Its about nitrilase production and its industrial application

1. Nitrile carboxylic acid +
ammonia
1

2. Nitrilase in drug and drug
intermediate production
Purification Application
Future
aspects
2

3. • Nitriles are highly toxic, mutagenic, and
carcinogenic due to their cyano group
• Bioremediation with nitrile-converting
enzymes
3

4. • Nitrile catabolism comprises two distinct
pathways
(1) Nitrilases directly convert nitriles to
corresponding carboxylic acids and NH3
(2) Nitrile hydratases catalyze the formation of
corresponding amides from nitriles
(3) Amidases subsequently hydrolyze amides to
carboxylic acids and NH3
4

5. Nitrilase sources
• Bacteria :
Rhodococcus, Acinetobacter, Pseudomonas,
Corynebacterium
• Filamentous fungi :
Fusarium, Gibberella, Aspergillus,
and Penicillium
• Yeasts :
Candida, Pichia, Saccharomyces, Debaryomyce
s, Geotrichum
5

6. Isolation
6
Conventional High throughput

7. Putative nitrilase active site [95]. The related residues of Glu54,
Lys127 and Cys169 were considered functionally important as a
catalytic triad in the nitrilase superfamily.
7

8. 8
Intracellular
enzyme Fragile
Susceptible to
outside
environment

9. Purification
9
Done at 40 C
Cell disruption
was performed
by
ultrasonication
high-pressure
homogenization
grinding in
mortar
disintegration
with glass
beads
to prepare
cell-free
extracts
The cell lysate
was centrifuged
to remove cell
debris

10. 10

11. In F. solani O1 nitrilase
• 90% of the whole-cell activity was recovered
• Using an extraction buffer supplemented with
0.8 M ammonium sulfate
• Contaminating cellular protein was removed
• Various types of column chromatography
techniques
11

12. Nirtrilase in Organic Acids
Synthesis
12

13. 13
Nicotinic acid
Feedstuff
additives Pharmaceutical
intermediates

14. 14
Mandelic acid
optical
resolving
agents
pharmaceutical
and agricultural
products

15. 15
Surface
coatings
Textiles
Adhesives
Paper
treatment
Polymeric
flocculants
Dispersants
Acrylic acid

16. 16
Household
cleaners
Industrial
and
Electronic
elements
Automotive
oil additives
pH
controllingCosmetics
Glycolic acid

17. 17

18. Reference
• Nitrilase in nitrile biocatalysis : recent progress
and forthcoming research
{PMCID:PMC3537687(Jin-Song Gong et.al)
• www.sciencedirect.com
• Journal of the American Chemical
Society(J.Am.Chem.Soc124(31),9024-
9025,2002)
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