BIOLOGICAL PROCESSING OF AGRO INDUSTRIAL WASTE (CORN STOVER) FOR INDUCED PRODUCTION OF PHYTASE BY Schizophylum commune AND ITS PRACTICAL APPLICATIONS
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Author:
HALEMA BIBI
Citable URI :
https://vspace.vu.edu.pk/detail.aspx?id=344
Publisher :
Virtual University
Date Issued:
7/4/2020 12:00:00 AM
Abstract
Phytase is an industrial important enzyme which has the ability to breakdown the phytic acid into
myo-inositol and inorganic phosphate. In this regard, present research was designed to produce
phytase enzyme from cost effective and abundant source like corn stover as a solid substrate in
the presence of fungus Schizophyllum commune. Furthermore, the phytase was produced and
optimized by using response surface methodology (RSM) using different factors such as
temperature (25, 35, 45°C), pH (5, 6, 7, 8, 9), time of incubation (1, 3, 5, 7 days), moisture level
(10, 30, 50, 70%), nitrogen level (0.25, 0.5, 1, 1.5, 2.183g) and inoculum size (1, 2, 3.5, 5,
7.049ml). The maximum phytase activity (498.98U/mL) was recorded with optimized conditions
like temperature 45°C, pH 6, time of incubation 1 day, moisture level 70%, nitrogen level 1.5g
and inoculum size 5mL. In addition to that enzyme was purified by ammonium sulphate
precipitation and gel filtration chromatography. For ammonium sulphate precipitation salt range
was kept from 20% to 90%. Maximum precipitation of phytase was observed at 50%. Dialysis of
protein was done in buffer of pH 6 for 6 hours altering buffer after time interval of two hours. For
further purification, column chromatography with Sephadex G-100 was used and out of 32
elutions, no.17 elution showed maximum absorbance on spectrophotometer at 820nm. Besides
that, characterization of crude enzyme for optimum temperature and pH was done. Range of pH
for characterization was 4-8 and temperature range was 25-50°C. Vmax and Km values were
determined by reciprocal graphs. To get the Vmax and Km values, substrate concentration range
was kept 0.25-1.25mL. As a result of characterization the phytase was observed with more
activity at pH 5 and temperature 45°C. The Km and Vmax values for phytase were recorded
0.25U/mL and 250 U/mL respectively. Finally, the phytase was used to check its effect on
fluffing of bread. The bread was fluffed upto3cm against control 2.5cm and increase in length
was observed upto15cm against the control 12.3cm.
URI :
https://vspace.vu.edu.pk/details.aspx?id=344
Citation:
Bibi, H(2019),BIOLOGICAL PROCESSING OF AGRO INDUSTRIAL WASTE (CORN STOVER) FOR INDUCED PRODUCTION OF PHYTASE BY Schizophylum commune AND ITS PRACTICAL APPLICATIONS.VIRTUAL UNIVERSITY OF PAKISTAN.(Lahore,Pakistan)
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Final Version
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