Title of Thesis
Exploitation of Agro-Industrial Wastes for the Microbial
Production on Intracellular and Extracellular Enzyme Penicillin-G-Acylase
Department Of Chemistry / Government College
|Number of Pages|
|Keywords (Extracted from title, table of contents and
abstract of thesis)|
Wastes, Microbial, Production, Intracellular, Extracellular, Enzyme,
Penicillin, G-Acylase, Fermentation, megaterium, soybean, sunflower,
deficient strains of E. coli and B. megaterium were isolated from
the local habitat for the production of intra-cellular and
extra-cellular enzyme penicillin-GAcylase (PGA), respectively. After
screening, the best enzyme producing strain of E. coli (PCSIR-102)
and B. megaterium (5-B) were mutagenized by UV, MNNG, SDS and EMS
reagents. The strain of E. coli (MNNG-37) gave higher enzyme PGA
activity (231 IU mg-1) in fermentation medium of composition:
Peptone 2%, KH2PO4 0.3%, K2HPO4 0.7%, MgSO4.7H2O 0.02%,
sodium-L-glutamate 0.5% and yeast extract 0.5%. at pH 6.0, 30 0C
temperature for 50 hours. The strain of B. megaterium (MNNG-9) gave
more higher enzyme PGA activity (329 IU mg-1) in fermentation medium
of composition (g/l): yeast extract 2.0; peptone 5.0; sodium
chloride 5.0 (Oxoid) at pH 7.0, temperature 37 0C for 45 hours.
Strain of B. megaterium (MNNG-9) after treatement with EMS was
further mutagenized with in MNNG (Two-stage mutagenization) which
gave higher enzyme activity of strain M-9 (525 IU mg-1) in molasses
medium of composition (g l-1): CaCl2.2H2O 0.05; K2HPO4 1; MgSO4.7H2O
0.5; phenyl acetic acid 10; molasses 12 and Soytone 30 at pH 7.0,
temperature 37 0C for 30 hours.
The specific activity of PGA by E. coli (PCSIR-102) and B.
megaterium (5-B) was also studied by using different substrates like
wheat bran, rice hulls and defatted oil seed cakes of soybean,
sunflower or cotton by solid substrate fermentation. Low activity of
PGA was found as compared to the submerged fermentation.
The cells of B. megaterium (M-9) were immobilized in calcium
alginate beads. The specific activity of PGA was (505 IU mg-1) after
15 hours of fermentation at 30 0C. However, the enzyme PGA produced
from B. megaterium (M-9) was immobilized giving better enzyme
activity as compared to the immobilized whole cells and free cells.
88% of the enzyme PGA from B. megaterium (M-9) was recovered from
the fermentation broth. The level of purification was also confirmed
with the help of Fast Performance Liquid Chromatography.