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Effect of compression force and corn starch on tablet disintegration time.

An unusual occurrence of decreasing tablet disintegration time with increasing tablet hardness was explored. Tablets were made with varying ratios of starch disintegrant to starch paste and compressed with eight different forces. This unusual disintegration pattern is modified by changes in the starch ratios. The reason for this phenomenon is ascribed to gain swelling as the mechanism by which starch acts as a disintegrant.

Chemistry, Pharmaceutical↗

Biodegradable microspheres. VI: Lysosomal release of covalently bound antiparasitic drugs from starch microparticles.

The possibilities of using polyacryl starch microparticles as a carrier for low molecular weight drugs have been investigated. Two drugs containing primary amino groups, primaquine and trimethoprim, have been covalently coupled to the starch microparticles via tri-, tetra-, and pentapeptide spacer arms. The drug-particle complexes were prepared by coupling different drug-peptide derivatives to the particles after activation of the starch with carbonyldiimidazole. The activation process with subsequent removal of activated groups did not change the degradability of the starch microparticles. The resulting drug-carrier complexes were stable in serum, while free drugs were released in a lysosome fraction. Thus, the microparticle-peptide-drug conjugates fulfill the basic requirements for a drug carrier used to target drugs to the lysosomes (e.g., for the treatment of lysosomal parasitic diseases).

Animals↗

Performance of a modified starch hydrophilic matrix for the sustained release of theophylline in healthy volunteers.

Two experimental formulations of theophylline with a hydrophilic starch matrix were evaluated for their sustained-release characteristics after single administration in healthy human volunteers. Theo-dur was chosen as a reference sustained-release formulation. In a first study, the extent of absorption was similar for a syrup, for Theo-dur, and for the experimental formulation of theophylline with 70% drum-dried corn starch as the sustained-release agent (DDCS-70). The maximal plasma concentration (Cmax) was significantly lower, and the time to reach Cmax as well as the time span during which the plasma concentration was at least 75% of the Cmax were significantly higher for Theo-dur than for the DDCS-70 formulation. A sustained-release profile, as for Theo-dur, was not reached for DDCS-70. In a second study the influence of the starch:drug ratio on the bioavailability was investigated. The decrease in starch content from 70 to 50% of the formulation did not improve the plasma concentration-time profile towards a sustained-release profile.

Adult↗

Analysis of purified maize starch synthases IIa and IIb: SS isoforms can be distinguished based on their kinetic properties.

Since starch synthases IIa (SSIIa) and SSIIb have not been purified from plant tissue, their structure-function relationships have not been well characterized. Therefore, we have expressed these SS genes in Escherichia coli, purified them to apparent homogeneity, and studied their kinetic properties. In addition, the N-terminally truncated forms of these enzymes were studied in an attempt to understand the function of the diverse N-terminal sequences in SS. Our results show that, like SSI, the N-terminal extensions of SSIIa and SSIIb are not essential for catalytic activity and no extensive changes in their kinetic properties are observed upon their N-terminal truncation. Each isoform of SS can be distinguished based on its kinetic properties. Maize SSI and maize SSIIb exhibit higher Vmax with glycogen as a primer, while the converse is true for SSIIa. However, the specific activity of SSIIb is at least two- to threefold higher than that for either SSI or SSIIa. Although SSIIb exhibits the highest maximal velocity of the isoforms compared, its apparent affinity for primer is twofold lower than the affinity of SSI and SSIIa for primer. Perhaps these differences in primer affinity, primer preference, and maximal velocities all contribute in some way to the different structure(s) of starch during its synthesis. Expression and purification of maize SS has now provided us a useful tool to address the role(s) of SS in starch synthesis and starch structure.

Adenosine Diphosphate Glucose↗

Increased carboplatin concentration in liver tumors through temporary flow retardation with starch microspheres (Spherex) and gelatin powder (Gelfoam): an experimental study in liver tumor-bearing rabbits.

Regional chemotherapy of primary and secondary malignant liver tumors is superior to systemic therapy. The regional advantage can be further increased by flow retardation. Absorbable gelatin powder (Gelfoam) and starch microspheres (Spherex) may serve as embolizing agents because of their particle size and embolization time. Carboplatin was for the first time applied as a cytostatic agent in regional chemotherapy. Embolization and flow retardation times were measured. The embolization time of Gelfoam was 27 min, and that of starch microspheres (Spherex), 7 min, on average. Mean flow retardation of Gelfoam was 153 min, and that of starch microspheres (Spherex) 38 min. The concentration differences in systemic and regional chemotherapy were determined in VX-2 liver tumor-bearing rabbits. In regional chemotherapy, the tumor concentration was increased by a factor of 3.6 compared with systemic therapy. Coapplication with an embolizing agent increased the tumor concentration of carboplatin by a factor of 44 to 47. Concentrations of absorbable gelatin powder (Gelfoam) and starch microspheres (Spherex) did not differ significantly.

Animals↗

The importance of starch biosynthesis in the wrinkled seed shape character of peas studied by Mendel.

The wrinkled-seed mutant (rr) of pea (Pisum sativum L.) arose through mutation of the gene encoding starch-branching enzyme isoform I (SBE1) by insertion of a transposon-like element into the coding sequence. Two isoforms of starch-branching enzyme have been documented in the developing pea embryo. The second isoform, SBEII, is expressed towards the later stages of embryo development while SBEI is expressed highly in the early stages. Due to mutation of SBEI the total amount of starch and the proportion of amylopectin, a branched starch polymer, are greatly reduced in the wrinkled (rr) line as compared to that in the wild-type, round (RR) line. Consequently, the level of sucrose in the rr line is nearly two fold that of the RR line. Increased sucrose concentration in the developing embryos of this mutant line causes increased uptake of water and thereby increases the cell size and fresh weight. During seed maturation in these mutant seeds a greater loss of water occurs. As a result, the wrinkled seed phenotype develops. Besides this morphological variation, the mutation also causes changes in the amount of lipid and of one storage protein, legumin. This review article discusses the role of the SBEI enzyme in causing such metabolic changes in the developing embryos with the implication that metabolism can play a central role in plant development.

1,4-alpha-Glucan Branching Enzyme↗

Transfer of genes for utilization of starch (sta2) and melibiose (mel) to industrial strains of Saccharomyces cerevisiae by single-chromosome transfer, using a kar1 mutant as vector.

A method has been developed for the transfer of genes from other yeast strains and species to industrial yeast strains, using a haploid, kar1-1 mutant strain of Saccharomyces cerevisiae as a vector. The sta2 gene, conferring the ability to metabolize starch was transferred from an auxotrophic haploid strain of S. cerevisiae (S. diastaticus) and the melibiose-metabolism (mel) gene(s), from S. kluyveri, to the kar1-1 mutant [K5-5A; (alpha ade2 his4 can1 gal) by normal mating and protoplast fusion. From this strain, the genes were transferred to baker's yeast and brewing yeast strains, which did not utilize starch, and to baker's yeast strains, which did not utilize melibiose, by protoplast fusion, spore-cell pairing, or rare-mating. Strains that utilized starch or melibiose were obtained by all three methods. Pulsed-field gel electrophoresis preparations showed little change in the mobility of the chromosomes of the hybrids. The most probable explanation for the results obtained is that single chromosomes were transferred, first, from the donor strains to the kar1-1 haploid mutant strain, and then from the kar1-1 vector to the recipient industrial strain of S. cerevisiae. The transfer of the genes is probably accomplished through formation of disomic strains and then, in the case of the hybrids that metabolize starch, by integration of the sta2 gene into the genome of the industrial yeast strains.

Chromosomes, Fungal↗

Synthesis of fructans in tubers of transgenic starch-deficient potato plants does not result in an increased allocation of carbohydrates.

Inhibition of starch biosynthesis in transgenic potato (Solanum tuberosum L. cv. Désirée) plants (by virtue of antisense inhibition of ADP-glucose pyrophosphorylase) has recently been reported to influence tuber formation and drastically reduce dry matter content of tubers, indicating a reduction in sink strength (Müller-Röber et al. 1992, EMBO J 11: 1229-1238). Transgenic tubers produced low levels of starch, but instead accumulated high levels of soluble sugars. We wanted to know whether these changes in tuber development/sink strength could be reversed by the production of a new high-molecular-weight polymer, i.e. fructan, that incorporates sucrose and thereby should reduce the level of osmotically active compounds. To this end the enzyme levan sucrase from the gram-negative bacterium Erwinia amylovora was expressed in tubers of transgenic potato plants inhibited for starch biosynthesis. Levan sucrase was targeted to different subcellular compartments (apoplasm, vacuole and cytosol). Only in the case of apoplastic and vacuolar targeting was significant accumulation of fructan observed, leading to fructan representing between 12% and 19% of the tuber dry weight. Gel filtration and 13C-nuclear magnetic resonance spectroscopy showed that the molecular weight and structure of the fructan produced in transgenic plants is identical to levan isolated from E. amylovora. Whereas apoplastic expression of levansucrase had deleterious effects on tuber development, tubers containing the levansucrase in the vacuole did not differ in phenotype from tubers of the starch-deficient plants used as starting material for transformation with the levansucrase. When tuber yield was analysed, no increase but rather a further decrease relative to ADP-glucose pyro-phosphorylase antisense plants was observed.

Carbohydrate Metabolism↗

Hormone treatment of roots causes not only a reversible loss of starch but also of structural polarity in statocytes.

Treatment of cress (Lepidium sativum L.) roots with phytohormones (4.3 x 10(-5) M gibberellic acid plus 4.3 x 10(-5) M kinetin, 30 h; T.H. Iversen, 1969, Physiol. Plant. 22, 1251-1262) caused not only complete destarching of amyloplasts but also destruction of the polar arrangement of cell organelles in statocytes. The nucleus was not positioned exclusively near the proximal cell pole as in the controls but was also found near the distal cell pole. The endoplasmic reticulum (ER) was no longer organized in parallel sheets at the distal cell pole but instead the ER-cisternae were randomly distributed. Additionally, the statocytes from hormone-treated roots contained a large central vacuole instead of numerous small ones as in the controls. The starch-free plastids had a reduced volume and an amoeboid shape. They did not sediment but were randomly distributed in the statocytes. The loss of structural polarity was accompanied by loss of graviresponsiveness although root growth still occurred. Twenty-two hours after removal of the hormones, structural polarity was restored and starch was resynthesized. The newly formed starch grains were smaller and more numerous per amyloplast compared to the controls. It is concluded that loss of gravisensitivity of roots after hormone treatment cannot be solely attributed to the loss of amyloplastic starch because there is a concomitant loss in the polar organisation of the statocyte.

Adenine↗

Glucan phosphorylases in Vicia faba L.: cloning, structural analysis and expression patterns of cytosolic and plastidic forms in relation to starch.

We have isolated and characterised cDNA sequences from a Vicia faba cotyledonary library encoding a plastidic isoform (VfPho1) and a cytosolic isoform (VfPho2) of an alpha-1,4-glucan phosphorylase (EC 2.4.1.1; Commission on Plant Gene Nomenclature 1994). The Pho1 isoform is characterized by the presence of a plastidial transit peptide and an 81-residue stretch of additional amino acids in the middle of the polypeptide which are not found in the Pho2 isoform. We define the position of this so-called insertion sequence differently from previous authors. The Pho1 transcripts were found predominantly in the early seed coat and in cotyledons, and accumulated until the late desiccation phase, whereas Pho2 transcripts were about equally abundant in all investigated tissues. Activity patterns of both enzymes in cotyledons roughly followed mRNA accumulation patterns, with the exception of the late desiccation phases when mRNAs were degraded but enzyme activities remained at high level, even in long-stored seeds. The distinct Pho1 and Pho2 gene expression patterns in seed coats coincided with the transient accumulation pattern of starch. Similarly, in-situ hybridisation revealed a relationship between Pho1 gene expression and starch granule formation in developing cotyledons. Expression data and enzyme activity patterns were associated with starch formation during seed development, and could simply reflect a continuous accumulation of enzyme protein, ensuring immediate participation in starch degradation during germination.

Amino Acid Sequence↗

Aspergillus nidulans nuclear proteins bind to a CCAAT element and the adjacent upstream sequence in the promoter region of the starch-inducible Taka-amylase A gene.

Aspergillus nidulans was used as an intermediate host to investigate the regulation of the Taka-amylase A (TAA) gene from Aspergillus oryzae. The induction of Taa by starch was confirmed to be regulated at the transcriptional level by analyzing the transcripts specific for Taa synthesized in vitro in nuclei from starch- and glucose-grown cells. A 55 bp DNA fragment containing a consensus CCAAT sequence from the promoter region of the Taa gene was shown to confer starch inducibility on the gene. A nuclear extract from starch-grown cells was assayed for proteins which bind to the promoter region of the Taa gene. A protein designated AnCP1 bound to the CCAAT sequence. A nuclear extract from glucose-grown cells contained two DNA-binding proteins designated AnCP2 and AnNP1. AnCP2 bound to the same CCAAT sequence as AnCP1, while AnNP1 bound to the 25 bp region just upstream of the AnCP2 binding site. Occupancy of the two binding sites appeared to be mutually exclusive, which is suggestive of a negative regulatory mechanism for gene expression.

Aspergillus nidulans↗

[Effect of isoenergetic replacement of starch by olive oil and fish oil concentrations of lipids in plasma and lipoprotein fractions in swine].

Two experiments with sows were performed to investigate the effect of isoenergetic replacement of starch by fish oil or olive oil on concentrations of lipids in plasma and lipoproteins. The first experiment was based on a cross-over design with three periods, each lasting 16 days. Each sow was fed during one of the periods a basal ration with isoenergetic addition of (1) starch (495 g/d), (2) olive oil (221 g/d), or (3) fish oil (223 g/d) based on energetic requirement for maintainance. The second experiment was based on a cross-over design with eight periods, each lasting 16 days. In the first and in the last periods, each sow was fed the basal ration. In the other six periods, each sow was fed the basal ration with addition of two different amounts of (1) starch (284/568 g/d), (2) olive oil (140/281 g/d), or (3) fish oil (141/282 g/d). The two different amounts of addition were selected to exceed the energetic requirement for maintainance by 25% or 50%. In both experiments blood samples were taken before each change of the ration. In both experiments olive oil elevated the concentration of cholesterol in plasma in comparison with starch. This elevation was due to a large elevation in high-density lipoproteins (HDL), and a slight elevation in low-density lipoproteins (LDL) and very-low density lipoproteins (VLDL). The ratio between HDL and LDL cholesterol was increased by feeding olive oil. The effect of olive oil on concentrations of cholesterol in plasma and lipoproteins was dose-dependent. In both experiments none of the two dietary oils significantly changed concentrations of triglycerides in plasma and lipoproteins. Concentrations of phospholipids in plasma, HDL, and LDL were elevated by olive oil. In both experiments addition of fish oil elevated concentration of cholesterol in plasma due to elevated cholesterol concentration in LDL. Concentration of HDL cholesterol was not changed by fish oil. Thus, the ratio between HDL cholesterol and LDL cholesterol was lowered by fish oil. The effect of fish oil on concentration of cholesterol in plasma and lipoproteins was also dose-dependent. Fish oil had no significant effect on phospholipid concentrations in plasma and lipoproteins. In conclusion, in the present experiment olive oil caused antiatherogenic changes of the lipoprotein profile, whereas fish oil caused proatherogenic changes of the lipoprotein profile.

Animals↗

Hepatic artery administration of degradable starch microspheres. I. Effect on energy charge and incorporation of precursors into rat liver nucleic acids.

The effects of hepatic artery administration of degradable starch microspheres on liver energy charge and nucleic acid anabolism were studied in rats. Liver energy charge was evaluated 20 and 60 min after the injection of degradable starch microspheres. As compared to controls the microspheres had no effect on liver energy charge. The incorporation of orotic acid, uracil, and thymidine into liver RNA or DNA was studied 1 h after hepatic artery injection of precursor alone or together with degradable starch microspheres. Orotic acid and uracil incorporation into RNA was studied in normal rats and the DNA incorporation of thymidine in animals with regenerating livers. Orotic acid and thymidine were given in trace amounts. Uracil was given in amounts corresponding to a therapeutic dose of 5-fluorouracil. The addition of microspheres had no effects on the incorporation of the nucleic acid precursors into RNA or DNA. Thus, in the normal liver degradable starch microspheres administered by the hepatic artery had no influence on liver energy charge or RNA anabolism in the liver. Also the microspheres had no negative effects on the DNA anabolism in proliferating liver cells.

Animals↗

Modified food starches in baby foods.

Modified food starches were developed as a stabilizer, providing desirable consistency, texture, and storage ability. They are used primarily in strained and junior foods and, to a minor extent, in infant formulas. However, despite the fact that there is an increasing tendency to introduce solid foods to infants at a very early age, there is few long-term studies to delineate the effect of starch feeding on the growth of young infants. Modified food starches used by the food industry for infants and young children are of concern and there is an urgent need for additional data regarding their bioavailability, effect on nutrient absorption, intestinal changes, and toxic, mutagenic, and carcinogenic effects. Therefore, the inclusion of modified food starches should be used prudently and sparingly.

Animals↗

Saccharification and alcohol fermentation in starch solution of steam-exploded potato.

Steam explosion pretreatment of potato for the efficient production of alcohol was experimentally studied. The amount of water-soluble starch increased with the increase of steam pressure, but the amounts of methanol-soluble material and Klason lignin remained insignificant, regardless of steam pressure. The potatoes exploded at high pressure were hydrolyzed into a low molecular liquid starch, and then easily converted into ethanol by simultaneous saccharification and fermentation using mixed microorganisms: an amylolytic microorganism, Aspergillus awamori, and a fermentation microorganism, Saccharomyces cerevisiae. The maximal ethanol concentration was 4.2 g/L in a batch culture at 15 g/L starch concentration, and 3.6 g/L in a continuous culture fed the same starch concentration. In the fed-batch culture, the maximal ethanol concentration increased more than twofold, compared to the batch culture.

Aspergillus↗

Effect of starch and inorganic nitrogen on microbial transformations of organic compounds in soil.

It was found in long-term incubation experiments (100 weeks) that in soil non amended with organic compounds, mineralization proceeded in a steady state after an initial reactive respiration and corresponded approximately to the rate of mineralization of soil organic matter under natural conditions. The addition of nitrogen influenced this mineralization process only very slightly. The addition of starch decreased the stability of organic compounds in the soil at first but the newly formed organic substances were gradually stabilized again. The addition of nitrogen to starch increased the extent and the rate of these alterations. The overall extent of mineralization of the substrate after the addition of starch alone exceeds the amount of the added substrate; a small accumulation of the substrate could be observed after the addition of starch together with nitrogen.

Biotransformation↗

Cross-linked starch microspheres: effect of cross-linking condition on the microsphere characteristics.

Cross-linked starch microspheres were prepared using different kinds of cross-linking agents. The influence of several parameters on morphology, size, swelling ratio and drug release rate from these microspheres were evaluated. These parameters included cross-linker type, concentration and the duration of cross-linking reaction. Microspheres cross-linked with glutaraldehyde had smooth surface compared with those prepared with epichlorhydrine or formaldehyde. The particle size increased with increasing the cross-linking time and increasing the drug loading. Swelling ratio of the particles was a function of cross-linker type but not the concentration or time of cross-linking. Drug release from starch microspheres was measured in phosphate buffer and also in phosphate buffer containing alpha-amylase. Results showed that microspheres cross-linked with epichlorhydrine released all their drug content in the first 30 minutes. However, cross-linking of the starch microspheres with glutaraldehyde or formaldehyde decreased drug release rate. SEM and drug release studies showed that cross-linked starch microspheres were susceptible to the enzymatic degradation under the influence of alpha-amylase. Changing the enzyme concentration from 5000 to 10,000 IU/L, increased drug release rate but higher concentration of enzyme (20,000 IU/L) caused no more acceleration.

Anti-Inflammatory Agents, Non-Steroidal↗

Starch-binding domains in the post-genome era.

Starch belongs to the most abundant biopolymers on Earth. As a source of energy, starch is degraded by a large number of various amylolytic enzymes. However, only about 10% of them are capable of binding and degrading raw starch. These enzymes usually possess a distinct sequence-structural module, the so-called starchbinding domain (SBD). In general, all carbohydrate-binding modules (CBMs) have been classified into the CBM families. In this sequence-based classification the individual types of SBDs have been placed into seven CBM families: CBM20, CBM21, CBM25, CBM26, CBM34, CBM41 and CBM45. The family CBM20, known also as a classical C-terminal SBD of microbial amylases, is the most thoroughly studied. The three-dimensional structures have already been determined by X-ray crystallography or nuclear magnetic resonance for SBDs from five CBM families (20, 25, 26, 34 and 41), and the structure of the CBM21 has been modelled. Despite differences among the amino acid sequences, the fold of a distorted beta-barrel seems to be conserved together with a similar way of substrate binding (mainly stacking interactions between aromatic residues and glucose rings). SBDs have recently been discovered in many non-amylolytic proteins. These may, for example, have regulatory functions in starch metabolism in plants or glycogen metabolism in mammals. SBDs have also found practical uses.

Amino Acid Sequence↗