A kind of method of Production by Enzymes pea peptide
Technical field
The inventive method relates to the deep process technology field of Semen Pisi sativi protein powder, be specifically related to a kind of method of Production by Enzymes pea peptide, be a kind of be raw material with the Semen Pisi sativi protein powder, utilize biological enzyme enzymolysis Semen Pisi sativi protein powder, utilize the biological enzyme modification technique that protein peptide is modified again, thereby extract the method for pea peptide.
Background technology
The pea seed contains protein 15.5%-39.7%, and fat is about 2%, also rich vitamin B1, B2 and nicotinic acid.The Semen Pisi sativi protein resource of China is extensive, low price, and Semen Pisi sativi protein is hydrolyzed can obtain the pea peptide, and it is neutral that the pH value is.Studies show that, contain in the pea peptide human body can not self synthetic 8 in amino acid, and their ratio approaches the recommendation pattern of FAO/WHO.
But, have a strong impact on absorption and the utilization of human body to them because these amino acid are gone up substantially with the polymeric form and are present in the protein.Studies show that mainly with the form absorption of small-molecular peptides, the specific absorption of hanging down peptide by evidence is bigger than amino acid whose specific absorption, is absorbed by the body sooner, utilizes than amino acid is easier after the digestive enzyme effect for protein.Based on this theory, the pea polypeptide that utilizes biotechnology pacemaker enzyme incision technology to develop is with a wide range of applications.
The pea polypeptide can not only provide the human body required nutritive substance that grows, and has good physical and chemical index and functional performance simultaneously.Through functional experiment to the pea polypeptide, the researchist finds that the pea polypeptide has better solvability, water-retentivity, oil absorbency, whipability, emulsifying property, gelation etc. than pea protein, and with regard to several factors the influence of pea polypeptide functional performance is proved, for the application of pea polypeptide provides theoretical foundation.
Pea polypeptide favorable effects and good functional property, make the pea polypeptide can be widely used in food and the healthcare products, the pea polypeptide has quite high water-retentivity, oil absorbency and good gel formation, can be used for meat products such as ham sausage, as good additive; The pea polypeptide has certain foaminess and froth stability, can partly replace eggs to add in the cake product; The pea polypeptide has extraordinary emulsifying property and emulsifying stability, can be used as the emulsifying agent of varieties of food items; The pea polypeptide is emulsifying fat rapidly, the good sausage of modulation emulsifying property, and the sausage that makes is very good to eat, is of high nutritive value; The pea peptide is used for biscuit can strengthen fragrance, strengthens protein, and can be developed into the protective foods of difference in functionality; The pea peptide can also be used for the Noodles goods, as can improve nutritive value, intensity and the biceps of noodles in the face with the pea peptide, can improve the outward appearance and the mouthfeel of cereal preparation.
From Semen Pisi sativi protein powder, extract the pea peptide and adopt following method mostly: Semen Pisi sativi protein powder → alkaline purification → high temperature steaming → enzymolysis → filtration → concentrate → spraying drying.Use a large amount of alkaline purifications in the leaching process, both environment was polluted and there is potential safety hazard in human body, destroyed proteinic composition again; And concentrate the product phase of destroying product easily, and do not use membrane separation technique in the prior art, influenced the quality of product.
Summary of the invention
At the deficiencies in the prior art, the object of the present invention is to provide a kind of method of Production by Enzymes pea peptide, this method is utilized biological enzyme enzymolysis Semen Pisi sativi protein powder, and further modifies the peptide bond debitterize, with membrane separation technique desalination and concentrated, finally extract highly purified pea peptide again.
To achieve these goals, the inventive method adopts following technical measures:
A kind of method of Production by Enzymes pea peptide, its step is as follows:
1, with the Semen Pisi sativi protein powder be raw material, cross earlier 40 mesh sieve removal of impurities, the 5-7 that adds raw material weight is doubly, distilled water, 95wt% ethanol and the sodium sulphite anhydrous 99.3 of 5-10% and 1-2% stir, at 90-95 ℃ of following back flow reaction 3-5h, and filtered while hot after react;
2, cooling filtrate is to 50-60 ℃, and adjust pH is 7-8, and the 2-4% that presses raw material weight adds Sumizyme MP, stirs enzymolysis 4-6h, keeps stablizing the pH value stabilization in the enzymolysis process; The 1-2% of raw material weight adds flavor protease again, stirs enzymolysis 5-8h, keeps stablizing the pH value stabilization in the enzymolysis process; The enzymolysis solution that obtains is warming up to 90-100 ℃, keeps the 10-20min enzyme that goes out;
3, with the centrifugal after-filtration of enzymolysis solution, remove proteolytic enzyme, add powdered active carbon and decolour, use the membrane sepn means to separate then, concentrate, promptly get pea peptide dry product after the concentrated solution drying.
The Sumizyme MP that uses in the inventive method and the vigor of flavor protease are respectively 25U/mg and 30U/mg.
Compared with prior art, advantage of the inventive method and beneficial effect are as follows:
1, adopt the purity height of the pea peptide product that the inventive method produces, the peptide content of molecular weight below 1000 dalton reaches more than 98% in its total protein, not only improved the nutritive value of pea, brought economic benefit for society simultaneously;
2, the pea peptide that adopts the inventive method to produce, molecular weight is little, and human body easily absorbs, and specific absorption reaches more than 90%;
3, the present invention adopts activated carbon decolorizing, residues in the product injury to human body when effectively having avoided using the hydrogen peroxide decolouring, and has guaranteed the natural colored degree of product;
4, adopt membrane separation technique of the present invention, the liquid towards material separates, concentrates and purifying, all operates, does not have phase-state change, energy-efficient at normal temperatures, and do not produce pollution in the production process;
5, the inventive method is with short production cycle, cost is low, do not produce any hazardous and noxious substances, and safety has no side effect, and can be widely used in fields such as makeup, food, medicine.
Embodiment
Below by specific embodiment the inventive method is described in further detail.
Embodiment 1:
A kind of method of Production by Enzymes pea peptide, its step is as follows:
Took by weighing commercially available Semen Pisi sativi protein powder behind 40 mesh sieves (crude protein>61wt% wherein, moisture content<10wt%, the 1kg of ash<3wt%), add 7kg distilled water, 50g 95wt% ethanol and 10g sodium sulphite anhydrous 99.3, after stirring, be warming up to 90 ℃ of backflow 5h, reacted the back filtered while hot; Cool off filtrate to 50 ℃ again, transfer pH to 7-8 with the food grade hydroxide flake sodium solution of 0.1M, add the 20g Sumizyme MP then, 50 ℃ are stirred enzymolysis 6h, in the enzymolysis process with the sodium hydroxide solution maintenance pH value of reaction system of above-mentioned 0.1M at 7-8; The flavor protease that adds 10g again, 50 ℃ of enzymolysis 8h, the sodium hydroxide solution with above-mentioned 0.1M in the enzymolysis process keeps pH value of reaction system at 7-8; Be warming up to 90 ℃ then, the insulation 20min enzyme that goes out; Then enzymolysis solution is filtered, get supernatant liquor; Add the 20g powdered active carbon in the supernatant liquor, 55 ℃ of whip attachment 1h decolourings, filtering gac; The filtrate via hole diameter is that the nanofiltration membrane separation system of 1nm separates, and desalination also concentrates; Concentrated solution is spray-dried, obtains the Powdered pea peptide of 386g white (adopt activated carbon decolorizing among the present invention, the color and luster of products obtained therefrom is a white, is better than the faint yellow of national Specification), and the examining report of products obtained therefrom sees Table 1.
The examining report of table 1 embodiment 1 products obtained therefrom
Test item |
National Specification |
Detected result |
Form |
Powdered, no caking phenomenon |
Powdered, there is not caking |
Color and luster |
Yellow or faint yellow |
White |
Flavour and smell |
The flavour and the smell that have product to have |
Do not have |
Impurity |
The visible tramp material of no twenty-twenty vision |
Inclusion-free |
Gross protein |
≥80.0% |
95.6% |
(the molecular weight 1000 of peptide in the total protein |
≥75.0% |
98.3% |
Dalton is following) content |
|
|
PH value (the 10wt% aqueous solution) |
7.0-8.0 |
7.3 |
Moisture |
≤7.0% |
1.5% |
Ash content |
≤10.0% |
2.1% |
Arsenic content (in the As element) |
≤0.5mg/kg |
0.22mg/kg |
Lead content (in the Pb element) |
≤0.5mg/kg |
0.16mg/kg |
Total plate count |
≤30000cfu/g |
1000cfu/g |
Coliform |
≤40MPN/100g |
<20MPN/100g |
Mould and yeast |
≤50cfu/g |
<10cfu/g |
Pathogenic bacterium |
Must not detect |
Do not detect |
Embodiment 2:
A kind of method of Production by Enzymes pea peptide, its step is as follows:
Took by weighing commercially available Semen Pisi sativi protein powder behind 40 mesh sieves (crude protein>61wt% wherein, moisture content<10wt%, the 2kg of ash<3wt%), add 10kg distilled water, 200g 95wt% ethanol and 30g sodium sulphite anhydrous 99.3, after stirring, be warming up to 92 ℃ of backflow 4h, reacted the back filtered while hot; Cool off filtrate to 55 ℃ again, transfer pH to 7-8 with the food grade hydroxide flake sodium solution of 0.1M, add the 60g Sumizyme MP then, 55 ℃ are stirred enzymolysis 5h, in the enzymolysis process with the sodium hydroxide solution maintenance pH value of reaction system of above-mentioned 0.1M at 7-8; Add the 30g flavor protease again, 55 ℃ of enzymolysis 7h, the sodium hydroxide solution with above-mentioned 0.1M in the enzymolysis process keeps pH value of reaction system at 7-8; Be warming up to 94 ℃ then, the insulation 16min enzyme that goes out; Then enzymolysis solution is filtered, get supernatant liquor; Add the 45g powdered active carbon in the supernatant liquor, 55 ℃ of whip attachment 1h decolourings, filtering gac; The filtrate via hole diameter is that the nanofiltration membrane separation system of 1nm separates, and desalination also concentrates; Concentrated solution is spray-dried, obtains 800g white powder pea peptide, and the examining report of products obtained therefrom sees Table 2.
The examining report of table 2 embodiment 2 products obtained therefroms
Embodiment 3:
A kind of method of Production by Enzymes pea peptide, its step is as follows:
Took by weighing commercially available Semen Pisi sativi protein powder behind 40 mesh sieves (crude protein>61wt% wherein, moisture content<10wt%, the 10kg of ash<3wt%), the ethanol and the 200g sodium sulphite anhydrous 99.3 that add 60kg distilled water, 700g 95wt%, after stirring, be warming up to 93 ℃ of backflow 4h, reacted the back filtered while hot; Cool off filtrate to 55 ℃ again, transfer pH to 7-8 with the food grade hydroxide flake sodium solution of 0.1M, add the 400g Sumizyme MP then, 55 ℃ are stirred enzymolysis 5h, in the enzymolysis process with the sodium hydroxide solution maintenance pH value of reaction system of above-mentioned 0.1M at 7-8; Add the 200g flavor protease again, 55 ℃ of enzymolysis 6h, the sodium hydroxide solution with above-mentioned 0.1M in the enzymolysis process keeps pH value of reaction system at 7-8; Be warming up to 96 ℃ then, the insulation 14min enzyme that goes out; Then enzymolysis solution is filtered, get supernatant liquor; Add the 250g powdered active carbon in the supernatant liquor, 55 ℃ of whip attachment 1h decolourings, filtering gac; The filtrate via hole diameter is that the nanofiltration membrane separation system of 1nm separates, and desalination also concentrates; Concentrated solution is spray-dried, obtains 4.01kg white powder pea peptide, and the examining report of products obtained therefrom sees Table 3.
The examining report of table 3 embodiment 3 products obtained therefroms
Embodiment 4:
A kind of method of Production by Enzymes pea peptide, its step is as follows:
Took by weighing commercially available Semen Pisi sativi protein powder behind 40 mesh sieves (crude protein>61wt% wherein, moisture content<10wt%, the 100kg of ash<3wt%), add 650kg distilled water, 8kg 95wt% ethanol and 1600g sodium sulphite anhydrous 99.3, after stirring, be warming up to 95 ℃ of backflow 3h, reacted the back filtered while hot; Cool off filtrate to 60 ℃ again, transfer pH to 7-8 with the food grade hydroxide flake sodium solution of 0.1M, add the 3000g Sumizyme MP then, 60 ℃ are stirred enzymolysis 4h, in the enzymolysis process with the sodium hydroxide solution maintenance pH value of reaction system of above-mentioned 0.1M at 7-8; Add the 1600g flavor protease again, 60 ℃ of enzymolysis 5h, the sodium hydroxide solution with above-mentioned 0.1M in the enzymolysis process keeps pH value of reaction system at 7-8; Be warming up to 100 ℃ then, the insulation 10min enzyme that goes out; Then enzymolysis solution is filtered, get supernatant liquor; Add the 2.2kg powdered active carbon in the supernatant liquor, 55 ℃ of whip attachment 1h decolourings, filtering gac; The filtrate via hole diameter is that the nanofiltration membrane separation system of 1nm separates, and desalination also concentrates; Concentrated solution is spray-dried, obtains 41kg white powder pea peptide, and the examining report of products obtained therefrom sees Table 4.
The examining report of table 4 embodiment 4 products obtained therefroms