Isolation, characterization and identification of cellulase (Endo-β-1,4-glucanase) producing bacteria from diverse locations


 The objective of the study was to isolate cellulase producing bacteria from natural sources and identify bacterial isolates that produce thermostable enzyme. Bacteria were isolated samples collected different locations like mango leaf litter, termite gut, composting yard (municipal solid waste, compost, soil), soil decaying paper wood, saline coastal belt by enrichment in broth containing 1% cellulose subsequent culturing on plates purified cultures stored slants solidified agar medium same composition as medium. Primary screening for (Endo β-1,4-glucanase) production done growing them Carboxy Methyl Cellulose (CMC) using indicator dye Congo red. Secondary positive showing cellulolytic Index > 4 crude enzyme extraction assay CMC 1N citrate buffer (pH 5.0) at pH values (4.8- 7.0) temperatures (35- 70℃). Cellulase activity (EA) U/mL calculated based absorbance change per min. which depends amount reducing sugar produced catalysed conversion sugar. Isolates characterized morphologically biochemically six (Gram -ve: T1, MSW, C1, S2 Gram +ve: W2, P) having significantly higher EA selected DNA extraction. quantified four (MSW, S2, sequence analysis. 16S rDNA sequencing carried out through PCR amplification Blast NCBI blast similarity search tool resulted identification W2 E.coli, Sulfitobacter pontiacus, Bacillus subtilis respectively P cereus Rhodococcus erythropolis strain e1. showed 6.2 varying temperatures. Bacterial C2 40℃, MSW 60℃ others 50℃ (EA ranging 0.0054 – 0.0604 U/mL). P, had 0.006 - 0.03 70℃. This producers could be grown (40 70℃) substrate. In this substrate specificity indicates is an endo- β-1,4-glucanase plays a prominent role initiating sustaining hydrolytic process randomly cleaves into glucose olygomeric polysaccharides. Hence selecting with cellulosic substance extreme environmental conditions high temperature, salinity it possible enzymes are stable processing other harsh importance industrial applications.
 Keywords: Endo-β-1,4-glucanase, Cellulolytic index, activity, Thermostable