Production and Purification of an Active endo-α-1 –> 6-D-mannanase Enzyme

The field of glycobiology heavily relies on enzymes capable of degrading glycoconjugates to determine their structure and analyze their structure-function relationship (e.g. to study the basis of their immunogenicity or toxic properties). Despite the prevalence of glycoconjugates containing alpha-1 –> 6-linked mannoses in nature, no endo-α-(1 –> 6)-D-mannanase is commercially available.

Previously, an endo-α-(1 –> 6)-D-mannanase from Bacillus circulans proved useful in characterizing the structure of two major lipopolysaccharides of mycobacteria known as lipomannan (LM) and lipoarabinomannan (LAM). Although first reported 45 years ago, no easily accessible form of this enzyme was available to the research community, a fact that may in part be explained by lack of knowledge of its complete gene sequence.