Purification and characterization of a chitosanase from Serratia marcescens TKU011.
A chitosanase was purified from the culture supernatant of Serratia marcescens TKU011 with shrimp shell wastes as the sole carbon/nitrogen source. Zymogram analysis revealed the presence of chitosanolytic activity corresponding to one protein, which was purified by a combination of ion-exchange and gel-filtration chromatography. The molecular weight of the chitosanase was 21 kDa and 18 kDa estimated by SDS-PAGE and gel-filtration, respectively. The optimum pH, optimum temperature, pH stability, and thermal stability of the chitosanase were 5, 50 degrees C, pH 4-8, and <50 degrees C, respectively. The chitosanase was inhibited completely by EDTA, Mn(2+), and Fe(2+). The results of peptide mass mapping showed that three tryptic peptides of the chitosanase were identical to a chitin-binding protein Cbp21 from S. marcescens (GenBank accession number gi58177632) with 63% sequence coverage.
Wang
San-Lang
SL
Graduate Institute of Life Sciences, Tamkang University, Tamsui, Taiwan. sabulo@mail.tku.edu.tw
Peng
Jo-Hua
JH
Liang
Tzu-Wen
TW
Liu
Kao-Cheng
KC
D000818:Animals; D000975:Antioxidants / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D001713:Biphenyl Compounds; D003386:Brachyura / Q000382:microbiology; D048271:Chitosan / Q000378:metabolism; D049832:Decapodiformes / Q000382:microbiology; D005609:Free Radicals / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D033561:Penaeidae / Q000382:microbiology; D010851:Picrates / Q000378:metabolism; D012706:Serratia marcescens / Q000201:enzymology* / Q000254:growth & development; D013379:Substrate Specificity
D000975:Antioxidants; D001713:Biphenyl Compounds; D005609:Free Radicals; D010851:Picrates; D048271:Chitosan; C004931:1,1-diphenyl-2-picrylhydrazyl; D006026:Glycoside Hydrolases; C019868:chitosanase