Production and Characterization of Recombinant Bacillus amyloliquefaciens ABBD Chitosanase (Csn 1) Expressed in Pichia pastoris GS115

• Main Author: Yasin, Ahmad
• Format: Final Project
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/38573
• Institution: Institut Teknologi Bandung

Chitosan is a linear polysaccharide detived from chi ti n, this compound is composed of D­ glucosamine and N-acetyl-D-glucosam in e which are connected via -I ,4-glycosidic bonds. Chitosan is hydrolyzed to produce a compound called kiitooligosaccharide (COS). COS compounds have a variety of impot1ant biological acti\ ities in the health sector such as antitumor. antibacteriaL and antifungal. One of the best ways to obtai n COS is by the enzymatic reactions using the chitosanase. The purpose of this study was to produce and characterize Csn I from the marine bacteria Bact/Ius amyloliquefacien'i ABBD expressed on J>JChra pa'itons GS J 15. The molecular mass of Csn I of - 35 kDa analyzed using sodium deodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The kinetic parameters \\ere detennined as follo\.\'s: the Michaelis-i\1enten constant (KM) with a value of 15.0-1- mg I mL. maximum velocity (Vma,) 4-1-.57 !llllOI s.mg, tumover number (kLal) of 1 559.83 s•'. and catalytic efficiency (kcm I Vma•J 103.71 mL I mg I s. The activ ity of Csn l significantly increased v. ith the presence of Mn2 howe,•er the acti\ i ty of Csn 1 is strongly inhibited by the addition of Cu.