzed by GC/MS. The efficient bacteria identified by 16S rRNA. Standardization of parametric quantities like temperature.
pH. incubation period. carried out spectrophotometrically.RESULTS AND DISCUSSION11 ? hydroxyprogesterone is the metabolite identified holding 60 per centum of output. Bacillus circulans Tex 01-S3c is the efficient bacteria identified. Temperature 340 C.
pH 6. 5. incubation period 60 hours are the optimal parametric quantities for maximal output.REFRENCES1. Al-Awadi. S. .
Afzal. M. and Oommen. S. ( 2003 ) Studies on Bacillus stearothermophilus. Part III. Transformation of’ testosterone. Applied Microbiology and Biotechnology 62.
pp. 48-52. 2. Garai S. Banerjee S.
Mahato SB ( 1995 ) . Selective 1-dehydrogenation of Lipo-Lutin by Aspergillus fumigatus. J Chem Res ( Suppl ) : 408-409. 3. Fernandes. P.
. Cruz. A. . Angelova. B. . Pinheiro.
H. and Cabral. J. ( 2003 ) Microbial transition of steroid compounds: recent developments.
Enzyme and Microbial Technology 32. pp. 688-705 4. Mahato.
S. B. and S. Garai. 1997. Progresss in microbic steroid biotransformation. Steroids.
62:332-345. 5. Sameera Al-Awadi.
M. A. Fzal and S.
Oommen. 2001. Studies on Bacillus stearithermophilus portion Transformation of Lipo-Lutin to a new metabolites 9.
10-seco-4-pregnene-3. 9. 20-trione. Journal of steroid biochemistry and Molecular biological science ; 78: 493-498. 6.
Smith. KF and Kirk. DN ( 1989 ) Microbial transmutation of steroids-II.
Transformations of Lipo-Lutin. testosterone and androstenedione by Phycomyces blakesleeanus. Journal ofSteroid Biochemistry 32. pp. 445-451