Start Submission Become a Reviewer

Reading: Isolation of a cellulase producing Bacillus cereus from cow dung and determination of the ki...

Download

A- A+
Alt. Display

Research Articles

Isolation of a cellulase producing Bacillus cereus from cow dung and determination of the kinetic properties of the crude enzyme

Authors:

Anthonymuthu Mary Brintha Croos,

University of Jaffna, Jaffna, LK
About Anthonymuthu Mary Brintha

Department of Botany, Faculty of Science

X close

Sarathadevi Rajendran ,

University of Jaffna, Jaffna, LK
About Sarathadevi

Department of Agricultural Chemistry, Faculty of Agriculture

X close

Kapilan Ranganathan

University of Jaffna, Jaffna, LK
About Kapilan

Department of Botany, Faculty of Science

X close

Abstract

The objective of the study was to isolate a thermostable alkaline cellulase producing bacterial strain from diverse natural sources such as goat excreta, cow dung, tropical soil, and organic matter. Opened hot cellulose agar plates were transferred on to the selective carboxy methyl cellulose (CMC) agar media and incubated for 2 days at 37 oC. The samples were cultured on Nutrient Agar in order to isolate cellulolytic bacteria. The bacteria isolated were screened for cellulolytic activity using serial dilution and pour-plate method after which they were characterised. The bacterial isolate with the highest CMC hydrolytic capacity which was obtained from cow dung was selected for further studies. Based on the morphological, biochemical and cultural analysis, the strain from cow dung was been identified as Bacillus sp. Pure culture of this bacterial strain was grown overnight, DNA was extracted and 16S rDNA was amplified by thermocycler using universal primers. When the amplified 16S rDNA PCR product was sequenced using automated sequencer and sequence similarity search was done for the 16S rDNA sequence using BLAST, the unknown organism was identified as Bacillus cereus with the GenBank accession no AF290555. Cellulase from Bacillus cereus showed zero order kinetics for 20 minutes. The optimum pH for the crude cellulase from Bacillus cereus was 9.0 at 45oC and the temperature optimum was 45oC at pH 9.0. Michaelis constant for the cellulase  from  this isolate to soluble cellulose was  38.60 g-1L and  Vmax was 3.32 µmol mL-1 min-1 at pH 9.0 and at 45oC. Since this cellulase enzyme was stable for more than an hour at 40oC and at alkaline pH values, it could be a good candidate for industrial applications.


 

How to Cite: Croos, A.M.B., Rajendran, S. and Ranganathan, K., 2019. Isolation of a cellulase producing Bacillus cereus from cow dung and determination of the kinetic properties of the crude enzyme. Journal of the National Science Foundation of Sri Lanka, 47(2), pp.261–267. DOI: http://doi.org/10.4038/jnsfsr.v47i2.9168
Published on 25 Jul 2019.
Peer Reviewed

Downloads

  • PDF (EN)

    comments powered by Disqus