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Adaptation of the rumen microbial population to native potato starch degradation determined with the gas production technique and the nylon bag technique.

Experiments were conducted to investigate the influence of the adaptation of rumen micro-organisms on the degradation of native potato starch (PS) in the rumen. Cows were fed with rations used for gas production (GP) analysis (dry cows, 1.6% starch) and for the nylon bag (NB) technique (lactating cows, 23% starch, mainly maize starch) and a ration containing 19% native PS (lactating cows). Fermentation characteristics of 13 samples were investigated with the GP technique using rumen fluid from cows fed each of the three rations. The same samples were investigated with the NB technique in the cows obtaining the NB ration and the PS ration. The results showed that the rate of GP was influenced by the source of the rumen fluid. The fermentation rate of PS was considerably enhanced by using rumen fluid adapted to the fermentation of native PS instead of using the other rumen fluids. Incubating in cows fed the PS ration, the rate of PS degradation determined with the NB technique, was higher compared with cows fed other rations. Using the PS ration the observed lag period for PS was shorter. The results show a clear influence of ration on the degradation characteristics of starch, determined with both the GP technique and the NB technique. However, these changes in behaviour did not explain observed differences in amounts of rumen escape PS measured in vivo in animal experiments and in situ, using the NB technique.

Adaptation, Physiological↗

Postoperative starch granuloma revealed as femoral herniation. A case report.

A rare case of postoperative starch granuloma in a 55-year-old woman, revealed as femoral herniation, is reported. A small, finger-tip-sized tumor showing femoral herniation was noticed 3 weeks after cholecystectomy for cholelithiasis, and no content in the hernia sac was noticed. Histologically, the granuloma situated in the wall of the hernia sac was diagnosed as a starch granuloma consisting of starch granules, which were clearly revealed by polarized light microscopy. These granules were detected in the cytoplasm of macrophages including multinucleated types, some showing a positive immune reaction with antibodies to the alpha-subunit of S-100 protein, vimentin and lysozyme. Starch granules from surgical gloves showed identical features by polarized light microscopy, thus proving the histogenesis of this granuloma. This is the first reported case in the Japanese literature of starch granuloma manifested as femoral herniation.

Cholecystectomy↗

Structure of the complex of a yeast glucoamylase with acarbose reveals the presence of a raw starch binding site on the catalytic domain.

Most glucoamylases (alpha-1,4-D-glucan glucohydrolase, EC 3.2.1.3) have structures consisting of both a catalytic and a starch binding domain. The structure of a glucoamylase from Saccharomycopsis fibuligera HUT 7212 (Glu), determined a few years ago, consists of a single catalytic domain. The structure of this enzyme with the resolution extended to 1.1 A and that of the enzyme-acarbose complex at 1.6 A resolution are presented here. The structure at atomic resolution, besides its high accuracy, shows clearly the influence of cryo-cooling, which is manifested in shrinkage of the molecule and lowering the volume of the unit cell. In the structure of the complex, two acarbose molecules are bound, one at the active site and the second at a site remote from the active site, curved around Tyr464 which resembles the inhibitor molecule in the 'sugar tongs' surface binding site in the structure of barley alpha-amylase isozyme 1 complexed with a thiomalto-oligosaccharide. Based on the close similarity in sequence of glucoamylase Glu, which does not degrade raw starch, to that of glucoamylase (Glm) from S. fibuligera IFO 0111, a raw starch-degrading enzyme, it is reasonable to expect the presence of the remote starch binding site at structurally equivalent positions in both enzymes. We propose the role of this site is to fix the enzyme onto the surface of a starch granule while the active site degrades the polysaccharide. This hypothesis is verified here by the preparation of mutants of glucoamylases Glu and Glm.

Acarbose↗

The granulation of ternary mixtures containing lactose and boric acid with different starches.

Ternary mixtures of lactose, boric acid and starch have been granulated by massing and screening. The properties of the granules have been compared with those resulting from granulation of blends of lactose and boric acid without starch. Potato, maize and rice starches have been studied as the third component at concentrations similar to those employed in tablet formulations. The starches caused a reduction in both mean granule size and granule strength. The effect is dependent on the proportions of the excipients. Increasing the amount of binder solution used gave bigger, stronger granules but did not compensate for the effects produced by the starches.

Boric Acids↗

Screening for and identification of starch-, amylopectin-, and pullulan-degrading activities in bifidobacterial strains.

Forty-two bifidobacterial strains were screened for alpha-amylase and/or pullulanase activity by investigating their capacities to utilize starch, amylopectin, or pullulan. Of the 42 bifidobacterial strains tested, 19 were capable of degrading potato starch. Of these 19 strains, 11 were able to degrade starch and amylopectin, as well as pullulan. These 11 strains, which were shown to produce extracellular starch-degrading activities, included 5 strains of Bifidobacterium breve, 1 B. dentium strain, 1 B. infantis strain, 3 strains of B. pseudolongum, and 1 strain of B. thermophilum. Quantitative and qualitative enzyme activities were determined by measuring the concentrations of released reducing sugars and by high-performance thin-layer chromatography, respectively. These analyses confirmed both the inducible nature and the extracellular nature of the starch- and pullulan-degrading enzyme activities and showed that the five B. breve strains produced an activity that is consistent with type II pullulanase (amylopullulanase) activity, while the remaining six strains produced an activity with properties that resemble those of type III pullulan hydrolase.

Amino Acid Sequence↗

Effect of regulatory protein levels on utilization of starch by Bacteroides thetaiotaomicron.

Bacteroides thetaiotaomicron, a gram-negative obligate anaerobe, appears to utilize starch by first binding the polymer to its surface and then translocating it into the periplasmic space. Several genes that encode enzymes or outer membrane proteins involved in starch utilization have been identified. These have been called sus genes, for starch utilization system. Previous studies have shown that sus structural genes are regulated at the transcriptional level and their expression is induced by maltose. We report here the identification and characterization of a gene, susR, which appears to be responsible for maltose-dependent regulation of the sus structural genes. The deduced amino acid sequence of SusR protein had a helix-turn-helix motif at its carboxy-terminal end, and this region had highest sequence similarity to the corresponding regions of known transcriptional activators. A disruption in susR eliminated the expression of all known sus structural genes, as expected if susR encoded an activator of sus gene expression. The expression of susR itself was not affected by the growth substrate and was not autoregulated, suggesting that binding of SusR to maltose might be the step that activates SusR. Three susR-controlled structural genes, susA, susB, and susC, are located immediately upstream of susR. These genes are organized into two transcriptional units, one containing susA and another containing susB and susC. susA was expressed at a lower level than susBC, and susA expression was more sensitive to the gene dosage of susR than was that of the susBC operon. An unexpected finding was that increasing the number of copies of susR in B. thetaiotaomicron increased the rate of growth on starch. This effect could be due to higher levels of susA expression. Whatever the explanation, the level of SusR in the cell appears to be a limiting factor for growth on starch.

Amino Acid Sequence↗

Characterization of four outer membrane proteins that play a role in utilization of starch by Bacteroides thetaiotaomicron.

Results of earlier work had suggested that utilization of polysaccharides by Bacteroides spp. did not proceed via breakdown by extracellular polysaccharide-degrading enzymes. Rather, it appeared that the polysaccharide was first bound to a putative outer membrane receptor complex and then translocated into the periplasm, where the degradative enzymes were located. In a recent article, we reported the cloning and sequencing of susC, a gene from Bacteroides thetaiotaomicron that encoded a 115-kDa outer membrane protein. SusC protein proved to be essential for utilization not only of starch but also of intermediate-sized maltooligosaccharides (maltose to maltoheptaose). In this paper, we report the sequencing of a 7-kbp region of the B. thetaiotaomicron chromosome that lies immediately downstream of susC. We found four genes in this region (susD, susE, susF, and susG). Transcription of these genes was maltose inducible, and the genes appeared to be part of the same operon as susC. Western blot (immunoblot) analysis using antisera raised against proteins encoded by each of the four genes showed that all four were outer membrane proteins. Protein database searches revealed that SusE had limited similarity to a glucanohydrolase from Clostridium acetobutylicum and SusG had high similarity to amylases from a variety of sources. SusD and SusF had no significant similarity to any proteins in the databases. Results of 14C-starch binding assays suggested that SusD makes a major contribution to binding. SusE and SusF also appear to contribute to binding but not to the same extent as SusD. SusG is essential for growth on starch but appears to contribute little to starch binding. Our results demonstrate that the binding of starch to the B. thetaiotaomicron surface involves at least four outer membrane proteins (SusC, SusD, SusE, and SusF), which may form a surface receptor complex. The role of SusG in binding is still unclear.

Bacterial Outer Membrane Proteins↗

Transcriptional and biochemical analysis of starch metabolism in the hyperthermophilic archaeon Pyrococcus furiosus.

Pyrococcus furiosus utilizes starch and its degradation products, such as maltose, as primary carbon sources, but the pathways by which these alpha-glucans are processed have yet to be defined. For example, its genome contains genes proposed to encode five amylolytic enzymes (including a cyclodextrin glucanotransferase [CGTase] and amylopullulanase), as well as two transporters for maltose and maltodextrins (Mal-I and Mal-II), and a range of intracellular enzymes have been purified that reportedly metabolize maltodextrins and maltose. However, precisely which of these enzymes are involved in starch processing is not clear. In this study, starch metabolism in P. furiosus was examined by biochemical analyses in conjunction with global transcriptional response data for cells grown on a variety of glucans. In addition, DNA sequencing led to the correction of two key errors in the genome sequence, and these change the predicted properties of amylopullulanase (now designated PF1935*) and CGTase (PF0478*). Based on all of these data, a pathway is proposed that is specific for starch utilization that involves one transporter (Mal-II [PF1933 to PF1939]) and only three enzymes, amylopullulanase (PF1935*), 4-alpha-glucanotransferase (PF0272), and maltodextrin phosphorylase (PF1535). Their expression is upregulated on starch, and together they generate glucose and glucose-1-phosphate, which then feed into the novel glycolytic pathway of this organism. In addition, the results indicate that several hypothetical proteins encoded by three gene clusters are also involved in the transport and processing of alpha-glucan substrates by P. furiosus.

Archaeal Proteins↗

Does super efficient starch absorption promote diverticular disease?

The amount of starch escaping absorption in the small intestine was measured in eight patients with symptomatic diverticular disease and eight controls. Unabsorbed starch was calculated from breath hydrogen measurements after a potato meal compared with the hydrogen response to lactulose. The proportion of unabsorbed starch was low in all the patients (mean 3.3%) and was only about a quarter of that in the controls (12.4%; p less than 0.01). These findings confirm that unabsorbed starch provides an important quantity of carbohydrate reaching the colon and suggest that super efficient starch absorption, by reducing this provision, may promote the development of diverticular disease.

Aged↗

Food-starch granulomatous peritonitis.

Two cases of peritoneal granulomatous reactions to food starch are described. They followed bowel perforation and clinically mimicked tuberculous and glove-powder starch peritonitis. Their histological differences from corn-starch peritonitis warrant attention in the absence of previous documentation of starch as a component of peritoneal food granulomas. Food-starch granules tend to be larger than those of glove powder, are often oval, and may be extremely resistant to salivary diastase digestion.

Aged↗

Granulomatous peritonitis caused by glove starch.

Corn starch particles are used as a surgical glove lubricant. At present there is no better alternative for this lubricant. Implantation of corn starch particles into the peritoneal cavity can induce foreign body reactions, starch peritonitis and starch granulomata, and may cause adhesions and intestinal obstruction. Starch peritonitis should be treated conservatively.

Gloves, Surgical↗

Composition, structure, functionality, and chemical modification of legume starches: a review.

The major carbohydrate of the legume seed is starch, which represents up to 45% of the total seed weight. In recent years, substantial progress has been made in understanding the relationship between starch structure and functionality. However, these studies have been mainly on cereal and tuber starches. The present status of knowledge on the composition, structure, functionality, digestibility, and chemical modification of legume starches is reviewed. In addition present concepts of granule structure, gelatinization, retrogradation, and rheology are also reviewed. Future research needs in the area of legume starch chemistry are discussed.

Fabaceae↗

Starch degradation.

Recent research reveals that starch degradation in Arabidopsis leaves at night is significantly different from the "textbook" version of this process. Although parts of the pathway are now understood, other parts remain to be discovered. Glucans derived from starch granules are hydrolyzed via beta-amylase to maltose, which is exported from the chloroplast. In the cytosol maltose is the substrate for a transglucosylation reaction, producing glucose and a glucosylated acceptor molecule. The enzyme that attacks the starch granule to release glucans is not known, nor is the nature of the cytosolic acceptor molecule. An Arabidopsis-type pathway may operate in leaves of other species, and in nonphotosynthetic organs that accumulate starch transiently. However, in starch-storing organs such as cereal endosperms and legume seeds, the process differs from that in Arabidopsis and may more closely resemble the textbook pathway. We discuss the differences in relation to the biology of each system.

Arabidopsis↗

Clearance and metabolism of starch foods in the oral cavity.

The presence of carbohydrates and organic acids was monitored in the oral cavity over a 3-hour period following the ingestion of six foods containing cooked starch (popcorn, potato chips, corn flakes, bread stick, hard pretzel and wheat cracker) and compared to a food containing sugar (chocolate-covered candy bar). Oral fluid samples were collected at 30-min intervals from five different tooth sites from 7 volunteers using absorbent paper points. Samples were analyzed for carbohydrates and organic acids using high-performance liquid chromatography. Analytical data for each food were pooled and compared to the results of the sugar food. The amount of lactic acid produced 30 min after ingestion was highest with the potato chips and lowest with the corn flakes. Potato starch contributed more readily to oral lactic acid production than wheat or corn starch. A direct linear relationship existed between lactic acid production and the presence of oral glucose produced from starch, which occurred via the metabolites maltotriose and maltose. Oral clearance of foods containing cooked starch proceeded significantly slower than that of the sugar food, thus contributing to a prolonged period of lactic acid production.

Acetic Acid↗

Demineralization potential of different concentrations of gelatinized wheat starch.

Six subjects wore intraoral devices carrying bovine enamel blocks covered with a layer of Streptococcus mutans. They swished solutions of 5% glucose or maltose, or sols or gels of 3, 5, 10, 15, or 20% gelatinized wheat starch in the mouth for 3 X 1 min. Demineralization was measured after 45 min by determining the change in iodide permeability (delta Ip) of the enamel. Spittings of the administered materials and samples of saliva, taken at intervals during the test, were analyzed for maltose, and the time of clearance was calculated. Demineralization was greatest for glucose followed by maltose, the starch gels, and the sols. The latter gave close to zero scores. The salivary clearance time increased with increase in concentration of the starch. Although the starch was hydrolyzed rapidly in the mouth, its oral retentiveness was greater than that of the maltose rinse. Demineralization was closely correlated with the final pH of the S. mutans cell layer. The data indicate that the starch in baked or cooked foods may have a significant demineralization potential and that it enhances oral retentiveness.

Animals↗

Changes in morphology of starch-based prothestic thermoplastic material during enzymatic degradation.

This work evaluates the structural changes of an interpenetrated starch thermoplastic blend withstanding different enzymatic alpha-amylase degradation periods (up to 200 days), and establishes the relationships between the kinetics degradation rate and the structure of the material. It characterises the different stages of the enzymatic degradation process on starch/ethylenevinyl-alcohol blends, based on the attack of the connected starch domains that can be accessed by the enzymatic solution. The completely encapsulated starch particles remain practically unchanged. Furthermore, it was also found that the enzymatic degradation process was limited after 100 days of immersion. In order to understand such phenomenon several techniques were used, namely differential scanning calorimetry, contact-angle measurements, high-performance liquid chromatography, Fourier transform infrared spectrometry, scanning electron microscopy and atomic force microscopy. The materials were evaluated with respect to the enzymatic degradation rate, surface morphology and degradation behaviour. The results show that the ethylene-vinylalcohol phase wraps the starch domains, preventing the respective degradation. Consequently, the degraded material in the solution comes only from the starch particles that could be reached by the enzyme.

Alcohols↗

In vitro caries-like lesion production by Streptococcus mutans and Actinomyces viscosus using sucrose and starch.

We investigated the formation of caries-like lesions on root and enamel sections by S. mutans and A. viscosus when grown in four different carbohydrate substrates. The substrates were: sucrose, starch + alpha-amylase, and a combination of starch and sucrose with and without alpha-amylase. Twenty-four sections with exposed windows on both the root and enamel surfaces were exposed to the individual bacterial species in each of the four substrates for three weeks. At three weeks, the sections were removed and lesion depths measured. When grown in sucrose alone, S. mutans produced significantly deeper lesions in both root and enamel surfaces when compared with A. viscosus. However, S. mutans failed to produce lesions when grown in "limited" sucrose with starch added, whereas A. viscosus in this same substrate produced lesions in the enamel and root surfaces. A viscosus also produced either similar or significantly deeper lesions in both root and enamel surfaces, when compared with S. mutans grown in the two substrates with amylase. Thus, under these in vitro conditions, it was demonstrated that A. viscosus could utilize starch, whereas utilization of starch by S. mutans was alpha-amylase-dependent.

Actinomyces↗

Starch digestion in young children with cystic fibrosis measured using a 13C breath test.

The study was designed to evaluate a non-invasive breath test using naturally 13C-rich corn (maize) as substrate to provide quantitative information about the digestion of starch by children in health and disease. The variability of background 13C:12C of young British children was investigated by collecting breath samples over 6 h from 17 healthy children, 6-35 mo old, and from seven children with cystic fibrosis, 25-48 mo old, in their homes. Background 13C enrichment was -26.6 and -25.4 delta per mil in the healthy and cystic fibrosis groups, respectively (p less than 0.01), and varied little during the day (SD = 0.4 delta per mil). Eight healthy children and five with cystic fibrosis were given a test breakfast of corn starch cooked in milk with sugar. The cystic fibrosis group repeated the test with the addition of an enzyme supplement containing alpha-amylase. Subjects accepted variable amounts of starch (range 0.2-2.8 g/kg body wt). The percentage of ingested 13C recovered during the 6 h after the meal increased with age in the healthy children (range 15-53%). Three children with cystic fibrosis had negligible 13C recoveries, and the other two had lower recoveries than expected for their age. The addition of enzymes did not consistently improve 13C recovery. We conclude that the 13C breath test based on corn has potential for investigating starch digestion in young children. Initial results suggest that starch digestion is impaired in some children with cystic fibrosis and that enzyme supplements do not improve digestion consistently or completely.

Breath Tests↗