Abstract
Three chitosanases designated PSC-I, PSC-II, and PSC-III were purified from commercial pepsin preparation by sequentially applying pepstatin A-agarose affinity chromotography, DEAE-Sephacel ion-exchange chromatography, Mono Q column chromatography, and Mono P chromatofocusing. With respect to chitosan hydrolysis, the optimal pHs were 5.0, 5.0, and 4.0 for PSC-I, PSC-II, and PSC-III, respectively; optimal temperatures were 40, 40, and 30 °C; and the Km's were 5.2, 4.0, and 5.6 mg/mL. The molecular masses of the three isozymes were ∼40 kDa, as estimated by both gel filtration and SDS-PAGE, and the isoelectric points were 4.9, 4.6, and 4.4, respectively, as estimated by isoelectrofocusing electrophoresis. All three chitosanase isozymes showed activity toward chitosan polymer and N,N′,N″-triacetylchitotriose oligomer. Most effectively hydrolyzed were chitosan polymers that were 68-88% deacetylated.
Original language | English |
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Pages (from-to) | 1042-1048 |
Number of pages | 7 |
Journal | Journal of Agricultural and Food Chemistry |
Volume | 51 |
Issue number | 4 |
DOIs | |
Publication status | Published - Feb 12 2003 |
Externally published | Yes |
Keywords
- Characterization
- Chitosanase
- Pepsin
- Purification
ASJC Scopus subject areas
- General Chemistry
- General Agricultural and Biological Sciences