Protein-sterol covalent complex as well as preparation method and application thereof
A compound and sterol technology, which is applied in the field of food processing, can solve the problems of low bioaccessibility and difficult addition of phytosterols, and achieve the effects of low bioaccessibility, enhanced oral bioaccessibility, and simple material composition
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Embodiment 1
[0040] 1. Preparation of protein-sterol covalent polymer
[0041] 1. Add whey protein isolate (WPI, CAS: 84082-51-9) with free carboxyl groups into deionized water to fully dissolve, stir at 4°C for 1.5 hours to obtain a protein stock solution; then add 0.7 moL per mole of carboxyl groups of the protein 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide (EDC), mixed with EDC in the protein stock solution, and stirred at 4°C for 2 hours to obtain a carboxyl-activated protein solution.
[0042] 2. Then add an equal volume of dimethyl sulfoxide solution (DMSO) containing phytosterol (β-Sitosterol, CAS: 83-46-5) to mix the sterol solution with the activated protein solution. In this process, ultrasonic treatment is combined. With a power of 200W and a 1s pulse mode, slowly add the phytosterol solution into the protein solution to obtain a 1:10 (w / w%) sterol:protein (w / w mixed) solution; react with magnetic stirring at 30°C for 24h.
[0043] 3. Then pour 2 times the volume of acetone, ...
Embodiment 2
[0059] 1. Preparation of stable O / W emulsion of whey protein isolate-phytosterol covalent complex
[0060] The sterol-whey protein isolate covalent complex was dissolved in phosphate buffer (10 mM) with a pH of 7.0 to obtain a protein-sterol covalent complex solution, so that the sterol concentration in the complex solution was 2 mg / mL. Medium-chain triglyceride (MCT) is used as the oil phase material and heated to 140°C. Using the shearing homogenization effect of the high-speed shearing machine, slowly add the oil phase into the water phase, the ratio of the oil phase to the water phase is 1:9 (w / w). Homogenize and shear for 2 minutes with a high-speed homogenizer to form a coarse emulsion. A high-pressure homogenizer is used to further homogenize to obtain a sterol-containing emulsion. Then use a high-pressure microfluidizer to further reduce the particle size of the emulsion 5 times under a pressure of 70 MPa, and the final sterol content is the same as in Example 1.
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Embodiment 3
[0086] Different kinds of protein-sterol covalent complexes
[0087] 1. When preparing the protein stock solution, the protein is selected from soybean protein isolate (SPI), corn protein (Zein), sodium caseinate (NaCAS), and the rest are the same as in Example 1.
[0088] Fully dissolve SPI and NaCAS in deionized water, fully dissolve Zein in ethanol, stir at 4°C for 1.5h to obtain a protein solution with a concentration of 5mg / mL; then mix the protein stock solution with 0.7moL of 1-(3 -Dimethylaminopropyl)-3-ethylcarbodiimide EDC mixed, stirred at 4°C for 2h to obtain carboxyl-activated protein solution.
[0089] 2. Then add an equal volume of phytosterol-containing dimethyl sulfoxide solution (DMSO) to mix the sterol solution with the activated protein solution; ultrasonic treatment, power 200W, 1s pulse mode, slowly add the phytosterol solution to the protein solution In 1:1, 1:5, 1:10, 1:15 sterol: protein (w / w) solution; 30 ℃ magnetic stirring reaction for 24h.
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