No description available.

No description available.

The complex structures of water-soluble wheat arabinoxylans have been mapped along individual molecules, and within populations, using the visualisation of the binding of inactivated enzymes by atomic force microscopy (AFM). It was demonstrated that site-directed mutagenesis (SDM) can be used to produce inactive enzymes as structural probes. For the SDM mutants AFM has been used to compare the binding of different xylanases to arabinoxylans. Xylanase mutant E386A, derived from the Xyn11A enzyme (Neocallimastrix patriciarium), was shown to bind randomly along arabinoxylan molecules. The xylanase binding was also monitored following Aspergillus niger arabinofuranosidase pre-treatment of samples. It was demonstrated that removal of arabinose side chains significantly altered the binding pattern of the inactivated enzyme. Xylanase mutant E246A, derived from the Xyn10A enzyme (Cellvibrio japonicus), was found to show deviations from random binding to the arabinoxylan chains. It is believed that this is due to the effect of a small residual catalytic activity of the enzyme that alters the binding pattern of the probe. Control procedures were developed and assessed to establish that the interactions between the modified xylanases and the arabinoxylans were specific interactions. The experimental data demonstrates the potential for using inactivated enzymes and AFM to probe the structural heterogeneity of individual polysaccharide molecules.

Tissue inhibitor of metalloproteinase-3 (TIMP-3) is a protein with multiple functions that include regulating the turnover of the extracellular matrix (ECM) by inhibiting members of the metzincin family. Extracellular levels of soluble TIMP-3 are low, reflecting its binding to components of the ECM including sulfated glycosaminoglycans (SGAGs) and its endocytosis by low density lipoprotein receptor-related protein 1. Because TIMP-3 inhibits ECM-degrading enzymes, the ability of SGAG mimetics to elevate extracellular concentrations of TIMP3 is of interest for osteoarthritis treatment. However, previous studies of such interactions have utilized immobilized forms of the protein or ligands. Here we have quantified the thermodynamics of the interactions of the inhibitory domain of TIMP-3 with chondroitin sulfate (CS), pentosan polysulfate (PPS) and suramin in solution using isothermal titration calorimetry. All three interactions are driven by a (favorable) negative enthalpy ychange combined with an unfavorable decrease in entropy. The heat capacity change (ΔCp) for the interaction of N-TIMP-3 with CS, PPS, or suramin is essentially zero, indicating an insignificant contribution from the hydrophobic effect. Based on the effects of ionic strength on the interaction of N-TIMP-3 with suramin, their interaction appears to be driven by electrostatic interactions. Modeling supports the view that the negatively charged sulfates of CS, PPS, and suramin interact with a cationic region on N-TIMP-3 that includes Lys -26, -27, -30, and -possibly 76 on the opposite face of TIMP-3 from its reactive site for metalloproteases. / Includes bibliography. / Dissertation (Ph.D.)--Florida Atlantic University, 2019. / FAU Electronic Theses and Dissertations Collection

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Effects of enzymatic modification of starch, proteins and pentosans on dough and tortilla properties were determined to establish the role of these wheat components in tortilla staling. Starch, protein and pentosans were respectively modified with a-amylase, protease and transglutaminase (TG), and xylanase. Tortillas were stored at 22oC and evaluated for at least three weeks. Amylase improved shelf-stability of tortillas, produced a significant amount of dextrins and sugars, retarded decrease in amylose solubility, and weakened starch granules. However, control and treated tortillas had similar degrees of amylopectin crystallinity. Staling of tortillas appears to involve starch that reassociates into an amorphous structure. Micrographs of control dough had thin protein strands forming a continuous matrix. Protease-treated dough had pieces of proteins in place of the continuous matrix, while TG-treated dough had thicker protein strands that were heterogeneously distributed. Both treatments resulted in shorter shelf-stability of tortillas. The organization of protein in dough is important for dough structure and appears to impact tortilla flexibility. Protein solubility and SDS-PAGE results did not differentiate control and treated dough or tortillas. The fractions or molecular weight distribution are not significant determinants of protein functionality. Tertiary and quaternary protein structures of gluten may be more related to tortilla shelf-stability. The 75 ppm xylanase treatment resulted in weaker tortilla structure and significantly higher amounts of low molecular weight saccharides and sugars. Control and the 25 ppm treatment sample had a similar shelf-stability and texture profile. Pentosans may affect staling indirectly through the effect on gluten development. Fresh tortillas have amylopectin in an amorphous state, while amylose is mostly retrograded. The gluten matrix provides additional structure and flexibility to the tortilla. Pentosans may or may not be attached to the gluten network. Upon storage, amylopectin retrogrades and recrystallizes, firming the starch granules, resulting in firmer tortillas. Starch hydrolysis decreased the rigid structure and plasticized polymers during storage. It also reduced the restriction imposed by retrograded starch on gluten and allowed it more flexibility. Thus, the flexibility of tortillas results from the combined functionalities of amylose gel, amylopectin solidifying the starch granules during storage, and the changed functionality of gluten after baking.

Thesis (Ph. D.)--Institute of Paper Chemistry, 1952. / Includes bibliographical references (p. 117-119).

Thesis (Ph. D.)--Institute of Paper Chemistry, 1975. / Includes bibliographical references (p. 60-67).

No description available.