Cloning and characterization of a small family 19 chitinase from moss (Bryum coronatum)

Chitinase-A (BcChi-A) was purified from a moss, Bryum coronatum, by several steps of column chromatography. The purified BcChi-A was found to be a molecular mass of 25 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an isoelectric point of 3.5. A cDNA encoding BcChi-A was cloned...

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Bibliographic Details
Published in:Glycobiology (Oxford) 2011-05, Vol.21 (5), p.644-654
Main Authors: Taira, Toki, Mahoe, Yoko, Kawamoto, Noriko, Onaga, Shoko, Iwasaki, Hironori, Ohnuma, Takayuki, Fukamizo, Tamo
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Antifungal Agents - chemistry
Antifungal Agents - metabolism
Antifungal Agents - pharmacology
Bryopsida - enzymology
Catalytic Domain
Chitinases - chemistry
Chitinases - metabolism
Chitinases - pharmacology
Cloning, Molecular
Enzyme Assays
Enzyme Stability
Hydrogen-Ion Concentration
Molecular Sequence Data
Phylogeny
Sequence Alignment
Trichoderma - drug effects
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Summary:Chitinase-A (BcChi-A) was purified from a moss, Bryum coronatum, by several steps of column chromatography. The purified BcChi-A was found to be a molecular mass of 25 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and an isoelectric point of 3.5. A cDNA encoding BcChi-A was cloned by rapid amplification of cDNA ends and polymerase chain reaction. It consisted of 1012 nucleotides and encoded an open reading frame of 228 amino acid residues. The predicted mature BcChi-A consists of 205 amino acid residues and has a molecular weight of 22,654. Sequence analysis indicated that BcChi-A is glycoside hydrolase family-19 (GH19) chitinase lacking loops I, II, IV and V, and a C-terminal loop, which are present in the catalytic domain of plant class I and II chitinases. BcChi-A is a compact chitinase that has the fewest loop regions of the GH19 chitinases. Enzymatic experiments using chitooligosaccharides showed that BcChi-A has higher activity toward shorter substrates than class II enzymes. This characteristic is likely due to the loss of the loop regions that are located at the end of the substrate-binding cleft and would be involved in substrate binding of class II enzymes. This is the first report of a chitinase from mosses, nonvascular plants.
ISSN:0959-6658
1460-2423
DOI:10.1093/glycob/cwq212