Application of immobilized proteinase in preparing cream protein peptide
A technology for immobilizing protease and milk casein, applied in the preparation of milk casein peptide, immobilized protease, immobilized enzyme and its application fields, can solve the problems that the research of milk casein peptide has not yet been reported, etc., and achieves convenient product separation, The effect of high peptide content and reduced production cost
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Embodiment 1
[0080] Example 1 Preparation of immobilized trypsin
[0081] (1) Stir 10% chitosan acid sol to disperse the sol droplets uniformly, and drop into 2.5% glutaraldehyde solution to make the final concentration (volume fraction) of glutaraldehyde solution reach 3% , Stir and add 3mol / L NaOH solution until the final concentration (volume fraction) of NaOH is 3%. Incubate at 70°C for 0.5% reaction. After standing and cooling, discard the upper layer, wash with acetone, absolute ethanol, and finally with distilled water Rinse and dry in vacuum to obtain immobilized carrier chitosan microspheres for use;
[0082] (2) Add immobilized carrier chitosan microspheres to 5% trypsin solution, where trypsin solution: immobilized carrier chitosan microspheres = 4:1; place it in a shaker shaker for 30 hours, then Dropping a 2.5% glutaraldehyde solution with a volume fraction to make the final concentration (volume fraction) of the glutaraldehyde solution reach 3%, and cross-linking for 0.2-10h to o...
Embodiment 2
[0083] Example 2 Preparation of immobilized pepsin
[0084] (1) Stir 0.5% chitosan acidic sol to uniformly disperse the sol droplets, and drop into 2.5% glutaraldehyde solution to make the final concentration (volume fraction) of the glutaraldehyde solution reach 0.2% , Add 0.5mol / L NaOH solution with stirring until the final concentration (volume fraction) of NaOH is 0.05%, keep at 40℃ for 5h, let stand and cool, discard the upper layer, wash with acetone and absolute ethanol in turn, and finally use Rinse with distilled water and dry in vacuum to obtain immobilized carrier chitosan microspheres for use;
[0085] (2) Add the prepared immobilized carrier chitosan microspheres to 0.1% pepsin solution, where pepsin solution: immobilized carrier chitosan microspheres = 1:1; placed in a shaker shaker for adsorption 2h, then drop in a 2.5% glutaraldehyde solution with a volume fraction to make the final concentration (volume fraction) of the glutaraldehyde solution reach 0.2%, and cros...
Embodiment 3
[0086] Example 3 Preparation of immobilized bromelain
[0087] (1) Stir 5% chitosan acidic sol to make the sol droplets evenly dispersed, and drop into 2.5% glutaraldehyde solution to make the final concentration (volume fraction) of the glutaraldehyde solution reach 0.5% , Add 0.8mol / L NaOH solution with stirring until the final concentration (volume fraction) of NaOH is 0.08%, keep at 50℃ and react for 2.5h, let it stand and cool, discard the upper layer, wash with acetone and absolute ethanol in turn, and finally Rinse with distilled water and dry in vacuum to obtain immobilized carrier chitosan microspheres for use;
[0088] (2) Add the prepared immobilized carrier chitosan microspheres accounting for 5% of the volume of the bromelain solution to the 1.5% bromelain solution, wherein the bromelain solution: immobilized carrier chitosan microspheres=3.5:1; Place it in a shaker shaker for 10 hours, then drop in a 2.5% glutaraldehyde solution to make the final concentration (volum...
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