Introduction of concept to use micro-organism as bio-catalyst for production of drugs bu reduction of chemical burden.

1. Selective Biocatalytic
Oxidation of Alcohols:
Developing a ‘Green’
Oxidizing Agent
NITU SINHA
STELLA MARIS COLLEGE
@ Indian institute of Madras

2.  INTRODUCTION
 WHY BIO OXIDATION
 OBJECTIVE
 METHODLOGY
 RESULT
CONTENTS:-
2

3. An enantiopure drug is a pharmaceutical that is
available in one specific enantiomeric form.
Most biological molecules (proteins, sugars, etc.) are
present in only one of many chiral forms, so different
enantiomers of a chiral drug molecule bind
differently (or not at all) to target receptors.
INTRODUCTION:-
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4. One enantiomer of a drug may have a desired
beneficial effect while the other may cause serious
and undesired side effects.
Ethambutol: One enantiomer is used to
treat tuberculosis, the other causes blindness.
Naproxen: One enantiomer is used to treat arthritis
pain, but the other causes liver poisoning with no
analgesic effect
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5. 5
Biocatalysts for oxidation ….Why?
1.Mild Reaction Conditions
25-35º C, near neutral pH and std. pressure.
2.High Specificity
Chemo specificity
Selectivity for a compound or group of
compounds (eg: Chemo selective bio-
oxidation of primary alcohol by Janibacter
terrae DSM 13953)

6. Regio specificity:-
Selectivity among
similar groups on
the same molecule
(eg: Selectivity
among hydroxyl
groups at different
positions in the
same molecule of
polyol)
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7. Stereo specificity :-
Selective for one
stereoisomer
Stereo-selectivity
between a (S)- and
(R) configured sec-
alcohol centers in a
meso-diol
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8. 8
3. Controllability (eg: bio-oxidation of benzyl alcohol
to the corresponding aldehyde avoiding over
oxidation to benzoic acid using oxidases, isolated
alcohol dehydrogenases (ADHs).
Janibacter terrae DSM 13953 )
4. Biodegradable waste management problems
reduced.
5. Primary and secondary alcohols are important in
the pharmaceutical, agricultural and food
Industries.
Orbegozoa. T, I. Lavanderab, W. M.F. Fabiana, B. Mautnerb, J. G. de Vriesc and W.
Kroutil. Tetrahedron 2009: 65, 346805-6809

9. - : Potentially– biological oxidation reactions
can provide feasible alternative to chemical
synthesis.
Fast gaining importance as eco friendly
“GREEN TECHNOLOGY”
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10. 1. Traditional alcohol oxidation has been performed
with heavy metals (eg. Cr,Mn)
2. Metal-free alcohol oxidation usually depends on
moisture – sensitive oxidants and environmentally
undesirable reaction media such as chlorinated
solvent. (eg: Dimethyl sulfoxide as oxidant in the
presence of an ‘activating’ reagent such as
N,N’-dicyclohexylcarbodimide)
3. Aldehydes are in general not stable in the
conventional chemical oxidation conditions of
primary alcohols
Why not CHEMICAL????????
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11. 11
Biocatalytic oxidation:-
1. Whole-Cell oxidation
Whole-cell oxidations are used when
- enzyme is intracellular.
- enzyme needs a cofactor to carry out the
catalysis.
- for multienzymatic process.

12. Enzymes as catalyst
-For the oxidation of alcohols two types of
oxidoreductase have been employed most
frequently Dehydrogenases and Oxidases.
(Peroxidase and mono oxygenases were employed to
lesser extent).
Chemo-enzymatic methods
- Example: TEMPO (2,2,6,6-tetramethylpiperidin-1-
yloxy) and enzymes (Laccase) for the oxidation of
primary alcohols.
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13. This is a chemical reduction of alpha tetralone
with ethanol.
In order of preparation of substrate.
It’s a overnight reduction over a magnetic
stirrer in round bottom flask.
Then work up process for reduction was done
with DCM in separating flask.
SODIUM BOROHYDRIDE REDUCTION
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14. 1. Silica gel with 100% hexane in column.
2. solvent :- 1% ,2%,3% ,4%,5%,6%,7% and 8%
of ethyl acetate and hexane.
3. with condition column was eluted and tlc was
check with every alternative tubes of column
fraction.
4. Then fraction having the desired compound is
concentrated by vacuum Rota vapor.
-By this compound is collected.
COLUMN CHROMATROGRAPHY
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15. 1.Biocatalytic oxidation of selected secondary
alcohol using candida parapsilosis ATCC 7330.
2.Optimization for Biocatalytic oxidation condition
such as
:-Biomass
:-pH
:-Substrate concentration
:-solvent concentration for dissolving substrate.
:-Reaction volume
:-Reaction time.
OBJECTIVE:-
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16. BIOTRASFORMATION PROCEDURES
Candida parapsilosis ATCC 7330 is grown in YMB
medium.(40hr)
Cells were harvested by centrifuge.(4.c, 6000 rpm,
15mins)
Wasted twice and suspended with distilled water
(7.6pH)
Reaction was done in orbital shaker at 150 rpm,
25.
c.
The product was isolated wit ethyl acetate by
vacuum rotator vapor.
METHDOLOGY:-
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17.  Conversion of substrate can determine by reverse
phase HPLC using C-18.
FTIR is done for, functional groups detection.
NMR is done for determination compound purity.
METHOD OF ANALYSIS:-
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18. Oxidation by candida parapsilosis was successfully
performed and the conversion was 30% in chrial
column.
RESULTS:-
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19. 1