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Comparison of Starch and ADP-Glucose Pyrophosphorylase Levels in Nonembryogenic Cells and Developing Embryos from Induced Carrot Cultures.

Cultures of carrot (Daucus carota L.) in a medium without added 2,4-dichlorophenoxyacetic acid were separated into fractions of embryos at different stages of development (large globular and heart, torpedo, and germinating) and nonembryogenic cells. The average starch content per cell in these fractions was similar. However, due to the smaller sizes of the cells of the embryos relative to the nonembryogenic cells, starch content per weight of tissue was higher in the embryos. The ADP-glucose pyrophosphorylase activity per cell in the nonembryogenic cells was double that of the embryo cells. Furthermore, the ratio of ADP-glucose pyrophosphorylase to starch was over 2-fold higher in the nonembryogenic cells, indicating that starch content is not simply determined by ADP-glucose pyrophosphorylase levels. ADP-glucose pyrophosphorylase activity of all culture fractions was directly proportional to the level of a single 50 kilodalton polypeptide detected by immunoblot analysis, using antiserum raised to the purified spinach leaf enzyme. In the same immunoblot analysis, novel polypeptides of 63 and 100 kilodalton were detected in embryos but were absent from nonembryogenic cells. This is one of the few reported examples of specific proteins which differentially accumulate in embryos and nonembryogenic cells.

Journal Article↗

Starch Phosphorylase Inhibitor Is beta-Amylase.

The proteinaceous noncompetitive inhibitor of starch phosphorylase isolated from the root of sweet potato (Ipomoea batatas [L.] Lam.) (TC Chang, JC Su 1986 Plant Physiol 80: 534-538) has been identified as a beta-amylase. The starch phosphorylase inhibitor and beta-amylase activities copurified to give a protein indistinguishable from commercial beta-amylase by electrophoretic and immunological methods, and the two activities showed parallel responses in pH, temperature, and inhibitor sensitivity tests. The amylolytic pattern of the inhibitor corresponded to that of beta-amylase and its inhibitory effect toward starch phosphorylase was due to neither deprivation of starch, the primer for the phosphorylase assay, nor the inhibitory effect of amylolytic products.

Journal Article↗

ADP-Glucose Transport by the Chloroplast Adenylate Translocator Is Linked to Starch Biosynthesis.

In organello starch biosynthesis was studied using intact chloroplasts isolated from spinach leaves (Spinacia oleracea). Immunoblot analysis using a specific antiserum against the mitochondrial adenylate (ADP/ATP) translocator of Neurospora crassa shows the presence of an adenylate translocator protein in the chloroplast envelope membranes, similar to that existing in mitochondria and amyloplasts from cultured cells of sycamore (Acer pseudoplatanus). The double silicone oil layer-filtering centrifugation technique was employed to study the kinetic properties of adenylate transport in the purified chloroplasts; ATP, ADP, AMP, and most importantly ADP-Glc were shown to be recognized by the adenylate translocator. Similar to the situation with sycamore amyloplasts, only ATP and ADP-Glc uptake was inhibited by carboxyatractyloside, an inhibitor of the mitochondrial adenylate translocator. Evidence is presented to show that the ADP-Glc transported into the chloroplast stroma is utilized for starch synthesis catalyzed by starch synthase (ADP-Glc:1,4-alpha-d-glucan 4-alpha-d-glucosyltransferase). The high activity of sucrose synthase producing ADP-Glc observed in the extrachloroplastic fractions suggests that starch biosynthesis in chloroplasts may be coupled with the direct import of ADP-Glc from the cytosol.

Journal Article↗

Fermentation of dietary starch in humans.

OBJECTIVE: Dietary starch that escapes digestion in the small intestine may be quantitatively more important than dietary fiber as a substrate for fermentation. The products of fermentation have important implications in the pathogenesis of colorectal cancer and other diseases of the large bowel, which are uncommon in Africans but have a high prevalence in Western populations. METHODS: Maize porridge is a staple of most blacks in South Africa. Stale maize porridge (high-resistant starch [HRS]) seems to induce greater fermentation in the large bowel than fresh maize porridge (low-resistant starch [LRS]). RESULTS: In the present study, healthy colostomy subjects fed stale maize porridge had significantly more production of short-chain fatty acids (SCFA) (mean SCFA, HRS = 182.6; mean SCFA, LRS = 116.1; p < 0.05) in their colostomy effluent together with a significant drop in stool pH (mean pH, HRS = 5.91; mean pH, LRS = 6.70; p < 0.001). The SCFA butyrate (mean, HRS = 35.1; mean, LRS = 17.6; p < 0.05) and acetate (mean, HRS = 93.9; mean, LRS = 65.8; p < 0.05) were significantly elevated on the stale maize porridge diet when compared with consumption of fresh maize porridge. SCFA propionate (mean, HRS = 43.1; mean, LRS = 24.8; p = 0.05), also increased with stale maize porridge, but was not statistically significant. CONCLUSION: A high-resistant starch diet and its resultant increase in fermentation products may be partly responsible for protecting the black population against colorectal cancers and other large bowel diseases.

Acetic Acid↗

Utilization of low concentrations of starch by a flavobacterium species isolated from tap water.

Experiments in well-cleaned glass flasks revealed that addition of starch in concentrations of 10 and 25 mug of substrate C per liter to the filtrate of slow sand filters stimulated the development of a yellow-pigmented bacterium which was identified as a Flavobacterium species. The isolate was able to multiply in tap water without substrates added, but addition of starch and glucose in amounts as low as 1 mug of substrate C per liter clearly enhanced growth. The substrate affinities of the Flavobacterium for these compounds were 3.9 mug of starch C and 3.3 mug of glucose C per liter. The results of this study indicate that microorganisms which rapidly utilize starch at a level of a few micrograms per liter commonly occur in water.

Journal Article↗

Improved method for detection of starch hydrolysis.

A new starch hydrolysis detection method which does not rely on iodine staining or the use of color-complexed starch is described. A linear relationship was obtained with agar-starch plates when net clearing zones around colonies of yeasts were plotted against enzyme levels (semilogarithm scale) produced by the same yeast strains in liquid medium. A similar relationship between starch clearing zones and alpha-amylase levels from three different sources was observed. These observations suggest that the method is useful in mutant isolations, strain improvement programs, and the prediction of alpha-amylase activities in culture filtrates or column effluents.

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Production of l-Phenylalanine from Starch by Analog-Resistant Mutants of Bacillus polymyxa.

p-Fluorophenylalanine-resistant mutants of starch-degrading Bacillus polymyxa ATCC 842, generated by ethyl methanesulfonate mutagenesis followed by incubation with caffeine, overproduced small amounts of l-phenylalanine (l-phe) from starch. A beta-2-thienylalanine-resistant mutant (BT-7) derived from p-fluorophenylalanine mutant (C-4000 FP-4) and resistant to both p-fluorophenylalanine and beta-2-thienylalanine produced 0.5 g of l-phe and 0.15 g of l-tyrosine per liter from 10 g of starch per liter when growing in a minimal medium. trans-Cinnamic acid (CA) was also excreted by both mutants, indicating the possibility of l-phenylalanine ammonia-lyase-induced deamination of l-phe to CA. The amount of l-phe-derived CA detected in BT-7 was less compared with mutant C-4000 FP-4. CA production was induced in the parent only when l-phe was used as a sole nitrogen source. Time of CA production in the two mutants could be delayed by addition of other nitrogen sources, an indication of possible l-phenylalanine ammonia-lyase inhibition or repression. The presence of l-phenylalanine ammonia-lyase in B. polymyxa mutant C-4000 FP-4 was confirmed by assays of cell-free extracts from cells grown in starch minimal medium containing l-phe as the sole nitrogen source. Preliminary studies of the regulation of deoxy-d-arabino-heptulosonate-7-phosphate synthase and prephenate dehydratase in the wild-type strain showed that deoxy-d-arabino-heptulosonate-7-phosphate synthase was subject to feedback inhibition by l-phe, l-tyrosine, and l-tryptophan. Inhibition by each amino acid was to a similar extent singly or in combination at a 0.5 mM level of each amino acid. Prephenate dehydratase was feedback inhibited by l-phe, but not by l-tyrosine or l-tryptophan or both. In the double analog-resistant mutant BT-7, deoxy-d-arabino-heptulosonate-7-phosphate synthase had specific activity similar to that in the wild type, and the enzyme was still subject to feedback inhibition. However, prephenate dehydratase had increased specific activity and it was also insensitive to feedback inhibition by l-phe. The overproduction of aromatic amino acids by BT-7 was thought to be due, at least in part, to deregulation of feedback inhibition of prephenate dehydratase. Chorismate mutase was not subject to feedback inhibition in the wild type and was unaffected in the mutant.

Journal Article↗

Degradation of degradable starch-polyethylene plastics in a compost environment.

The degradation performance of 11 types of commercially produced degradable starch-polyethylene plastic compost bags was evaluated in municipal yard waste compost sites at Iowa State University (Ames) and in Carroll, Dubuque, and Grinnell, Iowa. Masterbatches for plastic production were provided by Archer Daniels Midland Co. (Decatur, Ill.), St. Lawrence Starch Co. Ltd. (Mississauga, Ontario, Canada), and Fully Compounded Plastics (Decatur, Ill.). Bags differed in starch content (5 to 9%) and prooxidant additives (transition metals and a type of unsaturated vegetable oil). Chemical and photodegradation properties of each material were evaluated. Materials from St. Lawrence Starch Co. Ltd. and Fully Compounded Plastics photodegraded faster than did materials from Archer Daniels Midland Co., whereas all materials containing transition metals demonstrated rapid thermal oxidative degradation in 70 degrees C-oven (dry) and high-temperature, high-humidity (steam chamber) treatments. Each compost site was seeded with test strips (200 to 800 of each type) taped together, which were recovered periodically over an 8- to 12-month period. At each sampling date, the compost row temperature was measured (65 to 95 degrees C), the location of the recovered test strip was recorded (interior or exterior), and at least four strips were recovered for evaluation. Degradation was followed by measuring the change in polyethylene molecular weight distribution via high-temperature gel permeation chromatography. Our initial 8-month study indicated that materials recovered from the interior of the compost row demonstrated very little degradation, whereas materials recovered from the exterior degraded well. In the second-year study, however, degradation was observed in several plastic materials recovered from the interior of the compost row by month 5 at the Carroll site and almost every material by month 12 at the Grinnell site. The plastic bags collected from each community followed a similar degradation pattern. To our knowledge, this is the first scientific study demonstrating significant polyethylene degradation by these materials in a compost environment.

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THE SYNTHESIS OF THE STARCH GRANULE.

This review describes and discusses the implications of recent discoveries about how starch polymers are synthesized and organized to form a starch granule. Three issues are highlighted. 1. The role and importance of ADPglucose pyrophosphorylase in the generation of ADPglucose as the substrate for polymer synthesis. 2. The contributions of isoforms of starch-branching enzyme, starch synthase, and debranching enzyme to the synthesis and ordered packing of amylopectin molecules. 3. The requirements for and regulation of the synthesis of amylose.

Journal Article↗

Role of olfaction in starch and oil preference.

Two different techniques were used to assess the role of olfaction in the preference rats show for dilute nutrient suspensions and solutions. Removal of the olfactory bulbs reduced but did not abolish preference for 1% starch and oil emulsions. Removal of the olfactory bulbs also reduced preference for sucrose, Polycose, and saccharin. In another experiment, rats were rendered anosmic with intranasal zinc sulfate. The degree of anosmia was evaluated by measuring preference for an aqueous solution of citral and vanillin. Rats whose avoidance of citral plus vanillin had been abolished by zinc sulfate continued to prefer a 1% starch suspension over vehicle, although starch preference was significantly reduced. It is concluded that starch and oil preference is mediated by both olfactory and nonolfactory cues.

Animals↗

Microbial hydrogen production from sweet potato starch residue.

Clostridium butyricum could produce hydrogen from a sweet potato starch residue upon supplementation of nitrogen sources. A repeated batch culture using a mixed culture of C. butyricum and Enterobacter aerogenes produced hydrogen with a yield of 2.4 mol H2/mol glucose under a controlled culture pH of 5.25 in a medium consisting of the sweet potato starch residue and 0.1% Polypepton without addition of any reducing agents. Rhodobacter sp. M-19 produced hydrogen from the supernatant of the culture broth obtained in the repeated batch culture containing C. butyricum and E. aerogenes when 50 microg/l Na2MoO4.2H2O and 20 mg/l EDTA were added to the supernatant and it was cultured under a controlled culture pH of 7.5. A high yield of hydrogen of 7.0 mol H2/mol glucose from the starch remaining in the starch residue was attained in two-step repeated batch cultures containing C. butyricum and E. aerogenes, and by Rhodobacter sp. M-19.

Journal Article↗

[Effects of dietary fiber on the glucose tolerance in spontaneously diabetic rats--comparison among barley, rice, and corn starch].

OBJECTIVES: In this study, the effects of a barley diet containing high dietary fiber on the onset and development of diabetes mellitus in spontaneously diabetic rats was investigated by comparing with a rice diet containing low dietary fiber and an alpha-corn starch diet containing very low dietary fiber. METHODS: 30 male Goto-Kakizaki (GK) strain rats (8 weeks of age) were randomly assigned to 3 groups; high barley (HB) group on a barley diet (dietary fiber intake, 1.79 g/day/rat), rice (R) group on a rice diet (dietary fiber intake, 0.46 g/day/rat), and alpha-corn starch (CS) group on an alpha-corn starch diet (dietary fiber intake, 0.24 g/day/rat). The carbohydrate (70%), fat (10%), and protein (20%) contents of these 3 diets were equal, and the rats were pair-fed each diet for 3 months. RESULTS: Feeding for 3 months showed that fasting plasma glucose level in the HB group was significantly lower than in the R and CS groups; the glucose tolerance in the HB group was markedly improved. Moreover, the plasma cholesterol and triglyceride levels in the HB group were significantly lower than those of the R and CS groups. CONCLUSIONS: Our findings demonstrated that barley enabled glycemic control and improved glucose tolerance compared with rice or alpha-corn starch.

Animals↗

The effect of varying dietary starch and fat content on serum creatine kinase activity and substrate availability in equine polysaccharide storage myopathy.

The effect of dietary starch and fat content on serum creatine kinase (CK) activity and substrate availability was evaluated in 4 mares of Quarter Horse-related breeds with polysaccharide storage myopathy (PSSM). Four isocaloric diets ranging in digestible energy (DE) from 21.2% (diet A), 14.8% (B), 8.4% (C), to 3.9% (D) for starch, and 7.2% DE (diet A), 9.9% (B), to 12.7% DE (diet C and D) for fat were fed for 6-week periods (4 weeks with exercise) using a 4 X 4 Latin square design. Postprandial glucose and insulin responses were measured, and 4 hours postexercise, serum CK activity, glucose, insulin, free fatty acids (FFA), and beta-hydroxybutyrate (beta-HBA) were analyzed. Glycogen, glucose-6-phosphate, citrate synthase, 3-hydroxy-acyl-CoA dehydrogenase, lactate dehydrogenase as well as abnormal polysaccharide and lipid content were measured in middle gluteal muscle samples. Postprandial insulin and glucose response was higher for diet A versus D. Log CK activity was higher with diets A, B, and C versus D. Daily insulin was higher and FFA lower on diet A versus B, C, and D, whereas glucose varied only slightly with diet. Muscle oxidative capacity and lipid stores were low in PSSM horses and muscle glycogen and abnormal polysaccharide content high on both diets A and D. Individual variation occurred in the response of PSSM horses to diets differing in starch and fat content. However, for those horses with clinical manifestations of PSSM, a diet with <5% DE starch and >12% DE fat can reduce exertional rhabdomyolysis, potentially by increasing availability of FFA for muscle metabolism.

Animals↗

Supplemental cracked corn for steers fed fresh alfalfa: I. Effects on digestion of organic matter, fiber, and starch.

The effect of supplementation with different levels of cracked corn on the sites of OM, total dietary fiber (TDF), ADF, and starch digestion in steers fed fresh alfalfa indoors was determined. Six Angus steers (338 +/- 19 kg) fitted with cannulas in the rumen, duodenum, and ileum consumed 1) alfalfa (20.4% CP, 41.6% NDF) ad libitum (AALF); 2), 3), and 4) AALF supplemented (S) with .4, .8, or 1.2%, respectively, of BW of corn; or 5) alfalfa restricted at the average level of forage intake of S steers (RALF), in a 5 x 5 Latin square design. Total OM intake was lower (P < .01) in steers fed RALF than in those fed AALF but level of forage intake did not affect sites of OM, TDF, or starch digestion (P > .05). Forage OM intake decreased (P < .01) linearly (8,496 to 5,840 g/d) but total OM intake increased (P = .03) linearly (8,496 to 9,344 g/d) as corn increased from .4 to 1.2% BW. Ruminal apparent and true OM disappearance was not affected, but OM disappearing in the small intestine increased (P < .01) linearly with increasing levels of corn. Total tract OM digestibility (71.2 to 76.2%) and the proportion of OM intake that was digested in the small intestine (15.4 to 24.5%) increased (P < .01) linearly as corn increased. The TDF and ADF intakes decreased (P < .01) linearly as level of corn increased. Total tract TDF and ADF digestibilities were not different among treatments (average 62.9 and 57.8%, respectively). Starch intake and starch digested in the rumen and small and large intestine increased (P < .01) linearly with increasing corn level. Ruminal pH and VFA concentrations decreased and increased (P < .01), respectively, with increasing corn. Supplementation with corn increased OM intake, decreased forage OM intake, and increased the proportion of OM that was digested in the small intestine, but fiber digestion was not affected.

Animals↗

Effects of limestone on starch digestion in Holstein steers.

Eight calcitic and three dolomitic limestones were examined for particle size and reactivity with hydrochloric acid at pH 3.5 and 5.5. Reactivity was higher in calcitic than dolomitic limestone. Within calcitic limestones, reactivity was highly related to particle size. A highly reactive limestone was selected for use in two digestion trials with Holstein steers. In Trial 1, 4 steers (approximately 300 kg) were fed diets consisting of 40% corn silage and 60% concentrate (DM basis) with .62 or 2.34% limestone in a single reversal trial with 3-wk experimental periods. Added limestone affected only starch digestibility, which increased from 91.3 to 93.2%. Fecal pH increased from 5.7 to 6.4 by limestone addition. In trial 2, four rumen and duodenally cannulated steers (approximately 475 kg) were fed the same diets in the same design. Limestone addition increased fecal pH from 5.7 to 6.5. There were no effects of limestone on rumen or total tract digestion of starch or other nutrients. Limestone addition had no effect on rumen pH, buffering capacity, or VFA concentrations. Addition of a highly reactive limestone to diets containing high amounts of starch had little effect on starch digestion in either the rumen or lower gastrointestinal tract.

Animals↗

[Genetic analysis of starch branching enzyme activity in rice grain].

To make an genetic analysis of starch branching enzyme (Q enzyme) inheritance, which is important in catalyzing the formation of amylopectin and thereby raising the amylopectin/amylose ratio of starch in rice grains, a linkage map of 207 DNA markers were constructed by using 247 recombinant inbred lines derived from an indica-indica rice cross Zhenshan 97B/Milyang46. In 2002 and 2003, the activities of starch branching enzyme of the parents and 247 RILs were measured 10 d and 20 d respectively after flowering (Table 1). A total of 3 quantitative trait loci (QTLs) were detected to have significant additive effects on Q enzyme activities 10 d after flowering with 10% phenotypic variations explained for the three QTLs (Table 2, Fig.1). Meanwhile, qQ10-6 with significant QTL x environment interaction was detected. Epistasis analysis detected 5 and 2 significant additive-by-additive interactions for Q enzyme 10 d and 20 d respectively after flowering (Table 3), and three pairs of QTLs 10 d after flowering with significant epistasis x environment interactions were detected, explaining 3% to 12% of the phenotypic variation. The results showed that the environmental factors had obvious effect on the gene expression of Q enzyme activity in rice grain. Meanwhile, the location of qQ10-6 in relation to that of QTLs for gelatinization temperature, gene loci involved in ADPG pyrophosphorylase and soluble starch synthase were discussed with the help of a genetic map.

1,4-alpha-Glucan Branching Enzyme↗

Effect of starch malabsorption on fecal bile acids and neutral sterols in humans: possible implications for colonic carcinogenesis.

Epidemiological and experimental studies indicate a strong association between an elevated colon cancer risk and increased fecal excretion of secondary bile acids, neutral sterols, and prolonged gastrointestinal transit time. Starch malabsorption, on the other hand, has been reported to be a possible protective factor in colon carcinogenesis. To study the impact of starch malabsorption on these parameters, 12 healthy volunteers consumed a diet rich in starch for two 4-week periods. During a double-blind crossover trial they received the alpha-glucosidase inhibitor acarbose (BAY g 5421) in one of the study periods and placebo in the other. During acarbose treatment stool wet weight increased by 68%, stool dry weight by 57%, and gastrointestinal mean transit time by 30%. Fecal concentrations (mg/g dry weight) of the neutral sterols coprostanol, coprostanone, campesterol, 4-cholesten-3-one, and beta-sitosterol decreased by 36.8, 48.7, 42.1, 34.6, and 39.4%, respectively, under acarbose. Concentrations of the major secondary bile acids, deoxycholic and lithocholic acid, decreased by 59.9 and 52.2%, respectively. In spite of an increased stool weight, also daily excretion (mg/day) of these two bile acids was lower under acarbose (47.9 and 36.6%, respectively) compared to placebo, whereas excretion of the main primary bile acid, cholic acid, rose from 22.58 mg/day to 379.80 mg/day during the acarbose period. The changes in fecal bile acid and neutral sterol excretion found during acarbose treatment may explain a protective effect of starch malabsorption on colon cancer development.

Acarbose↗

Effects of cholestyramine on low density lipoprotein binding sites on liver membranes from rabbits with endogenous hypercholesterolemia induced by a wheat starch-casein diet.

Rabbits fed a wheat starch-casein diet develop a marked hypercholesterolemia with a lipoprotein distribution similar to that of humans. Approximately 76% of the total cholesterol is carried in the low density lipoprotein (LDL) fraction (1.006 less than d less than 1.063 g/ml). Inclusion of 1% cholestyramine in the diet prevents the increase in plasma cholesterol. The cholestyramine effect is mediated through an increased fractional catabolic rate of 125I-LDL. In order to determine the potential role of hepatic LDL receptors in the removal of LDL from the plasma, binding of 125I-LDL and 125I-beta-VLDL (beta-migrating very low density lipoproteins) to hepatic membranes prepared from livers of rabbits fed the wheat starch-casein diet with or without cholestyramine supplementation was investigated. Membranes from livers of the cholestyramine-supplemented animals exhibit high levels of specific EDTA-sensitive binding of either of the 125I-labeled lipoproteins. Very little EDTA-sensitive binding occurs on liver membranes from wheat starch-casein-fed rabbits that have not been treated with cholestyramine. These results indicate that the hypercholesterolemia in rabbits associated with the wheat starch-casein diet is wholly or partially the result of a decreased number of specific hepatic LDL receptors and thus a decreased catabolism of plasma cholesterol. The response of the liver to the inclusion in the diet of the bile acid sequestrant, cholestyramine, is to maintain or increase the number of specific LDL binding sites, thus promoting catabolism of plasma cholesterol.

Animals↗