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Active protein nanoparticle prepared by restrictive enzymatic hydrolysis and preparation method and application thereof

A technology of active protein and nanoparticle, which is applied in the direction of nanotechnology, nanotechnology, nanomedicine, etc., can solve the problems of high equipment requirements, inability to use industrial scale production, organic solvent residues, etc., and achieve good industrialization prospects and excellent resistance Effects of oxidation activity and mild reaction conditions

Pending Publication Date: 2019-08-16
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this method is an effective means to prepare protein nanoparticles, there is a risk of organic solvent residues due to the use of organic solvents such as acetone, dimethyl sulfoxide, ethanol, and tert-butanol.
Residues of organic solvents may make the prepared protein nanoparticles toxic, causing safety hazards
In addition, methods for preparing protein particles include high temperature and high pressure treatment, ultrasonic treatment, supercritical fluid treatment, etc. The preparation process or reaction conditions are relatively severe, or the requirements for equipment are high, and they cannot be used for industrial scale production. To some extent, the application of protein nanoparticles in food and medicine is limited

Method used

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  • Active protein nanoparticle prepared by restrictive enzymatic hydrolysis and preparation method and application thereof
  • Active protein nanoparticle prepared by restrictive enzymatic hydrolysis and preparation method and application thereof
  • Active protein nanoparticle prepared by restrictive enzymatic hydrolysis and preparation method and application thereof

Examples

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Effect test

Embodiment 1

[0040]A method for preparing active protein nanoparticles by restricted enzymolysis provided in Example 1, comprising the following steps: mixing 0.5g of rice protein with 50mL of deionized water, stirring with a glass rod for 0.5h, and adjusting with 1mol / L sodium hydroxide solution pH to 8.5, adjust the temperature of the constant temperature oscillator to 35 degrees Celsius, adjust the rotation speed of the constant temperature oscillator to 140rpm, add trypsin at 0.05% of the rice protein mass under the state of stirring with a magnetic stirrer to carry out restricted enzymolysis, and the enzymolysis time is 1h , control the degree of proteolysis at 3%. After the enzymatic hydrolysis, adjust the pH to 6.0 with 1mol / L hydrochloric acid solution, adjust the temperature to 80 degrees Celsius, incubate for 15 minutes with a glass rod stirring at 100 rpm to inactivate the enzyme, centrifuge at 6000 rpm and 25 degrees Celsius for 20 minutes, and take the The serum was freeze-dri...

Embodiment 2

[0042] A method for preparing active protein nanoparticles by restricted enzymolysis provided in Example 2, comprising the following steps: mixing 4 g of potato protein with 100 mL of deionized water, stirring with a stirring paddle for 1 h, adjusting the pH to 1.5 with 2 mol / L hydrochloric acid solution, Adjust the temperature of the constant temperature incubator to 37 degrees Celsius, adjust the speed of the constant temperature incubator to 150rpm, add pepsin at 0.25% of the potato protein content under the state of stirring with a magnetic stirrer for restrictive enzymolysis, and the enzymatic hydrolysis time is 2h to control proteolysis degree at 6%. After the enzymolysis, adjust the pH to 7.0 with 2mol / L sodium hydroxide solution, adjust the temperature to 90 degrees Celsius, incubate for 10 minutes with the stirring paddle stirring at a rate of 120 rpm to inactivate the enzyme, and centrifuge at 8000 rpm and 15 degrees Celsius for 15 minutes. The supernatant was taken ...

Embodiment 3

[0044] A method for preparing active protein nanoparticles by restricted enzymolysis provided in Example 3, comprising the following steps: mixing 12 g of soybean protein with 150 mL of deionized water, stirring with a magnetic stirrer for 2 h, and adjusting the pH with 3 mol / L sodium hydroxide solution to 7.0, adjust the temperature of the constant temperature oscillator to 50 degrees Celsius, adjust the speed of the constant temperature oscillator to 160rpm, and in the state of stirring with a magnetic stirrer, add flavor protease according to 0.5% of the protein amount for restriction enzymolysis, and the enzymatic hydrolysis time is 1.5h , control the degree of proteolysis at 7%. After enzymatic hydrolysis, adjust the pH to 7.5 with 3mol / L sodium hydroxide solution, adjust the temperature to 95 degrees Celsius, incubate for 10 minutes with a magnetic stirrer stirring at a rate of 130 rpm to inactivate the enzyme, and centrifuge at 10,000 rpm and 20 degrees Celsius for 8 min...

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Abstract

The invention discloses active protein nanoparticle prepared by restrictive enzymatic hydrolysis and a preparation method and an application thereof. The preparation method comprises the steps of: mixing protein and water in a mass to volume ratio of 1 to 10:100 (g / mL) to adjust the pH of the protein liquid, adding pancreatin, Alcalase, pepsin, papain, neutral protease, flavor protease, complex protease or bromelain for restrictive enzyme digestion, controlling the degree of proteolysis at 3 to 9%, performing steps of insulated enzyme deactivation and centrifugation, taking a supernatant, andperforming freeze drying or spray drying to obtain the active protein nanoparticle. The method has mild reaction conditions, simple process, and green safety, and no organic reagent is involved in theprocess; The prepared protein nanoparticle has small particle size, good stability, also has superior antioxidant activity, and has good development and application prospects.

Description

technical field [0001] The invention belongs to the field of functional nano-biological products, and in particular relates to an active protein nanoparticle prepared by restricted enzymolysis, a preparation method and application thereof. Background technique [0002] Nanoparticles are typical nanostructure units with unique structures and the ability to load active substances, and have been widely used as delivery vehicles for drugs or nutrients. [0003] At present, the nanoparticles used in the food and pharmaceutical industries are mostly inorganic particles or synthetic polymers, which have good processing performance, but most of them have poor biocompatibility, and have potential risks to human health to a certain extent. Therefore, how to research and develop new, safer and more reliable edible nanoparticles has become an urgent problem for scientific researchers. As biological macromolecules, protein nanoparticles not only have the scale effect and functional char...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06A61K47/42B82Y40/00B82Y5/00
CPCA61K47/42B82Y5/00B82Y40/00C12P21/06
Inventor 周非白赵谋明沈鹏辉张远红
Owner SOUTH CHINA UNIV OF TECH
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