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PMID:19258047
| Citation |
Li, Y, Yin, Q, Ding, M and Zhao, F (2009) Purification, characterization and molecular cloning of a novel endo-beta-1,4-glucanase AC-EG65 from the mollusc Ampullariacrossean. Comp. Biochem. Physiol. B, Biochem. Mol. Biol. 153:149-56 |
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| Abstract |
A novel endo-beta-1,4-glucanase, AC-EG65, with a molecular mass of 65 kDa, was purified from the gastric juice of the mollusc, Ampullaria crossean, by ammonium sulfate fractionation, anion exchange, gel filtration, hydrophobic interaction and a second round of anion exchange chromatography. AC-EG65 showed specific carboxymethyl cellulose hydrolytic activity of 13.3 U/mg protein and the optimal pH and temperature of the activity were pH 5.5-6.5 and 50-55 degrees C, respectively. From the cDNA library of A. crossean stomach tissue, eight endo-beta-1,4-glucanase genes with high similarity were successfully cloned based on the partial amino acid sequences of AC-EG65 and were classified into 3 groups: eg65-a, eg65-b, and eg65-c. The open reading frames of the groups eg65-a, eg65-b, and eg65-c were 2142 bp, 2171 bp, and 2169 bp in length, encoding 713, 723 and 722 amino acids, respectively. The eight deduced proteins consisted of a family II carbohydrate-binding module (CBM2) and a glycosyl hydrolase family 9 (GHF9) catalytic domain. More than 98% amino acid identities were shared within the same group and more than 87% sequence identities among the groups. The endogenous origins of these EGase genes were supported by PCR amplification using ovary genomic DNA as template. |
| Links |
PubMed Online version:10.1016/j.cbpb.2009.02.011 |
| Keywords |
Amino Acid Sequence; Animals; Catalytic Domain; Cellulase/chemistry; Cellulase/isolation & purification; Cellulase/metabolism; Cloning, Molecular; Gastric Juice/enzymology; Hydrogen-Ion Concentration; Molecular Sequence Data; Mollusca/enzymology; Phylogeny; Sequence Homology, Amino Acid; Substrate Specificity; Temperature |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0033903: endo-1,3(4)-beta-glucanase activity |
ECO:0000314: |
F |
Fig 2 |
complete | ||||
See also
References
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