ISOLATION AND CLONING GENE ENCODING ENDOGLUCANASE FROM MARINE BACTERIUM BACILLUS AMYLOLIQUEFACIENS PSM 3.1

• Main Author: DEWI KURNIASIH (NIM 10505051); Pembimbing: Dr. Zeily Nurachman, SARI
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/13110
• Institution: Institut Teknologi Bandung

Cellulases are cellulolytic enzymes degrading cellulose polimer into simple sugar glucose or oligosaccharide. The enzyme has been used in many industrial applications such as textile- and paper industries. Most of cellulases are produced by bacteria and some fungi. In the previous research, cellulases from marine bacteria has been screened, isolated, and characterized. According to the microbiology test result, this marine bacterium was identified as Bacillus amyloliquefaciens PSM 3.1. The objective of this research was to isolate and to clone gene encoding endoglucanase from marine bacterium B. amyloliquefaciens PSM 3.1. The result showed that a gene encoding an endoglucanase II (egII) -a celullase hydrolizing internal chain of sellulose- consists of 1500 bp. The gene has an identity of 97% to endoglucanase genes of either B. amyloliquefaciens strain UMAS1002 (GenBank No.AF363635.1) or B. subtilis DLG (GenBank No.16185.1B). In-silico translation analysis showed that endoglucanase II expressed by egII contained 2 domains, namely catalytic domain and substrate binding domain. The catalytic domain of the endoglucanase II belongs to glycoside hydrolase 5 (GH 5) families with the catalytic residues predicted on residues Glu169 (as a proton donor) and Glu257 (as a nucleophile). Meanwile, substrate binding domain of the enzyme is classified as cellulose binding module 3 (CBM 3), in which residues for substrate binding are Asn13, Asp52, and Tyr53.