Characterization of three chitosanase isozymes isolated from a commercial crude porcine pepsin preparation

Ju Yueh Fu, Sheng Ming Wu, Chen Tien Chang, Hsien Yi Sung

Research output: Contribution to journalArticle

24 Citations (Scopus)

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 languageEnglish
Pages (from-to)1042-1048
Number of pages7
JournalJournal of Agricultural and Food Chemistry
Volume51
Issue number4
DOIs
Publication statusPublished - Feb 12 2003
Externally publishedYes

Keywords

  • Characterization
  • Chitosanase
  • Pepsin
  • Purification

ASJC Scopus subject areas

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

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