Process for preparing phosphopeptide non-phosphopeptide by hydrolyzing protein through enzyme

Abstract

The invention discloses a method for preparing casein phosphopeptide and casein non-phosphopeptide through enzymatic hydrolysis casein. The method mainly uses the casein as raw material, uses single alkali protease Alcalase2.4L to control the hydrolysis, uses a trinitrobenzene sulfonic acid method to monitor a DH value, and uses an alcohol-calcium isolation method to simultaneously prepare CPP and CNPP from the casein hydrolyte. The mol ratio of nitrogen to phosphor is 20-25 to 1 with a yield above 35 percent. The obtaining ratio of CNPP reaches 45 percent. The amino acid sequences of two ACE inhibition bioactive peptides and an oxidation resistant bioactive peptides in the CNNP product are identified as follows: Tyr-Leu, Glu-Asp-Val-Pro and His-Ser. The method provided by the invention has a good enzyme hydrolysis effect, a proper price, a reasonable process route, a high material utilization rate and easy amplification. The products of CPP and CNPP can be widely applicable to functional food ingredient and health food.

Description

technical field [0001] The invention belongs to the technical field of dairy product development and functional health food, and relates to a method for preparing phosphopeptide (CPP) and non-phosphopeptide (CNPP) by enzymatically hydrolyzing casein. 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. The incidence of osteoporosis in the elderly in my country reaches 30%-50%, and the incidence of rickets in children is as high as 20%-50%. Current studies have shown that...

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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; researchers have also used the integrated process of enzymatic hydrolysis and membrane filtration including trypsin and alkaline protease Continuously prepare casein biologically active peptides, but the enzymes used are many and expensive (CN1546682); in addition, some researchers use bovine milk casein as raw material, use protease hydrolysis, and ion exchange resin desalination to produce hexapeptides to undecapeptides. ACE inhibitory peptide (CN1552891), but its process route is more complicated, and freeze-drying makes the cost expensive; there are also researchers who use compound protease to enzymatically hydrolyze casein to prepare casein phosphopeptide (CPP), but they do not use CNPP

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