“Evaluation of Physico-chemical Parameters for the Bio-reduction of Para chloro acetophenone by Aspergillus species”

SYNOPSIS FOR REGISTRATION

OF

M.PHARM DISSERTATION

Submitted to

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA

By

BIJJU.A

1ST M.PHARM

Department Of Pharmaceutical Chemistry

DAYANANDA SAGAR COLLEGE OF PHARMACY

2010

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE

KARNATAKA

ANNEXURE - II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name of the Candidate
and Address /
BIJJU.A S/O ANNAMALAI
#2143 ANANDI NILAYA VIDYANAGARA BEHIND GANESHA TALKIES H.B.ROAD
H.D.KOTE (TALUK) (POST) MYSORE(D)-571114
2. / Name of the Institution / Dayananda Sagar College of Pharmacy,
Kumaraswamy layout, Bangalore – 560 078.
3. / Course of Study and
Subject / M. Parma in Pharmaceutical Chemistry.
4. / Date of Admission / 7th JULY-2010
6.0 / Brief resume of the intended work:
6.1 – Need for the study:
Biocatalysis has become an increasingly valuable tool for the synthetic chemists. The development of novel biocatalytic methods is a continuously growing area of chemistry, microbiology and genetic engineering. Novel microorganism and or their enzymes are subject of intensive screening. Bio-transformation reactions are chemo-, regio-, stereo- and enantioselective producing a wide variety of fine chemicals that are intermediates of drugs, food ingredients and agro chemical intermediates1.
Under stringent guidelines set by the US Food and Drug Administration and potential industrial applications, the preparation of enantiomerically pure chiral drugs and drug intermediates is currently gaining momentum2.
Optically active alcohols have been increasingly recognized in the pharmaceutical field. Biocatalytic preparation of these chiral alcohols using microbial carbonyl reductase is one of the best methods for preparation of chiral auxiliaries from their corresponding prochiral ketones3.
The advantages of biocatalysts over chemical synthesis are that the reaction can be carried out at ambient temperature and atmospheric pressure thus avoiding the use of extreme conditions which could cause problems with isomerisation, recemisation, epimerization and rearrangement.
Using microbial whole cells as biocatalyst is advantageous for carrying out reduction reaction as they contain multiple dehydrogenases and thus they can accept a wide variety of unnatural substrates and they also contains necessary co-factor needed for bio-transformation4.
There are many studies on enantiomeric reduction of acetophenone and its derivatives. A preliminary study on the screening of micro organisms for the reduction of Para chloro acetophenone has been carried out and a potential organism for the purpose has been identified. In the present work, we are aiming to improve the yield of alcohol by optimizing the procedure.
7.0 / 6.2  – Review of the literature:
1. Keyller Bastos Borges et al1., reviews stereoselective reactions mediated by fungi, such as stereoselective hydroxylation, sulfoxidation, epoxidation, Baeyer-villiger oxidation, deracemization, and stereo- and enantioselective reduction of ketones.
2. A. Goswami et al2., have reported the use of surfactant for the microbial reduction of ω-bromoacetophenone. Several ω-bromoacetophenone derivative were reduced to (R)-(-) 2- bromo –1- phenyl / substituted phenyl ethanol derivatives with whole cell biocatalyst in good yield. The yield were more than 95% using anionic surfactant under an inert atmosphere in an aqueous medium. Among three organisms screened, i.e. S.cerevisiae, R.ribra, and C.tropicalies. In that R.rubra gave the best result.
3. Ashwini L. Kamble et al3., reported the enantioselective reduction of napthyl acetophenone to S(-)-1-(1-napthyl) ethanol by a novel isolate of Candida viswanathii, which is an important synthetic intermediate of mevinic acid analogue (potential inhibitor of 3- hydroxyl methyl coenzyme A reductase). They also reviews the optimization of initial pH, temperature for growth and effect of various carbon and nitrogen source, reaction time, reaction pH, reaction temperature and resting cell concentration for growth.
4. Ramesh N. Patel et al4., reported the enantioselective microbial reduction of substituted acetophenones. The chiral intermediate (S)-1-(2-bromo-4-fluoro phenyl) ethanol was prepared by the enantioselective microbial reduction of 2-bromo-4-fluoro acetophenone. Organisms from genus Candida, Rhodotorula, Baker’s yeast were used for microbial reduction.
5. Katja Goldberg et al5, have reviewed biocatalytic ketone reduction for the production of chiral alcohols by whole cell reduction. The review highlights biotechnological processes for the production of chiral alcohols by reducing prochiral ketones with whole cell for both laboratory and industries.
6. Eshabi B. Kubanoglu et al6, have carried out enantioselective reduction of substituted acetophenones to 1-phenylethanol derivatives by isolated EBK-9 Aspergillus niger as a effective biocatalyst. Under the optimized conditions substituted acetophenones were converted to corresponding optically active alcohol in up to 99% yield.
7. Kaoru Nakamura et al7 ., reviewed the recent development in asymmetric reduction of ketones with biocatalyst. The review includes methodologies to control enantioselectivities of biocatalytic asymmetric reduction with selected examples of ketones like aliphatic ketone, hydrogenated aromatic ketones, diketones, hydroxy ketones etc., The reviewers are of the opinion that significant progress has been made so that it is now possible to reduce various ketones to both (S) and (R) alcohols with excellent yield.
8. Lenilson C. Rocha et al8., carried out bioreduction of α-chloroacetophenone by using whole cells of seven strains of marine fungi to (S)-(-)-2-chloro-1-phenylethanol with enantiomeric excess of 50% and an isolated yield of 60%. They report that when fungi were grown in the absence of artificial sea water, no reduction was observed and it was more efficient at neutral rather than acidic pH.
9. Esabi B. Kurbanoglu et al9., carried out asymmetric reduction of acetophenone and its analogues using immobilized Rhodotorula glutinis cells. The performance and reaction parameters of the immobilized cells were also investigated with the effect temperature, buffer pH, buffer level and reaction time in the bioreduction were studied. The yield was found to be 77%. The authors report that the buffer level had a significant effect on the reaction.
10. Esabi B. Kurbanoglu et al10., carried out the reduction of acetophenone to (S)-isomer of 1-phenyl alcohol with a high enantiomeric excess by locally isolated ten strains o Alternaria alternata using ram horn peptone. The effect of incubation
time, pH, temperature on reduction were studied. The excellent enantiomeric excess up to 99% and the yield 86% were obtained.
11. Ramesh N. Patel et al11., carried out novel stereoselective process for preparation of (S)-1-arylehanol by reduction of substituted acetophenone by some selected microorganisms and they reported yield up to 99%.
12. Michel M. Chartrain et al12., carried out novel enantioselective bioreductive process using yeast to form intermediates in the syntesis of an endothelin antagonist.
6.3 – Objective of the Study:-
The present project focuses on the:-
* Optimization of medium composition for bio-reduction:-
a) By varying Carbon source
b) By varying Nitrogen source
* Optimization of initial pH for growth and Enzyme production.
* Optimization of Temperature.
* Selection of suitable co-substrate for bio-reduction.
* To study the effect of Anionic and Cationic surfactant on bio-reduction.
Materials and Methods:
7.1- Source of Data:
Review from chemical abstracts and journals like Tetrahedron Asymmetry, Journal of molecular catalysisB:Enzymatic, PANS, European Journal of Biochemistry, Journal of Antibiotics, Journal of medicinal Chemistry, Organic process R&D, Journal of Bioresource Technology, Internet browsing and standard books.
7.2 - Method of collection of data:
Microbial culture will be obtained from culture collection centers like MTCC, Chandigarh and NCL, Pune. All the chemicals and other reagents will be collected from standard companies. The reactions will monitored by Thin layer chromatography. Enantiopurity of the synthesized molecule will be confirmed by spectral data.
7.3 - Does the study require any investigations or interventions to be conducted on patients or other humans or animals? If so, Please describe briefly:
-No-
7.4 – Has ethical clearance been obtained from your Institution in case of 7.3?
Not applicable
8.0 / List of References:
1.  Keyller Bastos Borges, Warley de Souza Borges, Rosa Duran-Portan, Monica Tallarico Pupo, Pierina Sueli Bonato, Isidro Gonzalez Collado. Stereoselective biotransformations using fungi as biocatalysts. Tetrahedron: Asymmetry, 2009; 20: 385-397.
2.  A. Goswami, R.L Bezbaruah, J.Goswami, N.Borthakur, D.Dey and A.K.Hazarika. Microbial reduction of ω-bromoacetophenones in the presence of surfactants. Tetrahedron: Asymmetry, 2000; 11: 3701-09.
3.  Ashwini L. Kamble, Pankaj soni and Uttam C. Banerjee. Biocatalytic synthesis of S(-)-1-(1’-napthyl) ethanol by novel isolate of Candida viswanathii. Journal of molecular catalysis B: Enzymatic, 2005; 35(1-3): 1-6.
4.  Ramesh N. Patel, Animesh Goswami, Linda Chu, Mary Jo Donovan, Dana Cazzulino et al., Enantioselective microbial reduction of substituted acetophenone. Tetrahedron: Asymmetry, 2004; 15: 1247-1258.
5.  Katja Goldberg, Kirsten Schroer, Stephan Lutz and Andreas Liese. Biocatalytic ketone reduction—a powerful tool for production of chiral alcohols—part ΙΙ: whole-cell reductions. Appl.Microbiol.Biotechnol, 2007;76:249-55.
6.  Eshabi B. Kurbanoglu, Kani Zilbeyaz, Namudar I. Kurbanoglu and Hamdullah Kilic. Enantioselective reduction of substituted acetophenones by Aspergillus niger. Tetrahedron: Asymmetry, 2007; 18:1159-1162.
7.  Kaoru Nakamura, Rio Yamanaka, Tomoko Matsuda and Tadao Harada, Recent developments in asymmetric reduction of ketones with biocatalysts, Tetrahedron: Asymmetry, 2003, 14: 2659-81.
8.  Lenilson C. Rocha, Hercules V. Ferreira, Eli F. Pimenta, Roberto G.S. Berlinck, Mirna H.R. Seleghim, Darci C.D. Javaroti, Lara D. Sette, Rafaella C. Bonugli, Andre L.M.Porto. Bioreduction of α-chloroacetophenone by whole cells of marine fungi. Biotechnol Lett, 2009, 31: 1559-1563.
9.  Esabi B. Kurbanoglu, Kani Zilbeyaz, Murat Ozdal, Mesut Taskin, Namudar I. kurbanoglu. Asymmetric reduction of substituted acetophenones using once immobilized Rhodotorula glutinis cells. Bioresource Technology, 2010, 101: 3825-3829.
10.  Esabi B. Kurbanoglu, Kani Zilbeyaz, Namudar I. Kurbanoglu and Mesut Taskin. Highly enantioselective reduction of acetophenone by locally isolated Alternaria alternata using ram horn peptone. Tetrahedron: Asymmetry, 2007; 18: 1529-1532.
11.  Ramesh N. Patel, Animesh Goswami, Linda Chu, Mary Jo Donovan, et al., stereoselective reduction of substituted acetophenone. United States Patent no; US 6,800,477 B2, 2004.
12.  Michel M. Chartrain, Barbara A. Krulewicz, Paul N. Devine, David M. Tschaen. Enantioselective bioreduction using yeast. United States Patent no; 6,121,026, 2000.
9. / Signature of the candidate /
(BIJJU.A)
10. / Remarks of the Guide:
11. / Name and Designation of:
11.1 Guide:
11.2 Signature: / Smt. M.S. SANDHYAVALI.
Associate Professor
Dept of Pharmaceutical Chemistry
Dayananda Sagar College of Pharmacy,
Kumaraswamy layout, Bangalore – 78.
11.3 Co-Guide:
11.4 Signature: / Smt. BRAHAMANI PRIYADARSHINI.S.R
Associate Professor
Dayananda Sagar College of Pharmacy,
Kumaraswamy layout, Bangalore – 78.
11.5 Head of the Department:
11.6 Signature: / Dr. V. Murugan
Professor and principal
Dept of Pharmaceutical Chemistry
Dayananda Sagar College of Pharmacy,
Kumaraswamy layout, Bangalore – 78.
12. / 12.1 Remarks of the Chairman and Principal