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Pectin/ethylcellulose film coating formulations for colonic drug delivery.

PURPOSE: The purpose of the study was to investigate the potential of pectin, ethylcellulose combinations as a practical film coating for colonic delivery. METHODS: Combinations of pectin and ethylcellulose, in the form of an aqueous dispersion, were used as coating formulations. Paracetamol cores were used as the substrate. The coatings were assessed by a flow through dissolution system simulating in vivo conditions by changes in pH and residence time. Pectinolytic enzymes were used to simulate the bacterial flora of the colon. RESULTS: Drug release was controlled by the ratio of ethylcellulose to pectin in the film coat. Increasing the proportion of ethylcellulose and increasing the coat weight reduced drug release in pH1 and pH7.4 media. The addition of pectinolytic enzymes to pH6 media increased the release of drug. CONCLUSIONS: Combinations of ethylcellulose and pectin can provide protection to a drug in the upper g.i. tract while allowing enzymatic breakdown and drug release in the colon.

Cellulose↗

Calculation of gross energy in pet foods: new data on heat combustion and fibre analysis in a selection of foods for dogs and cats.

Seven pectin samples, six galactomannan sources, five carrageen samples, four alginate samples, one sample of gum traganth, agar agar and gum arabicum, two xanthan samples, two inulin samples and a galacto oligosaccharide, 22 cellulose samples, six lignin samples, four starch samples, nine protein samples, six isolated fats, three meat samples, two lung samples, two samples of skimmed milk powder, 12 prepared complete dry dog foods, 21 moist dog foods, nine dry and 25 moist cat foods and 10 faecal samples were analysed for heat combustion (adiabatic bomb calorimetry), crude nutrients, acid detergent fibre (ADF), and acid detergent lignin (ADL). Some of the non-starch polysaccharides which gave low levels of crude fibre and ADF were also analysed for total, insoluble and soluble fibre. The heat combustion of cellulose ranged between 17.0 and 17.5 kJ/g organic matter (OM). The variation was somewhat larger for other non-starch polysaccharides (pectin, galactomannan sources, carageen, alginate, gums, xanthan, inulin) where heat combustion ranged between 14.0 and 18.2 kJ/g OM. The heat combustion of lignin averaged 25.5 kJ/g OM with considerable variation (17.0-29.2 kJ/g OM). Starch had a narrow range (17.2-17.3 kJ/g OM). Heat combustion of protein samples varied between 22.0 and 24.6 kJ/g, and of fat samples varied between 38.0 and 39.6 kJ/g OM. When cellulose was analysed for crude fibre only between 62 and 85% OM was detected. ADF analyses of cellulose ranged between 75 and 93% OM. The crude fibre content of all other non-starch polysaccharides did not exceed 13% OM, with the exception of pectins (ADF 0.7-37% OM) and alginates (ADF 39-66% OM), the ADF content was also below 13% in these samples. In contrast the total fibre content was above 80% OM in all non-starch noncellulose polysaccharides and the percentage of soluble fibre was high (25-93% OM). Unprocessed lignin gave high readings for crude fibre (39-61% OM) and ADF (96-99% OM), while processed lignin had low crude fibre content (< 1% OM) and low ADF content (< 32%). ADL determined unprocessed lignin (78-91% OM), but again processed lignin was analysed incompletely (< 29%). Pectin and alginate gave false positive ADL readings of up to 31% OM, while all other non-starch polysaccharides were not determined by ADL. When gross energy was calculated with the factors (kJ/g) 24 for protein, 38 for fat and 17 for all carbohydrate, including fibre, there was a good correlation between calculated gross energy and heat combustion in 67 pet foods as well as in meat, lung and skimmed milk powder. In contrast to this the same factors underestimated heat combustion of faeces by around 8%.

Animal Feed↗

Hypoglycaemic effect of Coccinia indica: mechanism of action.

The hypoglycaemic effect of orally administered extracts of leaves and roots of Coccinia indica has been reported earlier. The oral administration of the pectin isolated from the fruit of the above plant at a dose of 200 mg/100 g BW/day showed a significant hypoglycaemic action in normal rats. Pectins isolated from many other plants have also been reported to have hypoglycaemic actions. The pectin administration resulted in a significant reduction in blood glucose and an increase in the liver glycogen. Glycogen synthetase activity was highly significant. Incorporation of labelled glucose into hepatic glycogen was also found to be higher. A significant reduction in phosphorylase activity was noted in the pectin-administered groups.

Animals↗

Wheat bran increases high-density-lipoprotein cholesterol in the rat.

1. Five groups of male Sprague Dawley rats were given for 6 months a diet with high protein and fat contents but a very low dietary fibre content (group B), and this diet mixed with (g/kg) 50 low-methoxyl pectin (group L), 50 high-methoxyl pectin (group H), 50 guar gum (group G) and 200 wheat bran (group WB, corresponding to 100 wheat fibre) respectively. 2. The weight increment was significantly lower in group G than in the other groups. Assuming no energy value of the dietary fibre, the weight increment (/kJ) was the same in groups B, L and H, lower in group G and higher in group WB, indicating that a proportion of the bran fibre might in fact be available as a source of energy. 3. Wheat bran increased total plasma cholesterol and high-density-lipoprotein cholesterol after 6, 12 and 26 weeks. Group G had significantly lowered plasma cholesterol after 12 weeks. Pectin on the other hand did not significantly influence total or HDL-cholesterol levels. It is therefore possible that the plasma cholesterol lowering effect of pectin previously demonstrated in the rat is dependent on the presence of significant amounts of dietary cholesterol as our diets did not contain added cholesterol. Plasma triglycerides decreased with age but were similar in all groups.

Animals↗

Effect of different sources of dietary fibre on growth performance, intestinal morphology and caecal carbohydrases of domestic geese.

1. The aim of this study was to examine the effects of dietary fibre sources on growth and on the development of the gastro-intestinal tract in growing geese. 2. Six-week-old female White Roman geese were divided at random into 6 groups with 6 dietary treatments. Diets were isoenergetic and isonitrogenous and contained alfalfa meal, barley bran, rice hulls, cellulose, pectin or lignin as the major dietary source of fibre. 3. Food intake was significantly higher in the barley bran group than in the other groups. Both daily weight gain and food conversion of the pectin and alfalfa meal groups were lower than those of the other groups. 4. In the 9-week-old geese, the relative weight and the length of the small intestine did not significantly differ among treatment groups. However, the relative weight was significantly lightest and the caecal length was significantly shortest (P < 0.05) in the pectin group. 5. Activities of amylase and cellulose hydrolases of the caecal contents were also highest in the pectin group. 6. From SEM micrographs, no morphological damage of the villi was observable in the different intestinal segments of the geese in any of the treatments.

Amylases↗

Effect of preheating on potato texture.

Preheating potatoes at 50 to 80 degrees C has a firming effect on the cooked potato tissue. This effect is particularly pronounced at a preheating temperature of 60 to 70 degrees C followed by cooling. Several theories have been presented in the literature to explain this firming effect: retrogradation of starch, leaching of amylose, stabilization of the middle lamellae and cell walls by the activation of the pectin methylesterase (PME) enzyme, and by the release of calcium from gelatinized starch and the formation of calcium bridges between pectin molecules. Most probably, none of these theories alone can explain the phenomenon and more than one mechanism seems to be involved. Some of these mechanisms seem to be interdependent. As an example, calcium could be considered as a link all the way through release after starch gelatinization to cross-linking pectin substances in the cell wall and the middle lamellae, which has been demethylated by the PME enzyme. More research and "clear cut" experiments are needed in order to elucidate the role of each mechanism, especially which of them is the main contributor to the process of firming. Most probably, the calcium-pectin-PME mechanism plays a secondary role, that is, it only retards the collapse of the tissue structure that would otherwise occur during the final heating without preheating, and it is not the main factor of firmness.

Carboxylic Ester Hydrolases↗

Dietary fiber and progression of atherosclerosis: the Los Angeles Atherosclerosis Study.

BACKGROUND: Several epidemiologic studies found weak protective relations between dietary fiber intake and the risk of cardiovascular disease events. However, few of the studies addressed possible mechanisms of the effect. OBJECTIVE: In the present study, we estimated relations between the progression of atherosclerosis and the intake of selective dietary fiber fractions. Mediation of the relations by serum lipids was also investigated. DESIGN: Participants who were free of heart disease and aged 40-60 y were recruited into the cohort (n = 573; 47% women). The intima-media thickness (IMT) of the common carotid arteries was measured ultrasonographically at the baseline examination and at 2 follow-up examinations (n = 500), dietary intakes were assessed with six 24-h recalls (3 at baseline and 3 at the first follow-up examination), and blood samples were analyzed at baseline and at both follow-up examinations. RESULTS: A significant inverse association was observed between IMT progression and the intakes of viscous fiber (P = 0.05) and pectin (P = 0.01). Correction for measurement error increased the magnitude of these estimated effects. The ratio of total to HDL cholesterol was inversely related to the intakes of total fiber (P = 0.01), viscous fiber (P = 0.05), and pectin (P = 0.01). The magnitude of the association between IMT progression and the intakes of viscous fiber and pectin was attenuated by adjustment for serum lipids. CONCLUSIONS: The intake of viscous fiber, especially pectin, appears to protect against IMT progression. Serum lipids may act as a mediator between dietary fiber intake and IMT progression.

Adult↗

Fermentable dietary fibers elevate urinary methylmalonate and decrease propionate oxidation in rats deprived of vitamin B-12.

This study examines the effect of dietary fiber supplements of different degrees of bacterial fermentability on biochemical indicators of vitamin B-12 deficiency in rats. Groups of rats were fed a fiber-free diet deficient in vitamin B-12 or the fiber-free diet diluted with 5% of a poorly fermentable dietary fiber (cellulose, lignin or alginic acid) or a highly fermentable fiber (pectin, guar gum or xylan). Poorly fermentable fibers had no significant effect on apparent B-12 status, whereas the highly fermentable fibers significantly increased urinary methylmalonic acid and depressed oxidation of [14C]propionate to 14CO2. Pectin consistently induced significantly greater effects than did xylan or guar gum. The data are consistent with the hypothesis that fermentable fibers stimulate bacterial propionate production and exaggerate certain biochemical indicators of B-12 deficiency. Since pectin had a more pronounced effect than did other fermentable fibers, the possibility that pectin may also interfere with B-12 absorption requires further study.

Animals↗

The effects of dietary fiber feeding on cholesterol metabolism in rats.

The flux through the sterol biosynthetic pathway was studied in hepatocytes isolated from male Sprague-Dawley rats fed diets containing one of four fiber sources: cellulose, pectin, oat bran and wheat bran. Sterol synthesis measured by the incorporation of tritiated water or [2-14C]mevalonic acid was not inhibited in hepatocytes isolated from animals fed diets containing cellulose, pectin, oat bran or wheat bran when compared to animals fed a fiber-free diet. Based on these results, it is concluded that the intake of fiber has no inhibitory effect on endogenous sterol synthesis. In fact, in comparison to that in fiber-free controls, sterol synthesis was markedly elevated in pectin- and wheat bran-fed animals. In the case of the pectin-treated animals, the higher synthetic rate corresponded to an increase in 3-hydroxy-3-methylglutaryl coenzyme A reductase activity.

Animals↗

The fermentation of different dietary fibers is associated with fecal clostridia levels in men.

Only a few reports have compared the fermentation of pectin and cellulose using the hydrogen-breath test, and no studies have examined the relation between the hydrogen breathing pattern and colonic microflora. Using breath-hydrogen measurements, we investigated whether different dietary fibers (DFs) were fermented differently and whether there were individual differences after ingestion of the same DF; we also examined the relation between individual fecal microflora and the fermentation of DF. Results of hydrogen tests in 14 men were compared after they had ingested 20 g of pectin, 20 g of cellulose, or 6 g of lactulose (a DF-like substance). We examined the relation between the breath hydrogen results and the subjects' fecal microflora. We defined significant fermentation (i.e., positive cases) as a continuous rise in hydrogen in the expiratory air of >19 ppm. The subjects were divided into 3 groups according to their hydrogen breath test pattern, i.e., positive for lactulose and pectin (Group LP, n = 4); positive for lactulose alone (Group L, n = 7); and negative for pectin, cellulose, and lactulose (Group N, n = 3). Individual differences were noted in subjects from Group LP and Group L. The detection frequency of lecithinase-negative clostridia was higher in Group LP than in the other groups (P < 0.05), and the detection frequency and the number of lecithinase-positive clostridia were higher in Groups LP and L than in Group N (P < 0.05). These findings suggest that the Clostridium species are associated with hydrogen production. The hydrogen breath test results of DFs depend on both the type of DF and the individual colonic microflora. The amount and constitution of colonic microflora might be predicted by the hydrogen-breath test using different DFs.

Adult↗

Effect of extracellular calcium, pH and borate on growth oscillations in Lilium formosanum pollen tubes.

Calcium ions (Ca(2+)), protons (H(+)), and borate (B(OH)(4)(-)) are essential ions in the control of tip growth of pollen tubes. All three ions may interact with pectins, a major component of the expanding pollen tube cell wall. Ca(2+ )is thought to bind acidic residues, and cross-link adjacent pectin chains, thereby strengthening the cell wall. Protons are loosening agents; in pollen tube walls they may act through the enzyme pectin methylesterase (PME), and either reduce demethylation or stimulate hydrolysis of pectin. Finally, borate cross-links monomers of rhamnogalacturonan II (RG-II), and thus stiffens the cell wall. It is demonstrated here that changing the extracellular concentrations of Ca(2+), H(+) and borate affect not only the average growth rate of lily pollen tubes, but also influence the period of growth rate oscillations. The most dramatic effects are observed with increasing concentrations of Ca(2+) and borate, both of which markedly reduce the rate of growth of oscillating pollen tubes. Protons are less active, except at pH 7.0 where growth is inhibited. It is noteworthy, especially with borate, that the faster growing tubes exhibit the shorter periods of oscillation. The results are consistent with the idea that binding of Ca(2+) and borate to the cell wall may act at a similar level to alter the mechanical properties of the apical cell wall, with optimal concentrations being high enough to impart sufficient rigidity to the wall so as to prevent bursting in the face of cell turgor, but low enough to allow the wall to stretch quickly during periods of accelerating growth.

Borates↗

Localization of aluminium in the maize root apex: can morin detect cell wall-bound aluminium?

Morin is a fluorochrome which forms a fluorescent complex with aluminium (Al) and is thus used to localize Al in plant tissues. However, reports about the cellular distribution of Al-apoplastic versus symplastic-based on morin staining are often conflicting. The objective of this work was to investigate whether Al localization with morin staining can show the proper cellular distribution of Al. Fresh root cross-sections were made from root apices of maize (cv. Lixis) treated with 25 muM Al for 6 h and stained with morin. Fluorescence microscopic investigation showed Al-morin fluorescence in the cytosol, but not in the cell wall. This is in contrast to the growing evidence which shows that Al mainly accumulates in the cell wall, especially bound to the pectin matrix. Therefore, in vitro analyses were carried out to study whether morin can form a fluorescent complex with Al, which is bound to pectin, cell wall, and other Al-binding ligands such as phosphate, galacturonate, DNA, and ATP. Compared with the control treatment without Al-binding ligands, fluorescence intensity was reduced by about 10-fold in the presence of pectin and isolated cell walls, but fairly unaffected in the presence of phosphate and galacturonate. Al associated with DNA and ATP also formed a fluorescent complex with morin. This implies that, although Al is mainly accumulated in the cell wall, it cannot be detected with morin as it is tightly bound to cell-wall pectin. Thus, morin staining should not be used to study the distribution of Al between cell compartments.

Aluminum↗

Comparative genomics of the KdgR regulon in Erwinia chrysanthemi 3937 and other gamma-proteobacteria.

In the plant-pathogenic enterobacterium Erwinia chrysanthemi, almost all known genes involved in pectin catabolism are controlled by the transcriptional regulator KdgR. In this study, the comparative genomics approach was used to analyse the KdgR regulon in completely sequenced genomes of eight enterobacteria, including Erw. chrysanthemi, and two Vibrio species. Application of a signal recognition procedure complemented by operon structure and protein sequence analysis allowed identification of new candidate genes of the KdgR regulon. Most of these genes were found to be controlled by the cAMP-receptor protein, a global regulator of catabolic genes. At the next step, regulation of these genes in Erw. chrysanthemi was experimentally verified using in vivo transcriptional fusions and an attempt was made to clarify the functional role of the predicted genes in pectin catabolism. Interestingly, it was found that the KdgR protein, previously known as a repressor, positively regulates expression of two new members of the regulon, phosphoenolpyruvate synthase gene ppsA and an adjacent gene, ydiA, of unknown function. Other predicted regulon members, namely chmX, dhfX, gntB, pykF, spiX, sotA, tpfX, yeeO and yjgK, were found to be subject to classical negative regulation by KdgR. Possible roles of newly identified members of the Erw. chrysanthemi KdgR regulon, chmX, dhfX, gntDBMNAC, spiX, tpfX, ydiA, yeeO, ygjV and yjgK, in pectin catabolism are discussed. Finally, complete reconstruction of the KdgR regulons in various gamma-proteobacteria yielded a metabolic map reflecting a globally conserved pathway for the catabolism of pectin and its derivatives with variability in transport and enzymic capabilities among species. In particular, possible non-orthologous substitutes of isomerase KduI and a new oligogalacturonide transporter in the Vibrio species were detected.

Artificial Gene Fusion↗

Overexpression of polygalacturonase in transgenic apple trees leads to a range of novel phenotypes involving changes in cell adhesion.

Polygalacturonases (PGs) cleave runs of unesterified GalUA that form homogalacturonan regions along the backbone of pectin. Homogalacturonan-rich pectin is commonly found in the middle lamella region of the wall where two adjacent cells abut and its integrity is important for cell adhesion. Transgenic apple (Malus domestica Borkh. cv Royal Gala) trees were produced that contained additional copies of a fruit-specific apple PG gene under a constitutive promoter. In contrast to previous studies in transgenic tobacco (Nicotiana tabacum) where PG overexpression had no effect on the plant (K.W. Osteryoung, K. Toenjes, B. Hall, V. Winkler, A.B. Bennett [1990] Plant Cell 2: 1239-1248), PG overexpression in transgenic apple led to a range of novel phenotypes. These phenotypes included silvery colored leaves and premature leaf shedding due to reduced cell adhesion in leaf abscission zones. Mature leaves had malformed and malfunctioning stomata that perturbed water relations and contributed to a brittle leaf phenotype. Chemical and ultrastructural analyses were used to relate the phenotypic changes to pectin changes in the leaf cell walls. The modification of apple trees by a single PG gene has offered a new and unexpected perspective on the role of pectin and cell wall adhesion in leaf morphology and stomatal development.

Cell Adhesion↗

Isolation and characterization of psychrophilic yeasts producing cold-adapted pectinolytic enzymes.

AIMS: The present study was conducted to screen for psychrophilic yeasts that are able to degrade pectin compounds at low temperature, and to examine the cold-active pectinolytic enzymes produced by the isolated psychrophilic yeasts. METHODS AND RESULTS: Psychrophilic yeasts, which grow on pectin as a sole carbon source, pectinolytic-psychrophilic yeast (PPY) strains PPY-3, 4, 5 and 6, were isolated from soil from Abashiri (Hokkaido, Japan). The sequences of 28S rDNA D1/D2 of strains PPY-3 and 4 indicated a taxonomic affiliation to Cryptococcus cylindricus and Mrakia frigida, respectively, strains PPY-5 and 6 belonged to Cystofilobasidium capitatum. The isolated strains were able to grow on pectin at below 5 degrees C, and showed the activities of several cold-active pectinolytic enzymes. CONCLUSION: The findings of this study indicate the possibility that the isolated strains produce novel pectinolytic enzymes that are able to degrade pectin compounds at low temperature. SIGNIFICANCE AND IMPACT OF THE STUDY: It is possible that the cold-active pectinolytic enzymes from the isolated strains can be applied to the food industry, e.g. the clarification of fruit juice below 5 degrees C.

Basidiomycota↗

Role of unstirred water layer in the exsorption of quinidine.

Intestinal ++exsorption of salicylic acid, thiopentone, theophylline, and quinidine was measured during perfusion of the intestinal lumen with Tyrode solution. The effect of pectin or bovine serum albumin added to the perfusate on intestinal clearance (CLi) was investigated. Increasing pectin concentration from 0.0 to 0.5, 1.0 and 1.5% gave CLi values for quinidine of 499 +/- 18, 363 +/- 35, 237 +/- 56, and 300 +/- 28 mL h-1 kg-1, respectively. One per cent of pectin in the perfusate also decreased the CLi of thiopentone, but had no effect on the CLi of salicylic acid or theophylline. Pectin may have increased the thickness of the unstirred water layer on the mucous membrane and the resistance of drug exsorption for some drugs. When bovine serum albumin was added, drug binding in the perfusate increased, and the CLi of salicylic acid, thiopentone, and theophylline increased; the CLi of quinidine was unaltered. Co-administration of theophylline with quinidine decreased the CLi of quinidine without affecting quinidine binding in serum or in the perfusate. The CLi theophylline was not affected by quinidine. These observations are consistent with the hypothesis that the exsorption of quinidine is rate-limited by diffusion through the unstirred water layer on the mucous membrane. The CLi of quinidine is affected by the microclimate-pH in the unstirred water layer. An alternative possibility is that quinidine exsorption is mediated by a carrier-transport pathway.

Animals↗

Adaptation of mesophilic anaerobic sewage fermentor populations to thermophilic temperatures.

Thermophilic (50 degrees C) and obligately thermophilic (60 degrees C) anaerobic carbohydrate- and protein-digesting and methanogenic bacterial populations were enumerated in a mesophilic (35 degrees C) fermentor anaerobically digesting municipal primary sludge. Of the total bacterial population in the mesophilic fementor, 9% were thermophiles (36 x 10(6)/ml) and 1% were obligate thermophiles (4.5 x 10(6)/ml). Of these 10%, the percentages of bacteria (thermophiles and obligate thermophiles, respectively) able to use specific substrates were further enumerated as follows: bacteria able to digest albumin, casein, starch, and mono- and disaccharides, 30 and 10%; pectin degraders, 10 and 0.2%; cellulose degraders, 2 and 0.06%; methanogens that grow with H2 and CO2, methanol, and dimethylamine, 9 and 1%; methanogens that grow with formate, 8 and 5%; and methanogens that grow with acetate, 25 and less than 0.8%. Shortly after the temperature was elevated from 35 to 50 or 60 degrees C, the digestion of albumin, casein, starch, and mono- and disaccharides was detected, and methane was produced from H2 and CO2. Methane produced from acetate was not delayed at 50 degrees C, but was delayed by 29 days at 60 degrees C. Methane produced from formate was delayed by 3 days, from methanol by 7 days, and from dimethylamine by 5 days at 50 and 60 degrees C. A 10- and 20-day acclimation period was required for hydrolysis of pectin and cellulose, respectively, at 50 degrees C. Digestion of pectin required 20 days and cellulose longer than 85 days when the temperature was elevated abruptly from 35 to 60 degrees C. The acclimation period for the digestion of pectin and cellulose at 60 degrees C was shortened to 3 and 15 days, respectively, by seeding with a small amount of a culture acclimated to 50 degrees C. The data suggest that enrichment of cellulolytic, pectinolytic, and acetate-utilizing bacteria is crucial for the digestion of sewage sludge at 60 degrees C.

Acetates↗

Bacteroides pectinophilus sp. nov. and Bacteroides galacturonicus sp. nov.: two pectinolytic bacteria from the human intestinal tract.

Studies on the physiological characteristics of two obligately anaerobic, rod-shaped bacteria from the human intestinal tract indicated that the organisms represented two previously undescribed species of Bacteroides, for which we propose the names Bacteroides pectinophilus (type strain, N3) and Bacteroides galacturonicus (type strain, N6). Both strains were pectinophilic; that is, they utilized as fermentable substrates for growth only pectin and a few related compounds. The two species differed significantly from each other in guanine plus cytosine content of the DNA, in substrate utilization patterns, and in other phenotypic characteristics. Both species deesterified pectin by means of an extracellular pectinesterase (EC 3.1.1.11) activity. Polygalacturonate (the main component of deesterified pectin) was depolymerized extracellularly with formation of unsaturated products by both species. The depolymerizing activity required Ca2+, functioned at a higher rate when polygalacturonate was the substrate as compared with pectin, and had an alkaline pH optimum. These data, as well as viscosity decrease studies and identification of products formed from polygalacturonate, indicated that the extracellular depolymerizing activity of either species was characteristic of an exopectate (exopolygalacturonate) lyase. The exopectate lyase activity had an unusual action pattern that resulted in terminal cleavage of unsaturated trigalacturonic acid units from polygalacturonate. An unsaturated trimer was the major product that accumulated in cell-free reaction mixtures, where it was not cleaved further. Growing cells of both Bacteroides species released the exopectate lyase into the external environment by processes that did not involve cell lysis to any significant extent.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteroides↗