HPLC ANALYSIS OF ANOMERIC FORMATION AND CLEAVAGE PATTERN BY CHITINOLYTIC ENZYME

The reactions of N-acetylchitooligosaccharides with chitinolytic enzym e were analyzed by HPLC using a Tosoh TSK-Gel amide-80 column with 70% acetonitrile as an eluent. We separated alpha and beta anomeric forms of N-acetyl-chitooligosaccharides, and obtain the following advantage s of this HPLC method. 1. We can easily identify the reaction mechanis m of chitinolytic enzymes by this method, distinguishing the inverting mechanism showing alpha anomer formation from the retaining mechanism showing beta anomer formation. 2. We can also estimate the cleavage p atterns of N-acetylchitooligosaccharides by chitinolytic enzymes by us ing natural substrates.