Novel plastics that are biodegradable and made from renewable natural resources are currently being researched as alternatives to traditional petroleum-based plastics. One such plastic, thermoplastic starch (TPS) is produced from starch processed at high temperatures in the presence of plasticizers, such as water and glycerol. However, because of its hydrophilic nature, TPS exhibits poor mechanical properties when exposed to environmental conditions, such as rain or humidity. The overall objective of this thesis was to produce a thermoplastic starch based material with low water absorption that may be used to replace petroleum-based plastics. Three different methods for reducing water absorption were investigated, including the following: extrusion of starch with hydrophobic polymers, starch modifying chemicals, and citric acid/sorbitol as plasticizers. It was found that all methods reduced the water absorption of TPS.

Novel plastics that are biodegradable and made from renewable natural resources are currently being researched as alternatives to traditional petroleum-based plastics. One such plastic, thermoplastic starch (TPS) is produced from starch processed at high temperatures in the presence of plasticizers, such as water and glycerol. However, because of its hydrophilic nature, TPS exhibits poor mechanical properties when exposed to environmental conditions, such as rain or humidity. The overall objective of this thesis was to produce a thermoplastic starch based material with low water absorption that may be used to replace petroleum-based plastics. Three different methods for reducing water absorption were investigated, including the following: extrusion of starch with hydrophobic polymers, starch modifying chemicals, and citric acid/sorbitol as plasticizers. It was found that all methods reduced the water absorption of TPS.

Most of the calories in cereal foods come from starch. Decreasing starch digestibility is fundamental to prevent obesity and diabetes. This study investigated interactions of condensed tannins (proanthocyanidins-PA) and other sorghum phenolic compounds with starch molecules and their effect on in vitro starch digestibility. High tannin (predominant in large molecular weight PA, 80%), black (monomeric polyphenols) and white (low in polyphenols) sorghum phenolic extracts were cooked with starches varying in amylose content. Starch pasting properties, polyphenol profile and in vitro starch digestibility were evaluated. Unlike other treatments, samples with tannin phenolic extracts had significantly (P ≤ 0.05) lower setback in the test using a Rapid Visco Analyser (RVA) compared to control. The same treatments had the least extractable phenol and PA contents after cooking with all starches. These evidences suggest interactions between starch molecules and PA. Furthermore, after mixing tannin phenolic extracts with pure amylose/amylopectin, extractable polymeric PA was in much lower concentration (62% less) in presence of amylose compared to amylopectin. This drop in concentration increased to 85% when purified tannin extract (90% polymeric PA) was used. This indicates a stronger interaction between amylose and large molecular weight PA. When high amylose starch was used in an autoclave cooking/cooling technique, the RS content of control (26.4%) was similar (P > 0.05) to samples with black phenolic extracts (27%); samples with tannin phenolic extracts increased RS to about 40%. The RS increased to 46% when purified tannin extract was used. All these evidences suggest that sorghum condensed tannins, specifically the polymeric PA, directly interacted with amylose, increasing RS content, whereas the monomeric polyphenols did not. This study opens opportunities to use tannin sorghum to develop products for diabetics and weight control, high in dietary fiber and natural dark color. In the other part of this project, polyphenols from black and tannin sorghum bran were extracted using an Accelerated Solvent Extractor (ASE) and eco-friendly solvents such as water, and mixtures ethanol/water. ASE at 120 and 150 degrees C using 50 and 70% ethanol/water was efficient in extracting as much phenols (45 mg GAE/g) and 12% more antioxidants (628 μmol TE/g) from black sorghum compared to conventional methods using aqueous acetone and acidified methanol. Therefore, ASE extracts from black sorghum could be used in beverages and in colorants containing high antioxidant content.

Ethanol can be used as a complete fuel or as an octane enhancer, and has the advantages of being renewable and environmentally friendly. Ethanol produced by a fermentation process, generally referred to as bioethanol, is considered to be a partial solution to the worldwide energy crisis. Traditionally, industrial bioethanol fermentation involves two major steps: starch hydrolysis and fermentation. Since the key microorganism, Saccharomyces cerevisiae, lacks amylolytic activity and is unable to directly utilize starch for proliferation and fermentation, it requires intensive amount of energy and pure starch hydrolyzing enzymes to gelatinize, liquefy and dextrinize the raw starch before fermentation. It has been suggested that genetically engineered yeast which expresses amylolytic enzymes could potentially perform simultaneous starch hydrolysis and fermentation. This improvement could greatly reduce the capital and energy costs in current bioethanol producing plants and make bioethanol production more economical. In this project, a novel yeast strain of Saccharomyces cerevisiae was genetically engineered in such a way that barley alpha-amylase was constitutively expressed and immobilized on the yeast cell surface. This particular alpha-amylase was selected based on its superior kinetic properties and its pH optimum which is compatible with the pH of yeast culture media. The cDNA encoding barley Ñ-amylase, with a secretion signal sequence, was fused to the cDNA encoding the C-terminal half of a cell wall anchoring protein, alpha-agglutinin. The fusion gene was cloned downstream of a constitutive promoter ADH1 in a yeast episomal plasmid pAMY. The pAMY harbouring yeast showed detectable amylolytic activity in a starch plate assay. In addition, alpha-amylase activity was detected only in the cell pellet fraction and not in the culture supernatant. In batch fermentation studies using soluble wheat starch as sole carbon source, even though pAMY harbouring yeast was able to hydrolyse soluble starch under fermentation conditions, no ethanol was produced. This was probably due to insufficient alpha-amylase activity which resulted from the enzyme being anchored on the cell wall by alpha-agglutinin. Further research using alternative cell surface anchoring system might be able to produce yeast with industrial applications.

Ethanol can be used as a complete fuel or as an octane enhancer, and has the advantages of being renewable and environmentally friendly. Ethanol produced by a fermentation process, generally referred to as bioethanol, is considered to be a partial solution to the worldwide energy crisis. Traditionally, industrial bioethanol fermentation involves two major steps: starch hydrolysis and fermentation. Since the key microorganism, Saccharomyces cerevisiae, lacks amylolytic activity and is unable to directly utilize starch for proliferation and fermentation, it requires intensive amount of energy and pure starch hydrolyzing enzymes to gelatinize, liquefy and dextrinize the raw starch before fermentation. It has been suggested that genetically engineered yeast which expresses amylolytic enzymes could potentially perform simultaneous starch hydrolysis and fermentation. This improvement could greatly reduce the capital and energy costs in current bioethanol producing plants and make bioethanol production more economical. In this project, a novel yeast strain of Saccharomyces cerevisiae was genetically engineered in such a way that barley alpha-amylase was constitutively expressed and immobilized on the yeast cell surface. This particular alpha-amylase was selected based on its superior kinetic properties and its pH optimum which is compatible with the pH of yeast culture media. The cDNA encoding barley Ñ-amylase, with a secretion signal sequence, was fused to the cDNA encoding the C-terminal half of a cell wall anchoring protein, alpha-agglutinin. The fusion gene was cloned downstream of a constitutive promoter ADH1 in a yeast episomal plasmid pAMY. The pAMY harbouring yeast showed detectable amylolytic activity in a starch plate assay. In addition, alpha-amylase activity was detected only in the cell pellet fraction and not in the culture supernatant. In batch fermentation studies using soluble wheat starch as sole carbon source, even though pAMY harbouring yeast was able to hydrolyse soluble starch under fermentation conditions, no ethanol was produced. This was probably due to insufficient alpha-amylase activity which resulted from the enzyme being anchored on the cell wall by alpha-agglutinin. Further research using alternative cell surface anchoring system might be able to produce yeast with industrial applications.

A common modification of starch is esterfication with anhydrous octenyl succinic acid (OSA).  The modification makes the polymer surface active and it also incorporates a carboxyl group to the starch, which can be negatively charged. The characteristics of OSA starch make it interesting for usage in combination with egg yolk proteins in food emulsions. It is not only the individual ingredients that affect the product; interactions between ingredients and ingredient-dispersion medium have a great impact on factors such as structure and stability. Knowledge about how the interactions affect emulsion properties would make it possible to predict the behavior of an emulsion, which would be a great advantage in the formulation of food emulsions. Therefore, this is a subject of interest. The purpose of this master thesis was to further investigate the interactions between OSA starch and α – β-livetin in solutions and in emulsions. First, the charges of the macromolecules were studied by titration. Interactions in solution were then analyzed through turbidity and solubility measurements. The adsorption of OSA starch onto livetin and the interfacial rheology were also studied. Finally, an emulsion stability experiment was made. Strong interactions between the two macromolecules were observed in solutions at pH 4.0. This was probably due to hydrophobic interaction; however it could also be explained by electrostatic interaction. In the emulsions the adsorption of starch onto livetin was highest at pH 4.5, and then decreased with increasing pH values. The absence of OSA starch adsorption at pH 4.0, despite the strong interaction in solution, could be explained by complex formation immediately in solution. Less starch would then be able to reach the interface and adsorb. In the interfacial rheology experiments, an indication of decreased complex dilational modulus of the interfacial layer, caused by OSA starch addition was seen at low pH values. This could be due to aggregation of the proteins and formation of an uneven interfacial layer. OSA starch would then be able to adsorb and disturb the elasticity. Some differences in the stability of an emulsion only containing livetin, and an emulsion with both livetin and OSA starch could be observed. However, more investigations are needed to be made to understand the underlying mechanisms.

Eight cecally fistulated geldings were used in a randomized 4 x 4 Latin square design to observe the effect varying levels of dietary starch had on cecal environment. The 4 treatment rations contained 2 g starch/kg BW (Diet 2), 4 g/kg BW (Diet 4), 6 g/kg BW (Diet 6), or 8 g/kg BW (Diet 8). The rations were comprised of a commercial pelleted feed to meet 2 g starch/kg BW in each treatment, with ground corn used to fulfill the remaining starch requirements in each diet. Soybean meal was added to ensure diets were iso-nitrogenous, and cottonseed hulls were used to equalize dry matter intake. A 21 day adaptation period was allowed before cecal contents were sampled. Samples were drawn 4 hours after the morning meal and were immediately tested for pH. Samples were used to count total anaerobic bacteria and lactic acid bacteria, as well as determine methane activity, ammonia activity, volatile fatty acids, and in vitro dry matter digestibility (IVDMD). Stoichiometric calculations were performed to give an indirect measure of fermented hexose, methane, and carbon dioxide. Diet did not influence dry matter intake (DMI), however it did have an effect on starch intake (P < 0.0001) and caused a linear increase in starch consumption as the amount of offered starch increased (P < 0.0001). Diet did not influence the pH of the cecum (P > 0.05), although a tendency for a linear decrease (P < 0.06) in pH from 6.92 ? 6.58 occurred when dietary starch increased. Total anaerobic bacteria and lactic acid bacteria were unaffected by treatment diets (P > 0.05). Propionate production was affected by dietary treatment (P < 0.05), causing a quadratic increase (P = 0.04) from 8.26 to 14.13 mM as starch in the diets increased. Diet did not affect the production of acetate, butyrate, or ammonia (P > 0.05). Results found that stoichiometric calculations and IVDMD values were not affected by diet (P > 0.05). These results show that starch intake influenced the production of fermentative by-products, which caused decreases in pH, although there was no observed increase in the bacterial populations of the cecum.

Fractions from white and tannin sorghums were processed into extrudates, thick porridges, and breads. The effects of sorghum type and fraction on the in vitro starch availability of the products were evaluated, and the estimated glycemic indexes (EGI) of the products were obtained. Sorghum extrudates were significantly more slowly digested than corn meal extrudates for all preparation methods (whole, cracked and decorticated kernels). Furthermore, tannin extrudates were less digestible than white sorghum extrudates. The soft endosperm nature of the tannin sorghum limited friction formation inside the extruder, reducing starch gelatinization. On the other hand, condensed tannins also interfered with the starch availability for digestion. White sorghum was more 'suitable' for extrusion, giving extrudates with higher starch degradation and expansion than the tannin sorghums. However, tannin sorghums also gave acceptable products offering the benefit of lower EGI values. Sorghum porridges were more slowly digested than a corn flour porridge when using whole and decorticated flours. In addition, tannin sorghum porridges had a lower starch digestibility compared to all the samples. Tannin sorghum flours produced soft porridges with enhanced initial starch digestibility. However, condensed tannins seemed to offset the starch digestion by limiting starch availability. All sorghum porridges had significantly lower EGI values than the corn porridge. Extrudates and porridges had reduced starch digestibilities and EGI values when using whole grains compared to using the decorticated fractions. This was observed in both the white and the tannin sorghum. Therefore, whole-grain products from sorghum have health benefits attributed to whole grain foods and slower digesting starches; for instance, prevention and treatment of diseases such as diabetes, insulin resistance, obesity, cardiovascular disease, and some types of cancer. When 12% of tannin bran was added to a wheat bread formulation, a slower rate of starch digestion was observed compared to a wheat bread. The high concentration of non-starch components of the bran (i.e. dietary fiber, condensed tannins) affected starch digestion. The addition of tannin sorghum bran significantly reduced the EGI value of wheat bread, besides being a natural source of brown color, and an excellent source of antioxidants and dietary fiber.

Two tannin sorghums, (high-tannin, black with tannin), high anthocyanin sorghum (black), and non-tannin sorghum (white) were used to investigate the effects of sorghum phenolic compounds on in vitro starch digestibility, estimated glycemic indices (EGI) and resistant starch contents (RS) of porridges. Sorghum varieties were chosen to have a wide range of total phenols (3-23 mg/g gallic acid) and tannin contents (0-34 mg catechin eq./g). Normal corn starch, enzyme resistant high amylose corn starch, and whole sorghum grains were cooked with the aqueous fraction of sorghum bran extracts obtained with 70% aqueous acetone. Endosperms of soft and hard sorghum varieties were mixed with sorghum brans and cooked into porridges with distilled water. Hi-tannin, black and black with tannin sorghum bran extracts significantly (p<0.05) decreased starch digestibility and estimated glycemic index (EGI) while they increased resistant starch (RS) contents of normal corn starch, enzyme resistant high amylose corn starch, and whole sorghum grain porridges. The highest reduction in starch digestibility of the porridges occurred with hi-tannin sorghum bran extracts, followed by black with tannin and black sorghum bran extracts. Double cooked corn starch porridges, which were cooked with these bran extracts had EGI values of 49-67 and RS contents of 9.1-57.7%. These RS values are higher than foods such as legumes, whole pasta and whole grain cereals which are considered health foods with low EGI (36-71) and high RS contents (2.9-6.8). Only brans of condensed tannin-containing sorghum varieties (tannin, black with tannin sorghums) significantly (p<0.05) decreased starch digestibility, and EGI, and increased RS contents of the endosperm porridges. When tannin sorghum bran extracts were cooked with zein added to corn starch porridges, starch digestibility of the porridges significantly (p<0.05) increased, while RS significantly (p<0.05) decreased because the zein reacted preferentially with the tannins. The cooking trials indicated that sorghums with tannins significantly reduced the activity of digestive enzymes, reduced EGI, and increased RS contents of porridges. Thus, specialty sorghum varieties have a potential to lower EGI and increase RS contents of starchy foods. Their aqueous bran extracts have potential use to reduce risk factors for type II diabetes and obesity.

Thesis (M.Sc.)--Memorial University of Newfoundland, 2000. / Restricted until November 2001. Bibliography: leaves 116-132.