fulltext.study @t Gmail

Purification, characterization, and molecular cloning of chitinases from the stomach of the threeline grunt Parapristipoma trilineatum

Paper ID Volume ID Publish Year Pages File Format Full-Text
34626 45037 2013 11 PDF Available
Title
Purification, characterization, and molecular cloning of chitinases from the stomach of the threeline grunt Parapristipoma trilineatum
Abstract

•Two chitinases (PtChiA and PtChiB) from the stomach of P. trilineatum were purified.•PtChiA and PtChiB showed different substrate specificities toward chitins.•Two chitinase cDNAs (PtChi-1 and PtChi-2) were cloned.•The fish stomach chitinases form unique 2 chtinase groups named as AFCase-1 and AFCase-2.

Two chitinase isozymes, PtChiA and PtChiB, were purified from the stomach of the threeline grunt, Parapristipoma trilineatum. The molecular masses of PtChiA and PtChiB were estimated to be 50 and 60 kDa by SDS-PAGE, respectively. Both chitinases were stable at pH 3.0–6.0 (acidic) and showed the optimum pH toward both short and long substrates in the acidic region (pH 2.5–5.0). PtChiA and PtChiB preferentially degraded the second and third glycosidic bonds from the non-reducing end of N-acetylchitooligosaccharides, respectively. PtChiA and PtChiB exhibited wide substrate specificities toward crystalline chitin. Moreover, 2 cDNAs encoding PtChiA and PtChiB, PtChi-1 and PtChi-2, respectively, were cloned. The deduced amino acid sequences of both chitinase cDNAs comprised N-terminal signal peptides, glycoside hydrolase 18 catalytic domains, linker regions, and C-terminal chitin-binding domains. Phylogenetic tree analysis of vertebrate chitinases revealed that fish stomach chitinases including PtChi-1 and PtChi-2 form unique chitinase groups, acidic fish chitinase-1 (AFCase-1) and acidic fish chitinase-2 (AFCase-2), which differ from the acidic mammalian chitinase (AMCase) group. The present results suggest that fish have a chitin-degrading enzymatic system in which 2 different chitinases, AFCase-1 and AFCase-2, with different degradation patterns are expressed in the stomach.

Keywords
(GlcNAc)n, N-acetylchitooligosaccharides; GlcNAc, N-acetyl-d-glucosamine; pNp-(GlcNAc)n, p-nitrophenyl N-acetylchitooligosaccharides; HPLC, high-performance liquid chromatography; SDS-PAGE, sodium dodecyl sulfate polyacrylamide electrophoresis; 5′ AAP, 5′
First Page Preview
Purification, characterization, and molecular cloning of chitinases from the stomach of the threeline grunt Parapristipoma trilineatum
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 48, Issue 9, September 2013, Pages 1324–1334
Authors
, , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us