Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “XYLAN”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 91 records · Page 5Linked to original sources
PubMed2000

Highly substituted glucuronoarabinoxylans (hsGAXs) and low-branched xylans show a distinct localization pattern in the tissues of Zea mays L.

Polyclonal antibodies which recognized highly substituted glucuronoarabinoxylans (hsGAXs) and low-branched xylans and did not cross-react with each other, were raised in order to examine localization of these epitopes in internodes of maize. Immunofluorescent labeling revealed different pattern between two succeeding developmental stages. The hsGAX epitope was localized evenly in primary walls in all tissue types, and strongly in unlignified secondary walls in phloem. However, lignified secondary walls in protoxylem, parenchyma and a part of fibers were faintly labeled with this epitope. Moreover, the epitope showed limited binding in lignified parenchyma and fiber walls at ultrastructural level. Low-branched xylan epitope was localized evenly throughout lignified walls in all tissue types. This epitope was also localized only in lignified walls of other organs such as leaf, root apex and dark-grown mesocotyl. Low-branched xylans are significantly related to lignification. Localization of hsGAX epitope in their organs was similar to that in internodes. The hsGAX epitope was distributed both in unlignified walls of all tissues and in lignified walls of parenchyma and annular thickening of protoxylem. We propose that hsGAX has separate functions in lignified and unlignified tissues. In conclusion, at tissue level, hsGAX is localized mainly in unlignified walls, and low-branched xylans in lignified walls.

Antibody Specificity↗
PubMed2005

Engineering of the gut commensal bacterium Bacteroides ovatus to produce and secrete biologically active murine interleukin-2 in response to xylan.

AIMS: The aim of this work was to engineer a gut commensal bacterium, Bacteroidesovatus, to produce and secrete a biologically active cytokine in a regulated manner as a basis for novel immunotherapies for chronic gut disorders. METHODS AND RESULTS: Bacteroides ovatus was engineered to produce murine interleukin-2 (MuIL2) intracellularly in response to xylan in culture media by inserting the MuIL2 gene into the xylanase operon of the organism. A second strain was engineered to secrete MuIL2 by adding Bacteroides fragilis enterotoxin secretion signal sequence to the protein. The recombinant strains produced MuIL2 only in the presence of xylan as determined by ELISA of cell lysates and culture supernatants. The IL2-dependent cell line CTLL-2 was used to demonstrate that MuIL2 produced by both B. ovatus strains was biologically active. This activity could be blocked by an anti-IL2 neutralizing antibody. The xylan-inducible nature of this system was demonstrated by RT-PCR. CONCLUSIONS: Bacteroides ovatus was successfully engineered to produce and secrete biologically active MuIL2 in a xylan-inducible manner. SIGNIFICANCE AND IMPACT OF THE STUDY: The production and secretion of a biologically active mammalian protein by a member of the gut microflora could lead to the development of new long-term immunotherapies for inflammatory gut diseases.

Animals↗
PubMed2003

Phenylacetic and phenylpropionic acids do not affect xylan degradation by Ruminococcus albus.

Since the addition of either ruminal fluid or a combination of phenylacetic and phenylpropionic acids (PAA/PPA) has previously been shown to dramatically improve cellulose degradation and growth of Ruminococcus albus, it was of interest to determine the effects of these additives on xylan-grown cultures. Although cell-bound xylanase activity increased when either PAA/PPA or ruminal fluid was added to the growth medium, total xylanase did not change, and neither of these supplements affected the growth or xylan-degrading capacity of R. albus 8. Similarly, neither PAA/PPA nor ruminal fluid affected xylan degradation by multiple strains of R. albus when xylan prepared from oat spelts was used as a carbohydrate source. These results show that the xylanolytic potential of R. albus is not conditional on the availability of PAA/PPA or other components of ruminal fluid.

Animals↗
PubMed1997

Hydrolysis of xylan by Aspergillus niger immobilized on non-woven fabrics.

Aspergillus niger, which produces xylan-degrading enzymes, was immobilized on non-woven fabrics. The maximum xylan hydrolysis activity (15 U/cm3-support) and the highest stability were obtained when the fungus was immobilized on non-woven fabric made of silk. The enzymatic properties of the immobilized preparation were similar to those of the free enzyme. Ten times repeated batch hydrolysis of birch-wood xylan was done over a period of 450 h. Hydrolysis of different xylan substrates such as oat spelts and rice bran by the immobilized mycelia was also investigated.

Aspergillus niger↗
PubMed1990

Physiology and genetics of xylan degradation by gastrointestinal tract bacteria.

Hemicelluloses or xylans are major components (35%) of plant materials. For ruminant animals, about 50% of the dietary xylans are degraded, but only small amounts of xylans are degraded in the lower gut of nonruminant animals and humans. In the rumen, the major xylanolytic species are Butyrivibrio fibrisolvens and Bacteroides ruminicola. In the human colon, Bacteroides ovatus and Bacteroides fragilis subspecies "a" are major xylanolytic bacteria. Xylans are chemically complex, and their degradation requires multiple enzymes. Expression of these enzymes by gut bacteria varies greatly among species. Butyrivibrio fibrisolvens makes extracellular xylanases but Bacteroides species have cell-bound xylanase activity. Biochemical characterization of xylanolytic enzymes from gut bacteria has not been done. A xylosidase gene has been cloned from B. fibrosolvens 113. The data from DNA hybridizations using a xylanase gene cloned from B. fibrisolvens 49 indicate this gene may be present in other B. fibrisolvens strains. A cloned xylanase from Bact. ruminicola was transferred to and highly expressed in Bact. fragilis and Bact. uniformis. Arabinosidase and xylosidase genes from Bact. ovatus have been cloned and both activities appear to be catalyzed by a single, bifunctional, novel enzyme. Continued research in genetic and biochemical areas will provide knowledge and insights for manipulation of digestion at the gut level and improved understanding of colonic fiber digestion.

Animals↗
PubMed1996

Molecular cloning of cDNAs encoding (1-->4)-beta-xylan endohydrolases from the aleurone layer of germinated barley (Hordeum vulgare).

Heteroxylans are major constituents of cell walls in the graminaceous monocotyledons. Degradation of walls in the starchy endosperm of germinated cereal grains is mediated, in part at least, by the action of (1-->4)-beta-xylan endohydrolases (EC 3.2.1.8). Complementary DNAs encoding (1-->4)-beta-xylan endohydrolases from the aleurone layer of germinated barley have been isolated and characterized. Southern blot analyses suggest that the enzymes are derived from a family of 3 or 4 genes, and cDNAs corresponding to two of these genes have been sequenced. The amino acid sequence deduced from one cDNA almost exactly matches the amino acid sequence determined previously from the purified enzyme. This enzyme is designated (1-->4)-beta-xylan endohydrolase isoenzyme X-I. The mature enzyme consists of 395 amino acid residues, has a calculated M(r) of ca. 44600 and an isoelectric point of 6.1, and is likely to adopt an (alpha/beta)8 barrel conformation. The amino acid sequence of the barley (1-->4)-beta-xylan endohydrolase encoded by the other cDNA, which is designated isoenzyme X-II, shows ca. 13% sequence divergence compared with isoenzyme X-I. Both enzymes exhibit sequence and structural similarities with microbial xylanases. Expression of the genes in germinated grain appears to be confined largely to the aleurone layer, and no mRNA transcripts could be detected in young vegetative tissues.

Amino Acid Sequence↗
PubMed2006

Penicillium purpurogenum produces a family 1 acetyl xylan esterase containing a carbohydrate-binding module: characterization of the protein and its gene.

At least three acetyl xylan esterases (AXE I, II and III) are secreted by Penicillium purpurogenum. This publication describes more detailed work on AXE I and its gene. AXE I binds cellulose but not xylan; it is glycosylated and inactivated by phenylmethylsulphonyl fluoride, showing that it is a serine esterase. The axe1 gene presents an open reading frame of 1278 bp, including two introns of 68 and 61 bp; it codes for a signal peptide of 31 residues and a mature protein of 351 amino acids (molecular weight 36,693). AXE I has a modular structure: a catalytic module at the amino terminus belonging to family 1 of the carbohydrate esterases, a linker rich in serines and threonines, and a family 1 carboxy terminal carbohydrate binding module (CBM). The CBM is similar to that of AXE from Trichoderma reesei, (with a family 5 catalytic module) indicating that the genes for catalytic modules and CBMs have evolved separately, and that they have been linked by gene fusion. The promoter sequence of axe1 contains several putative sequences for binding of gene expression regulators also found in other family 1 esterase gene promoters. It is proposed that AXE I and II act in succession in xylan degradation; first, xylan is attacked by AXE I and other xylanases possessing CBMs (which facilitate binding to lignocellulose), followed by other enzymes acting mainly on soluble substrates.

Acetylesterase↗
PubMed1997

Cellulose and Xylan Utilisation in the Lower Termite Reticulitermes speratus.

The distribution of the enzymes of cellulose and xylan metabolism namely endo-beta-1,4-glucanase, beta-glucosidase, endo-beta-1,4-xylanase and beta-xylosidase activities, in Reticulitermes speratus (Kolbe) was measured both in the salivary glands and in the major gut sections and along the length of the gut in freshly collected termites. The majority of the endo-beta-1,4-glucanase activity (77.8%) was found in the salivary glands which also contained 23.9% of the beta-glucosidase activity. At least 70% of the remaining activity was located in the anterior section of the hindgut. A small amount of endo-beta-1,4-xylanase activity (2.4%), but no beta-xylosidase activity, was present in the salivary glands. The majority of these activities were in the anterior section of the hindgut. The RQ of freshly collected termites at 25 degrees C was 1.03+/-0.01. Maintaining termites for 16 days on wood, cellulose and xylan showed that the RQ values of termites fed on wood or xylan were not significantly different from those of freshly collected termites but significantly increased when maintained on cellulose. The RQ of starved termites after 11 days was 0.81+/-0.02. There were three effects on protozoan populations of feeding termites xylan for 20 days. One species, Dinenympha parva was not affected, while five others, Pyrsonympha grandis, Holomastigotes elongatum, Dinenympha rugosa, Dinenympha leidy and Dinenympha porteri survived for 20 days but slowly decreased in numbers. The numbers of P. grandis and D. leidy surviving for 20 days were significantly different from those in starved termites. The third group comprising the two large species, Teratonympha mirabilis and Trichonympha agilis and three small species, Pyrsonympha modesta, Dinenympha exilis and Dinenympha nobilis disappeared within 15 days as in starved termites. It is suggested that protozoa in the first two groups are xylanolytic. Protozoan populations on wood and cellulose diets were not markedly affected. Selective removal of the protozoa by u.v. irradiation led to the loss of xylanolytic activity and a life span comparable to starved termites. Copyright 1997 Elsevier Science Ltd. All rights reserved

Journal Article↗
PubMed1989

Purification and characterization of three (1----4)-beta-D-xylan endohydrolases from germinated barley.

Three (1----4)-beta-D-xylan xylanohydrolases (xylan endohydrolases, EC 3.2.1.8) have been purified 1200-2800-fold from extracts of germinated barley (Hordeum vulgare L. cv. Clipper) by a sequence of ammonium sulphate fractionation, Procion-blue-dye chromatography, ion-exchange and gel filtration chromatography. The enzymes are likely to function in the depolymerization of cell wall arabinoxylans during mobilization of the starchy endosperm. They are classified as endohydrolases on the basis of analyses of products released during hydrolysis of a (1----4)-beta-xylan. The three xylan endohydrolases are monomeric proteins of apparent Mr 41,000 and all have isoelectric points of 5.2. The sequences of the 30 NH2-terminal amino acids of the three enzymes are the same, but it is not yet known whether they represent the products of separate genes or originate by differences in post-translational modification of a single gene product.

Amino Acid Sequence↗
PubMed1996

Acetyl xylan esterase from Trichoderma reesei contains an active-site serine residue and a cellulose-binding domain.

The axe1 gene encoding acetyl xylan esterase was isolated from an expression library of the filamentous fungus Trichoderma reesei using antibodies raised against the purified enzyme. Apparently axe1 codes for the two forms, pI 7 and pI 6.8, of acetyl xylan esterase previously characterized. The axe1 encodes 302 amino acids including a signal sequence and a putative propeptide. The catalytic domain has no amino acid similarity with the reported acetyl xylan esterases but has a clear similarity, especially in the active site, with fungal cutinases which are serine esterases. Similarly to serine esterases, the axe1 product was inactivated with phenylmethylsulfonyl fluoride. At its C-terminus it carries a cellulose binding domain of fungal type, which is separated from the catalytic domain by a region rich in serine, glycine, threonine and proline. The binding domain can be separated from the catalytic domain by limited proteolysis without affecting the activity of the enzyme towards acetylated xylan, but abolishing its capability to bind cellulose.

Acetylesterase↗
PubMed1984

Bioconversion of xylan to triglycerides by oil-rich yeasts.

A series of lipid-accumulating yeasts was examined for their potential to saccharify xylan and accumulate triglyceride. Of the genera tested, including Candida, Cryptococcus, Lipomyces, Rhodosporidium, Rhodotorula, and Trichosporon, only Cryptococcus and Trichosporon isolates saccharified xylan. All of the strains could assimilate xylose and accumuate triglyceride under nitrogen-limiting conditions. Strains of Cryptococcus albidus were found to be especially useful for a one-step saccharification of xylan coupled to triglyceride synthesis. Cryptococcus terricolus, a strain constitutive for lipid accumulation, lacked extracellular xylanase, but did assimilate xylose and xylobiose and was able to continuously convert xylan to triglyceride if the culture medium was supplemented with xylanase.

Journal Article↗
PubMed1986

Utilization of Xylan by Yeasts and Its Conversion to Ethanol by Pichia stipitis Strains.

Yeasts able to grow on d-xylose were screened for the ability to hydrolyze xylan. Xylanase activity was found to be rare; a total of only 19 of more than 250 strains yielded a positive test result. The activity was localized largely in the genus Cryptococcus and in Pichia stipitis and its anamorph Candida shehatae. The ability to hydrolyze xylan was generally uncoupled from that to hydrolyze cellulose; only three of the xylan-positive strains also yielded a positive test for cellulolytic activity. Of the 19 xylanolytic strains, 2, P. stipitis CBS 5773 and CBS 5775, converted xylan into ethanol, with about 60% of a theoretical yield computed on the basis of the amount of d-xylose present originally that could be released by acid hydrolysis.

Journal Article↗
PubMed1994

Purification and Characterization of Aeromonas caviae ME-1 Xylanase V, Which Produces Exclusively Xylobiose from Xylan.

A xylanase, which produces exclusively xylobiose from oat spelt and birch xylans, was isolated from the culture medium of Aeromonas caviae ME-1. The enzyme (xylanase V) was purified by ammonium sulfate fractionation, hydrophobic interaction, and ion-exchange and gel filtration chromatographies. The homogeneity of the final preparation was demonstrated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and agarose gel electrofocusing. The molecular mass and isoelectric point of the xylanase were 46 kDa and 5.4, respectively. Xylanase V had a maximum activity at a pH of 6.8 and at a temperature between 30 and 37 degrees C. It was relatively stable at a pH between 5.0 and 8.6 and a temperature between 25 and 37 degrees C. When soluble birch xylan was used as the substrate, the enzyme had a K(m) and V(max) of 2 mg/ml and 182 mumol of xylose equivalent liberated . min . mg of protein, respectively. By the action of xylanase V on xylans (from oat spelt and birch), only one product corresponding to xylobiose was observed by thin-layer chromatography. The xylanase V putative product was confirmed to be xylobiose by acid and enzymatic hydrolyses. The xylanase had neither beta-xylosidase, alpha-l-arabinofuranosidase, cellulase, nor beta-1,3-xylanase activities. Xylotriose was the shortest substrate which the enzyme could attack. These findings suggest that xylanase V is a novel enzyme that cleaves a xylobiose unit from one of the ends of xylans, probably by an exomechanism.

Journal Article↗
PubMed1999

Characterization of an acetyl xylan esterase from the anaerobic fungus Orpinomyces sp. strain PC-2.

A 1,067-bp cDNA, designated axeA, coding for an acetyl xylan esterase (AxeA) was cloned from the anaerobic rumen fungus Orpinomyces sp. strain PC-2. The gene had an open reading frame of 939 bp encoding a polypeptide of 313 amino acid residues with a calculated mass of 34,845 Da. An active esterase using the original start codon of the cDNA was synthesized in Escherichia coli. Two active forms of the esterase were purified from recombinant E. coli cultures. The size difference of 8 amino acids was a result of cleavages at two different sites within the signal peptide. The enzyme released acetate from several acetylated substrates, including acetylated xylan. The activity toward acetylated xylan was tripled in the presence of recombinant xylanase A from the same fungus. Using p-nitrophenyl acetate as a substrate, the enzyme had a K(m) of 0.9 mM and a V(max) of 785 micromol min(-1) mg(-1). It had temperature and pH optima of 30 degrees C and 9.0, respectively. AxeA had 56% amino acid identity with BnaA, an acetyl xylan esterase of Neocallimastix patriciarum, but the Orpinomyces AxeA was devoid of a noncatalytic repeated peptide domain (NCRPD) found at the carboxy terminus of the Neocallimastix BnaA. The NCRPD found in many glycosyl hydrolases and esterases of anaerobic fungi has been postulated to function as a docking domain for cellulase-hemicellulase complexes, similar to the dockerin of the cellulosome of Clostridium thermocellum. The difference in domain structures indicated that the two highly similar esterases of Orpinomyces and Neocallimastix may be differently located, the former being a free enzyme and the latter being a component of a cellulase-hemicellulase complex. Sequence data indicate that AxeA and BnaA might represent a new family of hydrolases.

Acetylesterase↗
PubMed1999

An acetylxylan esterase and a xylanase expressed from genes cloned from the ruminal fungus Neocallimastix patriciarum act synergistically to degrade acetylated xylans.

A Neocallimastit patriciarum acetylxylan esterase (BnaA) was expressed from the cloned gene in Escherichia coli. Purified recombinant BnaA efficiently released acetate from soluble acetylated birchwood xylan (ABX), with a specific activity of 76 U mg-1. In contrast, release of acetate was very inefficient from the insoluble substrates, spear grass and delignified spear grass. Addition of a recombinant xylanase, XynA, also expressed from a cloned N. patriciarum gene, had no effect on the release of acetate from ABX. However, the combination of recombinant BnaA and XynA released more acetate from spear grass and delignified spear grass than did BnaA alone. Significantly more reducing sugar was also released from all three substrates by the combination of recombinant XynA and BnaA than by XynA alone. Thus the extent of digestion of acetylated xylans by XynA appears to be limited by the acetylation. In this system BnaA does not appear to increase the rate of cleavage of insoluble substrates by XynA, but probably allows the release of shorter xylose oligomers from already solubilised acetylated xylan polymers.

Acetates↗
PubMed1995

Characterisation of 4-deoxy-beta-L-threo-hex-4-enopyranosyluronic acid attached to xylan in pine kraft pulp and pulping liquor by 1H and 13C NMR spectroscopy.

A new acidic sidegroup in xylans, from both kraft pulp and pulping liquor, was identified by NMR spectroscopy. Unmodified oligosaccharides from kraft pulp xylan were obtained by enzymatic hydrolysis with xylanase (Trichoderma reesei). The acidic oligosaccharides were separated from the natural forms on an anion exchange resin. The new acidic sidegroup was identified as 4-deoxy-beta-L-threo-hex-4-enopyranosyluronic acid (hexenuronic acid) by 1H and 13C NMR spectroscopy. Hexenuronic acid is a beta-elimination product of 4-O-methylglucuronic acid and is formed during kraft pulping. HMBC and NOESY experiments showed that hexenuronic acid is attached beta-(1 --> 2) to xylose. The NOESY data further indicated that hexenuronic acid protrudes from the main xylan chain. The pKa values for hexenuronic acid (3.03) and 4-O-methylglucuronic acid (3.14) attached (1 --> 2) to xylose were determined from pH-dependent chemical shifts.

Carbohydrate Sequence↗
PubMed2000

Novel sugar-binding specificity of the type XIII xylan-binding domain of a family F/10 xylanase from Streptomyces olivaceoviridis E-86.

The type XIII xylan-binding domain (XBD) of a family F/10 xylanase (FXYN) from Streptomyces olivaceoviridis E-86 was found to be structurally similar to the ricin B chain which recognizes the non-reducing end of galactose and specifically binds to galactose containing sugars. The crystal structure of XBD [Fujimoto, Z. et al. (2000) J. Mol. Biol. 300, 575-585] indicated that the whole structure of XBD is very similar to the ricin B chain and the amino acids which form the galactose-binding sites are highly conserved between the XBD and the ricin B chain. However, our investigation of the binding abilities of wt FXYN and its truncated mutants towards xylan demonstrated that the XBD bound xylose-based polysaccharides. Moreover, it was found that the sugar-binding unit of the XBD was a trimer, which was demonstrated in a releasing assay using sugar ranging in size from xylose to xyloheptaose. These results indicated that the binding specificity of the XBD was different from those of the same family lectins such as the ricin B chain. Somewhat surprisingly, it was found that lactose could release the XBD from insoluble xylan to a level half of that observed for xylobiose, indicating that the XBD also possessed the same galactose recognition site as the ricin B chain. It appears that the sugar-binding pocket of the XBD has evolved from the ancient ricin super family lectins to bind additional sugar targets, resulting in the differences observed in the sugar-binding specificities between the lectin group (containing the ricin B chain) and the enzyme group.

Binding Sites↗
PubMed1999

Xylan binding subsite mapping in the xylanase from Penicillium simplicissimum using xylooligosaccharides as cryo-protectant.

Following a recent low-temperature crystal structure analysis of the native xylanase from Penicillium simplicissimum [Schmidt et al. (1998) Protein Sci. 7, 2081-2088], where an array of glycerol molecules, diffused into the crystal during soaking in a cryoprotectant, was observed within the active-site cleft, we utilized monomeric xylose as well as a variety of linear (Xn, n = 2 to 5) and branched xylooligomers at high concentrations (typically 20% w/v) as cryoprotectant for low-temperature crystallographic experiments. Binding of the glycosidic moiety (or its hydrolysis products) to the enzyme's active-site cleft was observed after as little as 30 s soaking of a native enzyme crystal. The use of a substrate or substrate analogue as cryoprotectant therefore suggests itself as a simple and widely applicable alternative to the use of crystallographic flow-cells for substrate-saturation experiments. Short-chain xylooligomers, i.e., xylobiose (X2) and xylotriose (X3), were found to bind to the active-site cleft with its reducing end hydrogen-bonded to the catalytic acid-base catalyst Glu132. Xylotetraose (X4) and -pentaose (X5) had apparently been cleaved during the soaking time into a xylotriose plus a monomeric (X4) or dimeric (X5) sugar. While the trimeric hydrolysis product was always found to bind in the same way as xylotriose, the monomer or dimer yielded only weak and diffuse electron density within the xylan-binding cleft, at the opposite side of the active center. This suggests that the two catalytic residues divide the binding cleft into a "substrate recognition area" (from the active site toward the nonreducing end of a bound xylan chain), with strong and specific xylan binding and a "product release area" with considerably weaker and less specific binding. The size of the substrate recognition area (3-4 subsites for sugar rings) explains enzyme kinetic data, according to which short oligomers (X2 and X3) bind to the enzyme without being hydrolyzed.

Binding Sites↗