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Comparative susceptibility to amylases of starch granules of several single endosperm mutants representative of floury-opaque, starch-deficient, and modified starch types and their double-mutant combinations with opaque-2 in four inbred lines of maize.

Starch granules were prepared from kernels of eight single endosperm mutants, brittle-1, (bt1), brittle-2 (bt2), floury-1, floury-2, soft starch, opaque-1 (o1), shrunken-2 (sh2), and sugary-2 (su2), and their double-mutant combinations with opaque-2 (o2) of four inbred lines of maize (Zea mays L.), B37, C103, Oh43 and W64A. We compared the susceptibility of various starch granules to Rhizopus glucoamylase and pancreatin. Starch granules of the su2 and su2o2 mutants were digested by amylases much faster than those of the normal counterparts. Starch granules of the bt1, bt2, o1 and sh2 mutants tended to be digested by amylases faster than those of normal maize. Starch granules of double-mutant combinations with the o2 gene were, in general, digested to an extent very comparable to their respective non-opaque single mutant counterparts in each of their four inbred backgrounds. We followed the relative digestion of starch granules by using scanning electron microscopy. Starch granules of endosperm mutants susceptible to amylases showed numerous pin holes on the surface layer and the pores penetrated into the inner layers of the granules during the attack by amylases. In some of the granules the inner portion, which appeared terraced or step-shaped, could be seen. This may be indicative of layered internal structures of the granules.

Amylases

Starch degradation by the mould Trichoderma viride. I. The mechanism of starch degradation.

The mechanism of starch degradation by the fungus Trichoderma viride was studied in strain CBS 354.44, which utilizes glucose, starch and dextrins but is unable to assimilate maltose. It was shown that the amylolytic enzyme system is completely extracellular, equally well induced by starch, amylose or amylopectin and that it consists mainly of enzymes of the glucoamylase type which yield glucose as the main product of starch hydrolysis. Small amounts of alpha-amylase are produced also. The enzymes produced in starch cultures degrade starch, amylose and amylopectin equally well. Enzyme synthesis in starch media takes place to a considerable extent after exhaustion of the carbon source when maximum growth has been attained. Low-molecular dextrins are degraded by extracellular enzymes of the glucoamylase type. These enzymes are produced in media containing starch or dextrins. Maltotriose is consumed for only one third leaving maltose in the culture filtrate. Maltose is hardly attacked and hardly induces any amylolytic enzyme activity. No stable alpha-glucosidase appears to be produced.

Amylases

In vitro comparison of the erythrocyte sedimenting properties of dextran, hydroxyethyl starch and a new low-molecular-weight hydroxyethyl starch.

A low-molecular-weight preparation of hydroxyethyl starch (LMW-HES) may serve as a desirable substitute for the erythrocyte sedimenting agents presently employed to improve neutrophil yields in centrifugation and gravity leukapheresis. The cell-separating and erythrocyte-sedimenting properties of LMW-HES, assessed in vitro by the recovery of various blood cells in plasma supernatant fluids, were similar to those of the higher-molecular-weight hydroxyethyl starch in current use and to dextran under a variety of conditions. These data predict success for LMW-HES as a sedimenting agent in leukapheresis and recommend that it be evaluated in clinical trials.

Blood Platelets

Digestibility of raw and cooked starches from legume seeds using the laboratory rat.

The in vivo digestibility and growth responses to diets containing raw and cooked legume starches were determined using male Wistar rats. These starches were obtained from seven legumes including smooth- and wrinkled-seeded field peas, navy beans, kidney beans, garbanzo beans, mung beans and lentils. Wheat and potato starch diets were fed for comparative purposes. With the exception of potato and the field pea starches, rats fed uncooked starch diets had higher weight gains than those fed cooked starch diets. The starches from wheat and all legumes, excluding the high-amylose wrinkled pea, were nearly 100% digestible but the legume starches reduced the digestibility of casein protein by 3 to 4%. Raw potato starch was shown to be only 57% digestible. The starches from potato and wrinkled pea reduced the digestibility of casein protein to 87% and gave high cecum weights. These characteristics were not altered by cooking the starch of wrinkled pea. The starches of smooth-seeded peas, lentils and all beans were highly digestible, showed no adverse effects on growth and were comparable to wheat starch in this evaluation.

Animals

Integrated transcriptomic and metabolomic analysis reveals candidate regulatory networks associated with starch accumulation in tetraploid potato.

Potato (Solanum tuberosum L.) tuber starch is a major determinant of crop quality and industrial value, yet the regulatory mechanisms underlying starch accumulation in autotetraploid cultivars remain poorly resolved. Here, we performed integrated transcriptomic and metabolomic analyses using a segregating tetraploid population derived from parents with contrasting starch content. Extreme phenotypes were selected to systematically dissect the molecular basis of starch accumulation. Transcriptome profiling revealed extensive transcriptional reprogramming between high- and low-starch genotypes, with differentially expressed genes significantly enriched in carbohydrate metabolism, particularly the starch and sucrose metabolism pathway. Notably, multiple transcription factor families, including AP2/ERF, MYB, and bHLH, were prominently represented, suggesting coordinated regulatory control. Metabolomic analysis identified substantial metabolic divergence, with differentially accumulated metabolites predominantly enriched in starch and sucrose metabolism as well as secondary metabolic pathways. Most metabolites exhibited negative associations with starch content, indicating competitive carbon allocation between primary and secondary metabolism. Integrative multi-omics analysis further resolved a core regulatory module comprising key structural genes and transcription factors tightly associated with starch-related metabolites. In particular, genes involved in sucrose cleavage and ADP-glucose metabolism, together with trehalose-6-phosphate synthase (TPS) and UDP-glucose-associated pathways, emerged as critical nodes linking carbon flux to starch biosynthesis. Correlation network analysis suggested that AP2/ERF-, MYB-, and bHLH-type transcription factors modulate these pathways by coordinating structural gene expression and metabolic flux distribution. Collectively, our study establishes a transcriptional-metabolic framework for starch accumulation in tetraploid potato, highlighting the central role of carbon allocation and signaling intermediates in shaping starch content, and providing candidate targets for molecular breeding and genome editing.

Solanum tuberosum

Resistant starch types 2 and 4 induce distinct and reversible changes in the human gut microbiome.

Resistant starch (RS) can confer benefits for the gut microbiome and host cardiometabolic health. However, different types of resistant starch can differentially affect gut microbiome composition and functional capacity, especially given interindividual variability in responses, thus limiting the application of resistant starch in dietary strategies. We used shotgun metagenomics to perform a secondary analysis of samples collected during a previously reported randomized clinical trial to determine the effects of dietary supplementation with two types of resistant starch (RS2 and RS4) and a digestible starch (control) on the gut microbiome. Both resistant starch types induced distinct but transient alterations in the gut microbial community. RS2 enriched the keystone degrader, Ruminococcus bromii, and Blautia glucerasea, whereas RS4 favored Parabacteroides distasonis and known but uncharacterized microbial species such as a Lachnospiraceae bacterium. Moreover, we detected strain-level differences in the response of Bifidobacterium adolescentis to resistant starch. Microbial functional profiling revealed an enhanced capacity for complex carbohydrate utilization following resistant starch intake, including increased abundance of specific α-amylases, glycoside hydrolases, starch utilization systems, and other currently uncharacterized genes. Identifying the bacterial strains and genes that respond to different RS types will help to more accurately predict who will benefit from a given RS type. Our findings demonstrate that RS2 and RS4 differentially shape microbial ecology and metabolic capacity and provide a foundation for microbiome-informed personalization of resistant starch-based dietary interventions.IMPORTANCEDietary intake influences human health by modulating metabolism, partly by shaping the microbiota inhabiting the gut. Resistant starch (RS), a dietary fiber, is associated with metabolic improvements. While previous research has explored how RS alters the gut microbiome, RS comprises five types with differing physical and chemical characteristics, and the distinct impacts of each type on the microbiome and host health have not been fully characterized, particularly using high-resolution approaches such as shotgun metagenomics. In this secondary analysis of samples from a longitudinal crossover intervention study, we link dietary supplementation with RS2 and RS4 with distinct and transient changes in the composition and functional potential of the human gut microbiome. Specifically, we identify species that increase in abundance with each RS type, accompanied by increases in genes and pathways involved in complex carbohydrate utilization. The findings support the development of precision nutrition strategies utilizing RS supplementation to improve metabolic health.This study is registered with ClinicalTrials.gov as NCT05743790.

Humans

Relation of amylase to starch and Lycasin metabolism in human dental plaque in vitro.

Acid production activity (APA) in plaque suspensions from glucose, boiled soluble starch and hydrogenated starch hydrolysate (Lycasin) was studied in 11 subjects. Amylase (alpha-1,4-glucan 4-glucanohydrolase, EC 3.2.1.1) activity was measured in plaque and whole saliva samples from the same persons. Lycasin was found to be hydrolyzed by salivary amylase under the formation of di- and oligosaccharides, however, with a lower rate than starch. A high correlation was found between APA from glucose and from soluble starch and between APA from soluble starch and plaque amylase activity. No correlation was found between amylase activity in saliva and APA from soluble starch or between amylase activity in saliva or plaque and APA from Lycasin. APA from Lycasin was about 62% and from soluble starch about 76% of the APA from glucose. 0-25% of the total number of cultivable microorganisms from the plaque produced extracellular starch-degrading enzymes. No correlation was found between number of starch-degrading microorganisms and APA from soluble starch or between these numbers and the plaque amylase activity. By electrophoreses only amylase fractions of human origin were found in whole saliva, plaque supernatants and plaque suspensions, indicating that the microbial amylase activity in the plaque is low compared with that of salivary origin.

Acids

Integrating GWAS and Transcriptome Analysis Identifies Candidate Genes for Kernel Starch Quality Traits in Maize.

Maize (Zea mays L.) starch quality is a complex trait with significant implications for grain processing and industrial applications. However, the genetic basis underlying starch quality, particularly for gelatinization and thermodynamic properties, remains poorly understood. In this study, we evaluated 12 starch quality traits, including seven gelatinization characteristics, four thermodynamic traits, and kernel starch content (KSC) in a diverse panel of 335 maize inbred lines. Considerable phenotypic variation was observed for all traits. A total of 228 quantitative trait loci (QTLs) were significantly associated with 12 starch quality traits through genome-wide association studies (GWAS). By integrating a dynamic transcriptome analysis of two maize inbred lines with contrasting starch quality, we identified 60 candidate genes. One gene, waxy1, encoding a starch synthase, was found to be associated with enthalpy of gelatinization (ΔHgel) and pasting temperature (Ptemp). Six variants in waxy1 contributed to natural variation in ΔHgel and Ptemp, and a cost-effective InDel and two PARMS-based molecular markers were developed and validated in 144 maize inbred lines, enabling efficient marker-assisted selection. Our findings provide key genes and molecular markers for high-quality maize breeding with improved starch properties.

Zea mays

Effect of modified and unmodified tapioca starches on 59Fe retention in rats.

The effects of a chemically-modified tapioca starch hydroxypropyl distarch phosphate (HDP), and unmodified tapioca starch (UMS) on 59Fe retention by rats were compared. Three experimental variables were evaluated: 1) the type of starch in the diet, 2) cooking of either the starch alone or the entire diet, and 3) the iron status of the rats. There were no significant differences in 59Re retention between iron-adequate rats fed either UMS or HDP. 59Fe retention by iron-deficient rats was not affected by the type of starch in the diet when uncooked starch was used. However, if the starch was cooked, substitution of HDP for UMS resulted in a significant depression in iron retention by iron-deficient rats. Cooking the entire diet produced a similar but less marked effect. The results of these experiments suggest that the inclusion of one particular type of modified tapioca starch in the diet may affect iron utilization.

Animals

[Genetic change in the waxy trait of barley under the influence of wild type DNA. Analysis of the composition of starch and the electrophoretic spectrum of caryopsis hordein from altered plants and the retention of these changes to the fourth generation].

Injection of DNA isolated from the wild type of barley into grains of recipient mutant plants (waxy mutants) at the milk stage of maturity leads to a change in starch synthesis; type of spikes and hordein composition. In the first generation of injected plants the wild type starch synthesis was observed in some separate plants (these observations were made at a haploid level in pollen cells). In the second generation of transformed plants along with the change in starch and hordein synthesis a modification of the type of spikes was also revealed. Recipient plants had six-rowded (hexastichous) spikes, and donor plants--two rowded (distichous) spikes. Disc-electrophoresis of hordeins of the wild type barley (Yuzhny var.), hordeins of the waxy mutant (defected in synthesis of normal starch) and barley plants transformed under the action of wild type exogenous DNA reveals differences in the protein spectrum between donor, recipient and transformants. In the second generation in many of the transformed plants starch synthesis reverted to the recipient mutant type. Simultaneously a reversion of hordein composition to the initial mutant type was observed, and the distichous pikes became hexastichous. Analysis of the components of starch revealed that donor plant that have amilose and amilopectin in starch, and the recipient plants that lack amilose, can be distinguished by the spectra of light absorption of starch. For characterizing these differences the plot of absoprtions at 490 versus that at 590 nm was used. The tangens of angles of these curves for the waxy mutant were equal to 1.05 +/- 0.07 and 1.81 +/- 0.04 for the wild type barley. All transformants have a 1.78 ratio and for revertants this value was 1.02.

DNA

Comparative susceptibility of starch granules of double- and triple-mutants containing amylose-extender, waxy, sugary-1, sugary-2 and dull genes of maize inbred OH43 (Zea mays L.) to amylase.

Starch granules were prepared from 14 double- and 26 triple-mutants containing amylose-extender (ae), 14 double- and 18 triple-mutants containing waxy (wx), 15 double- and 20 triple-mutants containing sugary-1 (su1), 13 double- and 23 triple-mutants containing sugary-2 (su2), and 14 double- and 19 triple-mutants containing dull (du) of maize inbred Oh43 (Zea mays L.). The relative susceptibilities of these starch granules to fungal glucoamylase were determined and the starch granules were examined by scanning electron microscopy. A commercial normal maize starch was used as a control. Starch granules of the double- and triple-mutants containing su1 and su2 were digested two to eight times faster than normal. The ae gene reduced susceptibility and seems to be epistatic to su1 and su2. Starch granules of the double- and triple-mutants containing wx were digested about two times faster than normal and those containing shrunken-2 (sh2) were digested 1.2 to eight times faster than normal. Starch granules of triple-mutant combinations with opaque-2 (o2) showed digestion properties which were comparable to those of their respective monopaque double-mutant counterpart.

Glucan 1,4-alpha-Glucosidase

Inhibition of intestinal iron absorption by laundry starch.

The pathogenesis of iron deficiency anemia associated with amylophagia is usually attributed to dietary iron lack. However, large quantities of starch may inhibit intestinal iron absorption. Accordingly, studies were carried out to determine the effect of laundry starch on the intestinal absorption of inorganic and hemoglobin iron. In vitro, laundry starch bound 19 to 80% of the available 59FeSO4 and 34 to 68% of the available 59Fe-hemoglobin. Binding of both forms of iron was pH-dependent, with maximal binding at pH 7.0. In vivo, laundry starch significantly inhibited mucosal uptake of 59FeSO4 from isolated duodenal loops. In nonanemic rats, administration of laundry starch (100 mg) 1 hr before a 100-mug dose of 59FeSO4 significantly decreased the absorption of 59FeSO4, as compared to saline or low iron chow controls (6.2 +/- 2.0 versus 14.9 +/- 2.1 and -1.8 +/- 1.7, respectively, P less than 0.001). In anemic rats the absorption of either a 100-mug dose of 59FeSO4 or a 500-mug dose of 59Fe-hemoglobin was also significantly decreased by prior administration of laundry starch. The data obtained indicated that laundry starch (1) binds appreciable quantities of inorganic and hemoglobin iron in vitro; (2) inhibits the mucosal uptake or inorganic iron by isolated intestinal loops; (3) inhibits the intestinal absorption of inorganic iron in normal nonanemic rats, and (4) blunts the compensatory increase in inorganic and organic iron absorption in anemic rats.

Anemia, Hypochromic