Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Pectinases”

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 73 records · Page 4Linked to original sources

Polygalacturonase from Sitophilus oryzae: possible horizontal transfer of a pectinase gene from fungi to weevils.

Endo-polygalacturonase, one of the group of enzymes known collectively as pectinases, is widely distributed in bacteria, plants and fungi. The enzyme has also been found in several weevil species and a few other insects, such as aphids, but not in Drosophila melanogaster, Anopheles gambiae, or Caenorhabditis elegans or, as far as is known, in any more primitive animal species. What, then, is the genetic origin of the polygalacturonases in weevils? Since some weevil species harbor symbiotic microorganisms, it has been suggested, reasonably, that the symbionts' genomes of both aphids and weevils, rather than the insects' genomes, could encode polygalacturonase. We report here the cloning of a cDNA that encodes endo-polygalacturonase in the rice weevil, Sitophilus oryzae (L.), and investigations based on the cloned cDNA. Our results, which include analysis of genes in antibiotic-treated rice weevils, indicate that the enzyme is, in fact, encoded by the insect genome. Given the apparent absence of the gene in much of the rest of the animal kingdom, it is therefore likely that the rice weevil polygalacturonase gene was incorporated into the weevil's genome by horizontal transfer, possibly from a fungus.

Amino Acid Sequence↗

Comparison of the efficacies of a novel aspergillus niger mycelium with separate and combined effectiveness of phytase, acid phosphatase, and pectinase in dephosphorylation of wheat-based feeds fed to growing broilers.

Efficacies of phytase, phosphorolytic enzymes (phytase + acid phosphatase), an enzymic "cocktail" (phytase + acid phosphatase + pectinase + citric acid), a novel Aspergillus niger (fungal) mycelium (FM), and FM enriched in phytase and antioxidants were investigated in growing broilers (Days 1 to 21) fed wheat-based diets. Broilers were fed the following seven diets at 0.69% Ca: 1) a negative control diet, 0.17% nonphytate P (NPP); 2) Diet 1 + 750 phytase units/kg diet; 3) Diet 1 + 750 phytase units + 3,156 units acid phosphatase/kg diet; 4) Diet 1 + 750 phytase units + 3,156 acid phosphatase units + 1,900 units of pectinase/g diet + 3% citric acid; 5) Diet 1 + 4% FM; 6) Diet 1 + 4% FM + 1,300 phytase units + 2% ascorbic acid and 1% of glucose oxidase; and 7) a positive control diet (Diet 1 + 0.24% NPP from dicalcium phosphate). The dietary treatments were fed to four pen replicates of eight birds each. Prior to feed formulation, mycelium and antioxidants dosages were optimized on Diet 1 by an in vitro technique and an experimental design module of a statistical software package. Phytase addition increased BW gain (BWG), feed intake, and P retention. Subsequent addition of acid phosphatase resulted in further increases in BWG, feed intake, and toe ash and reduced digesta viscosity; however, neither P nor Ca retention were improved. Body weight gain and feed intakes superior to those found in chicks fed Diet 7 were observed in birds receiving the cocktail of enzymes (Diet 4) or FM. Chicken fed Diet 6 had the highest percentage of toe ash and retained 76 and 51% of P and Ca, respectively. Supplementation of wheat-based 0.17% NPP diets with FM increased bursa of Fabricius weights and reduced the intestinal surface covered by Peyer's patches.

6-Phytase↗

Regulatory systems modulating the transcription of the pectinase genes of Erwinia chrysanthemi are conserved in Escherichia coli.

To depolymerize plant pectin, the phytopathogenic enterobacterium Erwinia chrysanthemi produces five isoenzymes of pectate lyases encoded by the five genes pelA, pelB, pelC, pelD and pelE. In Er. chrysanthemi, all genes involved in pectin degradation are specifically controlled by the KdgR repressor and are induced in the presence of a pectin catabolic product, 2-keto-3-deoxygluconate (KDG). transcription of the pectinase genes is dependent on many environmental conditions. Transcriptional fusions present on low-copy-number plasmids were used to study the regulation of the pel genes in a heterologous host, Escherichia coli. Some physiological regulations that take place in Er. chrysanthemi are conserved in E. coli. The five pel fusions in E. coli are affected by growth phase, catabolite repression and anaerobic growth conditions and are induced in the presence of galacturonate, a sugar whose catabolism leads to the formation of KDG, the inducer of pel transcription in Er. chrysanthemi. Expression of pelE increased with the osmolarity of the culture medium. In contrast, the regulation of pel expression by temperature or nitrogen starvation, observed in Er. chrysanthemi, was not conserved in E. coli, suggesting that the mechanisms responsible for these regulations are specific to Er. chrysanthemi. Analysis of different E. coli mutants allowed some regulators affecting the transcription of the pel genes to be identified. In E. coli, the growth-phase regulation of the pel genes is not dependent on the RpoS sigma factor and the fnr gene is not involved in the increase of pel expression in oxygen-limited conditions. The gene hns, involved in the regulation of numerous genes, appears to affect pel expression but the effects of E. coli hns mutations are not related to osmoregulation. In contrast, this analysis clearly demonstrates the interchangeability of two regulatory systems of E. coli and Er. chrysanthemi: the global control exerted by the catabolite activator protein CAP and the specific regulation mediated by the KdgR repressor.

Anaerobiosis↗

Changes in cellulase and pectinase activities in fruit tissues and separation zones of citrus treated with cycloheximide.

Cellulase activity increased in separation-zone tissues 1 day after "Valencia" orange (Citrus sinensis [L.] Osbeck) was treated with 20 micrograms per milliliter cycloheximide. Exocellulase was detected only in the separation zones of treated fruit, whereas endocellulase was present in zones from both treated and control fruit. Endocellulase activity in separation-zone tissue of treated fruit was nearly three times as great as that in control tissues. Cellulase activity was restricted to separation-zone tissue. Pectinase and an albedo-macerating factor activity were very low and were not influenced by the treatment. The cycloheximide effect in these experiments was apparently caused by ethylene produced by wound tissue.

Journal Article↗

pecS: a locus controlling pectinase, cellulase and blue pigment production in Erwinia chrysanthemi.

Erwinia chrysanthemi mutants (designated as pecS) displaying derepressed pectate lyase and cellulase synthesis were isolated. In addition, the pecS mutation is responsible for production of an extracellular insoluble blue pigment whose synthesis is cryptic in the wild-type 3937 strain. Transduction analysis indicates that the phenotype is due to a single mutation located near the xyl marker on the strain 3937 chromosome. This mutation was complemented by an R-prime plasmid carrying the xyl and argG genes of E. chrysanthemi, suggesting that the pecS product acts in trans to modulate pectinase, cellulase and blue pigment production. Insertion mutagenesis of the cloned region and recombination of the corresponding mutations in the bacterial chromosome by marker exchange revealed the existence of two divergently transcribed genes, pecS and pecM, that are both involved in the pectate lyase and cellulase regulation. The nucleotide sequences of pecS and pecM were determined. The pecS gene encodes a 166 amino acid polypeptide that shows similarity to the MprA regulatory protein of Escherichia coli whereas the pecM gene encodes a 297 amino acid polypeptide that was shown to be an integral membrane protein. The possible functions of the PecS and PecM proteins derived from the mutant phenotype and sequence analysis are discussed in terms of signal transduction and transcription regulation.

Amino Acid Sequence↗

Pectinase Activity of Anaerobic and Facultatively Anaerobic Bacteria Associated with Soft Rot of Yam (Diascorea rotundata).

Anaerobic and facultatively anaerobic bacteria associated with soft rot of yam (Diascorea rotundata) were isolated by the looping-out method and found to consist of Clostridium (three isolates), Corynebacterium (three isolates), Vibrio (one isolate), and Bacillus lentus (one isolate). Enzyme assay for hydrolase, lyase, and pectinesterase activities by the cup-plate method showed that except for Vibrio sp., B. lentus, and two isolates of Corynebacterium no pectinase activity could be detected for organisms cultured on pectin medium. Most of the cultures on yam tissue, however, showed activities for the three enzymes. The viscometric assay for hydrolase and lyase enzymes indicated a significant level of hydrolase activity (a 40.90% decrease in viscosity for Vibrio sp. and Corynebacterium spp.), but no lyase activity for most of the isolates. Two isolates of Corynebacterium and B. lentus caused changes in fresh yams suggestive of soft rot.

Journal Article↗

Cloning and expression in Escherichia coli of pectinase genes of Erwinia carotovora subsp. carotovora.

Genes coding for an endo-pectate lyase, an exo-pectate lyase, and an endopolygalacturonase of Erwinia carotovora subsp. carotovora Ecc71 were cloned in Escherichia coli HB101, using the cosmid pHC79. The products of the cloned pectinase genes paralleled their counterparts in strain Ecc71 in isoelectric mobility, mode of substrate degradation, and ability to macerate potato tuber tissue.

Cloning, Molecular↗

kdgREcc negatively regulates genes for pectinases, cellulase, protease, HarpinEcc, and a global RNA regulator in Erwinia carotovora subsp. carotovora.

Erwinia carotovora subsp. carotovora produces extracellular pectate lyase (Pel), polygalacturonase (Peh), cellulase (Cel), and protease (Prt). The concerted actions of these enzymes largely determine the virulence of this plant-pathogenic bacterium. E. carotovora subsp. carotovora also produces HarpinEcc, the elicitor of the hypersensitive reaction. We document here that KdgREcc (Kdg, 2-keto-3-deoxygluconate; KdgR, general repressor of genes involved in pectin and galacturonate catabolism), a homolog of the E. chrysanthemi repressor, KdgREch and the Escherichia coli repressor, KdgREco, negatively controls not only the pectinases, Pel and Peh, but also Cel, Prt, and HarpinEcc production in E. carotovora subsp. carotovora. The levels of pel-1, peh-1, celV, and hrpNEcc transcripts are markedly affected by KdgREcc. The KdgREcc- mutant is more virulent than the KdgREcc+ parent. Thus, our data for the first time establish a global regulatory role for KdgREcc in E. carotovora subsp. carotovora. Another novel observation is the negative effect of KdgREcc on the transcription of rsmB (previously aepH), which specifies an RNA regulator controlling exoenzyme and HarpinEcc production. The levels of rsmB RNA are higher in the KdgREcc- mutant than in the KdgREcc+ parent. Moreover, by DNase I protection assays we determined that purified KdgREcc protected three 25-bp regions within the transcriptional unit of rsmB. Alignment of the protected sequences revealed the 21-mer consensus sequence of the KdgREcc-binding site as 5'-G/AA/TA/TGAAA[N6]TTTCAG/TG/TA-3'. Two such KdgREcc-binding sites occur in rsmB DNA in a close proximity to each other within nucleotides +79 and +139 and the third KdgREcc-binding site within nucleotides +207 and +231. Analysis of lacZ transcriptional fusions shows that the KdgR-binding sites negatively affect the expression of rsmB. KdgREcc also binds the operator DNAs of pel-1 and peh-1 genes and represses expression of a pel1-lacZ and a peh1-lacZ transcriptional fusions. We conclude that KdgREcc affects extracellular enzyme production by two ways: (i) directly, by inhibiting the transcription of exoenzyme genes; and (ii) indirectly, by preventing the production of a global RNA regulator. Our findings support the idea that KdgREcc affects transcription by promoter occlusion, i.e., preventing the initiation of transcription, and by a roadblock mechanism, i.e., by affecting the elongation of transcription.

Amino Acid Sequence↗

[Production of pectinases by Penicillium simplicissimum A3263 in an amaranth-seed flour medium].

The present work studies the production of pectinases using a strain of Penicillium simplicissimum A3263 and considering the influence of adding Amaranthus cruentus seed meal in a selected medium. We also considered the influence of aeration on enzyme production. Research was oriented towards the production of pectin lyase, the enzyme having the highest commercial value. This work was carried out in Erlenmeyer flasks in rotary shaker to select the medium and in a mechanically stirred fermentor to study aeration. The microorganism was developed as pellets of 1 mm diameter. Enzyme levels were of the order of 8216.21 pectin lyase units and 167.57 polygalacturonase units per gram of fungal biomass, respectively, using a medium containing 40 g/l of amaranth seed meal. As for the influence of aeration, it was determined that the higher values were obtained at 750 rpm corresponding to an oxygen absorption rate of 2691 ml O2/lh for an air flow of 1 l/l.min. The results obtained are considered very important in view of the fact that they exceeded in 550% those obtained by other authors.

Aerobiosis↗

[Studies of the chronic toxicity of mold pectinase].

Studied was the action of mould pectinase produced in this country, using Asp. niger. Rats and guinea pigs were subject to the testing through continuous oral treatment. The preparation was mixed with feed at 0.1 and 0.2 ratio percents (1000 and 2000 ppm)--with the rats for a period of 90 days, and with the guinea pigs--in the course of 180 days. It was found that the enzyme preparation stimulates the growth (more strongly expressed in rats), showing no adverse effect on the appetite and behaviour as well as on the tested clinical and biochemical indices of the blood and urine and the structure and development of viscera and the reproductive capacity of the test animals. The positive effect on the growth of animals is explained by the better utilization of the nutrient components of the ration, and the harmlessness of the preparation--by its weak resorption in the digestive tract (resorption index 13) and the fact that it is not able to attack animal tissues having no pectin substances.

Administration, Oral↗

[Allergic pathology due to enzymes: a case report of asthma due to pectinase (polygalacturonase)].

The paper described the case of a worker handling pectinase, an enzyme used in the manufacture of food additives. This enzyme is a carbohydrase (polygalacturonase) obtained by extraction, and consists of three active phases. After little more than a year at the job, the subject complained of bouts of asthma, especially when she had to weigh the enzyme in dust form. Symptoms appeared even when this operation was performed by others near her work station. An allergological examination revealed skin hypersensitivity to the enzyme, without atopy. The methods of preparing the tests performed are described. The authors stress the need to perform similar investigations on other exposed workers in order to assess the allergogenic potential of the substance and adjust environmental preventive measures accordingly.

Adult↗

Molecular cloning of Erwinia chrysanthemi pectinase and cellulase structural genes.

Erwinia chrysanthemi 3937 secretes four major pectate lyase isoenzymes (PL, EC 4.2.2.2) and one endocellulase (Cx, EC 3.2.1.4). A genomic library of this strain was constructed in the Lambda L47-1 vector, and screened for the presence of PL and Cx on pectate and caboxymethylcellulose agar. Among the seven Cx-positive phage clones, three were shown to encode an enzyme of the same mol. wt. as the one found in the culture supernatant of strain 3937. The 34 PL-positive phage clones were analyzed by electrofocusing and could, according to the PL they produced, be arranged in five classes. Phages from three classes produced three different single PL, named PLb, c and d. No common fragment was evidenced between the inserts of the phages of these three classes. This demonstrated that, in strain 3937, PLb, C, and d were encoded by three different genes called pelB, C, and D. Furthermore, our results suggest the existence of two additional genes encoding PLa and e. In addition, a pectin methylesterase gene was found closely linked to pelD.

Journal Article↗

Inducing properties of analogs of 2-keto-3-deoxygluconate on the expression of pectinase genes of Erwinia chrysanthemi.

In Erwinia chrysanthemi, all the genes involved in pectin degradation are controlled by the negative regulatory gene kdgR. 2-keto-3-deoxy-gluconate (KDG) is the inducing molecule that interacts with KdgR to allow the expression of all the genes of the kdg regulon. The inducing properties on the expression of genes regulated by kdgR of various analogs and derivatives of KDG were tested. All the inducers share the common moiety COOH-CO-CH2-CHOH-C-C included in a pyranic cycle. Our results show that esterification of C1 prevents induction. Presence of a ketone function on C2 and absence of hydroxyl on C3 are necessary for induction. The nature and the configuration of substituent on C5 has no influence on induction. Two compounds have interesting properties: 5-O-methyl-KDG is a gratuitous inducer, and gluconic acid can prevent induction.

Erwinia↗