Application of shear and cross-linking enzyme transglutaminase (Tgase) induced fibrous hierarchical structures in dense (30% w/w) calcium caseinate (Ca-caseinate) dispersions. Using Tgase was essential for the anisotropic structure formation. The fibrous materials showed anisotropy on both micro- and macroscale as determined with scanning electron microscopy (SEM) and mechanical analyses, respectively. SEM revealed protein fibers with a diameter of approximately 100-200 nm; visually, we observed fibers of about 1 mm. Both shear and Tgase affected the reinforcement of the fibers to a large extent, whereas the mechanical properties in the direction perpendicular to the shear flow remained constant. Shearing Ca-caseinate without Tgase yielded a slightly anisotropic layered structure. Both cross-linking in the absence of shear and cross-linking during mixing resulted in gels without alignment. The formation of shear- and enzyme-induced anisotropic structures was explained by aligning of protein aggregates due to shear and concurrent solidification of the aligned protein aggregates.
Rheological measurements of dense calcium caseinate and sodium caseinate dispersions (> or =15%) provided insight into the factors determining shear-induced structure formation in caseinates. Calcium caseinate at a sufficiently high concentration (30%) was shown to form highly anisotropic structures during shearing and concurrent enzymatic cross-linking. In contrast, sodium caseinate formed isotropic structures using similar processing conditions. The main difference between the two types of caseinates is the counterion present, and as a consequence, the size of structural elements and their interactions. The rheological behavior of calcium caseinate and sodium caseinate reflected these differences, yielding non-monotonic and shear thinning flow behavior for calcium caseinate whereas sodium caseinate behaved only slightly shear thinning. It appears that the intrinsic properties of the dense caseinate dispersions, which are reflected in their rheological behavior, affect the structure formation that was found after applying shear. Therefore, rheological measurements are useful to obtain an indication of the structure formation potential of caseinate dispersions.
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