Preparation of antibacterial chito‐oligosaccharide by altering the degree of deacetylation of β‐chitosan in a Trichoderma harzianum chitinase‐hydrolysing process

BACKGROUND: Chito‐oligosaccharide (COS) is generally known to possess many specific biological functions, especially antibacterial activity, depending on its size. To prepare a specific size range of COS, however, has proved difficult. The aim of this study was to establish a method for preparing a specific size range of antibacterially active COS by adjusting the degree of deacetylation (DD) of β‐chitosan in a Trichoderma harzianum chitinase‐hydrolysing process. RESULTS: The molecular weight spectrum, elucidated by viscosity‐average molecular weight, high‐performance liquid chromatography and thin layer chromatography, of COS in chitosan hydrolysate was significantly related to the DD of its original chitosan. Compared with the original form, COS produced at 90% DD showed superior activity against most Gram‐negative bacteria tested, with a minimum inhibition concentration (MIC) ranging from 55 ± 27 to 200 ± 122 µ g mL^?1. Conversely, most Gram‐positive strains tested were less sensitive to COS (MIC > 880 ± 438 µ g mL^?1) than to its original form. Among the Gram‐positive strains, Staphylococcus xylosus was the only exception in that it showed a high susceptibility to COS and had an MIC as low as 45 ± 11 µ g mL^?1. CONCLUSION: The results indicate that the production of a specific size range of COS product is possible by altering the DD of chitosan in the chitinase‐catalysed process. To produce various sizes of COS for versatile biological functions, as seen in this study to inhibit various types of bacteria, is made possible in this established process. Copyright © 2008 Society of Chemical Industry