Isolation and Purification of Recombinant Chitosanase (Csn1) from Bacillus amyloliquefaciens for Screening Antibacterial Chitooligosaccharides

• Main Author: AHMAD FALAQ (NIM : 10512082), GALIH
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/27439
• Institution: Institut Teknologi Bandung

<p align="justify">As a maritime nation, Indonesia has various marine resources. One of the greatest benefit is crustacean, such as shrimps and crabs. This commodity is a primary export sector of Indonesia in which 170 units of the shrimp processing industries exist approximately with the production capacity of 500,000 tons annually. Even though the shrimp processing industries generate more income, they produce shrimp shell waste. To optimize the utilization of the waste, it can be done by turning it into chitin, which is the main compound of shrimp shell, and chitosan, chitin derivatives. One of the hydrolysis product of chitosan, chitooligosaccharide, has various biological activities such as antibacterial, antifungal, and antiinflammatory. This study was projected to produce the recombinant chitosanase from Bacillus amyloliquefaciens used for hidrolysis of chitosan to chitooligosaccharide with antibacterial potency. The bacterium constitutively produced chitosanase in a Luria Bertani culture. The csn1 gene expression was induced by IPTG (isopropyl-&#946;-D-1-thiogalactopyranoside) 0.5 mM. The supernatant extract cells containing chitosanase was purified using the CM Toyopearl cationic exchange chromatography and was characterized according to its enzyme properties. The molecular weight of the purified enzyme was about 27 kDa. The enzyme specific activity was 81917.39 U/mg with a purification fold of 163. However, the activity toward chitosan has not produce antibacterial chitooligosaccharides yet.<p align="justify">