Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.2.1.15 (pectinase)
2,440 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Pectinases produced by Aspergillus flavus and A. parasiticus are believed to play a significant role in the ability of these fungi to spread in cotton bolls and other crops. Utilizing a DNA probe, generated by PCR, of the Aspergillus niger pgaII gene, we have isolated a novel, constitutively expressed polygalacturonase (PG)-encoding gene (pecA) from an A. parasiticus cDNA library. DNA sequence analysis and the deduced amino acid (aa) sequence of pecA demonstrated significant identity at the nucleotide and aa levels with other PG of fungal origin. Northern blot analysis of RNA isolated from A. parasiticus grown on either glucose or pectin as the sole carbon source showed that pecA was expressed during growth in both media.
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PMID:Cloning and characterization of a novel polygalacturonase-encoding gene from Aspergillus parasiticus. 788 76

Bacteroides thetaiotaomicron strain 217 can use pectins as a sole carbon source. Preliminary characterization of the pectinolytic enzymes revealed three complementary activities in this strain: a pectin methylesterase (PME), a pectate lyase (PL) and a polygalacturonase (PG), which were all inducible by pectin or polygalacturonate. Use of the lambdoid phage replacement vector lambda EMBL3 allowed a 13.2 kb insert mediating both PL and PME activities to be isolated. Subcloning of two EcoRI fragments in pBR325 led to the separate isolation of the pel and pme genes. They were expressed constitutively in Escherichia coli HB101, as proved by the activities observed even in mineral medium supplemented only with glucose. In addition, the pme gene was expressed in both orientations. These results suggest that each gene represents an individual transcriptional unit. Several properties of the cloned PL were different from those of the original strain: it was mainly associated to the outer membrane, its optimum pH was higher, and its stability at 50 degrees C was lost but partially preserved by CaCl2. In addition, the apparent specific PL activity in the E. coli membrane fraction was about 30-fold higher. On the other hand, most of the properties of the cloned PME were similar to those of the original. Despite an enhanced thermostability, the apparent specific activity of the cloned PME was about 6-fold lower, and was independent of the insert orientation.
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PMID:Molecular cloning and expression in Escherichia coli of genes encoding pectate lyase and pectin methylesterase activities from Bacteroides thetaiotaomicron. 802 7

The endo-alpha-(1-->4)-polygalacturonase-resistant fractions (PG-1, PG-2, and PG-3) from an antiulcer pectin (Bupleuran 2IIc), isolated from the roots of Bupleurum falcatum L., were further analysed by lithium degradation. The results indicated that PG-1 contained a small proportion of long, branched arabinosyl chains and a large proportion of short, neutral oligosaccharide chains. GLC-MS analysis showed that, after methylation the short, neutral oligosaccharide fraction consisted of at least 22 kinds of di- to tetra-saccharide alditols, such as Gal-(1-->4)-Rha-ol (a major component), Ara-(1-->4)-Rha-ol, Glc-(1-->4)-Rha-ol, Ara-->Ara-->Ara-ol, and Ara-->Ara-->Ara-->Ara-ol (minor components) in addition to heteroglycosyl alditols. After deesterification, PG-2 and PG-3 were digested with endo-alpha-(1-->4)-polygalacturonase again, and the enzyme-resistant intermediate size fraction (PG-2') was purified. Component sugar analysis indicated that PG-2' contained 2-Me-Fuc, 2-Me-Xyl, apiose (Api), aceric acid (AceA), 3-deoxy-D-lyxo-heptulosaric acid (Dha), and 3-deoxy-D-manno-2-octulosonic acid (Kdo) in addition to Rha, Fuc, Ara, Xyl, Man, Gal, Glc, GalA, and GlcA. Lithium degradation of PG-2' gave mainly a pentosyl-->6-deoxyhexosyl-->6-deoxyhexosyl-->pentitol fragment, with some neutral di- and tri-saccharide alditols, including a pentosyl-->deoxyhexitol. Methylation analysis of these degradation products indicated that they contained terminal Rha, Araf, Fuc, Xyl, and Gal, 4-linked Rha, 3-linked Fuc, 3-linked Ara, and 3'-linked Api. Bupleuran 2IIc was eluted as essentially a single peak on gel filtration on Sepharose CL-6B. The neutral sugar content of the successive fractions increased with increasing molecular weight, but each fraction also contained, in addition to Rha, Ara, and Gal, 2-Me-Fuc, 2-Me-Xyl, and Api.
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PMID:Structural studies of endopolygalacturonase-resistant fragments of an antiulcer pectin from the roots of Bupleurum falcatum L. 814 69

Production of polygalacturonases and pectinases from Sclerotinia sclerotiorum was induced in vitro by galacturonic acid. The inductive effect of galacturonic acid was abolished by the presence of glucose, leading to a basal enzyme production. Zymograms of extracellular enzymes showed that galacturonic acid induced the synthesis of six polygalacturonase and one pectin-methylesterase isoforms. Immunoblotting revealed that an exo-polygalacturonase and an exo-polymethylgalacturonase were secreted in all conditions. They are not glucose repressed and not regulated by galacturonic acid. These constitutive enzymes provide the pathogen with the inherent ability to release galacturonic acid from plant cell walls and to trigger inducible enzyme synthesis.
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PMID:Regulation by Galacturonic Acid of Pectinolytic Enzyme Production by Sclerotinia sclerotiorum 866 89

Orange peel, an abundant byproduct of the citrus processing industry, is converted to a mixture of glucose, galacturonic acid, fructose, arabinose, galactose, and xylose by hydrolysis with mixed pectinase and cellulase enzymes. All these sugars can be fermented to ethanol or ethanol and acetic acid by the recombinant bacterium Escherichia coli KO11. The fermentation efficiency is improved by the addition of yeast extract, tryptone, mixed amino acids, corn steep liquor, or by proteolytic digestion of endogenous proteins. Batch fermentations of supplemented peel hydrolysate containing 111 g/L of initial total sugars produced 35-38 g/L of ethanol in 48-72 h and a 75-85% yield.
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PMID:Fermentation of orange peel hydrolysates by ethanologenic Escherichia coli. Effects of nutritional supplements. 866 5

Northern-blot analysis of RNA isolated from Sclerotinia sclerotiorum grown on either glucose or polygalacturonate as the sole carbon source showed that pg1, encoding a neutral polygalacturonase, was not expressed during growth in both media. In contrast, transcripts of this gene were detected during infection of sunflower germlings. Analysis of the promoter sequence revealed a number of cis-acting sequences known to regulate the expression of many fungal promoters. Protein-DNA-binding experiments showed that proteins extracted from mycelia grown on polygalacturonate or glucose interacted with different regions of the promoter. The GST-CREA fusion protein, containing the two zinc fingers of the Aspergillus nidulans repressor CREA involved in carbon catabolite repression, forms several complexes with DNA fragments carrying the consensus 5'-SYGGRG-3'. These results suggest that a CREA homolog may be involved in the regulation of pg1.
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PMID:Expression of the Sclerotinia sclerotiorum polygalacturonase pg1 gene: possible involvement of CREA in glucose catabolite repression. 875 53

Two genes encoding trehalose-6-phosphate synthase were cloned from Aspergillus niger. tpsA was cloned using the Saccharomyces cerevisiae GGS1/TPS1 gene as a probe. It encodes a 517-amino acid polypeptide with 64-70% similarity to trehalose-6-phosphate synthase of S. cerevisiae, Kluyveromyces lactis, and Schizosaccharomyces pombe. Its transcription occurs constitutively and is enhanced on carbon-derepressing carbon sources, coinciding with the presence of a CreA-binding nucleotide motif in the 5'-noncoding region of tpsA. Disruption of tpsA only weakly reduces growth on glucose, and neither influences the glucose induction of a low affinity glucose permease nor interferes with the catabolite repression of a pectinase; it causes reduced the heat tolerance of conidia. tpsB was cloned by a polymerase chain reaction-based strategy. Its 480 amino acid sequence showed 76.5% identity to tpsA. Its transcription was hardly detectable at ambient temperatures but was enhanced strongly upon heat shock, which agrees with the presence of several copies of a C4T stress-responsive element in its 5'-upstream sequences. Hence the function of yeast GGS1/TPS1 has been split into two differentially regulated genes in A. niger, of which none appears to be involved in glucose sensing.
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PMID:The filamentous fungus Aspergillus niger contains two "differentially regulated" trehalose-6-phosphate synthase-encoding genes, tpsA and tpsB. 900 11

Following the previous isolation of CLPG1, a gene encoding an endopolygalacturonase (endoPG) secreted into the culture filtrate of Colletotrichum lindemuthianum, we have isolated and sequenced an additional endoPG gene, CLPG2. This gene is present as a single copy in the genome of the fungus. At the amino acid level, CLPG2 shows 61% identity to CLPG1 and between 37 to 59% identity to other fungal endoPGs. RNA blot analyses of endoPG gene expression were followed with specific probes during in vitro culture of the fungus. When conidia were used to inoculate a synthetic medium containing pectin as sole carbon source, only CLPG1 was found to be expressed after 3 days of culture. However, transferring the mycelium grown on glucose for 4 days to a pectin-containing medium allowed the detection of CLPG1 and CLPG2 transcripts as early as 12 h after transfer on this substrate. Expression of CLPG2 was transient while that of CLPG1 was more prolonged. Immunocytological localization of endoPG in C. lindemuthianum-infected bean tissues with antibodies against CLPG1 confirmed that the protein is produced in planta and is associated with extensive degradation of the host cell wall. Detection of endoPG transcripts by reverse transcription-polymerase chain reaction revealed that CLPG1, but not CLPG2, is expressed at the beginning of the necrotrophic stage of infection. These results show that the two endoPG genes are differentially expressed and that CLPG1 encodes the major secreted endoPG both during saprophytic growth and during plant infection.
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PMID:Endopolygalacturonase genes from Colletotrichum lindemuthianum: cloning of CLPG2 and comparison of its expression to that of CLPG1 during saprophytic and parasitic growth of the fungus. 924 38

Genetic control of polygalacturonase (PG) activity from Fusarium oxysporum f.sp. radicis lycopersici was analyzed on pectin and glucose cultures. One exopolygalacturonase from F. oxysporum f.sp. radicis lycopersici was strongly induced, in stationary culture, when the fungus was grown on apple pectin, while on glucose no extracellular PG activity could be detected. Although SDS-PAGE detected the presence of a putative PG band (66 kDa) in both conditions, specific antibodies obtained against the purified PG only detected it in PG-inducing conditions, that is to say, when apple pectin was used as the carbon source. Northern blot analysis of RNA of two isolates of F. oxysporum f.sp. radicis lycopersici (r6 and r2) confirmed that this regulation of PG synthesis was exerted at the transcriptional level. Only one single mRNA species of around 1400 nucleotides was detected on the cultures containing pectin and was absent in glucose-grown cultures. Southern blot analysis of genomic DNA indicated that pg gene seems to be present in a single copy in the genomes of F. oxysporum f.sp. radicis lycopersici r6 and r2 and Fusarium oxysporum f.sp. lycopersici, showing similar hybridization patterns in all species. The partial sequence of this pg gene from F. oxysporum f.sp. radicis lycopersici r6, which is also reported, showed high similarity to diverse PGs already reported. Exopolygalacturonase of F. oxysporum f.sp. radicis lycopersici r6 is heavily glycosylated; its deglycosylated form had a molecular mass of 50 kDa.
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PMID:Control of polygalacturonase synthesis in Fusarium oxysporum f.sp. radicis lycopersici. 943 11

pg1 encoding the major in vitro extracellular endopolygalacturonase of the tomato vascular wilt pathogen Fusarium oxysporum f. sp. lycopersici was cloned and sequenced. The deduced mature protein had a calculated molecular mass of 35.5 kDa and a pI of 6.2, and showed significant similarity with other fungal endoPGs. pg1 mRNA was induced in vitro by citrus pectin, tomato vascular tissue, 0.1% D-galacturonic acid, and polygalacturonic acid, and repressed by 1% D-galacturonic acid and 1% glucose. Reverse transcription-polymerase chain reaction revealed pg1 expression in roots and lower stems of tomato plants infected by F. oxysporum f. sp. lycopersici. Three naturally occurring F. oxysporum f. sp. melonis isolates deficient in PG1 were transformed with the cloned gene. The PG1 enzyme secreted by the transformants had the same molecular mass, pI, and glycosylation pattern as those of the donor isolate. Polygalacturonase activity in cultures of transformants grown in vitro on citrus pectin and on melon plants, but not on glucose, increased 10- to 20-fold, compared with the PG1-deficient wild-type isolate, whereas mycelial dry weight increased two- to three-fold. Transformants exhibited the same degree of virulence toward susceptible muskmelon cultivars as the wild-type isolate and were avirulent on a resistant cultivar.
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PMID:Cloning, expression, and role in pathogenicity of pg1 encoding the major extracellular endopolygalacturonase of the vascular wilt pathogen Fusarium oxysporum. 945 Mar 33


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