TY - JOUR
T1 - Characterization of three chitosanase isozymes isolated from a commercial crude porcine pepsin preparation
AU - Fu, Ju Yueh
AU - Wu, Sheng Ming
AU - Chang, Chen Tien
AU - Sung, Hsien Yi
PY - 2003/2/12
Y1 - 2003/2/12
N2 - 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.
AB - 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.
KW - Characterization
KW - Chitosanase
KW - Pepsin
KW - Purification
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U2 - 10.1021/jf020675g
DO - 10.1021/jf020675g
M3 - Article
C2 - 12568570
AN - SCOPUS:0037433115
VL - 51
SP - 1042
EP - 1048
JO - Journal of Agricultural and Food Chemistry
JF - Journal of Agricultural and Food Chemistry
SN - 0021-8561
IS - 4
ER -