ISOLATION AND CHARACTERIZATION OF COLLAGENASE PRODUCING BACTERIA FROM DIFFERENT NATURAL SOURCES.

1. Under the guidance of
Dr. Kirat Kumar Ganguly ,
Assistant Professor,
Department of Microbiology ,
Michael Madhusudan Memorial
College, Durgapur
Submitted by
Saikat Roy
M.Sc. Biotechnology 4th semester
Registration No: 02192 of 2020-2021
Roll-PG/VUOGP57/BIT-IVS No-047

2.  Collagenases are enzymes that break the peptide bonds in
collagen. They normally target the connective tissue in muscle
cells and other body organs.
 The Collagenases produced by Clostridium histolyticum ,
moreover Pseudomonas, Vibrio, Streptomyces sp also
produced collagenase .
 Collagenase 1, Collagenase 2, Collagenase 3 and Collagenase 4
are belongs to MMP-1,MMP-8,MMP-13,MMP-18.
 Collagen, a key component of the animal ECM, is made through
cleavage of pro-collagen by collagenase once it has secreted
from cell.
 Collagenase can be made by the body as part of this normal
immune response.

3.  MILK (from cow shed).
 CURD (from grocery store).
 MUTTON (from mutton shop)
 POULTRY WASTE (from poultry farm).
 DRAIN WATER (from drain behind college) .
 TAP WATER (college tap water ).
 SOIL ( from college campus).
 FISH ( from fish market).

4.  At first produced NB media plate with collagen.
 Inoculum was taken from sample plat and grew .
 Added filter papers into culture tube and incubated.
 Made agarose media plates(0.5% agarose,1%collagen present).
 Filter paper was taken from inoculum tube & put into agarose media
plates and incubated.
RESULT
After 24hrs incubation, very little amount of liquification zone was visible. For
clear results incubate next 48hrs.

5. 8 samples used, this 8 sample was equally given into plates .
This plates was made NA & 0.1%collagen. 48 hrs
incubation.
RESULTS
After 48hrs incubation zone was noticed in the plates.

6.  Neem, halud, tulsi were used as sample with PBS.
 8 NA plate was to be make and 8 sample( milk , curd, mutton, poultry
waste, drain water , tap water , soil, fish) spread well.
 3 well was doing in all plates and gave all this 3 sample ( neem, halud,
tulsi) paste.
RESULT
After 24 hrs incubation zone was found out in the 4 plates(mutton ,fish
,tapwater ,drain water).

7. Zymography is an electrophoretic method for enzyme
activity.
This technique is particularly useful for analyzing the
proteinase, composition of biological sample.
Coomassie blue staining of the gel reveals sites of
proteolysis as white bands on a dark blue background.

8.  Collagenases have been approved for medical uses
for:
1.Treatment of Dupuytren’s contracture and Peyronie's
disease
2.Wound healing.
3.Cellulite.