CN1246473C - Process for biological enzyme extraction of grape seed polypeptide protein - Google Patents

Abstract

The present invention relates to a technology for extracting plant protein, namely a technology for extracting polypeptide protein in grape seeds by a biological enzyme method. The technology of the present invention is characterized in that grape seeds are used as raw materials to be pulverized, softened by adding water, hydrolyzed by adding the mixed enzyme of interior contact subacid cellulose and papain, hydrolyzed by adding exterior contact aspergillus oryzae, separated, concentrated and sprayed by powder to obtain powdery grape seed polypeptide protein. The powdery grape seed polypeptide protein has the characteristics of good taste, no foreign taste, high protein extracting rate and low production cost, and the nutrition can be prevented from being lost in the process of manufacture of raw materials to the maximum.

Description

Biological enzyme extracts Semen Vitis viniferae polypeptide protein technology

Technical field

The present invention relates to a kind of vegetable-protein extraction process technology, promptly biological enzyme extracts Semen Vitis viniferae polypeptide protein technology.

Background technology

In prior art, Semen Vitis viniferae is the tankage in winery, the grape beverage factory production process, is wide material sources, the resourceful main resource of natural function food that is rich in nutrition.Contain rich in protein in the Semen Vitis viniferae benevolence, its content accounts for 28.38% of Semen Vitis viniferae, and quality is superior.Through inquiry, do not see that having with the Semen Vitis viniferae is the report that raw material extracts polypeptide protein.

Summary of the invention

The objective of the invention is to provide a kind of at above-mentioned deficiency is raw material with the Semen Vitis viniferae, technology advanced person, and the biological enzyme that the polypeptide protein quality is high extracts Semen Vitis viniferae polypeptide protein technology.

Technical solution of the present invention is: biological enzyme extracts Semen Vitis viniferae polypeptide protein technology:

(1) with the Semen Vitis viniferae is raw material;

(2) Semen Vitis viniferae after will selecting is crushed to the 20-40 order, and shell (shell is standby), benevolence (accounting for about 46%) are separated;

(3) Semen Vitis viniferae benevolence is ground into the 80-100 order again, adds 8-12 and doubly purifies softening water, gets material solution;

(4) material solution is heated to 95-100 ℃, after material solution is quickly cooled to 45-50 ℃;

(5) add the hydrolysis of inscribe mixed enzyme: add subacidity cellulase 4-6 ‰, add simultaneously any 2-4 ‰ in inscribe papoid, bromeline, aspartic protease or 2709 Sumizyme MPs again, hydrolysis temperature 45-50 ℃, hydrolysis time 1.5-2.5 hour; Be preferably papoid.

(6) add circumscribed protease hydrolysis: add that any 2-4 ‰ is hydrolyzed in circumscribed aspergillus oryzae, bacillus enzyme or the black mold, hydrolysis temperature 45-50 ℃, hydrolysis time 1.5-2.5 hour; Be preferably aspergillus oryzae.

(7) separate: behind the Semen Vitis viniferae benevolence proteolysis, isolate the albumen of molecular weight below 5000.

(8) concentrate and to dust: parting liquid through vaporizer concentrate, spraying drying, Powdered Semen Vitis viniferae polypeptide protein.Can approximately extract 170 gram Semen Vitis viniferae polypeptide proteins as 1000 gram Semen Vitis viniferaes.

The screening of enzyme of Semen Vitis viniferae proteolysis:

1, the screening of protein incision enzyme:

From existing protein incision enzyme: the screening that experimentizes of enzymes such as 1# papoid, 2# bromeline, 3# aspartic protease, 4#2709 Sumizyme MP, every kind of proteolytic enzyme all adds subacidity crude cellulase 5 ‰ respectively in experiment, help opening the vegetable-protein complex construction, make protein structure become soft, non-protein gene exposes out, proteolysis is accelerated in the effect of favourable proteolytic enzyme inscribe.From the hydrolysis effect analysis, see Table 1-2

1) do not adding crude cellulase, the single endo-protease results of hydrolysis that adds sees Table 1:

Table 1

	The 1# papoid	The 2# bromeline	The 3# aspartic protease	The 4#2709 Sumizyme MP
					AN(g/100ml)	0.56	0.45	0.55	0.53

2) add the inscribe mixed enzyme

In Semen Vitis viniferae benevolence protein solution (1: 10), add endo-protease 3 ‰ more respectively behind adding 5 ‰ acidic cellulases earlier, 1#, 2#, 3#, 4# test its protease hydrolysis result respectively, see the following form 2:

Table 2

	The 1# mixed enzyme	The 2# mixed enzyme	The 3# mixed enzyme	The 4# mixed enzyme
					AN(g/100ml)	0.69	0.58	0.63	0.60

Mix the papoid best results visible the adding from table 1,2, and taking second place is to mix aspartic protease.

2, circumscribed proteolytic enzyme screening

The vegetable-protein hydrolysis, the simple mixed protein endo-protease that adds does not reach practical application effect, enzymic hydrolysis is later above 3 hours will to produce the small molecules bitter peptides adding, make product produce bitter taste, so must shorten the inscribe lytic enzyme time, increase excision enzyme again and make the further hydrolysis of protein, excision enzyme makes after the hydrolysis hydrolyze protein molecules through after modifying again simultaneously, not only strengthen the proteolysis effect, also increased the protolysate local flavor simultaneously.

From aspergillus oryzae, extract the capable again vegetable-protein secondary hydrolysis of 1#, 2#, 3# excision enzyme in bacillus and the black mold,

See Table 3

Table 3

	The 1# excision enzyme	The 2# excision enzyme	The 3# excision enzyme
				AN(g/100ml)	1.20	0.91	1.10

As seen, through the secondary hydrolysis, vegetable-protein (Semen Vitis viniferae benevolence albumen) the hydrolyzed solution AN after inscribe is capable circumscribed again is up to 1.20g/100ml from table 3, and near the highest level of natural fermented AN more than month, this achievement provides favourable condition for actual production.

3, protein secondary hydrolysis application test aborning

Carrying out proteolysis under differing temps extracts protein the influence of production efficiency, production process energy consumption is seen Table 4

Table 4

Temperature ℃	AN (g/100ml)	Subsidence rate mm/30 branch	Accrual rate %	Parting liquid clarity T 100％1cm 550nm	Separation factor r	Remarks
							30	0.89	8.0	61.5	61.9	2900	Biological process 2 one-step hydrolysis
40	1.02	8.9	66	70.6	2850	Biological process 2 one-step hydrolysis
							50	1.18	9.3	73.5	86.0	2700	2 hydrolysis of biological process
50	0.56	3.0	58	29.8	3550	Nonbiological method

As seen utilizing the biological enzyme hydrolysis to extract Semen Vitis viniferae benevolence protein from table 4, is feasible.

Advantage of the present invention is: 1, utilize the two-step approach hydrolysis to extract vegetable-protein, solved proteinic bitter taste problem in the single stage method extraction, improved the solubleness of protein in water simultaneously, improved transmittance more than 55%, micromolecule polypeptide albumen obviously increases, improve the protein biological value, improved mouthfeel.2, improved extraction rate of protein 15.5% with this method, reduced cost, the extraction yield of polypeptide protein is higher than nonbiological method 10% after tested, is in first of the protein extracting ratio, just separates one and can save the energy 10%.3, amplify test with this method through industry, because solubleness increases in the protein liquid, viscosity degradation, the intramolecularly friction reduces, reduce liquid-solid isolating separating factor, helped proteic extraction and separate, kept raw material nutrition in the course of processing not suffer a loss to greatest extent.4, because protolysate fluid viscosity low flow is good, for concentrating, cryogenic vacuum in producing provides favourable condition, concentrate dry solids content up to 40%, can be spraying drying energy-conservation 20%.5, utilize biological process to extract the low temperature moisture evaporation concentration that albumen helps juice, the quick moisture evaporation drying that helps product, enhance productivity, reduce production costs, guarantee quality product, make high molecular weight protein be hydrolyzed into the biological activity protein of micromolecule polypeptide, soluble small molecular polypeptide active albumen, protein molecular weight 100,000 units before by hydrolysis become below 5,000 units after the hydrolysis, and content reaches more than 80%, help digesting and assimilating, improved proteinic biological value.

Below in conjunction with embodiment the present invention is described in further detail.

Embodiment

Embodiment

Biological enzyme extracts Semen Vitis viniferae polypeptide protein technology:

1, raw material: dried Semen Vitis viniferae raw material inclusion-free, nothing are gone mouldy, free from insect pests.

2, be crushed to the 20-40 order, shell, benevolence are separated.

3, Semen Vitis viniferae benevolence is ground into 100 orders again, adds 10 times and purifies softening water.

4, pre-treatment: the purpose of pre-treatment is by high temperature 95-100 ℃, and the material that enzyme is played unstable effect is destroyed, and helps the restriction endonuclease effect, accelerates proteolysis.When protein solution is quickly cooled to 45-50 ℃, adds enzyme again and be hydrolyzed standby.

5, add the hydrolysis of inscribe mixed enzyme: add subacidity cellulase (subacidity crude protein enzyme) 5 ‰, add inscribe papoid 3 ‰ simultaneously again, hydrolysis temperature under 45～50 ℃, hydrolysis 2 hours.

6, add circumscribed protease hydrolysis: add circumscribed proteolytic enzyme aspergillus oryzae 3 ‰, hydrolysis time 2 hours, 45～50 ℃ of hydrolysis temperatures, reaction end.

7, separate: behind the Semen Vitis viniferae benevolence proteolysis, separate through the vertical and high-speed whizzer, separating factor is greater than 2700,9600 rev/mins of rotating speeds, its transmittance of isolating clear liquor (T ^100%1cm 550nm) more than or equal to 86% o'clock, the molecular weight of small molecular protein this moment below 5000 reaches more than 80%, directly delivers to concentrated.

8, concentrate and to dust: parting liquid concentrates through the economic benefits and social benefits vacuum-evaporator, reaches at 40% o'clock and deliver to spraying drying in concentration, ultralow pressure (0.25～0.35Mpa) granulation of dusting is once finished, Powdered Semen Vitis viniferae polypeptide protein.

Claims (1)

1, a kind of biological enzyme extracts Semen Vitis viniferae polypeptide protein technology, it is characterized in that:

(1) with the Semen Vitis viniferae is raw material;

(2) Semen Vitis viniferae after will selecting is crushed to the 20-40 order, and shell, benevolence are separated;

(3) Semen Vitis viniferae benevolence is ground into the 80-100 order again, adds 8-12 and doubly purifies softening water, gets material solution;

(4) material solution is heated to 95-100 ℃, after material solution is quickly cooled to 45-50 ℃;

(5) add the hydrolysis of inscribe mixed enzyme: add subacidity cellulase 4-6 ‰, add simultaneously any 2-4 ‰ in inscribe papoid, bromeline, aspartic protease or 2709 Sumizyme MPs again, hydrolysis temperature 45-50 ℃, hydrolysis time 1.5-2.5 hour;

(6) add circumscribed protease hydrolysis: add that any 2-4 ‰ is hydrolyzed in circumscribed aspergillus oryzae, bacillus enzyme or the black mold, hydrolysis temperature 45-50 ℃, hydrolysis time 1.5-2.5 hour;

(7) separate: behind the Semen Vitis viniferae benevolence proteolysis, separate, contain the albumen of molecular weight below 5000 in the parting liquid;

(8) concentrate and to dust: parting liquid through vaporizer concentrate, spraying drying, Powdered Semen Vitis viniferae polypeptide protein.