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Enzymatic degradation of hydroxypropyltrimethylammonium wheat starches.

The enzymatic degradation of hydroxypropyltrimethylammonium modified starches synthesised by dry process was compared with that of hydroxypropyltrimethylammonium modified starches synthesised in glycerol-water plasticised molten medium. The enzymatic degradation rate of products from both origins decreased as the degree of substitution increased. However, two distinct enzymatic degradation profiles were obtained. Dry process products displayed a regular decrease pattern as DS increased. Molten medium synthesised cationic starches displayed a constant degradation level on a wide DS range with alpha,beta-amylase and amyloglucosidase, whereas isoamylase degradation rapidly reached its degradation limit at DSs 0.05. The various plasticising conditions used to synthesise cationic starch in molten medium show no influence on the enzymatic degradation. By measuring the affinity of alpha-amylase, beta-amylase and isoamylase for native, extruded non-modified and hydroxypropyltrimethylammonium-modified starches. It was evident that the enzymes' affinity for the substrate diminishes with increasing chemical modification, particularly in the case of alpha-amylase, suggesting that the location of cationic groups impairs the enzyme's recognition of the substrate. Structural elements of limit dextrins were analysed by (1)H NMR.

Amylases↗

Effect of different cultivation conditions on Lactobacillus manihotivorans OND32T, an amylolytic lactobacillus isolated from sour starch cassava fermentation.

Study of the cassava sour starch fermentation has led to the isolation of a new homofermentative amylolytic lactic acid bacterium, Lactobacillus manihotivorans OND32T, whose nutritional requirements have been investigated in this work. The main effect of deleting one of the substrate components of the MRS-starch medium was to reduce the amylase production. When starch fermentation with nitrogen as a gas phase was compared to fermentation under aerobic conditions, both growth and amylase production were reduced whereas lactic acid formation was not affected. Addition of carbon dioxide (> or = 20% v/v) to the nitrogen gas phase restored growth and amylase production. The amylase production was high with starch, maltose or cellobiose contrary to glucose, fructose and sucrose. During mixed fermentation of glucose and maltose, a diauxic growth was observed. The maltose consumption and the amylase production started after the glucose depletion. The presence of maltose altered the carbon assimilation from glucose, whereas the energetic pathway was not affected. It is concluded that the elimination of soluble sugars by the wet extraction of starch during the processing of cassava, together with the expected in situ CO2 production, are conditions favouring the growth and the amylase synthesis. However, these are likely to be limited by the low nitrogen content in cassava.

Amylases↗

Solid-state fermentation of cornmeal with the basidiomycete Hericium erinaceum for degrading starch and upgrading nutritional value.

The ability of the basidiomycete Hericium erinaceum to degrade starch and upgrade nutritional value of cornmeal during solid-state fermentation was studied. On the basal medium which consisted of cornmeal and salt solution, H. erinaceum produced a strong alpha-amylase on the 15th day after inoculation, which resulted in a 52% degradation of the starch. By supplementation with 5-15 g soybean meal per 100 g cornmeal the alpha-amylase activity and degradation rate of starch was raised significantly (P < 0.01). Prolongation of fermentation time from 15 to 30 days did not increase significantly the degradation rate of starch, though the alpha-amylase activity reached its maximum value of 179 U/g on the 20th day after inoculation. Under conditions close to the theoretical optimum fermentation conditions, that was after 25 days at 25 degrees C in the medium with added 15 g soybean meal per 100 g cornmeal, the starch content in the product decreased from 63% to 22% (P < 0.001) and protein content increased from 12% to 17% (P < 0.01). In the protein in the product, the lysine content was increased from 36 to 56 mg/ g and tryptophan from 9 to 13 mg/g. Using egg protein as a standard, an evaluation on the protein quality of the fermented product showed that it was superior to that of the nonfermented control and to other cereals, was close to that of soybean and chicken, but was inferior to that of milk and red meats.

Basidiomycota↗

Direct compressible polymethacrylic acid-starch compositions for site-specific drug delivery.

The purpose of this study was to generate and characterize a direct compressible pH-sensitive excipient composition for controlled drug delivery. In acidic aqueous solutions, polymethacrylic acid (PMAA) forms complexes and precipitates with starch. As the extent of the interaction between PMAA and starch reaches a maximum at a weight ratio of 1:1.38 (PMAA/starch), this composition was used for the direct compression of tablets. These tablets (30 mg) showed no disintegration even after 2 days in a disintegration test apparatus according to the USP XXIII, in simulated gastric fluid at pH 1.2. In contrast, in 100 mM phosphate buffer pH 7.0 they disintegrated within 40.25+/-8.42 min (mean+/-S.D., n=3). Control tablets of starch disintegrated within the first minute at both pH values. Dissolution studies with the model peptide peroxidase demonstrated no release within 120 min at pH 1.2, whereas at pH 7.0, 100% of the peptide was released within 330 min. Similar release profiles were obtained with the model drugs amoxicillin and rifampicin. In addition, the use of a PMAA-starch composition as a carrier matrix for peroxidase and amoxicillin provided a protective effect towards pepsin and hydrolytic degradation at pH 1.2, respectively. According to these results, the PMAA-starch composition may be a useful tool to overcome the very harsh environment of the stomach for future delivery systems.

Amoxicillin↗

Drug release from starch-acetate films.

The aim of the present work was to compare the drug release rates from the native and acetylated starches. The average degree of acetyl substitution per glucose residue of potato starch was either 1.9 (SA DS 1.9) or 2.6 (SA DS 2.6). Bovine serum albumin (BSA) (mol. wt. 68,000), FITC-dextran (mol. wt. 4400), timolol (mol. wt. 332, log P=1.91) and sotalol-HCl (mol. wt. 308, log P=-0.62) were used as model drugs. All of the model drugs were released rapidly from the potato starch film in PBS pH 7.4 with and without alpha-amylase in the dissolution medium (t50% varied from 0.17 to 3.37 h). When compared to the potato starch film, all of the studied drugs were released at a substantially slower rate from the SA films in the corresponding mediums. The release of the smaller drugs (sotalol, timolol) from the SA films was faster than that of the macromolecules (FITC-dextran, BSA). Furthermore, sotalol was released faster than the more lipohilic timolol from the SA films. Release of macromolecules from the SA films was biphasic with and without alpha-amylase in the dissolution medium: an initial fast release phase was followed by a slower release phase (SA DS 1.9) or no release occurred after the initial phase (SA DS 2.6). All of the drugs were released faster from the SA DS 1.9 film than the weight loss of the film itself. When compared to the SA DS 1.9 film, the model drugs (except sotalol) were released slower from the SA DS 2.6 film. The macromolecule release from the SA DS 2.6 film was erosion-controlled. The weight loss of the SA DS 2.6 film was slow with and without alpha-amylase in the incubation medium. The present results show that acetylation of potato starch can substantially retard drug release. The drug release profiles may be controlled by the degree of substitution, since drug release from the SA DS 1.9 film was faster than the corresponding release from the SA DS 2.6 film.

Acetates↗

Rye products in the diabetic diet. Postprandial glucose and hormonal responses in non-insulin-dependent diabetic patients as compared to starch availability in vitro and experiments in rats.

Rye flakes, rye bread and white wheat bread were given as suspensions to rats and in standardized breakfast meals to non-insulin-dependent diabetics. In both cases the postprandial glucose response was lower after rye bread than after wheat bread. A larger amount of starch remained in the stomach of the rats 15 min after ingesting rye bread compared to wheat bread, indicating that delayed gastric emptying may be one factor explaining the lower response after rye bread. Although the incremental postprandial glucose areas after rye flakes and wheat bread were similar, the rate of decrease of the glucose curve was slower after flaked rye. This would point to a prolonged absorption of some starch in the rye flakes, also indicated by higher late immunoreactive insulin (IRI) values after that product. In the rats the content of starch in the stomachs 15 min after feeding was higher after rye flakes compared to wheat bread. In vitro incubations with alpha-amylase showed lower availability of the starch in rye flakes than in the breads, indicating that several factors may contribute to the differential postprandial glucose response after the wheat and rye products. The levels of insulin, C-peptide, gastric inhibitory polypeptide (GIP), glucagon, somatostatin, triglyceride and glycerol were followed after the breakfast meals. No pronounced differences of these parameters were seen. However, wheat bread gave significantly higher glucagon and GIP responses than did rye flakes. In conclusion, the absorption pattern and metabolic response after rye bread seems preferable to that after wheat bread. The flaked rye on the other hand was not effective in reducing postprandial glycaemia despite a lower availability of starch in vitro.

Aged↗

Bioaffinity extraction of glucoamylase in aqueous two-phase systems using starch as free bioligand.

Aqueous two-phase systems with bioligands bound to the polymer, usually polyethylene glycol (PEG), have high selectivity. However, such kind of systems can be costly since some specific synthetic ligands are expensive, making them non-viable for bioaffinity extraction of low-cost proteins. The use of free bioligands is an alternative for decreasing the high cost of these systems. This work describes the use of starch as a free bioligand on the partition of glucoamylase in PEG 300-phosphate system. The results were separated into two parts. In the first part, analysis of the starch distribution between the phases showed one-sided distribution to the bottom phase. In the second part, filtered broth from submerged cultivation of Aspergillus awamori, containing glucoamylase and contaminants, was submitted to the extraction in the system with starch. In the system without starch, glucoamylase and contaminants are partitioned in the upper phase. Upon starch addition, the partition coefficient of glucoamylase was decreased nine-fold without altering the partition of contaminants. These results indicate the possibility of separation of enzymes with high-molecular-mass and hydrophilic substrates, like glucoamylase, cellulase and pullulanase, from their contaminants in a one-step extraction. Since systems made of low-molecular-mass PEG partitioned almost all of the proteins to the upper phase, the separation can be achieved by extraction of the target enzyme in the bottom phase, as in the case of the presented study.

Chromatography, Affinity↗

Microstructural characterization of starch systems by NMR relaxation and Q-space microscopy.

Nuclear magnetic resonance (NMR) relaxation and q-space diffusion measurements have been used to probe the microscopic water distribution in a variety of starch-based systems, including packed beds of native starch granules with varying water contents, starch gels, and freeze-dried starch gels. The q-space data for the granular beds is compared with a variety of theoretical models and conforms best to unbounded diffusion in a lower dimensional space. In contrast to some earlier reports, the data for the gelatinized samples are not anomalous and conform to simple unrestricted diffusion in a three-dimensional space. The paper concludes with a novel method for probing pore size distribution in freeze-dried starch gels by infusion of acetone.

Crystallization↗

Availabilities of calcium, iron, and zinc from dairy infant formulas is affected by soluble dietary fibers and modified starch fractions.

OBJECTIVE: Insoluble dietary fiber is a known inhibitor of mineral absorption, whereas the effects of soluble dietary fibers (including prebiotics) are less known. The aim was to study calcium, iron, and zinc availabilities from dairy infant formulas supplemented with soluble dietary fibers and modified starches in vitro. METHODS: Dairy infant formulas were supplemented with soluble dietary fibers (3%, dry wt) and modified starches (16% pregelatinized rice starch and 1.9% maltodextrin, dry wt) and kept in a well-controlled and defined environment in vitro. Pooled mature human milk was used as the reference standard. RESULTS: Calcium availability from standard formula was elevated by 30% after inulin supplementation (17.2%), whereas locust bean gum (11.9%) and high esterified pectin (11.7%) reduced availability by approximately 10%. Iron availability from standard formula was increased by pregelatinized rice starch (3.8%), whereas availability was reduced in the following order: high esterified pectin (2.3%), oligofructose (2.2%), and low esterified pectin (2.1%). Zinc availability was highest after the addition of pregelatinized rice starch (13.5%) but lowest with the addition of locust bean gum (6.8%) and maltodextrin (5.4%). CONCLUSIONS: This study showed that addition of soluble dietary fiber affects calcium, iron, and zinc availabilities in positive (inulin) and negative ways, depending on the type of the dietary fiber used.

Biological Availability↗

Determination of BOD-values of starch-containing waste water by a BOD-biosensor.

The control of waste water plants is difficult or even impossible using the classical determination method for biological oxygen demand (BOD), because of its high time consumption of five days. A determination within some minutes is possible by microbial BOD-sensors. However, high molecular weight substances cannot be detected, a problem which can be overcome by the use of additional enzymes. For the application in a flow-through system to analyse starch containing waste water, alpha-amylase and amyloglucosidase were immobilized by adsorption to polystyrene or polypropylene carriers followed by crosslinking. Furthermore, covalent coupling to different nylon carriers, derivatives of chitin, silanized glass beads and silanized beads of foamed glass was tried. Chitin and Lewatit were the best suited carriers for the immobilization of alpha-amylase and amyloglucosidase. Two glass columns were filled with the immobilized enzymes and inserted into a commercial BOD-sensor containing the yeast Trichosporon cutaneum as biological component. The system was stable for more than two months under storage and one month under working conditions. A comparison of different starch types resulted in a hydrolysis of more than 80% in case of potato starch whereas grain starch was hydrolized only for 40-50%. Sensor-determined BOD-values of waste water with potato starch were nearly identical with BOD5-values resulting from the classical method.

Biosensing Techniques↗

Starch-based plastic polymer degradation by the white rot fungus Phanerochaete chrysosporium grown on sugarcane bagasse pith: enzyme production.

In this study, starch metabolites and enzymes were determined during starch-based plastic polymer biodegradation by the white rot fungus Phanerochaete chrysosporium, grown in sugarcane bagasse pith in tubular reactors. Various metabolites, amylase, ligninase and cellulase production were measured during P. chrysosporium growth on sugarcane bagasse pith with added glucose and starch polymer. On-line respirometric analyses followed during 32 days confirmed the P. chrysosporium capability of growing on sugarcane bagasse pith with starch polymer degradation. Enzyme activity during secondary metabolism increased, and a 70% and 74% starch degradation was reached with and without glucose addition, generating low molecular weight metabolites (e.g.) dextrin, maltotriose, maltose and glucose that were detected by high performance liquid chromatography.

Amylases↗

Evidence for intermediate formation in the mechanism of potato starch -hosphorylase from exchange of the ester and phosphoryl oxygens of alpha-D-glucopyranose 1-phosphate.

We have examined under a variety of conditions the ability of potato starch phosphorylase to cause exchange of the ester and phosphoryl oxygens of alpha-D-glucopyranose 1-phosphate (Glc-1-P). In the presence of phosphorylase and strach, under conditions where 40-50% of the glc-1-P is consumed in starch elongation, little if any exchange occurs that cannot be accounted for by accompanying starch phosphorolysis. Nor are the oxygens scrambled in the same or longer times by enzyme only when no release of inorganic phosphate occurs. But when D-maltotriose is used as a primer, or during primerless synthesis, and in the presence of phosphorylase and alpha- or beta-cyclodextrin, a large degree of scrambling does occur. We conclude that under these latter conditions a glucosyl cation or covalent glycosyl-enzyme intermediate in formed. If this same intermediate is formed in the absence of starch or its analogue, then the phosphate counterion is not free to rotate; if the intermediate is formed with starch, then again one must assume that the rotation of phosphate ion is hindered, or that formation of the intermediate is rate determining.

Alkaline Phosphatase↗

Influence of fiber on the phase transformations in the starch-water system.

High-sensitivity, temperature-controlled DSC measurements at a low heating rate and creation of differential DSC traces scaled with respect to the reference material (completely dehydrated starch or completely dehydrated fiber, or their respective blends) permitted investigation of the influence of fiber on phase transformations in the wheat-starch-water system in the course of thermal gelatinization. Thermal effects associated with water interactions over the temperature range from 283 to 384 K under atmospheric pressure were determined. These thermal effects and previous structural studies permit us to make the following observations: (1) The main endothermic transition associated with melting of the crystalline part of the starch granule followed by a helix-coil transition in amylopectin occurs over the temperature range 319-333 K independent of the water and fiber contents. Adding fiber causes that transition to disappear both in the native blends and in water suspensions at low water contents. After adding more water and heating, recrystallization is observed and the transition reappears. (2) The fiber content has practically no influence on the slow exothermic transformation, which follows melting and helix-coil transition in amylopectin, proving that the slow transformation has a specific chemical character. In this reaction, the free ends of the unwound helices of amylopectin reassociate with parts of amylopectin molecules other than their original helix duplex partner, forming physical junctions and creating more general amorphous hydrogen bonded associations. (3) The high-temperature transition and small, but reproducible, distortions on the peaks of the main endothermic transition for water contents near 70-80 wt % are associated with smectic and nematic transitions, respectively. These are significantly influenced by the fiber content; higher fiber content causes an almost complete disappearance of these transitions. (4) The slow exothermic effect appearing almost from the very beginning of the heating in the starch-water system, associated with softening and uptake of water in the amorphous growth rings of the starch granule, is significantly hindered by added fiber.

Biopolymers↗

Immobilization of alpha-amylase to a composite temperature-sensitive membrane for starch hydrolysis.

A composite membrane was made by casting hydrogel onto a nonwoven polyester support and used for enzyme immobilization. The hydrogel consists of N-isopropylacrylamide, cross-linker N, N'-methylenebis(acrylamide), 2-hydroxyethyl methacrylate, soluble starch, and N-(acryloxy)succinimide (NAS). The composite membrane is temperature-sensitive with a lower critical solution temperature (LCST) around 35 degreesC. It responds to temperature change by swelling below the LCST and shrinking above the LCST, corresponding to opening and closing of the membrane pores. alpha-Amylase was immobilized to the membrane by covalent bonds through reacting with the high reactive ester groups in NAS. The membrane-immobilized enzyme retained 32% of specific activity toward soluble starch when compared with that of free enzyme, and its properties were characterized and compared with those of the free enzyme. The immobilized enzyme was more thermally stable than the free enzyme. Kinetic constants, (Km) and the activation energy of the immobilized enzyme were both larger than those of the free enzyme. Starch hydrolysis with the immobilized enzyme was investigated in two-compartment permeation cells with a composite membrane between the cells. Reaction was carried out by hydrolyzing soluble starch in the donor side and collecting the hydrolyzed products in the receptor side. This reactor could be operated with temperature cycling to enhance the reaction and facilitate separation of products from the substrate. The best operating condition is cycling the temperature between 50 and 20 degrees C every 5 min. The membrane reactor was operated up to eight times for successive starch hydrolysis.

Enzymes, Immobilized↗

Mobility characterization of waxy corn starch using wide-line (1)H nuclear magnetic resonance.

The molecular mobility of waxy corn starch was studied by using wide-line (1)H nuclear magnetic resonance (NMR) spectroscopy. A suite of NMR techniques was used to measure relaxation times (i.e., T(2), T(2), and T(1)) and to characterize water and solid (starch) mobility of waxy corn starch. It was observed that the spectrum of each sample includes a complex broad proton component upon which is superimposed a narrow proton component over water activity (a(w)) ranges from 0.33 to 0.97 (i.e., 10.-25.6% water content) at 25 degrees C. Line shape analysis and relaxation times of both broad and narrow components show that T(2) and T(2) values decrease (i.e., decreasing mobility) with increasing solid concentration and show a "break point" in a concentration range between 19.8 and 21.9% water content. The T(1) shows a "T(1) minimum" in the same concentration range. Starch samples change from the glassy to viscous rubbery state in this same concentration range. This demonstrates that wide-line (1)H NMR relaxation times (i.e., T(2), T(2), and T(1)) may be useful as indicators of glass transition for starch samples in the solid state. The results demonstrate that wide-line (1)H NMR spectroscopy is able to separate modes and quantitate the magnitude of molecular mobility in complex systems.

Hydrogen↗

Effect of various domestic processing methods on antinutrients and in vitro protein and starch digestibility of two indigenous varieties of Indian tribal pulse, Mucuna pruriens Var. utilis.

The effect of various domestic processing methods on antinutrients and starch fractions and in vitro protein and starch digestibilities of white and black varieties of Mucuna pruriens var. utilis was studied. Cooking or autoclaving of both raw seeds and presoaked seeds in different solutions (water, tamarind extract, sodium bicarbonate, and citric acid) significantly (p < 0.05) reduced the content of total phenolics, phytic acid, trypsin inhibitor and chymotrypsin inhibitor activities, and L-dopa compared to soaking or dry heating techniques. The germination processes (24 and 48 h) were also effective in the reduction of various antinutrients, although this reduction appeared to be more pronounced in a prolonged period of germination (72 h). Water soaking followed by dehusking was found to be ineffective in the reduction of trypsin and chymotrypsin inhibitor activities in both varieties. All of the treatments were effective in significantly (p < 0.05) reducing the resistant starch content in the presently investigated samples. Cooking as well as autoclaving brought about a more significant (p < 0.05) improvement in the digestibility of protein and starch compared to germination and dry heat treatment. Moreover, among the different processing techniques, soaking in sodium bicarbonate solution followed by cooking (29.6-34.8%) or autoclaving (33.0-37.2%) seemed to be the best method for improving starch digestibility.

Cooking↗

Retention of aroma compounds in starch matrices: competitions between aroma compounds toward amylose and amylopectin.

The retention of three aroma compounds-isoamyl acetate, ethyl hexanoate, and linalool--from starch-containing model food matrices was measured by headspace analysis, under equilibrium conditions. We studied systems containing standard or waxy corn starch with one or two aroma compounds. The three studied aroma compounds interact differently: ethyl hexanoate and linalool form complexes with amylose, and isoamyl acetate cannot. However, in systems containing one aroma compound, we observed with both starches a significant retention of the three molecules. These results indicate that amylopectin could play a role in the retention of aroma. In systems containing two aroma compounds in a blend, the retentions measured for isoamyl acetate and for linalool were either equal to or less than those in systems where they were added alone. This phenomenon was attributed to competition between aroma compounds to bind with starch. The retention of aroma compounds blended in starch-based systems gave us additional information which confirmed that interactions occur not only with amylose but also with amylopectin.

Acyclic Monoterpenes↗

Sorption and vapor transmission properties of uncompressed and compressed microcellular starch foam.

Microcellular starch foams (MCFs) are made by a solvent-exchange process and consist of a porous matrix with pores generally ranging from approximately 2 microm to submicrometer size. MCF may potentially be useful as a slow-release agent for volatile compounds because of its ability to sorb chemicals from the atmosphere and to absorb liquids into its porous structure, and because it can be compressed to form a starch plastic. MCF made of high-amylose corn and wheat starches was prepared with or without 2% (w/w) silicone oil (SO) or palmitic acid (PA). The MCF was loaded with 1% of various volatile compounds with vapor pressures ranging from 0.02 to 28 mm. The MCF depressed the vapor pressure from 0.37 to 37% compared to a control containing no MCF. Incorporating SO or PA in the matrix of the MCF had little effect on sorption of volatiles. Compressing MCF at 1.4, 6.9, and 69 MPa made a starch plastic with varying porosity. The vapor transmission rate of various volatile compounds through MCF was positively correlated to the vapor pressure of the test compound but was inversely proportional to the compression force used to form the starch plastic. The results indicate that uncompressed and compressed MCFs could be effective slow-release agents for a variety of volatile compounds, especially if used together.

Adsorption↗