Beta-lactoglobulin-vitamin conjugate comprising sugar alcohols and preparation method thereof
A lactoglobulin and vitamin technology, which is applied in the biological field, can solve the problems of reduced assembly effect, limited application surface, and β-lactoglobulin sensitization is not eliminated, and achieves the effect of improving the binding rate and reducing the sensitization.
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[0031] The invention also discloses a preparation method of the aforementioned sugar alcohol-containing β-lactoglobulin-vitamin conjugate, which comprises the following steps:
[0032] Step 1. The β-lactoglobulin is made into a 1mg / mL β-lactoglobulin solution with 0.01mol / L pH7.4 phosphate buffer solution.
[0033] Step two, place the β-lactoglobulin solution in a high-voltage electric field for high-voltage pulse treatment.
[0034] Step 3: Combine β-lactoglobulin after high-pressure pulse glycosylation with VE, sorbitol, and maltitol, and then freeze-dry the composition to obtain dry β-lactoglobulin containing sugar alcohol Vitamin conjugate products.
[0035] In step 2, the β-lactoglobulin solution is placed in a high-voltage electric field for a processing time of 80 μs, wherein the electric field intensity of the high-voltage electric field is 30 kV / cm, and the electric field waveform is a unipolar square wave.
[0036] In step three, the combination method includes: weighing β-la...
Embodiment 1
[0044] The first preparation method of β-lactoglobulin-vitamin conjugate containing sugar alcohol of the present invention includes the following steps:
[0045] (11) The β-lactoglobulin is made into a 1mg / mL β-lactoglobulin solution with 0.01mol / L, pH7.4 phosphate buffer solution.
[0046] (12) Put the β-lactoglobulin solution in a high-voltage electric field with an electric field strength of 30kV / cm and a unipolar square wave for 80μs.
[0047] (13). Mix the β-lactoglobulin with VE, sorbitol, and maltitol obtained in step (12) at a mass ratio of 1:0.01:2, and then freeze-dry it. Put the freeze-dried powder in a centrifuge tube and put it in an incubator. React in a 59% saturated potassium iodide solution with a temperature of 65°C and a relative humidity of 65% by mass for 3 hours. After the reaction, the centrifuge tube is immediately placed in an ice bath. Terminate the reaction. After the reaction is over, add pre-cooled ultrapure water for reconstitution, and then use an ultr...
Embodiment 2
[0050] The second preparation method of the β-lactoglobulin-vitamin conjugate containing sugar alcohol of the present invention includes the following steps:
[0051] (21) The β-lactoglobulin is made into a 1mg / mL β-lactoglobulin solution with 0.01mol / L, pH7.4 phosphate buffer solution.
[0052] (22) Place the β-lactoglobulin solution in a high-voltage electric field with an electric field strength of 30kV / cm and a unipolar square wave for 80μs.
[0053] (23). Mix the β-lactoglobulin with VE, sorbitol, and maltitol in a mass ratio of 1:0.01:2, and then freeze-dry the β-lactoglobulin after the first-level high-pressure pulse glycosylation in step (22). Put the freeze-dried powder in a centrifuge tube and put it in an incubator, and react in a 59% saturated potassium iodide solution with a temperature of 65°C and a relative humidity of 65% by mass for 3 hours. After the reaction, the centrifuge tube is immediately placed in an ice bath Terminate the reaction. After the reaction is ove...
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