cellulase production

1. BY SREEREMYA .S
M.phil

2. INTRODUCTION
 Cellulose are produced mainly in plants.The solid
cellulosic waste can be treated efficiently.
 Fungal cellulase is an abundant resources ,is a potential
raw material for the microbial production of food, fuel etc.
 Cellulase enzymes are isolated from fungi present in higher
termite soil.
 Cellulase is a multienzyme ,system having 3 types of
enzymes –
carboxymethylcellulase(CMC),cellobiohydrolase
cellobiase.
 The significance of using CMC it has highly efficient
hydrolytic activity compared to other enzymes

3. METHODOLOGY
Collection of termite soil sample
20 gm of eight different termite soil (fungus
comb)were collected asceptically.
Isolation of fungi by spread plating
Identification of fungi(by LCBstaining)

4. METHODOLOGY
Screenig for cellulolytic activity
Measurement of cellulolytic activity
Standardization of temp& pH
Mass production

5. METHODOLOGY (CONT…)
Preteatement of substrates
Cellulase induction (basal media)
Endoglucanase assay(CMC)

6. METHODOLOGY (CONT…)
Preteatement of substrates
Cellulase induction (basal media)
Endoglucanase assay(CMC)

7. RESULT
Identification of fungi
*By Lactophenol cotton blue staining (LCB)
Microscopically observed the spores,sporearrangement,
mycelial pattern of fungi & thus eight fungi were identified
Aspergillus sp,Trichoderma sp (green
colonies),Trichoderma sp (white colonies),Penicillin
sp,Acremonium sp,Cladisporium sp,Neurospora sp,Phoma
sp

8. TERMITE SOIL

9. TERMITE SOIL

10. TERMITE SOIL

11. TERMITE SOIL

12. ISOLATED FUNGI FROM HIGHER TERMITE SOIL
Aspergillus sp Trichoderma sp

13. Trichoderma sp Neurospora sp

14. Acremonium sp Cladisporium sp

15. Penicillin sp Phoma sp

16. Screening for cellulolytic activity

17. ESTMATION OF PROTEIN BY LOWRY’S METHOD
SL
N0
:
STANDARD
VOL ml
SOL
Conc
µg
VOL
OF
d.H2O
VOL
OF
ALKAL
INE
CUSO4
Incu
bati
on
VOL of
folins
reagen
t
Incub
ation
at
room
temp
OD at
660n
m
S1 0.2 40 0.8 5 0.5 0.03
S2 0.4 80 0.6 5 0.5 0.07
S3 0.6 120 0.4 5 0.5 0.12
S4 0.8 160 0.2 5 0.5 0.15
S5 1.0 2000 - 5 0.5 0.18
B - - 1 5 0.5
T1 0.5 - 0.5 5 0.5 0.15
T2 0.5 - 0.5 5 0.5 0.22
T3 0.5 - 0.5 5 0.5 0.35
T4 0.5 - 0.5 5 0.5 0.28

18. LOWRY’S TEST
 T1-Phoma sp
 T2-Penicillin
 T3-Aspergillus
 T4-Trichodema viridae
 T5-Trichoderma sp
 T6-Neurospora sp
 T7Acremonium
 T8-Cladisporium

19. PRETREATED SUBSTRATE
SUGARCANEBAGASSE

20. PRETREATED SUBSTRATES
SAWDUST GROUNDNUT SHELL

21. PRETREATED SUBSTRATED
COCONUT HUSK,COTTON NEWSPAPER

22. ENDOGLUCANASE ASSAY
Aspergillus sp Trichoderma viridae
• Substrate- Enzyme activity
• Sawdust-1.13
• Newspaper-0.68
• Cotton-0.86
• Sugarcane bagasse-1.91
*Substrate- Enzyme activity
*Sawdust -0.36
*Newspaper – 1.74
*Cotton-0.81
*Sugarcanebagasse-1.80

23. Aspergillus niger - Sawdust Culture filtrate

24. Aspergillus - Groundnut Culture filtrate

25. Aspergillus niger Trichoderma viridae

26. Culture filtrate –Coconut husk
Aspergillus niger Trichoderma viridae

27. Substrate – Groundnut shell
Aspergillus niger Culture filtrate

28. Trichoderma viridae Culture filtrate

29. Sugarcane bagasse - Substrate
Aspergillus niger Trichoderma viridae

30. News paper- Substrate
Aspergillus niger Trichoderma viridae

31. Cotton -Substrate
Aspergillus niger
 Cotton as substrate is used
except Aspergillus sp other
fungi had not produced any
result.