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At least 343 records · Page 19Linked to original sources
PubMed1994

The nodulation-signaling protein NodO from Rhizobium leguminosarum biovar viciae forms ion channels in membranes.

The secreted nodulation-signaling protein NodO was purified from the supernatant of cultures of Rhizobium leguminosarum biovar viciae. The native protein has a M(r) of approximately 67,000, suggesting that it exists as a dimer since the DNA sequence predicts a M(r) of 30,002. Pure NodO protein had no protease, pectinase, or cellulase activity, and no binding was observed to lipooligosaccharide nodulation factors. Although NodO is relatively hydrophilic, it appeared to insert into liposomes and was protected by liposomes from proteolytic cleavage. When added to planar lipid bilayers, NodO formed cation-selective channels that allowed the movement of monovalent cations (K+ and Na+) across the membrane. NodO is a Ca(2+)-binding protein; in the presence of high concentrations of Ca2+, channel activity was reduced. We hypothesize that NodO plays a role in nodulation signaling by stimulating uptake of nodulation factors or by forming cation-specific channels that function synergistically with the proposed lipooligosaccharide-induced depolarization of the plasma membrane of leguminous plants.

Bacterial Proteins↗
PubMed1997

The filamentous fungus Aspergillus niger contains two "differentially regulated" trehalose-6-phosphate synthase-encoding genes, tpsA and tpsB.

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.

Amino Acid Sequence↗
PubMed2001

Morphine metabolism in the opium poppy and its possible physiological function. Biochemical characterization of the morphine metabolite, bismorphine.

We identified a novel metabolic system of morphine in the opium poppy (Papaver somniferum L.). In response to stress, morphine is quickly metabolized to bismorphine consisting of two morphine units, followed by accumulation in the cell wall. This bismorphine binds predominantly to pectins, which possess high galacturonic acid residue contents, through ionical bonds. Our newly developed method using artificial polysaccharides demonstrated that bismorphine bridges are formed between the two amino groups of bismorphine and the carboxyl groups of galacturonic acid residues, resulting in cross-linking of galacturonic acid-containing polysaccharides to each other. The ability of bismorphine to cross-link pectins is much higher than that of Ca2+, which also acts as a cross-linker of these polysaccharides. Furthermore, we confirmed that cross-linking of pectins through bismorphine bridges leads to resistance against hydrolysis by pectinases. These results indicated that production of bismorphine is a defense response of the opium poppy. Bismorphine formation is catalyzed by anionic peroxidase that pre-exists in the capsules and leaves of opium poppies. The constitutive presence of morphine, together with bismorphine-forming peroxidase, enables the opium poppy to rapidly induce the defense system.

Carbohydrate Sequence↗
PubMed2002

Yeasts associated with fresh and frozen pulps of Brazilian tropical fruits.

The occurrence of yeasts on ripe fruits and frozen pulps of pitanga (Eugenia uniflora L), mangaba (Hancornia speciosa Gom.), umbu (Spondias tuberosa Avr. Cam.), and acerola (Malpighia glaba L) was verified. The incidence of proteolytic, pectinolytic, and mycocinogenic yeasts on these communities was also determined. A total of 480 colonies was isolated and grouped in 405 different strains. These corresponded to 42 ascomycetous and 28 basidiomycetous species. Candida sorbosivorans, Pseudozyma antarctica, C. spandovensis-like, C. spandovensis, Kloeckera apis, C. parapsilosis, Rhodotorula graminis, Kluyveromyces marxianus, Cryptococcus laurentii, Metchnikowia sp (isolated only from pitanga ripe fruits), Issatchenkia occidentalis and C. krusei (isolated only from mangaba frozen pulps), were the most frequent species. The yeast communities from pitanga ripe fruits exhibited the highest frequency of species, followed by communities from acerola ripe fruits and mangaba frozen pulps. Yeast communities from frozen pulp and ripe fruits of umbu had the lowest number of species. Except the yeasts from pitanga, yeast communities from frozen pulp exhibited higher number of yeasts than ripe fruit communities. Mycocinogenic yeasts were found in all of the substrates studied except in communities from umbu ripe fruits and pitanga frozen pulps. Most of the yeasts found to produce mycocins were basidiomycetes and included P. antarctica, Cryptococcus albidus, C. bhutanensis-like, R. graminis and R. mucilaginosa-like from pitanga ripe fruits as well as black yeasts from pitanga and acerola ripe fruits. The umbu frozen pulps community had the highest frequency of proteolytic species. Yeasts able to hydrolyse casein at pH 5.0 represented 38.5% of the species isolated. Thirty-seven percent of yeast isolates were able to hydrolyse casein at pH 7.0. Pectinolytic yeasts were found in all of the communities studied, excepted for those of umbu frozen pulps. The highest frequency of pectinolytic activity was found in mangaba frozen pulp communities. Around 30% of all isolates produced pectinases. The ability to split arbutin was observed in all communities ranging from 8% in yeasts from pitanga frozen pulps to 40.6% in acerola ripe fruit communities. Among 432 species tested, 125 were active for beta-glucosidase production, and Kloeckera apis, P. antarctica, C. sorbosivorans, and C. spandovensis-like were the most active species.

Arbutin↗
PubMed2002

Purification and partial characterization of exopolygalacturonase I from Penicillium frequentans.

A polygalacturonase with a molecular mass of 74 kDa, an isoelectric point around pH 4.2 and pH--and temperature optima of 3.9 and 50 degrees C, respectively, was purified from a culture fluid of Penicillium frequentans. The enzyme was characterized as an exo-alpha-1,4-polygalacturonase (exo-PG I). Km and Vmax for sodium polypectate hydrolysis were 0.68 g/l and 596.8 U x mg(-1), respectively. The enzyme, a glycoprotein with a carbohydrate content of 81%, is probably the main pectinase of Penicillium frequentans responsible for cleaving monomer units from the non-reducing end of pectin.

Brazil↗
PubMed2000

Effect of food enzymes on utilisation of lupin carbohydrates by broilers.

1. The effects of 3 commercial enzyme products on the nutritive value of 2 lupin species were investigated with the emphasis on changes in composition of non-starch polysaccharides (NSPs) along the digestive tract. Enzyme A contained primarily cellulase, beta-glucanase and xylanase activities, enzyme B primarily hemicellulase, pentosanase and xylanase activities, and enzyme C primarily hemicellulase, pectinase and beta-glucanase activities. 2. The enzymes were added to semi-purified diets based on sorghum and casein containing 35% whole seed lupins (Lupinus angustifolius cv Gungurru or Lupinus albus cv Kiev mutant). Control diets contained no lupins. 3. Food conversion ratio (FCR), excreta moisture content and apparent metabolisable energy (AME) were affected by lupin species but not by enzyme supplementation. 4. In diets with L. angustifolius, enzyme C significantly increased digesta viscosity and increased the concentration of soluble NSPs in all sections of the intestine. 5. Digestibility of protein and NSPs in the ileum and microbial fermentation in the ileum and caeca were not affected by adding enzymes to diets containing L. angustifolius. 6. Enzyme addition to diets with L. albus did not affect digesta viscosity nor concentration of soluble NSPs but caused a significantly (P<0.05) reduced concentration of insoluble NSP in the ileum. 7. Enzyme addition to L. albus significantly (P<0.05) increased NSP digestibility in the ileum but had no effects on protein digestibility and fermentation in the ileum and caeca.

Animal Feed↗
PubMed2003

Chemical composition and nutritive value of kapok seed meal for broiler chickens.

1. Mechanically extracted kapok (Ceiba pentandra) seed meal (KSM) contained 324 g crude protein, 97 g ether extract, 289 g fibre, 94 g ash, 128 g available carbohydrates, 3-8 g calcium, 11 g phosphorus, 10.4 g cyclopropenoid fatty acids and 15 g tannins per kg. 2. In a 4 x 2 factorial experiment, KSM was incorporated in broiler starter and finisher feeds at 0, 30, 60 and 90 g/kg, replacing sunflower meal (SFM) w/w; without and with multi-enzyme supplementation. 3. No significant differences were noticed between treatments in body weight gain, feed consumption, feed conversion efficiency, mortality or carcase yields. 4. Multi-enzyme (amylase, endoxylanase, hemicellulase, beta-glucanase, pectinase, phytase and protease) supplementation did not improve the growth performance of broilers fed on the KSM diets.

Animal Feed↗
PubMed2003

Supplementation of diets containing pea meal with exogenous enzymes: effects on weight gain, feed conversion, nutrient digestibility and gross morphology of the gastrointestinal tract of growing broiler chicks.

1. The potential for the nutritional improvement of pea-based diets by supplementation with a cocktail of exogenous carbohydrases was investigated using growing broiler chicks. 2. Pea meals (grown in the UK) were included in wheat-based diets at 300 g/kg as a partial replacement for an approximately isonitrogenous mixture of wheat and soybean meal. A wheat/soybean meal diet served as a control and each diet was supplemented with a cocktail of alpha-amylase, pectinase and cellulase. The diets were fed to 1-d-old broiler chicks for a period of 21 d. Weight gain and feed conversion were monitored weekly and excreta were collected during the final week in order to determine nutrient digestibility coefficients and metabolisable energy. On d 21, the gastrointestinal (GI) tract was excised and gross morphology measured. 3. Inclusion of pea meal reduced weight gain, feed conversion, nutrient digestibility and also increased the relative sizes of the distal sections of the GI tract. 4. Enzyme addition partially ameliorated the detrimental effects of pea meal inclusion although similar improvements were also noted for birds fed on the control diet. 5. It is concluded that the nutritive value of pea-meal-based diets can be improved by the addition of carbohydrases, and that some pea cultivars show considerable potential as vegetable protein sources for broiler chicks.

Animal Feed↗
PubMed1996

Effects of enzyme supplementation on the nutritive value of dehulled lupins.

1. Separate balance experiments were conducted to assess the potential of 2 commercial enzyme supplements to improve the nutritive value of dehulled lupin kernels. One supplement (enzyme A) contained primarily xylanase, pentosanase, hemicellulase activities and the other (enzyme B) primarily beta-glucanase, hemicellulase and pectinase activities. 2. The enzymes were added at 0, 0.25, 0.50, 0.75 and 1.00 g/kg in diets containing (g/kg) lupins 300, sorghum 543, casein 91, celite (as marker) 20, and vitamins and minerals 46. Control diets, with and without enzyme supplementation contained sorghum and casein at 800 and 134 g/kg, respectively, and no lupins. 3. Growth rates and food conversion ratios (FCR) of birds over 7 days were not affected by lupin inclusion or enzyme supplementation. FCR of broilers fed on the sorghum diet was improved by enzyme A but not by enzyme B. 4. Ileal starch digestibilities were slightly lower in birds fed on the lupin control diet (no enzyme) compared to the basal control diet. 5. Enzyme A increased the AME of the lupins from 10.01 MJ/kg DM to 11.65 MJ/kg DM when added at 0.5 g/kg. Higher rates of supplementation did not lead to further increases in AME values. 6. Enzyme A did not improve starch digestion in the diets but insoluble non-starch polysaccharides concentration in the digesta decreased (50.41-42.71 g/g acid insoluble ash marker) with increasing enzyme supplementation, suggesting that the improvement in AME was the result of increased fermentation of fibre in the hindgut. 7. Enzyme B did not affect the AME of lupins nor the ileal digestibility of nutrients, but caused an increase in the concentrations of soluble non-starch polysaccharides in the ileal digesta of chickens (19.21-35.77 mg/ml). This was accompanied by an increase in ileal digesta viscosity (11.4-34.2 m.Pa/s).

Analysis of Variance↗
PubMed2004

Ferulic acid: an antioxidant found naturally in plant cell walls and feruloyl esterases involved in its release and their applications.

Ferulic acid is the most abundant hydroxycinnamic acid in the plant world and maize bran with 3.1% (w/w) ferulic acid is one of the most promising sources of this antioxidant. The dehydrodimers of ferulic acid are important structural components in the plant cell wall and serve to enhance its rigidity and strength. Feruloyl esterases are a subclass of the carboxylic acid esterases that hydrolyze the ester bond between hydroxycinnamic acids and sugars present in plant cell walls and they have been isolated from a wide range of microorganisms, when grown on complex substrates such as cereal brans, sugar beet pulp, pectin and xylan. These enzymes perform a function similar to alkali in the deesterification of plant cell wall and differ in their specificities towards the methyl esters of cinnamic acids and ferulolylated oligosaccharides. They act synergistically with xylanases and pectinases and facilitate the access of hydrolases to the backbone of cell wall polymers. The applications of ferulic acid and feruloyl esterase enzymes are many and varied. Ferulic acid obtained from agricultural byproducts is a potential precursor for the production of natural vanillin, due to the lower production cost.

Antioxidants↗
PubMed2004

The microbiology of cocoa fermentation and its role in chocolate quality.

The first stage of chocolate production consists of a natural, seven-day microbial fermentation of the pectinaceous pulp surrounding beans of the tree Theobroma cacao. There is a microbial succession of a wide range of yeasts, lactic-acid, and acetic-acid bacteria during which high temperatures of up to 50 degrees C and microbial products, such as ethanol, lactic acid, and acetic acid, kill the beans and cause production of flavor precursors. Over-fermentation leads to a rise in bacilli and filamentous fungi that can cause off-flavors. The physiological roles of the predominant micro-organisms are now reasonably well understood and the crucial importance of a well-ordered microbial succession in cocoa aroma has been established. It has been possible to use a synthetic microbial cocktail inoculum of just 5 species, including members of the 3 principal groups, to mimic the natural fermentation process and yield good quality chocolate. Reduction of the amount of pectin by physical or mechanical means can also lead to an improved fermentation in reduced time and the juice can be used as a high-value byproduct. To improve the quality of the processed beans, more research is needed on pectinase production by yeasts, better depulping, fermenter design, and the use of starter cultures.

Acetic Acid↗
PubMedDate not supplied

Pectin/zein beads for potential colon-specific drug delivery: synthesis and in vitro evaluation.

Novel complex hydrogel beads were prepared from two edible polymers: pectin, a carbohydrate from citrus fruits, and zein, a protein from corn. The pectin/zein complex hydrogels did not swell in physiological environments, but hydrolyzed in the presence of pectinases. An in vitro study showed the capacity of the hydrogels to endure protease attack and residence time variation. The physical and biological properties of the new hydrogels were attributed to molecular entanglement of the two polymers. The pectin networks were stabilized by the bound zein molecules. In turn, the pectin networks shielded the bound zein from protease digestion.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed2006

Applications of alginate in bioseparation of proteins.

Alginate is a polysaccharide that is a block polymer consisting of block units of guluronic acid and mannuronic acid. It shows inherent biological affinity for a variety of enzymes such as pectinase, lipase, phospholipase D, a and ss amylases and glucoamylase. Taking advantage of its precipitation with Ca2+ and the above-mentioned property, alginate has been used for purification of these enzymes by affinity precipitation, aqueous two phase separation, macroaffinity ligand facilitated three phase partitioning, immobilized metal affinity chromatography and expanded bed affinity chromatography. Thus, this versatile marine resource has tremendous potential in bioseparation of proteins.

Alginates↗
PubMed2002

Inhibition of hydrolytic enzyme activities and plant pathogen growth by invertase inhibitors.

The invertase inhibitory protein isolated from Cyphomandra betacea Sendt and Solanum tuberosum inhibited the invertase activity from different species, genera and even plant family. Furthermore, proteinaceous inhibitors are not invertase specific; fungal, bacterial and higher plant enzymes including polygalacturonase, pectinase, pectin lyase, alpha-L-arabinofuranosidase and beta-glucosidase are also shown to be inhibited. Both inhibitors exhibited an in vitro antibacterial action against phytopathogenics strains of Xanthomonas campestris pvar vesicatoria CECT 792, Pseudomonas solanacearum CECT 125, Pseudomonas corrugata CECT 124, Pseudomonas syringae and Erwinia carotovora var carotovora.

Enzyme Inhibitors↗
PubMed2002

In situ fluorescent nick translation procedure for plant chromosomes.

Pectinase and cellulase, which are used to macerate plant material, always show traces of DNase activities that result in DNA nicking. Moreover, the DNA polymerase I usually applied in the in situ nick translation techniques shows both 5' to 3' and 3' to 5' exonuclease activities. As a result, significant nonspecific labeling appears in control preparations that are not digested by a restriction endonuclease. Our procedure includes blocking nonspecific nick labeling before incubation with restriction enzymes (HpaII and HaeIII). This is achieved by incorporation of ddGTP into DNA by the Taq polymerase which lacks 3' to 5' exonuclease activity. This method gives satisfactory results because it eliminates nonspecific nick translation signals that are present after applying the methods described for animal material.

Chromosomes↗
PubMed2003

Biodegradation studies of rosin-based polymers.

This study was designed to investigate two rosin-based polymers (R-1 and R-2) for their in vitro and in vivo biodegradation behavior. The in vitro hydrolytic degradation was carried out in buffer solutions of pH 4.4, 7.4, and 10.4 at 37 degrees C. Enzymatic degradation was studied using enzymes lipase, pancreatine, and pectinase. Free films of the two polymers were subcutaneously implanted in rabbits for the in vivo biodegradation. The extent of degradation was determined quantitatively by weight loss and was followed qualitatively by scanning electron microscopy. The extent and the rate of degradation was better in vivo than in vitro. The polymers showed poor enzymatic degradation and a highly pH-dependent hydrolytic degradation.

Animals↗
PubMed1989

RGD-dependent linkage between plant cell wall and plasma membrane: consequences for growth.

Soybean (Glycine max [L.] Merr. cv. Mandarin) root cells (SB-1 cell line) grown in suspension culture containing Glycyl-Arginyl-Glycyl-Aspartyl-Seryl-Proline (GRGDSP) (0.25 mg/ml), a synthetic peptide containing the RGD sequence found in many extracellular matrix adhesive proteins, demonstrated (a) significantly enhanced growth rate, and (b) aberrant cell wall/plasma membrane interactions and organization. Substitution of the Asp (D) by a Glu (E) amino acid in the hexapeptide, or inversion of the RGD sequence to GDR, abolished the morphological and growth effects observed for GRGDSP in plant cells. Immunoblots, which were prepared from beta-octylglucoside extracts of whole soybean cells and protoplasts, probed with polyclonal antibodies raised against human vitronectin receptor (hVNR) complex, demonstrated a single band with an apparent molecular mass of 70-72 kD. Chromatography of beta-octylglucoside extracts of SB-1 cells on a Gly-Arg-Gly-Asp-Ser-Pro-Lys-Sepharose affinity column demonstrated the retention of a single 70-72 kD polypeptide that reacted specifically with anti-hVNR antiserum. In contradistinction, no cross-reactivity was observed with antifibronectin receptor antiserum. Epifluorescence microscopy of whole soybean cells, after moderate treatment with pectinase, demonstrated punctate fluorescent patches at the cell membrane/wall boundary when probed with anti-hVNR and rhodamine-derivatized secondary antibodies. We propose that coordination and control of plant cell division and proper cell wall biosynthesis may be mediated by an RGD-dependent recognition system in which RGD binding protein(s) promote cell membrane-cell wall attachment.

Amino Acid Sequence↗
PubMed1971

Lignofibrils on the external cell wall surface of cultured plant cells.

Small strands and bundles of strands extend from the outside surface of suspension-cultured cells of Daucus, Ipomoea, and Phaseolus into the medium. This fibrous cell coat is present in all samples from various growth stages but appears to increase in quantity in the order Ipomoea < Phaseolus < Daucus. The bundles are often many microns in length and display great variation in frequency, size, and form. Identification of the composition of the strands and bundles as lignin is consistent with the following observations: alkaline nitrobenzene oxidation of the strands to compounds which resemble monomers of wood lignin; resistance of the strands to pronase, trypsin, pectinase, and lipase; strong irreversible adsorption of heavy metals; deposition of silver granules by treatment with silver nitrate-hexamine reagent; extraction of the bundles with aqueous dioxane (Björkman procedure); presence in quantity of a structured form of Klason lignin; and existence of material giving a positive test with the Wiesner reagent. Large individual strands (lignofibrils) from Phaseolus show the form of a flat ribbon with very thin branches at irregular intervals. This form does not vary with preparatory techniques, although its electron opacity does. Intercellular spaces display considerable structure and sometimes contain sheets of fibrillar material merging with both the middle lamella between the cells and the surface bundles facing the medium. These sheets are probably another form of association of the lignofibrils. It is suggested that natural fibrous lignin may be a much commoner component of plant tissue than suspected hitherto.

Cell Fractionation↗