Abstract
Aim:This study aims to produce and assay cellulolytic enzyme activity (endo-(1,4)-β-D-glucanase, exo-(1,4)-β-D-glucanase, and β-glucosidase, at optimum temperature and optimum pH) of Enterobacter cloacae WPL 214 isolated from bovine rumen fluid waste of Surabaya Abbatoir, Indonesia.Materials and Methods:To produce enzyme from a single colony of E. cloacae WPL 214, 98 × 1010 CFU/ml of isolates was put into 20 ml of liquid medium and incubated in a shaker incubator for 16 h at 35°C in accordance with growth time and optimum temperature of E. cloacae WPL 214. Further on, culture was centrifuged at 6000 rpm at 4°C for 15 min. Pellet was discarded while supernatant containing cellulose enzyme activity was withdrawn to assay endo-(1,4)-β-D-glucanase, exo-(1,4)-β-D-glucanase, and β-glucosidase.Results:Cellulase enzyme of E. cloacae WPL 214 isolates had endoglucanase activity of 0.09 U/ml, exoglucanase of 0.13 U/ml, and cellobiase of 0.10 U/ml at optimum temperature 35°C and optimum pH 5.Conclusion:E. cloacae WPL 214 isolated from bovine rumen fluid waste produced cellulose enzyme with activity as cellulolytic enzyme of endo-(1,4)-β-D-glucanase, exo-(1,4)-β-D-glucanase and β-glucosidase.
Highlights
Rumen is an excellent environment for microbial growth consisting of bacteria, fungi and protozoa which are widely known to play important role in the fermentation process of ruminant cattle feed [1]
E. cloacae WPL 214 isolated from bovine rumen fluid waste produced cellulose enzyme with activity as cellulolytic enzyme of endo-(1,4)-β-D-glucanase, exo-(1,4)-β-D-glucanase and β-glucosidase
Cellulase is an enzyme produced by cellulolytic microbes capable of hydrolizing β-1,4 glycoside bond in cellulose, a polysaccharide structure often found in plants [2]
Summary
Rumen is an excellent environment for microbial growth consisting of bacteria, fungi and protozoa which are widely known to play important role in the fermentation process of ruminant cattle feed [1]. Cellulase is an enzyme produced by cellulolytic microbes capable of hydrolizing β-1,4 glycoside bond in cellulose, a polysaccharide structure often found in plants [2]. Cellulose degradation by cellulolytic bacteria is a product of synergy in a group of cellulase enzymes. Cellulase enzyme system consists of three groups of hydrolytic enzymes, i.e. (1) endo-(1,4)-β-D-glucanase (endoglucanases), (2) exo-(1,4)-β-D-glucanase (exoglucanases), and (3) β -glucosidase [3]. Endo-(1,4)-β-D-glucanase enzyme hydrolyzes β bonds randomly in a morphous regions of cellulose fibers [4], generates oligosaccharides of different lengths, and can form a new chainend [5]. Exo-(1,4)β-D-glucanase enzyme works towards reducing and non-reducing end of polysaccharide chains, especially on crystalline cellulose region, and liberates glucose as the main product resulted by β-glucosidase
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