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Thesis (Ph. D.)--Columbia University. / Vita. Includes bibliographies.

Retrograded starches are commonly found in foods due to the production and/or processing conditions they have received prior to consumption. These resistant starches escape digestion in the small intestine and are fermented in the colon by the microflora present, to produce gases and SCFA in varying amounts. These are utilised by the host animal as an energy source, with a low gut pH being maintained by the production of SCFA. The fermentation of carbohydrates within the colon is beneficial to the health of the gut, as the beneficial bacterial species such as Lactobacillus and Bifidobacterium spp. are maintained, and a low pH reduces the activity of potentially harmful species such as the coliforms. The production of toxic metabolites from the breakdown of proteins will be reduced if these resistant starches persist further along the colon as a carbohydrate source. This is particularly important in the distal region of the colon, where the carbohydrate source usually becomes limited. The fermentation of both native and retrograded starches from various botanical sources containing varying amounts of the major components amylose and amylopectin, was examined. In particular, the effects on bacterial fermentation of variations in the ratios of amylose and amylopectin in starch, and of treatments such as retrogradation and/or pancreatin digestion was examined.

Antistaling properties of enzymes (xylanase, bacterial maltogenic and conventional a-amylases), CMC and vital wheat gluten on corn tortillas were evaluated during storage for up to 21 days. Effect of storage time (0-21 days) and temperature (-40, -20, 3, 10 and 21 oC) on tortilla staling was evaluated with or without additives. Addition of 275-1650 AU of ICS maltogenic amylase effectively reduced amylopectin retrogradation without reducing tortilla yields, but did not improve tortilla flexibility. The combination of 825 AU of ICS amylase (to interfere with intra-granular amylopectin re-crystallization) and 0.25% CMC (to create a more flexible inter-granular matrix than retrograded amylose) produced less stiff, equally flexible and less chewy tortillas than 0.5% CMC. Corn tortilla staling followed the basic laws that control aging in starch-based semi-crystalline systems such as starch gels, bread and other baked products. Amylopectin re-crystallization was the driving force behind the staling of corn tortillas. Increasing levels of re-crystallized amylopectin measured by DSC correlated significantly with increased tortilla stiffness and reduction in tortilla rollability, pliability and rupture distance during storage. Re-crystallization of amylopectin in fresh tortillas was not detected. It increased rapidly during the first 24 hr reaching a plateau after 7 days storage. The level of amylopectin re-crystallization on tortillas showed a bell-shaped trend along the evaluated storage temperature range with a maximum around 7 oC. However, a negative linear relationship of peak pasting viscosity with storage temperature of tortilla extracts without additives after 21 days suggests other compounds besides amylopectin affect tortilla staling. Thus, interfering with amylopectin re-crystallization is not the only way to retard staling. Further research is required to optimize the addition of maltogenic amylases in continuous processing lines that use fresh masa instead of nixtamalized corn flour, to determine how these amylases interfere with amylopectin re-crystallization and to elucidate if amylose retrogradation continues during storage and plays a role in tortilla staling.

Starch is the major constituent of matured wheat grain. The details of subtle localized differences in the evolution of the structure of starch are important for an understanding of starch biosynthesis. However, the distinct stages involved in the formation and transformation of the molecular structure of starch during starch biosynthesis are still not fully understood. In this study, starches extracted from wheat grains harvested at 3, 7, 14, 28, and 49 days after anthesis (DAA) were used as a means of examining the molecular structure of starch from developing wheat grain. Gel-permeation chromatography and high-performance anion-exchange chromatography were employed for the analysis of the structure of both whole starch and its isolated amylopectin (AP) component. Scanning electron microscopy of 3 DAA wheat grain cross-sections revealed the absence of endosperm but the presence of spherical transitory-type small starch granules in the pericarp. Endosperm was present at 7 DAA and contained lenticular-shaped developing large granules. From 14 DAA onward, spherical-shaped small granules coexisted with large granules in the endosperm. The structure of transitory pericarp starch (PS) was compared with that of matured endosperm storage starch (ES). The composition of PS and ES differed: PS granules contained 14 % apparent amylose (AAM), whereas ES granules contained 33 % AAM. The AAM fractions of PS showed characteristics similar to those of intermediate-type materials with short branches, whereas ES contained both linear and branched amylose (AM). Differences in the amylopectin component of PS and ES were also apparent, especially in their internal structures. PS amylopectin had longer chains and fewer A-chains, resulting in a structure less branched than that of ES amylopectin. Starches isolated from 7 DAA to 49 DAA were studied with respect to endosperm development. The AAM of both large and small granules increased with increasing maturity. The AAM fraction of starch granules at early maturity (7 DAA and 14 DAA) consisted of intermediate-type materials in addition to linear AM, whereas starch at later maturity stages (28 and 49 DAA) contained linear and branched AM. During granule development, the fine structure of AP varied with the maturity level as well as with the size of the granule. During the post-physiological maturity stage, when the net accumulation of sugars ceases, the grain dries out; however, structural changes occurred in AP at this stage, possibly due to the action of starch branching and debranching enzymes. In both large and small granules, the external AP structure was more organized at post-physiological maturity (49 DAA) than at pre-physiological maturity (7 DAA to 28 DAA). Compared to their characteristics at post-physiological maturity, at the pre-physiological maturity stage, isolated clusters of AP were larger with more branches and building blocks. In addition to the time-dependent discrepancies in the AP structure of developing starch, differences were also evident between large and small granules with regard to glucan trimming and the type of new chains produced. The clusters isolated from small starch granules were more tightly branched than those isolated from large granules

Neste trabalho, investigamos o microambiente da amilopectina na ausência e presença de tensoativos não-iônicos como o éter dodecil (7 e 23)-polioxietilênico (Unitol L-70 e L-230) e o nonilfenol (9,5)-polioxietilênico (Renex 95), por meio das propriedades físico-químicas de sondas de polaridade: pireno, xantona e Reichardt. Os estudos foram realizados através de técnicas espectroscópicas em estado estacionário (absorção de luz ultravioleta-visível e fluorescência) e resolvidas no tempo (baseadas na contagem de fótons únicos da emissão de fluorescência e na absorção triplete-triplete (T-T) e de intermediários reativos formados após a fotólise da amostra por pulso de laser). Estas técnicas forneceram os valores de: concentração de agregação crítica (cac), concentração micelar crítica (cmc), número de agregação (Nag) e polaridade (escala py e ET30) para os sistemas avaliados. Os dados obtidos foram comparados com as soluções contendo a presença ou não dos tensoativos não-iônicos e a carboximetil celulose (CMC) e 2-hidroxietilamido (2-HEAM). Por fim, a estabilidade do cátion 2-fenilbenzopirílio foi avaliada em diferentes solventes orgânicos, nas micelas dos tensoativos estudados, nos biopolímeros e nos complexos biopolímero-tensoativo. Dois procedimentos experimentais diferentes foram adotados para o preparo das soluções de amilopectina. As diferenças na preparação das soluções de amilopectina tornaram evidente que este biopolímero possui diferentes ambientes para a localização das sondas. Estas diferenças não foram observadas na presença de CMC e de 2-HEAM. Os resultados experimentais confirmaram a interação cooperativa entre a amilopectina e os tensoativos Unitol L-70 e L-230. Esta interação é menos pronunciada para os sistemas formados entre os tensoativos e 2-HEAM e desfavorecidos na presença de CMC. Nos experimentos onde foi avaliado o equilíbrio monômero-excímero do pireno, verificou-se que os complexos amilopectina-tensoativo agrupam um número maior de moléculas de pireno do que os clusters de amilopectina sozinhos. Estes ambientes favoreceram ambos os movimentos de rotação e translação entre as moléculas, para a formação do excímero. Os estudos resolvidos no tempo, utilizando a xantona como sonda fotofísica para as técnicas de fluorescência resolvida no tempo ou de fotólise por pulso de laser, demonstraram que na amilopectina, a sonda ocupa preferencialmente o ambiente aquoso, ao invés dos microambientes internos do polímero. Isto possivelmente é uma decorrência da proteção das cadeias ramificadas do seu esqueleto polimérico. Estes mesmos estudos indicaram que na presença de concentrações mais elevadas de CMC, a xantona migra do seio da solução aquosa para ambientes próximos à cadeia polimérica onde podem ocorrer sua protonação ou associação em sítios não protonados. Em relação à estabilização do cátion 2-fenilbenzopirílio, nas condições estudadas envolvendo tanto os biopolímeros quanto os sistemas biopolímero tensoativo, predominaram os isômeros da chalcona. Os maiores tempos de meia-vida foram observados em CMC, o que pode ser justificado pela maior viscosidade do meio. / In this work, the amylopectin microenvironment was studied in the presence and absence of the non-ionic surfactants polyoxyethylene (7 and 23) dodecyl ether (Unitol L-70, and L-230, respectively) and polyoxytethylene (9.5) nonylphenol ether (Renex 95), through the physical-chemical properties of the polarity probes: pyrene, xanthone, and Reichardt. Steady-state (UV-visible absorption, and fluorescence), and time-resolved (time-correlated single photon counting fluorometry, and laser flash photolysis) techniques were used to evaluate the critical aggregation concentration (cac), critical micellar concentration (cmc), average micellar aggregation numbers (Nag), and polarity (py, and ET30 scales) of the complexes of amylopectinnon-ionic surfactants. The amylopectin experimental results were compared with the obtained ones from aqueous solutions of carboxymethyl cellulose (CMC) and 2-hydroxyethyl starch (2-HEAM) in the absence and presence of nonionic surfactants. The stability of 2-phenylbenzopyrilium cation was also investigated in several organic solvents, in the non-ionic surfactant micelles, in the biopolymers and in the complexes of biopolymer-surfactants. The amylopectin aqueous solutions were prepared by two different experimental procedures. The determined results of the relative intensities of pyrene fluorescence emission bands 1 to 3, and excimer and monomer ratio have pointed out that the amylopectin presents different microenvironments for photophysical probes relocation. Such differences were not observed in the presence of CMC and 2-HEAM. The cooperative interaction between the amylopectin, and the Unitol-L70 and L- 230 surfactants were comproved by the experimental results. The complexes of 2- HEAMnon-ionic surfactant were less stable than the amylopectin ones, while CMC does not form complexes of biopolymer-surfactants. The single-photon timing and laser flash photolysis techniques employing xanthone as probe have shown that this probe remains at the aqueous bulk of the amylopectin solutions. There is no interaction between xanthone and the amylopectin backbone, probably due to the great extent of the polymeric branching. On the other hand, in presence of higher concentrations of CMC solutions, xanthone can relocate from the aqueous solution to the polymeric microenvironment, where the probe can be or not protonated. The evaluation of the stability of 2-phenylbenzopyrilium cation in the presence of biopolymer or complexes of biopolymer-surfactants has shown the predominance of the chalcone isomers. The higher half-life times were determined in the CMC because of the higher viscosity of the medium.

Wheat grain quality and consumption is influenced by its constituents structure and concentrations. In the first part of the dissertation, six Canadian bread wheat cultivars; four (CDC Teal, AC Superb, AC Barrie, AC Splendor) belonging to the Canada Western Red Spring (CWRS), and two (AC Foremost, and AC Crystal) to the Canada Prairie Spring Red (CPSR) market classes were characterized for the relationship between their starch constituents and starch in vitro enzymatic hydrolysis. CPSR cultivars with relatively longer amylopectin chains of DP 37-45, reduced chain lengths of DP 15-18, and a low volume percent of small C-type starch granules, had reduced starch in vitro enzymatic hydrolysis rates. In the second part of the dissertation, near-isogenic wheat lines differing at the Waxy locus were analyzed for the influence of genome-specific granule-bound starch synthase I (GBSSI/Waxy; Wx-A, Wx-B, Wx-D) on starch composition, structure and starch in vitro enzymatic hydrolysis. Amylose concentration was more severely affected in genotypes with GBSSI missing from two genomes (double nulls) than from one genome (single nulls) of wheat, indicating dosage dependent amylose synthesis. Subtle differences in amylopectin chain length distribution were observed among non-waxy, partial and completely waxy starches, suggesting a non-limiting role of genome-specific GBSSI for amylopectin synthesis. A suppressive role of Wx-D on the short chain phenotype of wheat amylopectin was observed. In addition, Wx-D increased the volume percentage of large A-type starch granules and reduced starch hydrolysis index. Thus, among the waxy isoproteins, Wx-D might be the major contributor for reducing the rate of in vitro starch enzymatic hydrolysis in wheat. In the third part of the dissertation, endosperm starch’s physicochemical properties and structure during grain development in wheat waxy-null genotypes were analyzed. The study was conducted with pure starch isolated from wheat grains at 3-30 days post anthesis (DPA), at three day intervals. Changes in amylopectin structure were observed until 12 DPA, suggesting the formation of a basic amylopectin skeleton by this stage. A differential influence of waxy isoproteins on amylopectin structure formation has been suggested, with Wx-B and Wx-D affecting short glucan chains of DP 6-8 at 3 and 6 DPA, Wx-A being effective at 9 and 12 DPA, and Wx-D affecting DP 18-25 chains from 18-30 DPA.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2004. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2004. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2004. / Härtill 5 uppsatser.