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Decreased-activity mutants of phosphoglucose isomerase in the cytosol and chloroplast of Clarkia xantiana. Impact on mass-action ratios and fluxes to sucrose and starch, and estimation of Flux Control Coefficients and Elasticity Coefficients.

1. Subcellular-compartment-specific decreased-activity mutants of phosphoglucose isomerase in Clarkia xantiana were used to analyse the control of sucrose and starch synthesis during photosynthesis. Mutants were available in which the plastid phosphoglucose isomerase complement is decreased to 75% or 50% of the wild-type level, and the cytosol complement to 64%, 36% or 18% of the wild-type level. 2. The effects on the [product]/[substrate] ratio and on fluxes to sucrose or starch and the rate of photosynthesis were studied with the use of saturating or limiting light intensity to impose a high or low flux through these pathways. 3. Removal of a small fraction of either phosphoglucose isomerase leads to a significant shift of the [product]/[substrate] ratio away, from equilibrium. We conclude that there is no 'excess' of enzyme over that needed to maintain its reactants reasonably close to equilibrium. 4. Decreased phosphoglucose isomerase activity can also alter the fluxes to starch or sucrose. However, the effect on flux does not correlate with the extent of disequilibrium, and also varies depending on the subcellular compartment and on the conditions. 5. The results were used to estimate Flux Control Coefficients for the chloroplast and cytosolic phosphoglucose isomerases. The chloroplast isoenzyme exerts control on the rate of starch synthesis and on photosynthesis in saturating light intensity and CO2, but not at low light intensity. The cytosolic enzyme only exerts significant control when its complement is decreased 3-5-fold, and differs from the plastid isoenzyme in exerting more control in low light intensity. It has a positive Control Coefficient for sucrose synthesis, and a negative Control Coefficient for starch synthesis. 6. The Elasticity Coefficients in vivo of the cytosolic phosphoglucose isomerase were estimated to lie between 5 and 8 in the wild-type. They decrease in mutants with a lowered complement of cytosolic phosphoglucose isomerase. 7. The implications of these results for regulation and for evolution are discussed.

Chloroplasts↗

Exploring and exploiting starch-modifying amylomaltases from thermophiles.

Starch is a staple food present in water-insoluble granules in many economically important crops. It is composed of two glucose polymers: the linear alpha-1,4-linked amylose and amylopectin with a backbone of alpha-1,4-glycosidic bonds and alpha-1,6-linked side chains. To dissolve starch completely in water it needs to be heated; when it cools down too much the starch solution forms a thermo-irreversible gel. Amylomaltases (EC 2.4.1.25) are enzymes that transfer a segment of an alpha-1,4-D-glucan to a new 4-position in an acceptor, which may be glucose or another alpha-1,4-D-glucan. Acting upon starch, amylomaltases can produce cycloamylose or a thermoreversible starch gel, both of which are of commercial interest.

Amylopectin↗

Wheat-starch-based gluten-free products in the treatment of newly detected coeliac disease: prospective and randomized study.

BACKGROUND: The safety of wheat-starch-based gluten-free products in the treatment of coeliac disease is debatable. Prospective studies are lacking. AIM: : To compare the clinical, histological and serological response to a wheat-starch-based or natural gluten-free diet in patients with newly detected coeliac disease. METHODS: Fifty-seven consecutive adults with untreated coeliac disease were randomized to a wheat-starch-based or natural gluten-free diet. Clinical response, small bowel mucosal morphology, CD3+, alphabeta+ and gammadelta+ intra-epithelial lymphocytes, mucosal human leucocyte antigen-DR expression and serum endomysial, transglutaminase and gliadin antibodies were investigated before and 12 months after the introduction of the gluten-free diet. Quality of life measurements were performed by standardized questionnaires and the bone mineral density was analysed. RESULTS: In both groups, abdominal symptoms were alleviated equally by a strict diet. There were no differences between the groups in mucosal morphology, the density of intra-epithelial lymphocytes, serum antibodies, bone mineral density or quality of life tests at the end of the study. Four patients on a natural gluten-free diet and two on a wheat-starch-based gluten-free diet had dietary lapses; as a result, inadequate mucosal, serological and clinical recovery was observed. CONCLUSIONS: The dietary response to a wheat-starch-based gluten-free diet was as good as that to a natural gluten-free diet in patients with newly detected coeliac disease.

Adult↗

Effect of some nutritional and environmental parameters on the production of diacetyl and on starch consumption by Pediococcus pentosaceus and Lactobacillus acidophilus in submerged cultures.

Three series of 5-day submerged cultures with Pediococcus pentosaceus MITJ-10 and Lactobacillus acidophilus Hansen 1748 were carried out in starch-based media, and the effect of cultural factors on the changes of starch, diacetyl and amylase activity determined. In axenic cultures, Ped. pentosaceus MITJ-10 produced more diacetyl (63.27 mg l(-1)) by adding glucose, yeast extract and CaCO3 (P < 0.01), at 28 degrees C (P < 0.05); but more starch was consumed (18.4 g l(-1)) in the absence of glucose (P < 0.01). Lact. acidophilus Hansen 1748 consumed more starch (26.56 g l(-1)) at 28 degrees C, with CaCO3, glucose (P < 0.01) and yeast extract (P < 0.05); however, the amylolytic activity (10077U l(-1)) was favoured at 35 degrees C (P < 0.01). Little starch was consumed in mixed cultures due to the low pH; nevertheless, diacetyl content rose to 135.76 mg l(-1) at 32 degrees C (P < 0.01). Therefore, both studied strains might be useful to produce aromatic extensors from starchy substrates. These natural aromatic extensors are of interest to the food industry.

Amylases↗

Characterization of factors affecting attachment of Bifidobacterium species to amylomaize starch granules.

AIMS: Two human-derived Bifidobacterium strains, PL1 and PL2, were tested for their ability to attach to amylomaize starch granules, and factors affecting binding were assessed. METHODS AND RESULTS: Good binding to granules was observed when the strains were grown on maltose or amylomaize starch, but not on glucose. Binding activity was localized to cell wall components and was sensitive to treatment with proteolytic enzymes. Several methodologies were employed to confirm these observations, including studies using radiolabelled cells, dot blot assays and scanning electron microscopy (SEM) analysis. CONCLUSION: Results from this study indicated that binding of strains PL1 and PL2 to amylomaize starch granules was mediated by a cell wall-associated proteinaceous factor that was induced when the strains were grown on starch or a related substrate, but not glucose. SIGNIFICANCE AND IMPACT OF THE STUDY: Attachment of probiotic strains to starch or other dietary fibres is believed to offer a selective advantage in the host intestine and may even prolong viability in adverse food environments. Therefore, characterizing the mechanisms of attachment has commercial implications in the design of synbiotic products.

Bacterial Adhesion↗

Starch granule initiation and growth are altered in barley mutants that lack isoamylase activity.

Two mutant lines of barley, Risø 17 and Notch-2, were found to accumulate phytoglycogen in the grain. Like the sugary mutants of maize and rice, these phytoglycogen-accumulating mutants of barley lack isoamylase activity in the developing endosperm. The mutants were shown to be allelic, and to have lesions in the isoamylase gene, isa1 that account for the absence of this enzyme. As well as causing a reduction in endosperm starch content, the mutations have a profound effect on the structure, number and timing of initiation of starch granules. There are no normal A-type or B-type granules in the mutants. The mutants have a greater number of starch granules per plastid than the wild-type and, particularly in Risø 17, this leads to the appearance of compound starch granules. These results suggest that, as well as suppressing phytoglycogen synthesis, isoamylase in the wild-type endosperm plays a role in determining the number, and hence the form, of starch granules.

Alleles↗

The action of starch synthase II on 6"'-alpha-maltotriosyl-maltohexaose comprising the branch point of amylopectin.

The principle of using a chemically synthesized, well-defined branched oligosaccharide to provide a more detailed knowledge of the substrate specificity of starch synthase II (SSII) is demonstrated. The branched nonasaccharide, 6"'-alpha-maltotriosyl-maltohexaose, was investigated as a primer for particulate SSII using starch granules prepared from the low-amylose pea mutant lam as the enzyme source. The starch granule preparation from the lam pea mutant contains no starch synthases other than SSII and is devoid of alpha-amylase, beta-amylase and phosphorylase activity. SSII was demonstrated to catalyse a specific nonprocessive elongation of the nonreducing end of the shortest unit chain of 6"'-alpha-maltotriosyl-maltohexaose, i.e. the maltotriose chain. Maltotriose and maltohexaose, representing the two linear building units of the branched nonasaccharide, were also tested as primers for SSII. Maltotriose was elongated more efficiently than 6"'-alpha-maltotriosyl-maltohexaose and maltohexaose was used less efficiently. Compared to the surface exposed alpha-glucan chains of the granule bound amylopectin molecules, all three soluble oligosaccharides tested were poor primers for SSII. This indicates that in vivo, the soluble oligosaccharides supposedly released as result of amylopectin trimming reactions are not re-introduced into starch biosynthetic reactions via the action of the granule bound fraction of SSII.

Amylopectin↗

Hydroxyethyl starch and change in renal function in patients undergoing coronary artery bypass graft surgery.

BACKGROUND: Several case reports and clinical lore have suggested that exposure to the colloid hydroxyethyl starch may impair renal function, but few studies have systematically addressed this issue, and several have produced conflicting results. We sought to study the question in a formal analysis of postoperative change in renal function in patients undergoing coronary artery bypass graft (CABG) surgery. METHODS: We identified 238 consecutive patients who underwent CABG surgery at a large academic medical center. Glomerular filtration rate (GFR) was estimated using the Cockroft-Gault formula at baseline as well as on postoperative days 3 and 5. Linear regression analysis was used to study the relation between changes in GFR and intraoperative hydroxyethyl starch administration. Multivariate models controlled for potential demographic, clinical, and surgery-related confounders. RESULTS: Hydroxyethyl starch use was independently associated with a reduction in GFR on both postoperative days 3 and 5, with GFR declining by 7.2 mL/min/1.73 m2 on day 3 per unit of hydroxyethyl starch administered (95% CI, 1.7 to 12.7; P = 0.012), and by 6.6 mL/min/1.73 m2 on day 5 (95% CI, 1.2 to 11.9; P = 0.018). CONCLUSION: Intraoperative use of hydroxyethyl starch may be associated with modest impairment in renal function in patients undergoing CABG surgery. Randomized clinical trials will be necessary to confirm these findings and to further investigate their clinical implications.

Adult↗

In situ and in vitro ruminal starch degradation of untreated and formaldehyde-treated wheat and maize.

Ruminal starch degradation of untreated and formaldehyde-treated wheat and maize was measured in situ (trial 1) and in vitro (trial 2). The in situ starch degradability was higher for wheat than for maize (82.1 vs 52.3%), for untreated cereals than for cereals treated with 1% formaldehyde (77.3 vs 67.0%) and for cereals treated with 1% formaldehyde than those treated with 5% formaldehyde (67.0 vs 57.2%). The in vitro results were similar. The treatment of cereals by formaldehyde decreased starch degradability more for wheat than for maize, suggesting that the treatment was more efficient when cereal starch and/or nitrogen was highly degradable. Formadehyde treatment of wheat was more effective at decreasing the rate of wheat protein degradability than starch degradability. The difference of response to treatment between the two cereals may be due to differences in properties of the protein matrix of these two cereals.

Animal Feed↗

Extrusion cooking of a high-fibre cereal product. 1. Effects on digestibility and absorption of protein, fat, starch, dietary fibre and phytate in the small intestine.

1. The effect of extrusion cooking of a high-fibre cereal product on digestibility of starch, fibre components and phytate in the stomach and small intestine was studied by in vivo digestion in ileostomy subjects, as well as its effect on ileostomy losses of fat, nitrogen, sodium and potassium. 2. Seven ileostomy subjects were studied during two periods (each of 4 d) while on a constant low-fibre diet supplemented with 54 g/d of a bran-gluten-starch mixture (period A) or the corresponding extruded product (period B). 3. Extrusion cooking, using mild conditions, did not change the content of starch, dietary fibre components or phytate of the bran product, but the phytase (EC 3.1.3.26) activity was lost. During the period using the extruded bran product, there was a significant increase in recovery of phytate-phosphorus (period A, 44% of intake; period B, 73% of intake). The amount of fibre components, fat, fatty acids, N, Na, K, water and the ash weight of the ileostomy contents did not differ between the two periods. Only 0.6 and 0.7% respectively of ingested starch was recovered in ileostomy contents in periods A and B, while the fibre components were almost completely recovered. 4. Extrusion cooking, using even mild conditions, may lead to a considerable impairment in the digestion of phytate, probably due to a qualitative change in phytate and a loss of phytase activity. Starch, before and after extrusion cooking, is almost completely digested in the stomach and small intestine while fibre components are digested to a very small extent.

Absorption↗

Evaluation of the net energy value of glucose (cerelose) and maize starch in diets for rainbow trout (Salmo gairdneri).

1. Quadruplicate groups of rainbow trout (Salmo gairdneri) (mean body-weight 24.9 g) were reared on six dietary treatments (practical-type diets) in a modified paired-feeding experiment for 12 weeks at 15 degrees to determine the net energy (NE) value of starch and glucose to rainbow trout. 2. Three test diets were prepared to contain (g/kg): 0 supplemented carbohydrate (diet 1), 250 maize starch (diet 2) and 250 glucose (diet 3) and were given ad lib. to the trout with the feeding rate of the glucose- and starch-fed groups being monitored after each feeding. The remaining three treatments involved controlled feeding of the trout with diet 1 at 75% of the feed intake of trout reared on diets 2 and 3, so as to provide the same levels of protein and lipids without carbohydrate, and with diet 2 at 100% of the feed intake of trout reared on diet 3. 3. The difference in the final carcass energy of the ad lib.-fed group and the respective controlled-fed group divided by the amount of dietary glucose or starch energy consumed by the trout is the NE value for that carbohydrate. 4. The determined NE value of glucose was 3.99 kJ/g and starch 2.17 kJ/g, which is 24.6 and 12.6% respectively of the gross energy values of these carbohydrates in rainbow trout. 5. The results indicate that digestible energy and calculated metabolizable energy values for carbohydrates in rainbow trout overestimate the utilizable energy content of the diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The inhibitory effects of hull polysaccharides and tannins of field beans (Vicia faba L.) on the digestion of amino acids, starch and lipid and on digestive enzyme activities in young chicks.

The effects of polysaccharides and tannins present in the hulls of field beans (Vicia faba L.) on the digestion of amino acids, starch and lipid were studied in poultry. A control diet without hulls and the same diet substituted with 400 g hulls/kg diet from three different varieties of beans were fed to 3-week-old chicks for 4 d. Digestibility coefficients for amino acids, starch and lipid were calculated from measurements made of these nutrients in the diets and the freeze-dried excreta with the aid of titanium dioxide as a marker. Activities of trypsin (EC 3.4.21.4), alpha-amylase (EC 3.2.1.1), and lipase (EC 3.1.1.3) in digesta removed from the upper jejunum, sucrase (EC 3.2.1.48) in the gut mucosa from the upper jejunum, and alpha-amylase and lipase in the pancreas were measured. The hulls were analysed for their polysaccharide and tannin contents. Results showed that the hulls were mostly carbohydrate in composition, with cellulose the predominant polysaccharide. Tannins present in the hulls of two coloured-flowering varieties (Brunette and Minica) were of the condensed type. The diet with tannin-free hulls (white-flowering variety Medes) lowered slightly the digestion of amino acids, starch and lipid compared with the control diet. This effect was believed to be due to inhibition of digestive enzymes, possibly through their adsorption onto the hulls. Diets with tannin-rich hulls (varieties Brunette and Minica) caused a large reduction in the digestion of amino acids, starch and lipid compared with the control diet mainly due to inactivation of digestive enzymes by the formation of tannin-enzyme complexes in the digestive tract. Enzyme activities could be partially restored by the addition of polyvinylpyrrolidone to the digesta. Tannins inactivated trypsin the most, alpha-amylase to a lesser extent and lipase the least and as a consequence lowered the digestion of amino acids the most, starch to a lesser extent and lipid the least. Tannins did not induce an increased pancreatic production of digestive enzymes, nor did they affect activity of jejunum mucosal sucrase. Condensed tannins from Brunette and Minica hulls were partially extractable in methanol alone, but required acidic methanol for fuller extraction. The vanillin:anthocyanidin ratio suggested that tannins were polymerized to the same degree in the Brunette and Minica varieties, both in the methanol and acidic methanol extracts. Hulls from the variety Minica contained a greater amount of methanol-extractable tannins, the quantity of remaining tannins that required acidic methanol for extraction being the same for both varieties.

Amino Acids↗

Digestion and physiological properties of resistant starch in the human large bowel.

The digestion of four sources of resistant starch (RS) has been studied in twelve healthy volunteers who ate controlled diets for 15 d periods. RS from potato, banana, wheat and maize (17-30 g/d) was compared with a starch-free diet, a diet containing wheat starch that was fully digested in the small intestine, and with 18.4 g NSP from brand/d. RS increased stool wet weight by 1.6 g/d per g RS fed for potato, 1.7 for banana, 2.5 for wheat and 2.7 for maize, but this was significantly less than bran NSP at 4.9 g/g. RS was extensively digested in twenty-seven of thirty-four diet periods but five subjects were unable to break down one or two of the RS sources. Faecal N and energy excretion were increased. RS decreased NSP breakdown and RS2 (resistant starch granules) tended to prolong transit time. All forms of RS increased faecal total short-chain fatty acid excretion. RS2 (from potato and banana) gave greater proportions of acetate in faeces, and RS3 (retrograded starch from wheat and maize) more propionate. We have concluded that RS2 and RS3 are broken down in the human gut, probably in the colon although in 26% of cases this breakdown was impaired. RS exerts mild laxative properties, predominantly through stimulation of biomass excretion but also through some sparing of NSP breakdown.

Adult↗

Preference for wheat straw by lambs conditioned with intraruminal infusions of starch.

We hypothesized that feed preference depends on the interplay between flavour and postingestive effects, and we tested two predictions based on this hypothesis: (1) lambs acquire preferences for poorly nutritious feeds paired with starch; and (2) preferences persist when starch is no longer administered. Twenty lambs were randomly allocated to two groups and conditioned as follows: on odd-numbered days, lambs in group 1 received onion-flavoured wheat straw and lambs in group 2 received oregano-flavoured wheat straw. On even-numbered days, the flavours were switched and starch (2.5-9.4% of the digestible energy received/d) was infused into the rumen of all animals during straw consumption. Four periods of 8 d of conditioning were performed and on the 9th day of each period all animals were offered a choice between onion- and oregano-flavoured straw. After conditioning, starch administration was suspended and lambs were offered onion- and oregano-flavoured straw at weekly intervals for 8 weeks (extinction). Lambs strongly preferred the flavoured straw paired with starch, and this preference persisted during extinction. Thus, these results suggest that the postingestive effects of energy play an important role in the development of feed preferences of ruminants.

Allium↗

Effect of starch on the cariogenic potential of sucrose.

Since in vitro and animal studies suggest that the combination of starch with sucrose may be more cariogenic than sucrose alone, the study assessed in situ the effects of this association applied in vitro on the acidogenicity, biochemical and microbiological composition of dental biofilm, as well as on enamel demineralization. During two phases of 14 d each, fifteen volunteers wore palatal appliances containing blocks of human deciduous enamel, which were extra-orally submitted to four groups of treatments: water (negative control, T1); 2 % starch (T2); 10 % sucrose (T3); and 2 % starch+10 % sucrose (T4). The solutions were dripped onto the blocks eight times per day. The biofilm formed on the blocks was analysed with regard to amylase activity, acidogenicity, and biochemical and microbiological composition. Demineralization was determined on enamel by cross-sectional microhardness. The greatest mineral loss was observed for the association starch+sucrose (P<0.05). Also, this association resulted in the highest lactobacillus count in the biofilm formed (P<0.05). In conclusion, the findings suggest that a small amount of added starch increases the cariogenic potential of sucrose.

Actinomyces↗

Effect of site of starch digestion on portal nutrient net fluxes in steers.

Processing of maize grain is known to modulate the site of starch digestion, thus the nature and amount of nutrients delivered for absorption. We assessed the effect of site of starch digestion on nutrient net fluxes across portal-drained viscera (PDV). Three steers, fitted with permanent digestive cannulas and blood catheters, successively received two diets containing 35 % starch as dent maize grain. Diets differed according to maize presentation: dry and cracked (by-pass, BP) v. wet and ground (control, C). Ruminal physicochemical parameters were not significantly affected. Between C and BP, the decrease in ruminal starch digestion was compensated by an increase in starch digestion in the small intestine. The amount of glucose and soluble alpha-glucoside reaching the ileum was not affected. The amount of glucose disappearing in the small intestine increased from 238 to 531 g/d between C and BP, but portal net flux of glucose remained unchanged (-97 g/d). The portal O2 consumption and net energy release were not significantly affected, averaging 16 % and 57 % of metabolizable energy intake, respectively. The whole-body glucose appearance rate, measured by jugular infusion of [6,6-2H2]glucose, averaged 916 g/d. The present study shows that the increase in the amount of glucose disappearing in the small intestine of conventionally fed cattle at a moderate intake level induces no change in portal net flux of glucose, reflecting an increase in glucose utilization by PDV. That could contribute to the low response of whole-body glucose appearance rate observed at this moderate level of intestinal glucose supply.

Absorption↗

Effect of DHA supplementation on digestible starch utilization by rainbow trout.

Rainbow trout has a limited ability to utilize digestible carbohydrates efficiently. Trout feeds generally contain high levels of DHA, a fatty acid known to inhibit a number of glycolytic and lipogenic enzymes in animals. A study was conducted to determine whether carbohydrate utilization by rainbow trout might be affected by dietary DHA level. Two low-carbohydrate (<4 % digestible carbohydrate) basal diets were formulated to contain 1 (adequate) or 4 (excess) g/100 g DHA diet respectively. The two basal diets were diluted with increasing levels of digestible starch (0 %, 10 %, 20 % and 30 %, respectively) to produce eight diets. These diets were fed to fish for 12 weeks at 15 degrees C according to a pair-fed protocol that consisted of feeding the same amount of basal diet but different amounts of starch. Live weight, N and lipid gains, hepatic glycogen and plasma glucose values significantly increased, whereas feed efficiency (gain:feed) significantly decreased, with increasing starch intake (P<0.05). The retention efficiency of N (N gain/digestible N intake) improved with starch supplementation but was not affected by DHA level (P>0.05). Starch increased the activity of glucokinase, pyruvate kinase, glucose 6-phosphate dehydrogenase and fatty acid synthase (P<0.05) but did not affect hexokinase and malic enzyme activity. DHA had no effect on growth but increased plasma glucose and reduced carcass lipid and liver glycogen contents (P<0.05). Glycolytic and lipogenic enzymes were not affected by DHA level, except for pyruvate kinase, which was reduced by increasing DHA level. These results suggest only a marginal effect of dietary DHA on the ability of fish to utilize carbohydrate.

Animal Feed↗

Starch-encapsulated chlorpyrifos: release rate, insecticidal activity and degradation in soil.

Incorporation of chlorpyrifos into starch matrices was achieved by thermal gelatinization. The level of amylose content in the starch matrices (10 and 52%) and the addition of auxiliary agents, i.e. solvent, emulsifiers or both, were varied and eight different formulations were prepared. According to differential scanning calorimetry (DSC) and X-ray diffraction measurements, chlorpyrifos was partly present in a crystalline form in all the starch formulations. The formulations had controlled release properties, and the release rate into water could be described by a linear model. In a bioassay, the formulations killed all larvae of the cabbage root fly at a dosing of 1.4 kg a.i. per ha. The degradation of chlorpyrifos in soil from the starch formulations could be described in a non-linear logistic model and the half-life was predicted to be 88 days. Differences in the amount of amylose in the starch as well as the addition of solvent and emulsifiers in the preparation procedure had no systematic influence on the release rate, the insecticidal effect and the degradation rate. No correlation between release rate into water and degradation in soil could be established. Two commercial chlorpyrifos formulations Lorsban 15 G (granular) and Cyfos 500 gl(-1) (emulsifiable concentrate), were included in the study for comparison.

Algorithms↗