Characterization of Actinomycetes and Trichoderma spp. for cellulase production utilizing crude substrates by response surface methodology.
Laboratory bench scaling was done and an average of 1.85 fold increase by Response Surface Methodology (RSM) optimization was obtained. It was found that the predicted value (4.96 IU/ml) obtained by RSM is in close accordance with observed activity 5.14 IU/ml. Endoglucanases are mainly induced by CMC while Wheat bran (natural substrate) exoglucanase is more active when induced by avicel and cellulose. Addition of substrate beyond a level caused inhibition of cellulase production. The molecular weight of protein as determined by SDS-PAGE is very similar to molecular weight of cellulase of Trichoderma viride (T. viride) cellulase and Trichoderma reesei (T. reesei) endoglucanase. T. reesei β-glucosidase has high enzymatic activity on CMC substrate when compared with T. viride β-glucosidase. Secondary structure analysed by using Circular Dichroism confirmed that composition of celluase system is very similar to other analysed species. The cellulase was found to be active in pH range of 4.8-5.5; while temperature range varied from 50°C to 70°C. Although the enzymatic activity produced by mutants were lesser than the parent, but in one case mutants of Trichoderma reesei's BGL has shown higher activity on cellulose.
Department of Bioresources, University of Kashmir, Srinagar, 190006 Jammu & Kashmir India.
Tanveer
T
Pirzadah
Department of Microbiology, School of Biotechnology and Biosciences Lovely Professional University, Punjab, India.
Shashank
S
Garg
Department of Microbiology, School of Biotechnology and Biosciences Lovely Professional University, Punjab, India.
Joginder
J
Singh
Department of Microbiology, School of Biotechnology and Biosciences Lovely Professional University, Punjab, India.
Ashish
A
Vyas
AIRF, Jawaharlal Nehru University, New Delhi 67, India.
Manish
M
Kumar
International Centre for Genetic Engineering and Biotechnology (ICGEB), New Delhi 67, India.
Naseem
N
Gaur
Institute of Nuclear Medicine and Allied Sciences (INMAS), DRDO, New Delhi 54, India.
Madhu
M
Bala
Department of Bioresources, University of Kashmir, Srinagar, 190006 Jammu & Kashmir India.
Reiaz
R
Rehman
Amity Institute of Microbial Technology, Amity University Uttar Pradesh, Noida, 201303 India.
Ajit
A
Varma
Amity Institute of Microbial Technology, Amity University Uttar Pradesh, Noida, 201303 India.
Vivek
V
Kumar
Department of Bioresources, University of Kashmir, Srinagar, 190006 Jammu & Kashmir India ; Amity Institute of Microbial Technology, Amity University Uttar Pradesh, Noida, 201303 India.
Manoj
M
Kumar
Avicel; CMC; Circular dichroism; Response surface methodology; Wheat bran; Zymogram