Determination of Mode of Chitosanase Action by Matrix Associated Laser Desorption Ionization - Time of Flight Mass Spectrometry Analysis

Abstract

A detailed knowledge about the mode of enzyme action is critical to get better understanding the productivity of target molecules. The present study was then performed to elucidate the mode of enzyme action and hydrolysis pattern of chitosanase toward high molecular weight chitosan. Hydrolysates derived from the reaction condition were then analyzed by Matrix Associated Laser Desorption Ionization - Time of Flight mass spectrometry (MALDI-TOF MS) analysis to define the real size of hydrolysates. Also, the sequence of linking pattern of hydrolysates derived from the action of chitosanase was determined based on the identification of both glucosamine (GlcN) and N-acetylglucosamine (GlcNAc). The results demonstrate that both homogeneous and heterogeneous chitooligosaccharides, designated as active molecular chitosan (AMC) can be produced without such a big differences. According to these results, we suggest that GlcNAc may not be a terminator for enzymatic hydrolysis processing as a key molecule in chitosanase reaction to determine the length of AMC. Upon identifying the type of linkage and size of AMC, nevertheless, these can be used as potent materials in various research fields including cosmeceutical and pharmaceutical applications.