FEBS Letters | |
Purification, characterization, cDNA cloning, and expression of a xyloglucan endoglucanase from Geotrichum sp. M128 | |
Yaoi, Katsuro1  Mitsuishi, Yasushi1  | |
[1] Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan | |
关键词: Xyloglucanase; Xyloglucan; Endo-β-1; 4-glucanase; XEG; xyloglucan-specific endo-β-1; 4-glucanase; OXG-RCBH; oligoxyloglucan reducing end-specific cellobiohydrolase; SDS–PAGE; sodium dodecyl sulfate–polyacrylamide gel electrophoresis; MALDI-TOF; matrix-assisted laser desorption/ionization time-of-flight; CMC; carboxymethylcellulose; RACE; rapid amplification of cDNA ends; CBM; carbohydrate-binding module; | |
DOI : 10.1016/S0014-5793(04)00068-7 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
A novel xyloglucan-specific endo-β-1,4-glucanase (XEG), xyloglucanase, with a molecular mass of 80 kDa and a pI of 4.8, was isolated from the fungus Geotrichum sp. M128. It was found to be an endoglucanase active toward xyloglucan and not active toward carboxymethylcellulose, Avicel, or barley 1,3-1,4-β-glucan. Analysis of the precise substrate specificity using various xyloglucan oligosaccharide structures revealed that XEG has at least four subsites (−2 to +2) and specifically recognizes xylose branching at the +1 and +2 sites. The full-length cDNA encoding XEG was cloned and sequenced. It consists of a 2436-bp open reading frame encoding a 776-amino acid protein. From its deduced amino acid sequence, XEG can be classified as a family 74 glycosyl hydrolase. The cDNA encoding XEG was then expressed in Escherichia coli, and enzymatically active recombinant XEG was obtained.
【 授权许可】
Unknown
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