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Bruce R Hamaker

Publications and source records attributed to Bruce R Hamaker.

12 recordsLinked to original sources

Cys155 of 27 kDa maize gamma-zein is a key amino acid to improve its in vitro digestibility.

Twenty-seven kilodalton gamma-zein is a subclass of the maize zein storage proteins and, due to its localization at the protein body periphery, is critical to digestibility characteristics of all zeins. This protein had low in vitro digestibility, presumably due to its high Cys content (7.35 mol%) that is similar to the hard-to-digest analogous sorghum protein, gamma-kafirin. Therefore, each of the conserved disulfide-bonded Cys' was mutated to create C144A, C148A, C155A, and C156A maize gamma-zein mutants. The C155A showed a remarkable increase in digestibility to proteases - pepsin, chymotrypsin, and trypsin. A high conservation of this Cys among cereal gamma-prolamins indicates the utility of this finding.

Amino Acid Sequence↗

Development of a low glycemic maize starch: preparation and characterization.

A low glycemic index starch was developed by partial alpha-amylase treatment, and its fine structure responsible for slowly digestible and resistant properties was investigated. Different digestion rates were obtained for gelatinized, retrograded starch by varying the enzyme dosage and reaction time. Analysis by high performance size-exclusion chromatography (HPSEC) coupled with multiangle laser-light scattering indicated that the molecular weighs of amylopectin and amylose were reduced during the digestion, to less than 100 kDa. A debranched chain length study using high performance anion-exchange chromatography equipped with an amyloglucosidase reactor and a pulsed amperometric detector and HPSEC revealed that short chains of amylopectin and noncrystalline amylose were rapidly digested, while DPn 121 chains showed resistance, followed by DPn 46 chains. X-ray diffraction analysis revealed that the crystalline structure in the treated starches survived cooking. These starches not only have slowly digestible and resistant character, but also retain some branched structure for adequate functionality.

Administration, Oral↗

Slow digestion property of native cereal starches.

The slow digestion property of native cereal starches, represented by normal maize starch, was investigated. The in vitro Englyst test showed that 53.0% of the maize starch is slowly digestible starch (SDS), and scanning electron microscopy (SEM) revealed that SDS starts from an increase of pore size until almost complete fragmentation of starch granules. However, similar amounts of SDS ( approximately 50%) were shown for partially digested fragmented starch residuals, which would normally be considered resistant to digestion based on the Englyst assay. Molecularly, both amylopectin (AP) and amylose (AM) contributed to the amount of SDS as evidenced by a similar ratio of AP to AM at different digestion times. Consistently, similar degrees of crystallinity, comparable gelatinization behavior, and similar debranched profiles of starch residuals following different digestion times indicated that the crystalline and amorphous regions of starch granules were evenly digested through a mechanism of side-by-side digestion of concentric layers of semicrystalline shells of native starch granules.

Crystallization↗

Structural basis for the slow digestion property of native cereal starches.

Native cereal starches are ideal slowly digestible starches (SDS), and the structural basis for their slow digestion property was investigated. The shape, size, surface pores and channels, and degree of crystallinity of starch granules were not related to the proportion of SDS, while semicrystalline structure was critical to the slow digestion property as evidenced by loss of SDS after cooking. The high proportion of SDS in cereal starches, as compared to potato starch, was related to their A-type crystalline structure with a lower degree of perfection as indicated by a higher amount of shortest A chains with a degree of polymerization (DP) of 5-10. The A-type amorphous lamellae, an important component of crystalline regions of native cereal starches, also affect the amount of SDS as shown by a reduction of SDS in lintnerized maize starches. These observations demonstrate that the supramolecular A-type crystalline structure, including the distribution and perfection of crystalline regions (both crystalline and amorphous lamellae), determines the slow digestion property of native cereal starches.

Amylopectin↗

A 50 kDa maize gamma-zein has marked cross-reactivity with the almond major protein.

Cross-reactivity of antibodies against almond major protein (AMP, a legumin), the major almond allergen, with cereal proteins may cause problems in detecting almond contaminants in cereal products when antibody-based assays are used. Rabbit polyclonal IgG antiserum produced against AMP was used to test cross-reactivity with protein extracts from maize, a cereal commonly found in breakfast and snack foods. Gradient SDS-PAGE followed by Western blotting was performed, and two cross-reactive proteins were detected by chemiluminescence. A fraction of maize proteins purified by elution from an IgG anti-AMP affinity column followed by electrophoreseis and immunoblotting showed a high degree of cross-reactivity with a minor 50 kDa protein of maize, as well as low cross-reactivity with the 27 kDa gamma-zein. The 50 kDa cross-reactive protein was identified as the 50 kDa gamma-zein by immunoreaction with anti-50 kDa gamma-zein antiserum. Notably, the 50 kDa maize gamma-zein also reacted with IgE from pooled human sera from patients with self-reported severe almond allergies. The high immunoreactivity of the 50 kDa gamma-zein should be considered in maize quality improvement programs, and such notable cross-reactivity is of relevance in the design of antibody-based assays for almond allergen detection.

Allergens↗

A three component interaction among starch, protein, and free fatty acids revealed by pasting profiles.

A three way interaction among starch, protein, and lipid that affects the Rapid Viscoanalyzer (RVA) paste viscosity profile was revealed using a model system composed of isolated sorghum starch, whey protein isolate, and free fatty acids (FFAs) (20:2:1, w/w/w). A prominent cooling stage viscosity peak in the RVA profile was produced when all three components were present in the system, while there was no viscosity peak when either protein or FFA alone was combined with starch. The magnitude of the cooling stage viscosity peak differed with addition of palmitic, oleic, or linoleic acids to starch and protein. Amylose was the major functional molecule of the starch component. Addition of both protein and FFA to starch substantially reduced starch solubility after gelatinization, while solubility was less affected by single addition of FFA and was not affected by protein. Nonspecificity of this interaction phenomenon was demonstrated by similar results using maize starch and other soluble proteins.

Absorption↗

Detection of a novel three component complex consisting of starch, protein, and free fatty acids.

A water soluble three way complex composed of starch, whey protein, and free fatty acid (FFA) was detected in a dilute three component system after heating. In high-performance size exclusion chromatography (HPSEC) profiles of the starch-protein-FFA system, the three way complex eluted between amylopectin and amylose. The molecular mass of the complex, based on multiangle laser light scattering/HPSEC and pullulan standards, was estimated to be approximately (6-7) x 10(6) Da. Carbohydrate measurement by the phenol-sulfuric acid method clearly showed that the starch amylose fraction shifted to a higher molecular weight elution volume following complexation. Whey protein existed as large disulfide-linked aggregates and is speculated to be the organizer of the three way complex. Differential scanning calorimetry of the freeze-dried complex showed the presence of an amylose-FFA melting endotherm, thus proving that FFA was the third component in the three way complex and that the amylose-FFA complex was one of the structural components of the complex. The complexation mechanism and its relationship with changes in starch functionality were discussed.

Amylopectin↗

Isolation, purification, and biochemical characterization of a novel water soluble protein from Inca peanut (Plukenetia volubilis L.).

A water soluble storage albumin from Inca peanut (IPA) accounted for approximately 25% (w/w) of defatted seed flour weight, representing 31% of the total seed protein. IPA is a 3S storage protein composed of two glycosylated polypeptides, with estimated molecular weights (MW) of 32800 and 34800 Da, respectively. IPA has an estimated sugar content of 4.8% +/- 0.92% (n = 6). IPA is a basic protein (pI of approximately 9.4) and contains all of the essential amino acids in adequate amounts when compared to the FAO/WHO recommended pattern for a human adult. The tryptophan content of IPA is unusually high (44 mg/g of protein), whereas the phenylalanine content is low (9 mg/g of protein). IPA is a highly digestible protein in vitro.

Amino Acid Sequence↗

Nutritional value of a highly digestible sorghum cultivar for meat-type chickens.

The nutritional value of a newly discovered sorghum mutant cultivar (P851171), with high in vitro protein digestibility, was compared to those of corn and two normal sorghums (P721N and 611Y) in two chick feeding trials. Although 8-20 day protein efficiency ratios and net protein ratios of all three sorghums were inferior to those of corn, P851171 and 611Y had markedly greater mean true amino acid digestibilities (TAAD) than either corn or P721N. In a subsequent 42-day experiment, all three sorghums supported weight gains equal to those of the corn-fed chicks. Feeding suboptimal levels of dietary protein resulted in reduced weight gains and no observed benefits of P851171 or 611Y. Furthermore, chicks fed P851171 exhibited poorer feed/gain values as compared to those fed the other cereals. It is possible that the starch content/carbohydrate profile of P851171 was inferior to that of the other sorghums, which offset its superior TAAD and resulted in poorer broiler performance.

Amino Acids↗

Maize opaque endosperm mutations create extensive changes in patterns of gene expression.

Maize starchy endosperm mutants have kernel phenotypes that include a brittle texture, susceptibility to insect pests, and inferior functional characteristics of products made from their flour. At least 18 such mutants have been identified, but only in the cases of opaque2 (o2) and floury2 (fl2), which affect different aspects of storage protein synthesis, is the molecular basis of the mutation known. To better understand the relationship between the phenotypes of these mutants and their biochemical bases, we characterized the protein and amino acid composition, as well as the mRNA transcript profiles, of nearly isogenic inbred lines of W64A o1, o2, o5, o9, o11, Mucuronate (Mc), Defective endosperm B30 (DeB30), and fl2. The largest reductions in zein protein synthesis occur in the W64A o2, DeB30, and fl2 mutants, which have approximately 35 to 55% of the wild-type level of storage proteins. Zeins in W64A o5, o9, o11, and Mc are within 80 to 90% of the amount found in the wild type. Only in the cases of o5 and Mc were significant qualitative changes in zein synthesis observed. The pattern of gene expression in normal and mutant genotypes was assayed by profiling endosperm mRNA transcripts at 18 days after pollination with an Affymetrix GeneChip containing >1400 selected maize gene sequences. Compared with W64A sugary1, a mutant defective in starch synthesis, alterations in the gene expression patterns of the opaque mutants are very pleiotropic. Increased expression of genes associated with physiological stress, and the unfolded protein response, are common features of the opaque mutants. Based on global patterns of gene expression, these mutants were categorized in four phenotypic groups as follows: W64A+ and o1; o2; o5/o9/o11; and Mc and fl2.

DNA-Binding Proteins↗

Interaction of maize zein with wheat gluten in composite dough and bread as determined by confocal laser scanning microscopy.

Protein body-free maize zein, when mixed at 35 degrees C (above its glass transition temperature range), significantly (p < 0.01) improved the rheological and leavening properties of sorghum-wheat composite flour dough, resulting in improved loaf volume. Confocal laser scanning microscopy was used to observe the structure of zein fibrils and the interaction between zein and gluten proteins in the composite dough and bread systems. Autofluorescence and immunolocalization techniques were used to locate gluten and zein, respectively. Optical sections were collected every 0.4 microm through the samples and digitally processed to produce reconstructed three-dimensional images. Results showed that zein fibrils form an outer layer that intermittently coats the gluten networks, thereby strengthening them. This type of microstructure is able to withstand the pressure exerted by gas cell expansion during yeast fermentation to increase loaf volume.

Bread↗

Role of the pericarp cellulose matrix as a moisture barrier in microwaveable popcorn.

Since moisture loss of popcorn can cause an increase in the number of unpopped kernels, pericarp properties of popcorn hybrids were analyzed to understand factors affecting moisture loss rate during microwave heating. Differential scanning calorimetry profiles of ground pericarp displayed a notable exothermal event, and hybrids with superior microwave popping performance (fewer unpopped kernels) exhibited significantly higher enthalpies. The number of unpopped kernels was highly correlated (r = 0.826, p = 0.011) with pericarp enthalpy values. X-ray analysis confirmed that cellulose and arabinoxylan are the major structural components of the pericarp. Structural changes in cellulose were induced by moisture and heat, and considerable enhancement in crystallinity occurred when the pericarp was heated in the presence of water. Results of this study indicate that the cellulose component of the pericarp is responsible for the development of exothermal events and increased crystallinity. Thus, the propensity of cellulose to form crystalline structures in the popcorn pericarp during microwave heating improves moisture retention and hence popping performance.

Cellulose↗