Purification and Characterization of Recombinant Endoglucanases from the Pine Wood NematodeBursaphelenchus xylophilus
| Autor: | Taisei Kikuchi, Hajime Shibuya |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Tylenchida
Molecular Sequence Data Bursaphelenchus xylophilus Cellulase Applied Microbiology and Biotechnology Biochemistry Pichia Substrate Specificity Analytical Chemistry Microbiology Pichia pastoris Cell wall chemistry.chemical_compound Gene Expression Regulation Fungal Animals Hemicellulose Glycoside hydrolase Amino Acid Sequence Cloning Molecular Cellulose Molecular Biology biology Organic Chemistry Temperature General Medicine Hydrogen-Ion Concentration biology.organism_classification Recombinant Proteins chemistry Xylophilus biology.protein Glycoside hydrolase family 45 Adsorption Biotechnology |
| Zdroj: | Bioscience, Biotechnology, and Biochemistry. 72:1325-1332 |
| ISSN: | 1347-6947 0916-8451 |
| Popis: | A family of endoglucanases belonging to glycoside hydrolase family (GHF) 45 have been isolated from the pine wood nematode Bursaphelenchus xylophilus. Here we describe the purification and characterization of the recombinant enzymes, named Bx-ENG-1, 2, and 3, expressed in Pichia pastoris. The respective molecular masses of purified Bx-ENG-1, 2, and 3 were estimated to be 18, 33-39, and 100-140 kDa by SDS-PAGE, and 18, 67, and 252 kDa by gel filtration, suggesting that Bx-ENG-1 existed in an unglycosylated monomeric form and Bx-ENG-2 and Bx-ENG-3 in a glycosylated dimeric form. The enzymatic properties of the recombinant enzymes were similar to each other: optimal activity at 60 degrees C at about pH 6.0, like other endoglucanases of GHF45. The recombinant enzymes displayed the highest activity toward lichenan, and lower activities were observed on carboxymethyl cellulose and amorphous cellulose. Nematode enzymes also hydrolyzed glucomannan, the most abundant hemicellulose in the cell walls of softwood. These substrate specificities suggest that B. xylophilus endoglucanases acted on the cellulose-hemicellulose complex in the cell walls, resulting in a weakening of the mechanical strength of the cell walls to facilitate the nematode's feeding on plant cells. |
| Databáze: | OpenAIRE |
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