SELECTIVITY OF PLANT AND MICROBIAL MEDIATED REDUCTION OF KETONES

SYNOPSIS FOR REGISTRATION

Of

M.PHARM DISSERTATION

SUBMITTED TO

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES,

KARNATAKA

By

SUSHMA K. R.

1ST M.PHARM

Department Of Pharmaceutical Chemistry

DAYANANDA SAGAR COLLEGE OF PHARMACY

2009

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, KARNATAKA

BANGALORE

ANNEXURE - II

PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION

1. / Name of the Candidate
and Address /
Sushma K.R.
1st M.Pharm,
Department of Pharmaceutical Chemistry,
Dayananda Sagar College of Pharmacy,
Kumaraswamy layout,
Bangalore – 560078
PERMANENT ADDRESS
D/O K. V. Ramesh Babu
#276, Ground floor,
7th main road,
Srinagar,
Bangalore- 560050
2. / Name of the Institution / Dayananda Sagar College of Pharmacy,
Kumaraswamy layout, Bangalore – 560 078.
3. / Course of Study and
Subject / M.Pharm in Pharmaceutical Chemistry
4. / Date of Admission / 19th JUNE-2009
5.  Title of The topic :

“Selectivity of plant and microbial mediated reduction of ketones”

6.0 / Brief resume of the intended work:
6.1 – Need for the study:
Chirality is a key factor in the efficacy of many drug products and thus the production of single enantiomers of chiral intermediates has become increasingly important in the pharmaceutical industry. One of the most important starting materials for synthesizing various biologically active compounds are achiral alcohols. One of the latest and useful ways to obtain chiral alcohols is by biocatalytic reduction of prochiral ketone. Biocatalysis is an important alternative that organic chemists may prefer for synthesis of certain intermediates and finished pharmaceutical and other commercial products.
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, recemization, 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 contain necessary co-factors needed for biotransformation.
Plant tissues are also whole cells with large number of enzymes and they express multitude of enzymatic activities. Using plant tissues is highly advantageous because of their easy and surplus availability and thus economical. They are also environment friendly and help in going green. In this project an attempt will be made to carryout reduction of certain ketone substrates using microbial cells and plant tissue. Efficiency of both the methods will be compared.
7.0 / 6.2 – Review of the literature:
1. Takamitsu Utsukihara et al., 1 reported the stereoselective reduction of ketones by various vegetables. Various vegetable reductions gave a new idea of a biotechnological process. Reduction of (+)-and (−)-camphorquinones (1a, 1b) by various vegetables (carrot, potato, sweet potato, apple, japanese radish, cucumber, burdock and onion) gave α-hydroxycamphor selectively.
2. Kiyoko Matsuo et al., 2 have worked on asymmetric reduction of ketones with germinated plant. The biocatalyst is easily obtainable from commercially available vegetable seeds and is very easy to use. A germinated radish sprout was used as a novel type of biocatalyst for the asymmetric reduction of ketones. The reactions proceeded with high enantioselectivities (>99% ee). The biocatalyst is easily obtainable from commercially available vegetable seeds and is very easy to use.
3. Luciana L. Machado et al., 3 reported the bioreduction of aldehydes and ketones using Manihot species. The reduced products were typically obtained in excellent yields and with excellent enantiomeric excess, except for vanillin.
4. J. S. Yadav et al., 4 carried out an efficient enantioselective reduction of ketones with Daucus carota root. A novel and efficient reduction of various prochiral ketones such as acetopehones, α-azido aryl ketones, β-ketoesters, and aliphatic acyclic and cyclic ketones to the corresponding optically active secondary alcohols with moderate to excellent chemical yield was achieved by using Daucus carota, root plant cells.
5. Alejandro A. Orden et al5 carried out asymmetric bioreduction of natural xenobiotic diketones by Brassica napus hairy roots. Enantioselective bioreduction of natural prochiral diketones and some chemical derivatives were mediated by Brassica napus hairy root cultures. The natural bioactive diketones 1-(5-acetyl-2-hydroxyphenyl)-3-methylbut-2-en-1-one and 6-acetyl-2,2-dimethyl-2,3-dihydro-4H-chromen-4-one were both transformed into 6-(1(S)-hydroxyethyl)-2,2-dimethyl-2,3-dihydro-4H-chromen-4- one in high enantiomeric excesses (>97%) and with good biotransformation rates (>78%).
6. Yang Zhonghua et.al.,6 carried out isolation of microbe for asymmetric reduction of prochiral aromatic ketone and its reaction characters. The microbes Pichia pastoris and Geotrichum candidum had shown higher catalytic activity than that of bakers yeast.
7. Mai kawa meto et.al.,7 reported biotransformation of ( +) -2-methyl cyclohexanone by fungi. Furarium species was one of the effective biocatalysts for oxidoreduction of 2-metyl cyclohexanone.
8. Elzbieta Pekala et al.,8 carried out Preparation of enantiopure (R)-hydroxy metabolite of denbufylline using immobilized Lactobacillus kefiri DSM 20587 as a catalyst. The maximum biotransformation yield (96-98%) and highest enantioselectivity (96% ee) for the obtained metabolite were reached using DMSO as a cosolvent at a concentration of 7.5% (v/v) in the presence of L. kefiri immobilized either in calcium alginate or in carrageenan.
9. Kaoru Nakamura et al.,9 carried out Cyanobacterium-catalyzed asymmetric reduction of ketones. Synechococcus sp. PCC 7942, a Cyanobacterium, acted as a biocatalyst to reduce aryl methyl ketones into the corresponding (S)-alcohols with excellent enantioselectivities under illumination.
10. KatjaGoldberg et al.,10 have reviewed, biocatalytic ketone reduction. They are the opinion that it is a powerful tool for the production of chiral alcohols. Enzymes are able to perform reactions under mild conditions, e.g., pH and temperature, with remarkable chemo-, regio-, and stereoselectivity. Enzymes that are used in whole-cell biotransformations are often more stable due to the presence of their natural environment inside the cell. Because reductase-catalyzed reactions are dependent on cofactors, one major task in process development is to provide an effective method for regeneration of the consumed cofactors.
6.3 – Objective of the Study:-
Ø  Screening of different plant tissues for bioreduction of selected ketone substrate.
Ø  Reduction of the same ketone substrates with the selected microorganism.
Ø  Identification of microbe and plant reduced product with chemically reduced standard.
Ø  Characterization and quantification of reduced product.
Ø  Comparative evaluation of the bioreduction.
In the proposed scheme selected prochiral ketones are subjected for reduction using various plant tissues like carrot cucumber etc; to get reduced products. The same ketones are taken up for bioreduction using selected microorganisms. The products so obtained are compared with chemically reduced standard products. A comparative evaluation of the two methodologies will be carried out.
Materials and Methods:
7.1- Source of Data:
Review of literature from chemical abstract and journal like Tetrahedron asymmetry, Journal of Molecular catalysis B: Enzymatic, PANS , European Journal of Biochemistry, Journal of Antibiotics, Journal of Medicinal Chemistry, Organic process R&D, Internet browsing and standard books.
7.2 - Method of collection of data:
Microbial culture will be obtained from culture collection centres like MTCC, Chandigarh, and NCL, Pune. All the chemicals and other reagents will be collected from standard companies. The reactions will be monitored by thin layer chromatography. Enantio purity 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. Zuhair Takamitsu Utsukihara, Satoshi Watanabe, Atsushi Tomiyama, Wen Chai and
C.Akira Horiuchi. Stereoselective reduction of ketones by various vegetables. J. Mol.
Catal. B: Enzym. 2006; 41(3-4):103-9.
2. Kiyoko Matsero, Sei-ichiro Kawabe, Yosuke Tokuda, Takashi Eguchi, Kaoru
Nakamura. Asymmetric reduction of krtones with germinated plant. Tetrahedron:
Asymmetry. 2008; 19(2):157-9.
3. Luciana L.Machoda, Joao Sammy N. Sauza, Marcos Carlos de Mattor, Solange K. Sakota, Geoffrey A. Cordell and Telma L.G. Lemos. Bioreduction of aldehydes and ketones using Manihot species. Phytochemistry. 2006; 67(15):1637-43.
4. Yadav J.S, Nanda S, Thirupathi Reddy P, and Bhaskar Rao A. Efficient Enantioselective Reduction of Ketones with Daucus carota Root. J. Org. Chem. 2002; 67(11):3900-3.
5. Alejandro A. Orden, Fabricio R. Bisogno, Diego A. Cifuente, Oscar S. Giordano and Marcela Kurina Sanz. Asymmetric bioreduction of natural xenobiotic diketones by Brassica napus hairy roots. J. Mol. Catal. B: Enzym. 2006; 42(3-4):71-7.
6. Yang Zhonghua, Zeng Rong, Wang Yu, Wang Guanghui, Yao Shanjing, Isolation of microbe for asymmetric reduction of prochiral aromatic ketone and its reaction characters. Front. Chem. Eng. Chin. 2007; 1(4): 416-20.
7. Mai Kawamota, Takamistu Utsukihara, Chika Abe, Michiko Sato, Masahero Koshimura, Nakahide Kato and C.Akira Horiuchi. Biotransformation of ( +)-2-methyl cyclohexanone by fungi. Biotechnol. Lett. 2008; 30(9):1655-60.
8. Elzbieta Pekala, Joachim C. Burbiel, Christa E. Muller. Preparation of enantiopure (R)-hydroxy metabolite of denbufylline using immobilized Lactobacillus kefiri DSM 20587 as a catalyst. Chirality. 2008; 21(8):713-8.
9. Kaoru Nakamura, Rio Yamanaka, Keiko Tohi and Hiroki Hamada. Cyanobacterium-catalyzed asymmetric reduction of ketones. Tetrahedron Lett. 2000; 41(35):6799-802.
10.KatjaGoldberg, KirstenSchroer, StephanLutz and AndreasLiese. Biocatalytic ketone reduction—a powerful tool for the production of chiral alcohols—part II: whole-cell reductions. Appl. Microbiol. Biotechnol.2007; 76(2):249-55.
9. / Signature of the candidate /
(SUSHMA)
10. / Remarks of the Guide:
11. / Name and Designation of:
11.1 Guide:
11.2 Signature: / Mrs. Kalpana Divekar
Associate Professor
Dept of Pharmaceutical Chemistry
Dayananda Sagar College of Pharmacy,
Kumaraswamy layout, Bangalore – 78.
11.3 Co-Guide:
11.4 Signature / Mrs.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
12.2 Signature
/ Dr. V. Murugan
Professor and principal
Dept of Pharmaceutical Chemistry
Dayananda Sagar College of Pharmacy,
Kumaraswamy layout, Bangalore – 78.