| Vitamin E(V_E) is one of essential nutrients for human and is the most effective antioxidant in human’s body. However, V_E can not be dissolved in water, and it can be destroyed easily by oxygen, ultraviolet radiation and alkali, so it is difficult to apply V_E in food industry. Microencapsulation can protect the sensitive components from the destruction of environment. β-lactoglobulin(β-Lg) is one of the important water-soluble proteins, and it has strong ability to bind some ligand and improve their stability and bioavailability. In this paper, inclusion complex of β-Lg and V_E was prepared, and the antioxidant activity, transshipment and absorption of complex have also been studied.1. Optimization for the preparation of combination of protein and V_E by response surface methodologyThe best processing conditions of combination of V_E combined by β-Lg,crude-extracted β-Lg and whey protein were determined by response surface methodolgy,and the results were as follows:(1)The best processing conditions of V_E combined by β-Lg were as follows: the quality ratio 5:1, magnetic stirring time 45 min, heat-shock temperature 49℃, p H6.27(pure water),and under the conditions the combined rate was 43.47%(2)The best processing conditions of V_E combined by crude-extracted β-Lg were as follows:the quality ratio 19:1, magnetic stirring time 49 min, heat-shock temperature 72℃, and under the conditions the combined rate was 68.5%.(3)The best processing conditions of V_E combined by whey protein were as follows: the quality ratio 14:1, magnetic stirring time 54 min, heat-shock temperature 83℃, and under the conditions the combined rate was 57.54%.Transmission electron microscopy photos indicated that, the inclusion complexes were spherical and dispersed uniform, and there are bright and inerratic material in the complexes. The grain size of combinations of V_E combined by β-Lg, crude-extracted β-Lg and whey protein were 0.17~3μm, 0.25~0.35μm and 0.25~0.45μm, respectively. The infrared spectra indicated that, the IR spectra of complexes and package proteins are almost same. There was no new peak in the IR spectra of complexes, so the leak of V_E was covered by the leaks of proteins.2. The effect of the complex of β-Lg and V_E on the antioxidant capacity in pubertal male miceBased on the vivo tests, the effects of different forms of V_E on the serum testosterone,serum total antioxidant capacity, cerebral superoxide dismutase(sod), liver superoxide dismutase(sod), catalase and testicular hydrogen peroxide enzyme in pubertal male mice were studied. The results indicated that, the six indexes of inclusion complex group were both remarkably higher than those of blank group, V_E group and zymolytic group(P<0.05);with the increase of feeding time, the content of serum testosterone in the mice of inclusion complex group increased, but there was no remarkably difference(P>0.05). Conversely, the other five indexes showed remarkably difference(P<0.05). The antioxidant activity of V_E group and zymolytic group also increased, but they did not present remarkable difference(P>0.05).3. The expression of ABCA1, SR-BI and PepT1 m RNA in the small intestine tissue and intestinal epithelial cell of miceBased on the vivo experiments(feeding mice) and vitro tests(cell culture), the levels of the expressions of ABCA1, SR-BI and Pep T1 in the small intestine tissues of the mice were compared by the quantitative PCR method, and the way of transportation and absorption of inclusion complex in the small intestine tissue of mice was also analysized.The results indicated that, the amount of expressed ABCA1 in inclusion complex group was higher than that of other groups significantly(P<0.05); the amount of expressed SR-BI m RNA in inclusion complex group was higher than that of blank group and V_E group significantly(P<0.01) and was higher than that of zymolytic inclusion complex group; The Pe PT1 m RNA of inclusion complex group was significantly higher than that of zymolytic inclusion complex group in the first and second week(P<0.01) and was significantly higher than that of the other groups(P<0.01). With the increasing of feeding time, the expressed amount of ABCA1, SR-BI and Pe PT1 m RNA in the intestine tissue did not present remarkable difference(P>0.05). But the expressed amounts of ABCA1, SR-BI and Pe PT1 m RNA in the intestinal epithelial cell were remarkably higher than that of blank group and V_E group(P<0.05). |
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