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Starch branching enzymes belonging to distinct enzyme families are differentially expressed during pea embryo development.

cDNA clones for two isoforms of starch branching enzyme (SBEI and SBEII) have been isolated from pea embryos and sequenced. The deduced amino acid sequences of pea SBEI and SBEII are closely related to starch branching enzymes of maize, rice, potato and cassava and a number of glycogen branching enzymes from yeast, mammals and several prokaryotic species. In comparison with SBEI, the deduced amino acid sequence of SBEII lacks a flexible domain at the N-terminus of the mature protein. This domain is also present in maize SBEII and rice SBEIII and resembles one previously reported for pea granule-bound starch synthase II (GBSSII). However, in each case it is missing from the other isoform of SBE from the same species. On the basis of this structural feature (which exists in some isoforms from both monocots and dicots) and other differences in sequence, SBEs from plants may be divided into two distinct enzyme families. There is strong evidence from our own and other work that the amylopectin products of the enzymes from these two families are qualitatively different. Pea SBEI and SBEII are differentially expressed during embryo development. SBEI is relatively highly expressed in young embryos whilst maximum expression of SBEII occurs in older embryos. The differential expression of isoforms which have distinct catalytic properties means that the contribution of each SBE isoform to starch biosynthesis changes during embryo development. Qualitative measurement of amylopectin from developing and maturing embryos confirms that the nature of amylopectin changes during pea embryo development and that this correlates with the differential expression of SBE isoforms.

1,4-alpha-Glucan Branching Enzyme↗

Diurnal changes in sucrose, nucleotides, starch synthesis and AGPS transcript in growing potato tubers that are suppressed by decreased expression of sucrose phosphate synthase.

Sucrose export from leaves is high during the day and lower at night, when it depends on starch remobilisation. We have investigated the consequences of diurnal changes of photoassimilate supply for starch synthesis and other metabolic processes in growing potato tubers. Sucrose, the levels of the transcripts for SUS and AGPS, the levels of key metabolites, and the rate of synthesis of starch and other major end products, including protein and cell wall polysaccharides, increased twofold or more between the start and end of the light period. The stimulation of starch synthesis was accompanied by an increase of UDPglucose and ADPglucose, whereas glycolytic intermediates remained unaltered, revealing that sucrose synthase and ADP-glucose pyrophosphorylase are being co-ordinately regulated. Sucrose synthase is stimulated via an increase of its substrates, UDP and sucrose. UDP increases due to an increase of the overall uridine nucleotide pool, and a decrease of the UTP/UDP ratio that occurs in parallel with a decrease of the ATP/ADP ratio and adenylate energy charge when biosynthetic fluxes are high at the end of the day. Within the time frame of the diurnal changes, the changes in the SUS and AGPS transcript levels do not lead to significant changes in the encoded enzymes. Transformants with a progressive decrease of sucrose phosphate synthase expression, where diurnal changes in leaf sugar levels were damped, exhibited a progressive attenuation of the diurnal changes of sucrose, nucleotide sugars and nucleotides, and fluxes in their tubers. It is concluded that metabolic processes in tubers are tightly linked to the momentary supply of sucrose.

Circadian Rhythm↗

Utilization of raw or heat-treated starch fed in liquid diet to pre-ruminants. 1. Calves.

Five 7-day-old Frisian bull calves, raised in Israel received twice a day a mixture of 40 g of soybean concentrate (65% protein) and 80 g of one of the following carbohydrates: glucose (G), expanded (heat-treated) (ES) or untreated (US) corn starch. In the afternoon the calves received in addition 400 g of milk replacer. Until weaning at experimental day 25, ES calves showed better growth and food utilization than their counterparts. Daily weight gain for the entire growing period up to slaughter was higher for the starch than for the glucose-fed group (P less than 0.05 for ES vs G). Heat treatment of starch increased its in vitro availability to amylase and its in vivo digestibility. Bypass of the rumen was complete in all G calves. In the ES and US groups, partial diversion of the liquid feed into the rumen was evident. The G group showed hyperglycemia after meals, while almost no increase in blood glucose level was observed after soy-starch meals (either US or ES).

Amylases↗

Digestion of starch and glycaemic response to mixed meals in pigs.

The digestion in the proximal intestine of mixed meals (5,160 kJ) containing either native (NS) or pregelatinized (PS) maize starches (approximately 200 g), and the postprandial glycaemic responses they induced were compared in pigs. For both meals, approximately 25% of the ingested starch was assimilated above the duodenal cannula (positioned 75 cm beyond the pylorus). Larger amounts of starch were collected for NS than for PS during the first 30 min. The glycaemic responses, however, indicated a higher rate of glucose absorption for PS during the first 30 min, which could be explained by the higher susceptibility of PS to hydrolysis, as we observed in vivo. Indeed, malto-oligosaccharides (G1-G3) represented almost 80% of the total alpha-glucans collected at 150 min in the duodenum after the PS meal. At that time, after the NS meal, only 30% of the alpha-glucans were malto-oligosaccharides. Thus, even after a mixed meal, the starch digestion rate can alter the observed postprandial glycaemic response.

Animals↗

[The accumulation of different substituted hydroxyethyl starches (HES) following repeated infusions in healthy volunteers].

AIM OF THE STUDY: Accumulation of hydroxyethyl starch (HES) after repeated applications of starches with different molar substitution and similar molecular weight was investigated. METHODS: Treatment with five consecutive infusions of hydroxyethyl starch was carried out using two medium molecular weight hydroxyethyl starches with a molar substitution of 0.5 and 0.62. Healthy volunteers received 500 ml 6% HES 200/0.62 (30 g) or 500 ml 10% HES 200/0.5 per day over a period of five consecutive days. Blood samples were taken in the morning before infusions were started (7.A.M.) and at each hour during the infusion period of 4 hours post infusionem until 4 hours after the infusions. During the first 10 days and on the 20th and on the 30th day after the last infusion blood samples were taken. RESULTS: Both HES solutions were subjectively and objectively well tolerated by healthy volunteers. No side effects were observed. However, pharmacokinetics of the investigated HES-formulations were significantly different. The model-independent calculated elimination half life time (T1/2) increased from day to day. During the five days T1/2 was prolonged for 20 h by high substituted HES (200/0.62) and for 2.5 h for medium substituted HES (200/0.5). The half life times related to the three compartment model calculation were with 0.6 h, 11.6 h and 211 h for HES 200/0.62 two fold higher than the times for HES 200/0.5 with 0.39 h, 6.98 h and 113 h. Plasma clearance for HES 200/0.5 (4.86 ml/min) was five fold higher than that from HES 200/0.62 with 0.98 ml/min. With the exception of the first day of infusion serum concentrations of HES 200/0.5 although only 30 g HES 200/0.6 versus 50 g HES 200/0.5 were infused. No difference of the hemodilution effects between the two HES-formulations were observed. The hemorheologic parameters were similar in both groups with the exception of plasma viscosity which was significantly higher after infusion of HES 200/0.62. CONCLUSION: High substituted HES accumulate in serum more than medium substituted HES. Especially when HES must be applied in multiple doses, high substituted HES should not be used or the infusion interval must be adapted to the elimination half life time of the used HES.

Adult↗

Trehalose 6-phosphate regulates starch synthesis via posttranslational redox activation of ADP-glucose pyrophosphorylase.

Trehalose is the most widespread disaccharide in nature, occurring in bacteria, fungi, insects, and plants. Its precursor, trehalose 6-phosphate (T6P), is also indispensable for the regulation of sugar utilization and growth, but the sites of action are largely unresolved. Here we use genetic and biochemical approaches to investigate whether T6P acts to regulate starch synthesis in plastids of higher plants. Feeding of trehalose to Arabidopsis leaves led to stimulation of starch synthesis within 30 min, accompanied by activation of ADP-glucose pyrophosphorylase (AGPase) via posttranslational redox modification. The response resembled sucrose but not glucose feeding and depended on the expression of SNF1-related kinase. We also analyzed transgenic Arabidopsis plants with T6P levels increased by expression of T6P synthase or decreased by expression of T6P phosphatase (TPP) in the cytosol. Compared with wild type, leaves of T6P synthase-expressing plants had increased redox activation of AGPase and increased starch, whereas TPP-expressing plants showed the opposite. Moreover, TPP expression prevented the increase in AGPase activation in response to sucrose or trehalose feeding. Incubation of intact isolated chloroplasts with 100 muM T6P significantly and specifically increased reductive activation of AGPase within 15 min. Results provide evidence that T6P is synthesized in the cytosol and acts on plastidial metabolism by promoting thioredoxin-mediated redox transfer to AGPase in response to cytosolic sugar levels, thereby allowing starch synthesis to be regulated independently of light. The discovery informs about the evolution of plant metabolism and how chloroplasts of prokaryotic origin use an intermediate of the ancient trehalose pathway to report the metabolic status of the cytosol.

Arabidopsis↗

Similar protein phosphatases control starch metabolism in plants and glycogen metabolism in mammals.

We report that protein phosphorylation is involved in the control of starch metabolism in Arabidopsis leaves at night. sex4 (starch excess 4) mutants, which have strongly reduced rates of starch metabolism, lack a protein predicted to be a dual specificity protein phosphatase. We have shown that this protein is chloroplastic and can bind to glucans and have presented evidence that it acts to regulate the initial steps of starch degradation at the granule surface. Remarkably, the most closely related protein to SEX4 outside the plant kingdom is laforin, a glucan-binding protein phosphatase required for the metabolism of the mammalian storage carbohydrate glycogen and implicated in a severe form of epilepsy (Lafora disease) in humans.

Animals↗

Influence of starches of low digestibility on the rat caecal microflora.

1. Male Sprague-Dawley rats were fed on either a purified, fibre-free diet or a diet in which half the maize starch was replaced with uncooked amylomaize or potato starch (equivalent to 100 or 200 g amylase-resistant starch (ARS)/kg diet respectively). Changes in short-chain fatty acids (SCFA), pH, ammonia and a number of bacterial variables in caecal contents were then assessed. 2. Both ARS supplements decreased caecal content pH by approximately 1-2 units, with an associated reduction in ammonia concentration. Potato starch significantly decreased the concentration of SCFA in the hindgut, while amylomaize supplementation increased propionic and butyric acids but decreased the occurrence of minor, branched-chain fatty acids. 3. Caecal bacterial biotransformation activities (beta-glucosidase (EC 3.2.1.21), beta-glucuronidase (EC 3.2.1.31), reduction of p-nitrobenzoic acid, apparent ammonia formation) were consistently decreased by both ARS sources. 4. The results demonstrate that amylase-resistant carbohydrate altered toxicologically important functions in the large-intestinal flora of the rat.

Ammonia↗

The effect of resistant starch on colon function in humans.

Starch that is resistant to human amylases forms during the cooking and subsequent cooling of some foods, and may therefore be a substrate for the bacterial flora of the colon. It is thus possible that resistant starch (RS) will affect colon function in a similar manner to non-starch polysaccharides. To test this theory, a group of eight volunteers took two diet supplements for 1 week each in a random order with a 1 week separation. One supplement comprised mainly 350 g Cornflakes/d and the other 380 g Rice Krispies/d, providing 10.33 and 0.86 g RS/d respectively. The amounts of amylase-digestible starch, nonstarch polysaccharides, total carbohydrate, energy, protein and fat were balanced between the two periods by giving small amounts of Casilan, wheat bran, butter and boiled sweets. The volunteers made faecal collections during day 3 to day 7 of each period. Whole-gut transit time was calculated using the continuous method. Stool consistency and ease of defaecation were assessed by the volunteers. All episodes of flatulence noticed were recorded in a diary, along with food intake. Serial breath hydrogen measurements were made at 15 min intervals for 8 h on day 1 of each supplement. Questionnaires regarding colon function were completed at the end of each dietary period. There were no significant differences in the stool mass, frequency or consistency, ease of defaecations, transit time or flatulence experienced during the two supplements (P greater than 0.05). Significantly more H2 (area under curve) was produced while eating Cornflakes than Rice Krispies (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Amylases↗

Alpha-amylase (EC 3.2.1.1) susceptibility rather than viscosity or gastric emptying rate controls plasma responses to starch in healthy humans.

The relationship between starch alpha-amylase (EC 3.2.1.1) susceptibility, plasma responses and gastric emptying rates has been investigated in humans. Nine randomly chosen healthy subjects were given three carbohydrate test meals (25 g starch or equivalent glucose units): two maize starch pastes with (a) 240 (S24) or (b) 500 (S50) g amylose/kg, and a glucose solution (GS). At 30 min, in vitro starch alpha-amylolysis was 48 (SD 4)% for S24 and 35 (SD 4)% for S50. Test meals differed in viscosity (mPa x s: S24, 54,000; S50, 190; GS, 4). Carbohydrates were labelled with 99mTechnetium and isotope gastric emptying was measured by external gamma counting. Carbohydrate isotopic gastric emptying patterns were exponential. Half gastric emptying time (min) was significantly (P less than 0.05) shorter for S50 (19(SD 2] than for GS (26(SD 2] or S24 (29(SD 2]. No correlation was found between half gastric emptying time and plasma response values. Values for peak insulin (pmol/l) above fasting were significantly (P less than 0.05) different: GS, 306 (SD 11); S24, 227 (SD 11); S50, 187 (SD 11). It is concluded that alpha-amylase susceptibility of the test carbohydrates is a determining factor in the insulin response of healthy subjects, while viscosity of the test meals and gastric emptying rate have no effect.

Adult↗

Effects of two fermentable carbohydrates (inulin and resistant starch) and their combination on calcium and magnesium balance in rats.

Resistant starch and inulin are complex carbohydrates that are fermented by the microflora and known to increase colonic absorption of minerals in animals. The fermentation of these substrates in the large bowel to short-chain fatty acids is the main reason for this increase in mineral absorption. The purpose of the present study was to examine the potential synergistic effect of a combination of these two fermentable carbohydrates. For this purpose, thirty-two adult male Wistar rats weighing 200 g were used in the present study. The rats were distributed into four groups, and fed for 21 d a fibre-free basal purified diet or diet containing 100 g inulin, or 150 g resistant starch (raw potato starch)/kg diet or a blend of 50 g inulin and 75 g resistant starch/kg diet. After an adaptation period of 14 d, the rats were then transferred to metabolic cages and dietary intake, faeces and urine were monitored for 5 d. The animals were then anaesthetized and caecal Ca and Mg absorption were measured. Finally, the rats were killed and blood, caecum and tissues were sampled. Ca and Mg levels were assessed in diets, faeces, urine, caecum and plasma by atomic absorption spectrometry. Our results confirmed that inulin and resistant starch ingestion led to considerable caecal fermentation in the three experimental groups compared with the control group diet. Moreover, both carbohydrates significantly increased the intestinal absorption and balance of Ca and Mg, without altering the plasma level of these two minerals. Interestingly, the combination of the studied carbohydrates increased significantly the caecal soluble Ca and Mg concentrations, the apparent intestinal absorption and balance of Ca, and non-significantly the plasma Mg level. In conclusion, a combination of different carbohydrates showed synergistic effects on intestinal Ca absorption and balance in rats. Further studies with other types of carbohydrate combinations should be carried out to extend these findings.

Animals↗

Inter-relationship of microbial activity, digestion and gut health in the rabbit: effect of substituting fibre by starch in diets having a high proportion of rapidly fermentable polysaccharides.

Caecal microbial activity, digestion and gut health were analysed in the young rabbit, in response to fibre substitution by starch, in diets with high proportions of rapidly fermentable polysaccharides (pectins+hemicelluloses:acid-detergent fibre (ADF) ratio of 1.7). A range of five diets corresponding to a 60 % linear reduction of the ADF level (230 to 92 g ADF/kg) without changes in the fibre quality, and to a corresponding linear increase in dietary starch, was given ad libitum to young rabbits from 18 d until 70 d of age. A one half reduction of the ADF level resulted in a sharp increase in energy digestibility (+25 units), associated with a lower feed intake (-35 %) and to an increase of the mean retention time (+6 h) in the whole digestive tract. Despite large variations in the fibre intake (20 to 59 g ADF/d), the fibre digestive efficiency remained similar among the five diets. Starch ileal concentrations were low after 4 weeks of age (<5 %), and variations with age were significant when the dietary starch level was over 19 %. A 65 % lower biomass production was measured when the ADF level progressed from 230 to 165 g/kg, and no precise relationship was found with fermentative activity. Reducing the fibre intake led to a linear decrease of caecal volatile fatty acids concentrations, and to higher pH and NH3 levels. An increased occurrence of mortality by diarrhoea was registered with the lowest fibre intake. It can be concluded that a sufficient supply of fibre, with high proportions in rapidly fermentable polysaccharides, stimulates the maturation of microbial activity and reduces the occurrence of diarrhoea.

Animal Feed↗

Ileal starch digestibility in growing broiler chickens fed on a wheat-based diet is improved by mash feeding, dilution with cellulose or whole wheat inclusion.

1. An experiment was conducted to study causes of low ileal starch digestibilities when broiler chickens were fed on wheat-based diets. Each of 5 cold-pelleted diets containing 771 g/kg DM wheat and one cold-pelleted diet containing 694 g/kg DM wheat were fed to 24 male broiler chickens in 8 cages from 10 to 21 d of age. 2. Feed intake and weight gain were significantly reduced when the wheat diet was crushed and fed in a mash form. 3. Ileal starch digestibility increased significantly from 0.79 to 0.95, 0.93 and 0.91, respectively, when the diet was crushed and fed in a mash form, was diluted with cellulose prior to pelleting, or when parts of the wheat were fed as whole grains. Correspondingly, random variation between individual birds was reduced. 4. These results indicate that an overload of wheat starch in the digestive tract may be the cause of poor digestibility for some broilers in a flock. Grinding of the wheat may also influence starch digestibility.

Animal Feed↗

Soft wheat instead of hard wheat in pelleted diets results in high starch digestibility in broiler chickens.

(1) The aim of the experiment was to re-examine variations in digestibilities of food components in 3-week-old broiler chickens fed on pelleted diets based on wheats differing in lipase activity and hardness. Fourteen wheat (Triticum aestivum) samples, each from a different cultivar, were included at 550 g/kg in 14 different diets given to male Ross broiler chicks from 7 d of age. The other main ingredients consisted of soyabean meal (353 g/kg) and rapeseed oil (55 g/kg). A 15th diet containing durum wheat (Triticum durum) was also tested. (2) Hardness of wheats (Triticum aestivum) varied between 14 (very soft) and 88 (very hard), and lipase activity of wheats varied from 1 to 13.1 (relative scale). No significant correlation was found between in vitro viscosities and other parameters such as hardness, particle size of wheat flours and lipase. Hardness was correlated with the mean particle size of wheat flours and durability of pellets. (3) Individual lipid digestibilities were negatively correlated with in vitro viscosities of wheats. (4) Individual starch digestibilities were negatively correlated with wheat hardness, particle size of wheat flour before pelleting, and pellet durability. The ratio of measured AME(N) to predicted AME(N) was also negatively correlated with wheat hardness. Simple regression calculation showed that a 100-point increase in wheat hardness resulted in a 3% decrease in the AME(N) value of diets. Multiple regression calculation showed the food/gain ratio (d 10 to d 21) to be positively related to wheat hardness and negatively related to pellet durability. (5) Wheat lipase activity was positively correlated with individual starch digestibility, which was the reverse of a result obtained in a previous experiment. Thus, wheat lipase activity did not seem consistent for predicting starch digestibility and AME(N) values. (6) Among all wheat samples, durum wheat showed the highest protein content and the lowest content of water-insoluble cell-wall. Starch digestibility of durum wheat tended to be lower than that of other wheats (0.916 vs 0.936). However, no significant difference in AME(N) was observed between the durum wheat sample and other wheats. (7) Gut morphometric data measured at d 24 did not show significant differences between dietary treatments.

Animal Feed↗

The ability of enteric diarrhoeal pathogens to ferment starch to short-chain fatty acids in vitro.

BACKGROUND: Short-chain fatty acids (SCFA), produced in the normal colon by bacterial fermentation, are decreased in acute diarrhoea. This may have deleterious effects on epithelial function in the colon. METHODS: The ability of several diarrhoeal pathogens to produce SCFA when incubated with starch in vitro was studied. Isolated pathogens were incubated for 24 h with either no added substrate, glucose, or starch under anaerobic conditions, and SCFA were quantitated by gas-liquid chromatography. RESULTS: Unlike the normal colonic flora, the pathogens produced acetate but not propionate or butyrate. D-Lactate was also produced by all the pathogens studied. When the pathogens were incubated in anaerobic medium containing starch, significantly greater amounts of acetate and significantly lesser amounts of lactate were produced. CONCLUSIONS: The inability of enteric pathogens to produce butyrate may impair epithelial cell function, whereas production of D-lactate may enhance mucosal damage in diarrhoeal disease. The presence of luminal starch may be helpful in shifting the fermentation profile to a more favourable pattern.

Butyrates↗

Development of suspending agent from sodium carboxymethyl mungbean starches.

Three sodium carboxymethyl mungbean starches (SCMMSs) were selected, based on the physicochemical profiles and evaluated as potential pharmaceutical suspending agent in comparison with a sodium carboxymethyl tapioca starch and other five commercial suspending agents. Ibuprofen suspension was employed as a model formulation with 0.5, 1.0, and 2.0% w/v of suspending agents. Evaluation parameters included the determination of sedimentation volume ratio, redispersibility, viscosity and rheological properties and content uniformity studies. The results revealed that a high-viscosity modified mungbean starch, MMS-M-04, possessed suitable properties as a suspending agent and, at 1% concentration, was as effective as sodium carboxymethylcellulose and xanthan gum, two most-commonly used suspending agents. This modified starch could be further developed and employed as a new commercial suspending agent in the pharmaceutical industry.

Excipients↗

Influence of time, temperature, moisture, ingredients, and processing conditions on starch gelatinization.

Starch gelatinization phenomena is extremely important in many food systems. This review focuses on factors affecting gelatinization characteristics of starch. Important variables which must be considered in design of processes in which starch undergoes gelatinization are heat of gelatinization and temperature of gelatinization. Major interactions are reviewed for the effects of lipids, moisture content, nonionic constituents and electrolytes on these characteristics. Furthermore, treatment of starch-containing systems prior to heating into the gelatinization temperature range can have a significant effect on ultimate gelatinization characteristics.

Amylose↗

Whole grains--impact of consuming whole grains on physiological effects of dietary fiber and starch.

Much of the present research on the physiological effects of dietary fiber and starch has been done on sources isolated from the parent material, and it is not clear whether they have the same effects if fed in the intact or whole grain. For dietary fiber, physiological effect depends on extent of fermentation in the large intestine, and this is influenced by chemical composition, solubility, physical form, and presence of lignin or other compounds. All of these factors are altered by isolation of a fiber source from the whole grain, and hence effects of eating fiber vary. Similarly, physical form and presence in the whole grain will affect digestibility of starch in the small intestine, which in turn influences the glycemic response and colonic effects determined by the extent of malabsorption and entry into the colon. Starch that enters the colon is fermented and produces short-chain fatty acids, particularly butyrate, which is necessary to maintain a healthy mucosa. Hence, their presence within the whole grain may have important implications for health for both dietary fiber and starch. Evidence indicates that such effects are beneficial and that whole-grain consumption should be encouraged.

Dietary Carbohydrates↗