Enzyme method for hydrolyzing casein and synchronous preparation of phosphopeptide and non-phosphopeptide

A casein phosphopeptide, hydrolyzed casein technology, applied in protein food processing, protein food ingredients, food science and other directions, can solve the problems of functional component structure elucidation, complex process route, many types of enzymes, etc. Conducive to popularization and utilization, clear structure, and the effect of reducing ash

Inactive Publication Date: 2007-08-01
JIANGNAN UNIV
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Problems solved by technology

[0005] At present, researchers have reported the separation and purification of casein-derived ACE inhibitory peptides, but have not elucidated the structure of their functional components; The process continuously prepares casein biologically active polypeptides, but the enzymes used are many and expensive (CN1546682); another researcher uses cow milk casein as raw material, uses protease hydrolysis, and ion exchange resin desalination to produce hexapeptides to undecapeptides Composed of ACE inhibitory peptide (CN1552891), but its process route is relatively complicated, and freeze-drying makes the cost expensive; there are also researchers who use complex protease to enzymatically hydrolyze casein to prepare casein phosphopeptide (CPP), but they do not use CNPP

Method used

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  • Enzyme method for hydrolyzing casein and synchronous preparation of phosphopeptide and non-phosphopeptide
  • Enzyme method for hydrolyzing casein and synchronous preparation of phosphopeptide and non-phosphopeptide
  • Enzyme method for hydrolyzing casein and synchronous preparation of phosphopeptide and non-phosphopeptide

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Effect test

Embodiment 1

[0024] Enzyme reactions were performed in a 10 L sandwich reactor. Add 5000ml of water into the reactor, raise the temperature to 55°C, then weigh 1000g of casein, and slowly add it into the water while stirring. Adjust the pH value to 8.0 with 2mol / L NaOH, dissolve all the casein at 55°C, add 11.5mL Alcalase 2.4L, and monitor DH with TNBS. Add 60g anhydrous CaCl 2 , stirred until completely dissolved, then slowly added edible ethanol under stirring until the final concentration reached 70% (v / v), left for 30min, centrifuged (3000rpm, 20min), and the precipitate was vacuum-dried (40°C) to obtain 354g of CPP product . The supernatant enters the nanofiltration device (the relative molecular mass cut off by the nanofiltration membrane is 300D) to concentrate, and the permeate is used to recover ethanol, and the concentrated (concentration is 60mg / ml) liquid is put on a DA201-C macroporous adsorption resin chromatographic column, washed with water After desalting, use 70% ethan...

Embodiment 2

[0027] Enzyme reactions were performed in a 25L sandwich reactor. Add 15000ml of water into the reactor, raise the temperature to 65°C, then weigh 3750g of casein, and slowly add it into the water while stirring. Adjust the pH value to 8.5 with 2mol / L NaOH, dissolve all the casein at 65°C, add 2.4L of 55mL Alcalase, and monitor DH with TNBS. Add 180g anhydrous CaCl 2, stirred until completely dissolved, then slowly added edible ethanol under stirring until the final concentration reached 80% (v / v), left it overnight, centrifuged (3000rpm, 15min) after siphoning the supernatant, and the precipitate was vacuum-dried (45°C ) to obtain CPP product 1480g. The supernatant enters the nanofiltration device (the relative molecular mass cut off by the nanofiltration membrane is 300D) to concentrate, and the permeate is used to recover ethanol, and the concentrated (concentration is 150mg / ml) liquid is put on a DA201-C macroporous adsorption resin chromatographic column, washed with wa...

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Abstract

The invention relates to a enzymatical hydrolyzed casein and the method for preparing casein phosphopeptide (CPP) and casein non phosphopeptide (CNPP), belonging to dairy further processing and functional health care food field. The invention takes casein as raw material, takes single alkali prolease Alcalase 2.4L for controlling hydrolysis, takes nitrobenzene-sulfonic acid for pH value monitoring, preparing CPP and CNPP from casein hydrolysate with ethanol- calcium separation method. The nitrogen / phosphorus molar ratio in CPP is 20- 25: 1, the productivity is above 35%; the productivity of CNPP reaches 45%; the determined amine aicd squence of two ACE inhibiting active peptide and one antioxidant active peptide in CNPP product is Tyr-Leu,Glu-Asp-Val-Pro and His-Ser. The invention is characterized by good enzymolysis effect, low price, rational process route, high raw material vailability ratio, and wide application of CPP and CNPP in functional food ingredient or health care food.

Description

technical field [0001] The invention discloses a method for simultaneously preparing phosphopeptide (CPP) and non-phosphopeptide (CNPP) by enzymatically hydrolyzing casein, which belongs to the technical field of dairy product resource development and functional health food. Background technique [0002] In recent years, the problem of human calcium insufficiency has received extensive attention. Calcium deficiency has long been a common phenomenon all over the world, especially in developing countries, and the most serious ones are pregnant women, children and the elderly. The recommended amount of calcium for normal people in my country is 800mg / d, but the results of nutritional surveys show that the per capita calcium intake is only 400mg / d, and the actual intake of children and pregnant women is less than half of the intake. In my country, the incidence rate of osteoporosis in the elderly reaches 30%-50%, and the incidence rate of rickets in children is as high as 20%-5...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02A23J3/34
Inventor 王璋许时婴张文斌杨瑞金袁博蔡健季万兰
Owner JIANGNAN UNIV
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