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Antioxidant peptide active protection in protein enzymolysis process and preparation method thereof

An antioxidant peptide and enzymatic hydrolysis technology, applied in the field of animal and plant protein development and utilization, can solve the problems of poor protein dispersibility and reduced antioxidant activity of products, and achieve the effects of strong antioxidant activity, improved antioxidant activity and high activity.

Inactive Publication Date: 2010-12-15
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to overcome technical problems such as the protease used in the process of protein enzymolysis and the oxidation of the antioxidative peptide produced, resulting in the reduction of the antioxidant activity of the product and the poor dispersion of some proteins, thereby providing an antioxidative peptide in the process of enzymatic hydrolysis and protease activity protection and preparation method thereof

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] 1. Preparation of zein suspension: Weigh 80g zein powder in a 3L beaker, add 2L of 0.03% sodium sulfite solution through ultrasonic degassing, stir and add alkali (1N NaOH) at room temperature, adjust and Maintain a stable pH around 9.0.

[0013] 2. Protected enzymolysis: put the prepared zein suspension into a 5L reaction kettle equipped with a stirrer, pH meter, thermometer and dissolved oxygen electrode, and quickly add 1.6mL alkaline protease (enzyme activity 1.6 *10 5 U / mL) immediately seal the reaction kettle; start the vacuum pump to remove the air in the reaction kettle (until the pressure in the reaction kettle drops to 10mm / Hg), then feed nitrogen to normal pressure, start the stirrer to stir, and start to heat up to 55 ℃, keep for 4h. During the enzymatic hydrolysis, the automatic injection port automatically adds alkali to keep the pH stable at 9.0. After the hydrolysis, the enzyme was inactivated in a boiling water bath for 10 min. Take out the hydrolyz...

Embodiment 2

[0017] 1. Collagen suspension preparation: Weigh 50g of collagen and dissolve it in 1L of ultrasonic degassed 0.005% mercaptoethanol solution, use HCL to adjust and maintain pH 3.0 until stable, and prepare 5% (W:V) protein suspension.

[0018] 2. Protected enzymatic hydrolysis: move the substrate suspension to the reaction kettle, with [E] / [S]=5% (enzyme activity 8.0*10 4 U / g) After the enzyme is added, the reactor is closed, and after vacuuming, carbon dioxide is introduced to normal pressure; the stirrer is started to stir, and the temperature is raised and maintained at 55° C., pH 3.0. Keep for 4h. After the hydrolysis, the reaction was completed by killing the enzyme in a boiling water bath for 10 minutes, taking out the hydrolyzate and cooling it rapidly in an ice bath, centrifuging at 12,000 r / min at 4°C for 15 minutes, and taking the supernatant.

[0019] 3. Antioxidant peptide product preparation: The supernatant obtained in the previous step is ultrafiltered, evapo...

Embodiment 3

[0022] 1. Red deer antler blood sample preparation: Weigh 50 g of red deer antler blood, add 0.005% dithiothreitol 500 mL, and prepare a red deer antler blood sample solution with a concentration of 10% (W:V).

[0023] 2. Protected enzymatic hydrolysis: put the sample solution in a water bath at 100°C for 10 minutes, then transfer it into the reaction kettle, adjust the pH to 7 with sodium hydroxide solution, and add neutral protease (enzyme activity 10.82× 10 4 U / g), after adding the enzyme, close the reaction vessel, and after vacuuming, introduce helium to normal pressure; start the stirrer to stir, react at 50°C for 2h, boil for 10min to inactivate the enzyme, rapidly cool to below 45°C, and [E ] / [S]=9% add papain (enzyme activity 14.10×10 4 u / g), vacuumize again and pass helium to normal pressure; react for 4h (pH 6.8), boil for 10min and cool down, centrifuge the hydrolyzate at 12000rpm / min for 10min, and collect the supernatant.

[0024] 3. Antioxidant peptide product...

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PUM

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Abstract

The invention discloses antioxidant peptide active protection in a protein enzymolysis process and a preparation method thereof, and belongs to the development and utilization field of plant and animal proteins. Animal protein, plant protein and animal blood are taken as raw materials; and protein dissolution and enzymolysis efficiency are improved, protease activity and antioxidant peptide activity are protected and a high-activity antioxidant peptide product is prepared by adopting a treatment method of inflating protective gas and adding a mercapto protective agent in the enzymolysis process. The preparation time, antioxidant activity and the like of the antioxidant peptide product prepared by the method are obviously improved compared with a product prepared by using the traditional method. The antioxidant peptide product has extremely good antioxidant activity, and is suitable to be applied in the fields of health-care products, advanced nutritional food, cosmetics, animal feed, animal food and the like.

Description

technical field [0001] The invention relates to the field of development and utilization of animal and plant proteins, in particular to a preparation method for protecting the anti-oxidation activity and protease catalytic activity of products by filling protective gas and isolating oxygen during the preparation of antioxidant peptides by proteolysis. Background technique [0002] Many studies have confirmed that oxidation is related to the pathogenesis of many diseases in humans and other animals, such as cancer, aging, arteriosclerosis, etc. Antioxidant active peptides are a class of biologically active peptides that can inhibit the peroxidation of biological macromolecules or scavenge free radicals in the body. [0003] In recent years, many scholars at home and abroad have used plant protein or animal protein as raw materials to conduct research on the preparation of antioxidant peptides. Many food source proteins can produce peptides with strong antioxidant activity af...

Claims

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Application Information

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IPC IPC(8): C12P21/06
Inventor 施用晖乐国伟马淑凤田许
Owner JIANGNAN UNIV
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