Polygalacturonase treatment affects carotenoid absorption from veggie juice.

The present study was conducted to investigate the effects of polygalacturonase (PG) treatment on carotenoid absorption upon digestion of HPH-treated combined peach and carrot juice (CJ) with or without the presence of lipids. Results showed that PG treatment reduced median particle diameter (D50) and viscosity of CJ, and increased total carotenoid bioaccessibility by 41%. In the presence of emulsion, the bioaccessibility of carotenoids was higher and it was not significantly affected by PG treatment. Xanthophylls (lutein and zeaxanthin) had higher bioaccessibility than the more lipophilic carotenes (ß-carotene and α-carotene); also, uptake in Caco-2 cells and transport of lutein and zeaxanthin were higher than for ß-carotene and α-carotene. Individual carotenoids bioaccessibility was negatively correlated with their transport. All together data showed digestion and absorption processes were two independent processes: factors improving carotenoid bioaccessibility did not necessarily affect their bioavailability.

The exogenous application of AtPGLR, an endo-polygalacturonase, triggers pollen tube burst and repair.

Plant cell wall remodeling plays a key role in the control of cell elongation and differentiation. In particular, fine-tuning of the degree of methylesterification of pectins was previously reported to control developmental processes as diverse as pollen germination, pollen tube elongation, emergence of primordia or elongation of dark-grown hypocotyls. However, how pectin degradation can modulate plant development has remained elusive. Here we report the characterization of a polygalacturonase (PG), AtPGLR, the gene for which is highly expressed at the onset of lateral root emergence in Arabidopsis. Due to gene compensation mechanisms, mutant approaches failed to determine the involvement of AtPGLR in plant growth. To overcome this issue, AtPGLR has been expressed heterologously in the yeast Pichia pastoris and biochemically characterized. We showed that AtPGLR is an endo-PG that preferentially releases non-methylesterified oligogalacturonides with a short degree of polymerization (< 8) at acidic pH. The application of the purified recombinant protein on Amaryllis pollen tubes, an excellent model for studying cell wall remodeling at acidic pH, induced abnormal pollen tubes or cytoplasmic leakage in the subapical dome of the pollen tube tip, where non-methylesterified pectin epitopes are detected. Those leaks could either be repaired by new ß-glucan deposits (mostly callose) in the cell wall or promoted dramatic burst of the pollen tube. Our work presents the full biochemical characterization of an Arabidopsis PG and highlights the importance of pectin integrity in pollen tube elongation.

Fermentation and addition of enzymes to a diet based on high-moisture corn, rapeseed cake, and peas improve digestibility of nonstarch polysaccharides, crude protein, and phosphorus in pigs.

Fluctuating prices of cereals have led to an interest in alternative ingredients for feed. The aim of this study was to evaluate the effect of fermentation and the addition of nonstarch polysaccharide (NSP)-degrading enzymes on the ileal and total tract digestibility of nutrients of a diet based on locally grown crops. Four diets were fed including a nonfermented liquid standard grower diet (Control) and 3 experimental diets based on high-moisture corn, rapeseed cake, and peas fed as nonfermented liquid feed (nFLF), fermented liquid feed (FLF), or FLF supplemented with an enzyme mixture of ß-glucanase + xylanase + pectinase (FLF+Enz). The FLF was prepared by mixing feed and water (1:2.5, wt/wt) and, once daily, replacing 50% of the mixture with an equal amount of fresh feed and water. The diets were fed to 8 ileal cannulated barrows in a double Latin square design. Ileal digesta and feces were collected after an adaption period of 10 d. Results showed microbiologically good-quality fermented diets. The levels of Enterobacteriaceae were 5.1 to 5.4 log cfu/g in FLF and FLF+Enz vs. 6.3 log cfu/g in nFLF in the ileum and 5.1 to 5.2 log cfu/g in FLF and FLF+Enz vs. 6.3 log cfu/g in nFLF in the feces. Apparent total tract digestibility (ATTD) of CP was increased by fermentation (73.2% in FLF vs. 69.0% in nFLF; P = 0.033), and digestibility of P showed a tendency (P = 0.073) toward an increase. Addition of the enzyme mixture resulted in a pronounced reduction of dietary NSP compared with FLF (12.8% total NSP in FLF+Enz vs. 15.9% total NSP in FLF; P< 0.001), which also led to increased apparent ileal digestibility (AID) of total and insoluble NSP (total NSP, 31.1% in FLF+Enz vs. 13.6% in FLF; P = 0.002). The Control did not, in general, show higher digestibility values than the experimental diet. However, in the cases were it did, fermentation and enzyme addition brought the digestibility to the level of the Control. In conclusion, fermentation increased the ATTD of CP and the AID of P, with the same tendency (P ≤ 0.07) for the ATTD. Addition of NSP-degrading enzymes resulted in a pronounced reduction in the concentration of NSP in the feed along with increased AID of NSP. Hence, the experimental diet seems to be a possible alternative to a traditional diet for pigs.

Effects of drought-affected corn and nonstarch polysaccharide enzyme inclusion on nursery pig growth performance.

The effectiveness of carbohydrase enzymes has been inconsistent in corn-based swine diets; however, the increased substrate of nonstarch polysaccharides in drought-affected corn may provide an economic model for enzyme inclusion, but this has not been evaluated. A total of 360 barrows (PIC 1050 × 337, initially 5.85 kg BW) were used to determine the effects of drought-affected corn inclusion with or without supplementation of commercial carbohydrases on growth performance and nutrient digestibility of nursery pigs. Initially, 34 corn samples were collected to find representatives of normal and drought-affected corn. The lot selected to represent the normal corn had a test weight of 719.4 kg/m3, 15.0% moisture, and 4.2% xylan. The lot selected to represent drought-affected corn had a test weight of 698.8 kg/m3, 14.3% moisture, and 4.7% xylan. After a 10-d acclimation period postweaning, nursery pigs were randomly allotted to 1 of 8 dietary treatments in a completely randomized design. Treatments were arranged in a 2 × 4 factorial with main effects of corn (normal vs. drought affected) and enzyme inclusion (none vs. 100 mg/kg Enzyme A vs. 250 mg/kg Enzyme B vs. 100 mg/kg Enzyme A + 250 mg/kg Enzyme B). Both enzymes were included blends of ß-glucanase, cellulose, and xylanase (Enzyme A) or hemicellulase and pectinases (Enzyme B). Pigs were fed treatment diets from d 10 to 35 postweaning in 2 phases. Feed and fecal samples were collected on d 30 postweaning to determine apparent total tract digestibility of nutrients. The nutrient concentrations of normal and drought-affected corn were similar, which resulted in few treatment or main effects differences of corn type or enzyme inclusion. No interactions were observed (P > 0.10) between corn source and enzyme inclusion. Overall (d 10 to 35), treatments had no effect on ADG or ADFI, but enzyme A inclusion tended to improve (P < 0.10; 0.74 vs. 0.69) G:F, which was primarily driven by the improved feed efficiency (0.76 vs. 0.72; P < 0.05) of pigs fed Enzyme A in Phase 2 (d 10 to 25 postweaning) and was likely a result of improved xylan utilization. In conclusion, drought stress did not alter the nonstarch polysaccharide concentration of corn beyond xylan concentration, so it was not surprising that enzyme inclusion showed little benefit to nursery pig growth performance. However, improved feed efficiency of pigs fed diets containing Enzyme A from d 10 to 25 postweaning warrants further investigation

Aeromonas punctata derived depolymerase that disrupts the integrity of Klebsiella pneumoniae capsule: optimization of depolymerase production.

Formation of dense, highly hydrated biofilm structures pose a risk for public and environmental health. Extracellular polymeric substances encompassing biofilms offer 1000-fold greater resistance as compared to the planktonic cells. Using enzymes as anti-biofouling agents, will improve penetration of antimicrobials and increase susceptibility of biofilms to components of immune system. The challenge of using enzymes derived from unrelated bacteria for the degradation of capsular matrix of Klebsiella pneumoniae has not been dealt in the past. Thus, statistical optimization was done to enhance depolymerase production by Aeromonas punctata, directed against the exopolysaccharide matrix of Klebsiella pneumoniae B5055, capable of substituting the available phage borne depolymerase enzyme. Optimization via central composite design (CCD) resulted in 16-fold enhancement in depolymerase yield (166.65 µmoles ml(-1) min(-1) ) over unoptimized medium. Out of the 19 variables, media composition giving maximum expression levels of the enzyme consisted of 1 mg ml(-1) galactose and ammonium chloride, 1.5 mg ml(-1) each of capsular polysaccharide (CPS) and magnesium sulfate. Tryptic peptide analysis of the purified 29 kDa band by Matrix assisted laser desorption ionization-time of flight (MALDI-TOF) showed a high homology with a protein of unknown function from Aeromonas cavaie Ae398. Further improvements in the enzyme can lead to its successful development as prophylactic and/or a therapeutic agent.

Albariño wine aroma enhancement through the use of a recombinant polygalacturonase from Kluyveromyces marxianus.

The possible biotechnological application of a recombinant endopolygalacturonase of Kluyveromyces marxianus (KMPG) for the aroma enhancement of Albariño wine was studied. The addition of this enzyme to the must gives rise to a significant increase of the total compounds responsible for the aroma as opposed to the effect when using a commercial pectic enzyme. This increase also results in a significant rise of the odoriferous aglycones which are direct determinants of the aroma. Wines made by using the KMPG enzyme are characterised by a greater richness and diversity with regard to the number of aromatic compounds present, clearly differing from those obtained with a commercial pectic preparation. Based on compounds with odour activity values (OAV)>1, the wines obtained with the enzyme KMPG are richer in citric, balsamic, spicy and above all floral (violet and rose) aromas than untreated wines or wines supplemented with a commercial enzyme.

Immobilization of pectinase on oxidized pulp fiber and its application in whitewater treatment.

Modified pulp fiber was originally used as a new type of carrier for pectinase immobilization. Pulp fiber was oxidized by sodium periodate to produce aldehyde groups for covalently binding with amino groups of pectinase. Results showed that the enzymatic activity of immobilized pectinase on pulp fiber reached 65 µgg(-1)min(-1) when immobilization pH value, temperature and time were of 7.0, 20 °C and 15 min, respectively. The immobilized pectinase showed higher thermo stability in a wider temperature range of 40-70 °C than its free type and its optimal pH shifted from 8.0 to 8.8. Furthermore, the immobilized pectinase exhibited good operational stability. When employed in whitewater treatment of papermaking industry, it still efficiently decreased the cationic demand after operating repeatedly for six batches. The results obtained demonstrate a promising route to prepare available, cheap and biodegradable carrier for immobilizing enzymes with potential application in wastewater treatment in papermaking industry.

[Growth inhibition effect of immobilized pectinase on Microcystis aeruginosa].

To confirm the growth inhibition effect of immobilized pectinase on algae, co-cultivation method was used to investigate the effect of immobilized pectinase on the growth of Microcystis aeruginosa. After co-cultivation, the damage status of the algae was observed through electron microscope, and the effect of immobilized pectase on the physiological and biochemical characteristics of the algae was also measured. The results showed that the algae and immobilized pectase co-cultivated solution etiolated distinctly on the third day and there was a significantly positive correlation between the extent of etiolation and the dosage as well as the treating time of the immobilized pectinase. Under electron microscope, plasmolysis was found in the slightly damaged cells, and the cell surface of these cells was rough, uneven and irregular; the severely damaged cells were collapsed or disintegrated completely. The algal yield and the chlorophyll a content decreased significantly with the increase of the treating time. The measurement of the malondiadehyde (MDA) value showed that the antioxidation system of the treated algal cells was destroyed, and their membrane lipid was severely peroxidated. The study indicated that the immobilized pectinase could efficiently inhibit the growth of M. aeruginosa, and the inhibitory rate reached up to 96%.

Studies on calcium release and H2O2 level produced by the elicitor induced plant cell by fluorescence probing.

Using fluorescence probing technology, we studied the mechanism and interrelations of calcium release and H(2)O(2) production in situ in living tissues of tobacco and cotton plants which were induced by pathogen elicitor, salicylic acid (SA) and pectinase respectively. Results showed that (1) pathogen elicitors could induced H(2)O(2) response in epidermis cells regardless of environmental calcium, but in mesophyll protoplast, H(2)O(2) response could only be induced at calcium condition. Similarly, SA and pectinase induced H(2)O(2) response could only be observed at calcium condition; (2) pathogen elicitors could induce calcium response in both epidermis cells and protoplasts regardless of environmental calcium, while calcium response couldn't be induced at non-calcium condition by SA and pectinase; (3) H(2)O(2) response and calcium response in protoplast were faster than that in the whole cell. These results indicated that pathogen elicitors can induce the release of cell wall calcium and the cell wall calcium release is independent to pectinase. And it is concluded that free calcium influx is necessary for the oxidative burst and cell wall calcium has an irreplaceable role in defense signal transduction.

Molecular and biochemical basis for stress-induced accumulation of free and bound p-coumaraldehyde in cucumber.

To elucidate the genetic and biochemical regulation of elicitor-induced p-coumaraldehyde accumulation in plants, we undertook a multifaceted approach to characterize the metabolic flux through the phenylpropanoid pathway via the characterization and chemical analysis of the metabolites in the p-coumaryl, coniferyl, and sinapyl alcohol branches of this pathway. Here, we report the identification and characterization of four cinnamyl alcohol dehydrogenases (CADs) from cucumber (Cucumis sativus) with low activity toward p-coumaraldehyde yet exhibiting significant activity toward other phenylpropanoid hydroxycinnamaldehydes. As part of this analysis, we identified and characterized the activity of a hydroxycinnamoyl-coenzyme A:shikimate hydroxycinnamoyl transferase (HCT) capable of utilizing shikimate and p-coumaroyl-coenzyme A to generate p-coumaroyl shikimate. Following pectinase treatment of cucumber, we observed the rapid accumulation of p-coumaraldehyde, likely the result of low aldehyde reductase activity (i.e. alcohol dehydrogenase in the reverse reaction) of CsCAD enzymes on p-coumaraldehyde. In parallel, we noted a concomitant reduction in the activity of CsHCT. Taken together, our findings support the hypothesis that the up-regulation of the phenylpropanoid pathway upon abiotic stress greatly enhances the overall p-coumaryl alcohol branch of the pathway. The data presented here point to a role for CsHCT (as well as, presumably, p-coumarate 3-hydroxylase) as a control point in the regulation of the coniferyl and sinapyl alcohol branches of this pathway. This mechanism represents a potentially evolutionarily conserved process to efficiently and quickly respond to biotic and abiotic stresses in cucurbit plants, resulting in the rapid lignification of affected tissues.

Nutraceutical composition of Zizyphus mauritiana Lamk (Indian ber): effect of enzyme-assisted processing.

Zizyphus (Indian ber) is an excellent source of several phenolic compounds. The effect of two cell wall degrading enzymes, namely pectinase and viscozyme, on the nutraceutical composition of Zizyphus juice was investigated in the present study. Enzyme assisted processing significantly (P < 0.05) improved the juice yield, total soluble solids, total phenolics and total antioxidant activity (AOX). There was significant increase in recovery of antioxidants, to the tune of 70.51%, 66%, and 45% respectively in ascorbic acid, total phenolics and total flavonoids through viscozyme. The in-vitro total AOX of juice extracted via enzyme-assisted processing was 20.9 and 15.59 µmol Trolox/ml in ferric-reducing antioxidant power and cupric-reducing antioxidant capacity assays, respectively. There was 41% increase in AOX of juice extracted with enzyme over straight pressed juice. Results indicate that enzyme-assisted processing can significantly improve the functional properties of the Zizyphus juice.

Means to improve the nutritive value of flaxseed for broiler chickens: the effect of particle size, enzyme addition, and feed pelleting.

Three experiments were conducted to explore dietary means (particle size, enzyme addition, bile salts addition, and feed pelleting) of minimizing the antinutritive effects of cell wall-nonstarch polysaccharides of flaxseed. Broiler chickens were fed corn-soybean meal-based diets containing 15% of full-fat flaxseed from 5 to 18 d. The effects of 2 enzyme preparations containing viscosity-reducing or cell wall-degrading activities on growth performance and nutrient digestibility were evaluated in experiment 1. Enzyme addition had beneficial effects (P<0.05) in increasing nonstarch polysaccharide digestibility and reducing intestinal viscosity. However, no differences in growth performance or fat digestibility were observed between the enzyme types. Therefore, the enzyme supplement containing both viscosity-reducing and cell wall-degrading activities was used in subsequent studies. A 2x2x2 factorial arrangement was used in experiment 2 to investigate the effects of particle size (coarse vs. fine), enzyme supplementation, and bile salt addition on the nutritive value of flaxseed for broiler chickens. In experiment 3, a 4x2 factorial arrangement was used to further investigate the effects of feed processing (whole seed, coarsely ground seed, and finely ground seed in pelleted diets or finely ground seed in mash diets) and enzyme addition on growth performance and fat utilization. Bile salt addition did not improve fat digestibility. Particle size reduction via grinding had no significant effect on growth performance no matter if present in the mash or pelleted diets. When compared with grinding, feed pelleting showed more pronounced and beneficial effects on growth performance particularly when whole, intact seeds were used, indicating a potential for using whole flaxseed in the pelleted diets. Enzyme addition resulted in an increase in total tract fat digestibility by 3 to 6%, which was reflected in an improved feed conversion ratio by 1 to 3%, regardless of the processing method used (P<0.05). In conclusion, enzyme addition and feed pelleting offer practical solutions to improve the nutritive value of flaxseed for broiler chickens.

Composite microparticle drug delivery systems based on chitosan, alginate and pectin with improved pH-sensitive drug release property.

Composite microparticle drug delivery systems based on chitosan, alginate and pectin with improved pH sensitivity were developed for oral delivery of protein drugs, using bovine serum albumin (BSA) as a model drug. The composite drug-loaded microparticles with a mean particle size less than 200mum were prepared by a convenient shredding method. Since the microparticles were formed by tripolyphosphate cross-linking, electrostatic complexation by alginate and/or pectin, as well as ionotropic gelation with calcium ions, the microparticles exhibited an improved pH-sensitive drug release property. The in vitro drug release behaviors of the microparticles were studied in simulated gastric (pH 1.2 and pH 5.0), intestinal (pH 7.4) and colonic (pH 6.0 and pH 6.8 with enzyme) media. For the composite microparticles with suitable compositions, the releases of BSA at pH 1.2 and pH 5.0 could be effectively sustained, while the releases at pH 7.4, pH 6.8 and pH 6.0 increased significantly, especially in the presence of pectinase. These results clearly suggested that the microparticles had potential for site-specific protein drug delivery through oral administration.

Restoration of mature etiolated cucumber hypocotyl cell wall susceptibility to expansin by pretreatment with fungal pectinases and EGTA in vitro.

Mature plant cell walls lose their ability to expand and become unresponsive to expansin. This phenomenon is believed to be due to cross-linking of hemicellulose, pectin, or phenolic groups in the wall. By screening various hydrolytic enzymes, we found that pretreatment of nongrowing, heat-inactivated, basal cucumber (Cucumis sativus) hypocotyls with pectin lyase (Pel1) from Aspergillus japonicus could restore reconstituted exogenous expansin-induced extension in mature cell walls in vitro. Recombinant pectate lyase A (PelA) and polygalacturonase (PG) from Aspergillus spp. exhibited similar capacity to Pel1. Pel1, PelA, and PG also enhanced the reconstituted expansin-induced extension of the apical (elongating) segments of cucumber hypocotyls. However, the effective concentrations of PelA and PG for enhancing the reconstituted expansin-induced extension were greater in the apical segments than in the basal segments, whereas Pel1 behaved in the opposite manner. These data are consistent with distribution of more methyl-esterified pectin in cell walls of the apical segments and less esterified pectin in the basal segments. Associated with the degree of esterification of pectin, more calcium was found in cell walls of basal segments compared to apical segments. Pretreatment of the calcium chelator EGTA could also restore mature cell walls' susceptibility to expansin by removing calcium from mature cell walls. Because recombinant pectinases do not hydrolyze other wall polysaccharides, and endoglucanase, xylanase, and protease cannot restore the mature wall's extensibility, we can conclude that the pectin network, especially calcium-pectate bridges, may be the primary factor that determines cucumber hypocotyl mature cell walls' unresponsiveness to expansin.

Enzyme-assisted processing increases antimicrobial and antioxidant activity of bilberry.

The effects of nine cell wall-degrading enzymes on the antimicrobial and antioxidant activities of bilberry were studied. Antimicrobial activity was measured using the human pathogens Salmonella enterica sv. Typhimurium and Staphylococcus aureus as test strains. Enzyme treatments liberated phenolics from the cell wall matrix, which clearly increased the antimicrobial activity of berry juices, press cakes, and berry mashes on the basis of plate counts. Antibacterial effects were stronger against Salmonella than against Staphylococcus bacteria. In general, the increase in activity measured as colony-forming units per milliliter was 3-5 logarithmic units against Salmonella and 1-2 units against Staphylococcus bacteria. Increase in antimicrobial activity was observed only in acidic conditions, which is also the natural environment in various berry products, such as juices. The activity profile of the pectinase preparation affected the chemistry of the phenolics due to the presence of deglycosylating activities in some preparations. The difference in phenolic profiles was reflected in the antimicrobial effects. Bilberry mashes treated with Pectinex Ultra SP-L, Pectinex 3 XL, and Pectinex BE XXL were most efficient against Salmonella bacteria, whereas mashes treated with Pectinex Smash, Pectinex BE 3-L, and Biopectinase CCM showed the strongest antimicrobial activity against Staphylococcus bacteria. Due to the liberation of phenolics from the cell wall matrix the antioxidant activity measured as radical scavenging activity was also increased on average about 30% by the enzymatic treatments. The highest increase in phenolic compounds was about 40%. Highest increases in anthocyanins and in antioxidant activity were observed in berry mash treated with Pectinex Smash XXL enzyme, and the lowest increase was observed after treatment with Pectinex BE 3-L. Enzyme-assisted processing is traditionally used to improve berry and fruit juice yields. However, enzymatic treatments also have an impact on the functional properties of the products. The increased liberation of phenolics from the cell wall matrix can prolong the shelf life of berry products by limiting the growth of contaminants during processing or storage. The increased amount of phenolic compounds may also have a positive effect on gut well-being.

Antimicrobial activity of flavonoids extracted from bergamot (Citrus bergamia Risso) peel, a byproduct of the essential oil industry.

AIMS: To evaluate the antimicrobial properties of flavonoid-rich fractions derived from bergamot peel, a byproduct from the Citrus fruit processing industry and the influence of enzymatic deglycosylation on their activity against different bacteria and yeast. METHODS AND RESULTS: Bergamot ethanolic fractions were tested against Gram-negative bacteria (Escherichia coli, Pseudomonas putida, Salmonella enterica), Gram-positive bacteria (Listeria innocua, Bacillus subtilis, Staphylococcus aureus, Lactococcus lactis) and the yeast Saccharomyces cerevisiae. Bergamot fractions were found to be active against all the Gram-negative bacteria tested, and their antimicrobial potency increased after enzymatic deglycosylation. The minimum inhibitory concentrations of the fractions and the pure flavonoids, neohesperidin, hesperetin (aglycone), neoeriocitrin, eriodictyol (aglycone), naringin and naringenin (aglycone), were found to be in the range 200 to 800 microg ml(-1). The interactions between three bergamot flavonoids were also evaluated. CONCLUSION: The enzyme preparation Pectinase 62L efficiently converted common glycosides into their aglycones from bergamot extracts, and this deglycosylation increased the antimicrobial potency of Citrus flavonoids. Pairwise combinations of eriodictyol, naringenin and hesperetin showed both synergistic and indifferent interactions that were dependent on the test indicator organism. SIGNIFICANCE AND IMPACT OF THE STUDY: Bergamot peel is a potential source of natural antimicrobials that are active against Gram-negative bacteria.

A mixture of pure cellulase, hemicellulase and pectinase improves broiler performance.

1. The effects of a mixture of pure enzymes (cellulase, hemicellulase and pectinase) and a commercial enzyme, Energex, were examined on performance and metabolisabilities in broiler chicks given a maize-soybean meal diet. Composition of the mixed enzyme was similar to Energex except that protease was not present. 2. Chicks were divided into three groups: control, mixed enzyme and Energex with 7 replicates per group. Male broiler chicks were raised at 25 degrees C in wire-floored cages for 12 d from 15 d of age. Feed and water were offered ad libitum. 3. The Energex group gained significantly more weight and the mixed enzyme group tended to gain more than the control. Feed intakes were similar and thus the feed conversion ratio of Energex was significantly improved while it tended to be improved by the mixed enzyme. 4. The mixed enzyme group showed significant improvement in carcase and muscle weight when compared with the control group. The mixed enzyme group also showed significant improvement in organic matter and crude protein metabolisabilities. In the groups given enzyme, abdominal fat weight tended to decrease. 5. It is concluded that a combination of cellulase, hemicellulase and pectinase is effective in improving organic matter and crude protein metabolisabilities and carcase yield of broilers on a maize-soybean meal diet.

Membrane-wall attachments in plasmolysed plant cells.

Field emission scanning electron microscopy of plasmolysed Tradescantia virginiana leaf epidermal cells gave novel insights into the three-dimensional architecture of Hechtian strands, Hechtian reticulum, and the inner surface of the cell wall without the need for extraction. At high magnification, we observed fibres that pin the plasma membrane to the cell wall after plasmolysis. Treatment with cellulase caused these connecting fibres to be lost and the pinned out plasma membrane of the Hechtian reticulum to disintegrate into vesicles with diameters of 100-250 nm. This suggests that the fibres may be cellulose. After 4 h of plasmolysis, a fibrous meshwork that labelled with anti-callose antibodies was observed within the space between the plasmolysed protoplast and the cell wall by field emission scanning electron microscopy. Interestingly, macerase-pectinase treatment resulted in the loss of this meshwork, suggesting that it was stabilised by pectins. We suggest that cellulose microfibrils extending from strands of the Hechtian reticulum and entwining into the cell wall matrix act as anchors for the plasma membrane as it moves away from the wall during plasmolysis.

Pectinase production by Aspergillus niger using wastewater in solid state fermentation for eliciting plant disease resistance.

An elicitor of plant disease resistance, pectinase, was produced by solid state fermentation with Aspergillus niger. Sugar beet pulp was used as carbon source and the wastewater from monosodium glutamate production was used as nitrogen and water source. The composition of the fermentation medium was: 11 ml concentrated wastewater (containing NH3-N 38.2 mg/ml), sugar beet pulp 10 g, Na2HPO4.12H2O 0.2 g, KH2PO4 0.04 g in a 500 ml Erlenmeyer flask. The fermentation temperature was 30 degrees C and the relative humidity of the air was 75-90%. The maximum production of pectinase was reached after 96 h cultivation. The crude pectinase extracted from the fermented materials could elicit disease resistance in cucumber and tomato seedlings.

[Changes in lectin activity in plants treated with resistance inducers]. / Izmenenie aktivnosti lektinov v rasteniiakh, obrabotannykh induktorami ustoichivosti.

We studied the changes in lectin activity in tobacco leaf disc and potato tubers treated with polysaccharides (chitosan, glucomannan, and dextran sulfate), enzymes (cellulase and pectinase), or monosaccharides (glucose and glucosamine). All studied substances changed lectin activity to a certain extent(significantly or as a trend). The number of membrane lectins in the chloroplasts (tobacco leaf discs) usually considerably decreased immediately after the treatment (1-2 days) but increased later (2-4 days). Generally, an increased lectin activity predominated in potato tubers treated with the inducers. The enzymes increased lectin activity during the whole observation period (5 days). A pronounced antiviral activity was observed in the hypersensitive tobacco-tobacco mosaic virus system only after treatment with chitosan and glucomannan.