ISOLATION AND IDENTIFICATION OF TWO NOVEL SDS-RESISTANT SECRETED CHITINASES

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have isolated two SDS-resistant endochitinases, designated as ASCHI53 and ASCHI61, from Aeromonas schubertii in the soil of southern Taiwan. Mass measurement with MALDI-TOF indicates that the molecular weights are 53,527 Da for ASCHI53 and 61,202 Da for ASCHI61. N-terminal sequencing of these two chitinases suggests that thye are novel proteins. Degredation of chitin with these two endochitinases gives rise to hexameric chitin oligosaccharide, a compound known to have various potent biomedical functions. In the presence of 5% SDS, ASCHI53 and ASCHI61 retain 65% and 75% chitinase activities, respectively while the chitinolytic activities completely retain even in the presence of 10% beta-mercaptoethanol.