Purification and characterization of (Acidic, alkaline and neutral) cellulase enzymes from thermophilic bacillus sp.

In this study, Bacillus sp. strains which isolated from soil and compost were used for production and characterization of acidic, alkaline and neutral cellulase enzymesAcidic CMCase of Bacillus sp.SC61 showed maximal activity at pH 3.0 and 60°C. The enzyme retained 42% and 72% of original activity at pH2.0-6.0 and temprature 80-100°C, respectively. MnCl2 and ß-mercaptoethanol induced activity. The end product of the CMCase was maltose. Molecular weight estimated as 88 kDa. CMCase inhibited with EDTA(52%) and completely inhibited with PMSF, these results shows the enzyme is a metalloenzyme and had serin residuesNeutral CMCase from Bacillus sp.SC6 presented optimal activity at pH 6.0 and 70°C , this enzyme showed 93% and 97% of its optimal activity at pH 4.0-8.0 and 50-90°C, respectively. Tween-20, ß-merkaptoetanol, EDTA, MnCl2 and NaCl induced enzyme activity. The end product of neutral CMCase was maltotriose. Molecular weight of the enzyme was detected as 121 kDa. The enzyme not inhibited with PMSF or EDTA, thus enzyme is not a metalloenzyme and serine protein.For SC41 alkaline CMCase optimal activity pH and temprature detected as pH 12.0 and 70°C, respectively. The end product of enzymatic hydrolysis was determined as maltose. The enzyme was %87 stable at 30-100°C and 87% stable at pH 9.0-12.0. Molecular weight estimated as 76.1 kDa. 35% inhibition by EDTA shows that the enzyme is a metalloenzyme, 31% inhibition with PMSF shows that SC41 CMCase has serine residues.These results show that the isolated enzymes can be used for biostoning and biofinishing in textile industry, biological deinking of wastepapers, fruit juice industry, bioethanol and biofuel production.