| 1. |
- Puchar, Vladimir, et al.
(författare)
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Substrate and positional specificity of feruloyl esterases for monoferuloylated and monoacetylated 4-nitrophenyl glycosides
- 2007
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Ingår i: Journal of Biotechnology. - : Elsevier BV. - 0168-1656 .- 1873-4863. ; 127:2, s. 235-243
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Tidskriftsartikel (refereegranskat)abstract
- 4-Nitrophenyl glycosides of 2-, 3-, and 5-O-(E)-feruloyl- and 2- and 5-O-acetyl-α-l-arabinofuranosides and of 2-, 3-, and 4-O-(E)-feruloyl- and 2-, 3- and 4-O-acetyl-β-d-xylopyranosides, compounds mimicking natural substrates, were used to investigate substrate and positional specificity of type-A, -B, and -C feruloyl esterases. All the feruloyl esterases behave as true feruloyl esterases showing negligible activity on sugar acetates. Type-A enzymes, represented by AnFaeA from Aspergillus niger and FoFaeII from Fusarium oxysporum, are specialized for deferuloylation of primary hydroxyl groups, with a very strong preference for hydrolyzing 5-O-feruloyl-α-l-arabinofuranoside. On the contrary, type-B and -C feruloyl esterases, represented by FoFaeI from F. oxysporum and TsFaeC from Talaromyces stipitatus, acted on almost all ferulates with exception of 4- and 3-O-feruloyl-β-d-xylopyranoside. 5-O-Feruloyl-α-l-arabinofuranoside was the best substrate for both TsFaeC and FoFaeI, although catalytic efficiency of the latter enzyme toward 2-O-feruloyl-α-l-arabinofuranoside was comparable. In comparison with acetates, the corresponding ferulates served as poor substrates for the carbohydrate esterase family 1 feruloyl esterase from Aspergillus oryzae. The enzyme hydrolyzed all α-l-arabinofuranoside and β-d-xylopyranoside acetates. It behaved as a non-specific acetyl esterase rather than a feruloyl esterase, with a preference for 2-O-acetyl-β-d-xylopyranoside.
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| 2. |
- Bennett, Neil A., et al.
(författare)
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Biochemical and catalytic properties of an endoxylanase purified from the culture filtrate of Thermomyces lanuginosus ATCC 46882
- 1998
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Ingår i: Carbohydrate Research. - 0008-6215 .- 1873-426X. ; 306:3, s. 445-455
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Tidskriftsartikel (refereegranskat)abstract
- An endoxylanase (1,4-beta-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 mumol 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 degrees C. The enzyme showed > 50% of its original activity between pH 5.5-9.0 and at 85 degrees 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 degrees C for 5 h and at pH 10.0 up to 55 degrees 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 beta-(1-->3)-beta(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 6 h, but showed some activity toward pNPX at 100 mM after 20-24 h. The results suggested that the endoxylanase from T. lanuginosus belongs to family 11.
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| 3. |
- Katapodis, Petros, et al.
(författare)
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Biochemical and catalytic properties of an endoxylanase purified from the culture filtrate of Sporotrichum thermophile
- 2003
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Ingår i: Carbohydrate Research. - 0008-6215 .- 1873-426X. ; 338:18, s. 1881-1890
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Tidskriftsartikel (refereegranskat)abstract
- An endo-β-1,4-xylanase (1,4-β-d-xylan xylanoxydrolase, EC 3.2.1.8) present in culture filtrates of Sporotrichum thermophile ATCC 34628 was purified to homogeneity by Q-Sepharose and Sephacryl S-200 column chromatographies. The enzyme has a molecular mass of 25,000 Da, an isoelectric point of 6.7, and is optimally active at pH 5 and at 70 °C. Thin-layer chromatography (TLC) analysis showed that endo-xylanase liberates mainly xylose (Xyl) and xylobiose (Xyl2) from beechwood 4-O-methyl-d-glucuronoxylan, O-acetyl-4-O-methylglucuronoxylan and rhodymenan (a β-(1→4)-β(1→3)-xylan). Also, the enzyme releases an acidic xylo-oligosaccharide from 4-O-methyl-d-glucuronoxylan, and an isomeric xylotetraose and an isomeric xylopentaose from rhodymenan. Analysis of reaction mixtures by high performance liquid chromatography (HPLC) revealed that the enzyme cleaves preferentially the internal glycosidic bonds of xylooligosaccharides, [1-3H]-xylooligosaccharides and xylan. The enzyme also hydrolyses the 4-methylumbelliferyl glycosides of β-xylobiose and β-xylotriose at the second glycosidic bond adjacent to the aglycon. The endoxylanase is not active on pNPX and pNPC. The enzyme mediates a decrease in the viscosity of xylan associated with a release of only small amounts of reducing sugar. The enzyme is irreversibly inhibited by series of ω-epoxyalkyl glycosides of d-xylopyranose. The results suggest that the endoxylanase from S. thermophile has catalytic properties similar to the enzymes belonging to family 11.
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| 4. |
- Puchart, Vladimı́r, et al.
(författare)
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Production of xylanases, mannanases, and pectinases by the thermophilic fungus Thermomyces lanuginosus
- 1999
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Ingår i: Enzyme and microbial technology. - 0141-0229 .- 1879-0909. ; 24:5-6, s. 355-361
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Tidskriftsartikel (refereegranskat)abstract
- A group of 17 strains of the thermophilic fungus Thermomyces lanuginosus was examined for the production of xylanases, β-mannanases, arabinanases, and pectinases. All strains were found to be xylanolytic, and several were proven to be outstanding producers of microbial xylanase on glucuronoxylan and corn cobs. The strains hyperproducing xylanase secreted low amounts of xylan-debranching enzymes and did not produce β-mannan and arabinan-degrading enzyme systems. Only the strains showing lower xylanase production exhibited a higher degree of xylan utilization and also the ability to produce a mannanolytic enzyme system. One of the mannanolytic strains was found to be capable of producing arabinan-degrading enzymes. This strain also showed the best production of pectinolytic enzymes during growth on citrus pectin or sugar beet pulp. Some of the strains have good potential for use as sources of important industrial enzymes of high thermal stability.
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