By Pooja Walke
Feb 22, 2017
"Lipases (try acryl glycerol acylhydrolase ) are
the enzymes which catalyze the hydrolysis and
the synthesis of ester formed from glycerol and
long chain fatty acid.
During hydrolysis lipases pick acryl group from
glycosides forming lipase acyl complex which
then transfers its acyl group to OH group water.
Enzyme production can be enhanced by
improving the medium. But the use of bacillus
subtilis in solid state fermentation is improved
maximum yield in the production of lipase.
Isolation of bacillus subtilis from soil
collected from the Mahalaxmi oil industry,
Nanded.
Lipase production by using four different oil
samples by submerged fermentation process.
Comparative study of lipase production on
different oil samples i.e groundnut oil,
coconut oil, soybean oil,gingelly oil etc.
SDS-PAGE of proteins was carried out in the
presence of sodium dodecyl sulphate – an
anionic detergent that readily binds and
dissociates oligomeric proteins in the
presence of reducing agent, 2-mercapto
ethanol into their subunits.
The protein SDS complex carries negative
charges, hence move towards the anode and
the separation is based on the size of the
protein.
REAGENTS Acrylamide 30% [W/V] Ammonium per sulfate (APS) 10% [2/V] Sodium dodecyl sulphate (SDS) 10% [W/V] Separating gel buffer 1.5M Tris Hcl (pH
8.8) Stacking gel buffer 1M Tris Hcl (pH6.8)
Sample Buffer Staining solution Destaining Solution Storage Solution Running Buffer
Lipase activity calculated by titrating against 0.1 N NaOH,
phenolphthalein as indicator was 0.0095µg/ml.
BAND OF SAMPLES
Bovine serum albumin (66.2 kids)
Effect of temperature on enzyme production:
Thus the maximum activity was reported at temperature 40˚C.
0
0.001
0.002
0.003
0.004
0.005
0.006
0.007
0.008
0.009
0.01
5 10 20 25 30 35 40 45
LIPASE ACTIVITY
Effect of pH on enzyme production:
Thus the maximum activity was reported at
pH 8.5.
0
0.001
0.002
0.003
0.004
0.005
0.006
0.007
0.008
0.009
0.01
5 6 6.5 7 7.5 8 8.5 9
LIPASE ACTIVITY
Lipases are extremely versatile enzyme, showing many interesting properties of industrial applications.
The production of lipase is significantly influenced by carbon sources (substrate), temperature, pH
The maximum production of lipase was obtained by substrate soya bean oil having pH 8.5 and temperature 40˚C.
Fermentation in shake flask improved the lipase yield with an activity 0.0095 µg/ml/min in 48 h using soybean oil as substrate by Bacillus subtillis with moisture content of 70%.
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