Glucosamine is widely used because of its diverse biological activity. However, its industrial production requires the hydrolysis reaction with concentrated acid at high temperature. Therefore, an environmentally friendly biocatalytic method is needed to replace the deacetylation of glucosamine by acid hydrolysis. In this study, the deacetylase (CqCBDA) from Cyclobacterium qasimii was screened and was successfully expressed heterologously in the Escherichia coli, which had a high level of soluble protein expression. The purified recombinant CqCBDA had the N-acetylglucosamine deacetylation activity and the specific enzyme activity was 11.5U/mg. The optimal temperature and pH value of CqCBDA was 40℃ and 7.6,while CqCBDA showed high thermal stability below 40℃. CqCBDA has great potential to be applied to the production of glucosamine by enzymatic hydrolysis. This study provides theoretical reference for green manufacturing of glucosamine by the enzymatic approach.