Characterization of an alkalophilic extracellular chitosanase from Bacillus cereus GU-02.

An alkalophilic extracellular chitosanase (ACTase) was characterized from the culture supernatant of Bacillus cereus GU-02. Kinetic properties of ACTase produced from B. cereus GU-02 after cultivation in anaerobic condition, alkaline medium (pH 10) at 37°C for 3 days were investigated. ACTase was found to be stable in alkaline pH range from 8 to 10. Interestingly, optimum pH and temperature were estimated to be 10 and 37°C, respectively, where ACTase showed chitosan degrading activity (87%), which was enhanced by 15% in the presence of calcium ions (8 mM). The ACTase produced from B. cereus GU-02 was partially purified from the culture supernatant, and its enzymatic activity was kinetically characterized. The Vmax and Km were estimated with a chitosan (degree of deacetylation, DD 92% as substrate) as 0.038 U/min/μg protein and 0.327 μM, respectively. A combination of the TLC and MALDI-TOF MS results showed that the chitosan oligosaccharides obtained from the hydrolysis of high molecular weight chitosan (HMWC) by ACTase of the B. cereus GU-2 comprise oligomers with degree of polymerization (DP) mainly from dimers to pentamers. High production of ACTase and chitooligosaccharides may be useful for various industrial and biological applications.