Biochemical and catalytic properties of an endoxylanase purified from the culture filtrate of Thermomyces lanuginosus ATCC 46882

 
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1998 (EN)
Biochemical and catalytic properties of an endoxylanase purified from the culture filtrate of Thermomyces lanuginosus ATCC 46882 (EN)

Ryan, J (EN)
Nerinckx, W (EN)
Vrsanska, M (EN)
Kremnicky, L (EN)
Kekos, D (EN)
Biely, P (EN)
Christakopoulos, P (EN)
Katapodis, P (EN)
Bhat, MK (EN)
Macris, BJ (EN)
Ntauma, P (EN)
Claeyssens, M (EN)
Bennett, NA (EN)

N/A (EN)

An endoxylanase (1,4-β-D-xylan xylanohydrolase, EC 3.2.1.8) from the culture filtrates of T. lanuginosus ATCC 46882 was purified to homogeneity by DEAE-Sepharose and Bio-Gel P-30 column chromatographies. The purified endoxylanase had a specific activity of 888.8 μmol min-1 mg-1 protein and accounted for approximately 30% of the total protein secreted by this fungus. The molecular mass of native (non-denatured) and denatured endoxylanase were 26.3 and 25.7 kD as, respectively. Endoxylanase had a pI of 3.7 and was optimally active between pH 6.0-6.5 and at 75°C. The enzyme showed > 50% of its original activity between pH 5.5-9.0 and at 85 °C. The pH and temperature stability studies revealed that this endoxylanase was almost completely stable between pH 5.0-9.0 and up to 60°C for 5h and at pH 10.0 up to 55°C for 5 h. Thin-layer chromatography (TLC) analysis showed that endoxylanase released mainly xylose (Xyl) and xylobiose (Xyl2) from beechwood 4-O-methyl-D-glucuronoxylan, O-acetyl-4-Omethyl-D-glucuronoxylan and rhodymenan (a β-(1→3)-β(1→4)-xylan). Also, the enzyme released an acidic xylo-oligosaccharide from 4-O-methyl-D-glucuronoxylan, and an isomeric xylotetraose and an isomeric xylopentaose from rhodymenan. The enzyme hydrolysed [1-3H]-xylo-oligosaccharides in an endofashion, but the hydrolysis of [1-3H]-xylotriose appeared to proceed via transglycosylation, since the xylobiose was the predominant product. Endoxylanase was not active on pNPX and pNPC at 40 and 100 mM for up to 6h, but showed some activity towards pNPX at 100 nM after 20-24 h. The results suggested that the endoxylanase from T. lanuginosus belongs to family 11.Microbial endoxylanases have attracted considerable research interest in recent years mainly due to their potential application in food, animal feed, paper and pulp industries. An endoxylanase (1,4,-β-D-xylan xylanohydrolase, EC 3.2.1.8) from the culture filtrates of T. lanuginosus ATCC 46882 was purified to homogeneity by DEAE-Sepharose and Bio-Gel P-30 column chromatographies. The purified endoxylanase had a specific activity of 888.8 μmol min-1 mg-1 protein and accounted for approximately 30% of the total protein secreted by this fungus. The molecular mass of native (non-denatured) and denatured endoxylanase were 26.3 and 25.7 kD as, respectively. Endoxylanase had a pI of 3.7 and was optimally active between pH 6.0-6.5 and at 75°C. The enzyme showed > 50% of its original activity between pH 5.5-9.0 and at 85°C. The pH and temperature stability studies revealed that this endoxylanase was almost completely stable between pH 5.0-9.0 and up to 60°C for 5 h and at pH 10.0 up to 55°C for 5 h. Thin-layer chromatography (TLC) analysis showed that endoxylanase released mainly xylose (Xyl) and xylobiose (Xyl2) from beechwood 4-O-methyl-D-glucuronoxylan, O-acetyl-4-O-methyl-D-glucuronoxylan and rhodymenan (a β-(1→3)-β(1→4)-xylan). Also, the enzyme released an acidic xylo-oligosaccharide from 4-O-methyl-D-glucuronoxylan, and an isomeric xylotetraose and an isomeric xylopentaose from rhodymenan. The enzyme hydrolysed [1-3H]-xylo-oligosaccharides in an endofashion, but the hydrolysis of [1-3H]-xylotriose appeared to proceed via transglycosylation, since the xylobiose was the predominant product. Endoxylanase was not active on pNPX and pNPC at 40 and 100 mM for up to 6h, but showed some activity towards pNPX at 100 mM after 20-24h. The results suggested that the endoxylanase from T. lanuginosus belongs to family 11. (EN)

journalArticle

Culture filtrate (EN)
unclassified drug (EN)
Temperature (EN)
structure analysis (EN)
Substrate Specificity (EN)
Catalytic properties (EN)
catalysis (EN)
Oligomers (EN)
Proteins (EN)
Fungi (EN)
Polysaccharides (EN)
enzyme purification (EN)
fungus (EN)
Plant cell culture (EN)
enzyme activity (EN)
Isoelectric Point (EN)
Xylosidases (EN)
pH (EN)
Mitosporic Fungi (EN)
Catalysis (EN)
priority journal (EN)
Xylans (EN)
thin layer chromatography (EN)
enzyme analysis (EN)
Filtration (EN)
Disaccharides (EN)
Thermomyces lanuginosus (EN)
Enzyme Stability (EN)
Fungal Proteins (EN)
molecular weight (EN)
xylan (EN)
Oligosaccharides (EN)
article (EN)
Xylose (EN)
Thin layer chromatography (EN)
xylose (EN)
Endoxylanase (EN)
ph (EN)
nonhuman (EN)
T. lanuginosus (EN)
Enzyme kinetics (EN)
Endo-1,4-beta Xylanases (EN)
Column chromatography (EN)
hydrolysis (EN)
Hydrogen-Ion Concentration (EN)

Εθνικό Μετσόβιο Πολυτεχνείο (EL)
National Technical University of Athens (EN)

Carbohydrate Research (EN)

1998


Elsevier Sci B.V., Amsterdam, Netherlands (EN)



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