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Purification and properties of extracellular chitinases from the parasitic fungus Isaria japonica.
MedLine Citation:
PMID:  16233143     Owner:  NLM     Status:  PubMed-not-MEDLINE    
Abstract/OtherAbstract:
Two chitinases (P-1 and P-2) induced with colloidal chitin were purified from the culture supernatant of Isaria japonica by chromatography on DEAE Bio-Gel, chromatofocusing and gel filtration with Superdex 75 pg. The enzymes were electrophoretically homogeneous and estimated to have a molecular mass of 43,273 (+/-5) for P-1 and 31,134 (+/-6) for P-2 by MALDI-MS. The optimum pH and temperature was 3.5-4.0 and 50 degrees C for P-1 and 4.0-4.5 and 40 degrees C for P-2. P-1 acted against chitosan 7B (degree of deacetylation, 65-74%) = glycol chitin > colloidal chitin = chitosan 10B (degree of deacetylation, above 99%) and P-2 against chitosan 7B > glycol chitin = chitosan 10B > colloidal chitin in order of activity. The products of hydrolysis of chitin and chitosan hexamer were analyzed by MALDI-MS. The products from the chitin hexamer obtained with P-1 were almost all dimers with only a small amount of trimer whereas those obtained with P-2 were mainly trimers with some dimer and tetramer. No hydrolysis of chitosan hexamer was observed. High homology in the amino-terminal sequence for chitinase P-1 was exhibited by chitinases from Trichoderma harzianum, Candida albicans and Saccharomyces cerevisiae in the range of 48-39%. The highest homology for Chitinase P-2 was shown by an endochitinase from Metarhizium anisopliae of 66%, while 44% homology was exhibited by chitinases of Leguminosae plants.
Authors:
I Kawachi; T Fujieda; M Ujita; Y Ishii; K Yamagishi; H Sato; T Funaguma; A Hara
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of bioscience and bioengineering     Volume:  92     ISSN:  1389-1723     ISO Abbreviation:  J. Biosci. Bioeng.     Publication Date:  2001  
Date Detail:
Created Date:  2005-10-19     Completed Date:  2005-11-03     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  100888800     Medline TA:  J Biosci Bioeng     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  544-9     Citation Subset:  -    
Affiliation:
Faculty of Agriculture, Laboratory of Biological Chemistry, Meijo University, 1-501 Shiogamaguchi, Tempaku-ku, Nagoya 468-8502, Japan.
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