Summary of the invention
The objective of the invention is to overcome the prior art above shortcomings, the preparation method of anti-oxidation peptide is provided.
The preparation method of anti-oxidation peptide of the present invention comprises the steps:
(1) control enzymolysis; With the dregs of beans is raw material, adds water and makes protein content reach 8-12% (w/w), 100 ℃ of sterilization 20min, be cooled to 45-65 ℃, add among Alcalase or the Protamex one or both, enzymolysis to degree of hydrolysis is 10-15%, the 85-95 ℃ of enzyme 15-25min that goes out gets enzymolysis liquid;
(2) high molecular weight protein is dispeled in ultrafiltration; The enzymolysis residue is dispeled in the enzymolysis liquid filtration, and crossing molecular weight is the milipore filter of 10-50KDa, gets through liquid;
(3) ion exchange resin: will pass through 717 strong basic type anion-exchange resins through liquid, be eluant, eluent with the pure water, collects eluent;
(4) Maillard reaction: it is 20-25% (w/w) that eluent is concentrated into solid content, adds 2-3% (w/w) monose, and 110-121 ℃ of reaction 30-60min gets maillard reaction product;
(5) concentrated, spray-drying, get anti-oxidation peptide: vacuum concentrates maillard reaction product, and spray-drying gets anti-oxidation peptide.
The present invention compared with prior art has following advantage:
1, the oxidation resistance of the anti-oxidation peptide announced of the present invention is synthetized oxidation preventive agent such as BHA, BHT and bamboo extractive, Vc, V
EDeng 2-10 times of natural, have typical sauce fragrant breeze flavor, and safety non-toxic, can be used as functional food ingredient and be widely used in the saline taste food service industry.
2, among the present invention the hydrolysate of soybean protein product through behind ion exchange resin and the Maillard reaction, its antioxidation activity common antioxidant in the present food industry.
3, the preparation technology of anti-oxidation peptide disclosed by the invention is simple, and product safety is produced on a large scale.
The specific embodiment
The ABTS radical scavenging activity of anti-oxidation peptide is measured in the embodiment:
ABTS
+Storing solution preparation: with deionized water with ABTS and K
2S
2O
8Dissolve respectively and mix, make its final concentration be respectively 7mmol/L and 2.45mmol/L, leave standstill 12~16h under room temperature, lucifuge condition, this storing solution can be stablized 3~4 days.During mensuration, with ABTS
+(10mmol/L, pH7.4) dilution makes its absorbance reach 0.700 ± 0.020 (under the 734nm wavelength) to storing solution, forms ABTS with phosphate buffer
+Measure liquid.Get 4.0mlABTS
+Measure liquid, add 40l Trolox or sample diluting liquid, the 30s that accurately vibrates, the light absorption value at 734nm place behind the assaying reaction 6min.Trolox concentration has the better linearity relation when 0~4mmol/L.Each sample is measured three concentration and linear at least, calculates average T rolox equivalent (Trolox Equivalent Antioxidant Capacity), i.e. TEAC value.
The assay method of peroxide value in the Guangdong style sausage:
Guangdong style sausage is made prescription and technology: lean meat: fat meat=7: 3; White granulated sugar 6%; Salt 3%, bent wine 4%; Monosodium glutamate 0.2%, nitrite 0.20g/Kg; Water 18%; Add the anti-oxidation peptide products substitution part lean meat of certain 0.05% (w/w) and guarantee material balance, blank for not adding any additives; Drying condition: two sections heated-air dryings, preceding 3 hours, 52 ℃, relative humidity 40%, wind speed 0.2m/s; Back 49 hours, 45 ℃, relative humidity 40%, wind speed 0.2m/s.After the vacuum packaging of Guangdong style sausage sample, be positioned over 37 ℃ of storages.Press the described method of GB/T5009.44-2003 and extract fat, press GB/T5009.37-2003 and measure peroxide value (POV).POV-0, POV-10, POV-30 represent respectively stored 0 day, 10 days and 30 days Guangdong style sausages in the peroxide value of fat.
The present invention is described further below in conjunction with specific embodiment.
Embodiment 1
(1) control enzymolysis; With the dregs of beans is raw material, adds water and makes protein content reach 12%, 100 ℃ of sterilization 20min, is cooled to 55 ℃, and adding Alcalase enzymolysis to degree of hydrolysis is 14.7%, 95 ℃ of enzyme 25min that goes out, and gets enzymolysis liquid;
(2) high molecular weight protein is dispeled in ultrafiltration; The enzymolysis residue is dispeled in the enzymolysis liquid filtration, and crossing molecular weight is the milipore filter of 10KDa, gets through liquid;
(3) ion exchange resin: will pass through 717 strong basic type anion-exchange resins through liquid, be the eluant, eluent wash-out with the pure water, collects eluent;
(4) Maillard reaction: it is 25% (w/w) that eluent is concentrated into solid content, adds 3% (w/w) glucose, and 121 ℃ of reaction 30min get maillard reaction product;
(5) concentrated, spray-drying, get anti-oxidation peptide: vacuum concentrates maillard reaction product, and spray-drying gets anti-oxidation peptide product 1.
Embodiment 2
(1) control enzymolysis; With the dregs of beans is raw material, adds water and makes protein content reach 8.3%, 100 ℃ of sterilization 20min, is cooled to 65 ℃, and adding Protamex enzymolysis to degree of hydrolysis is 10.6%, 88 ℃ of enzyme 25min that goes out, and gets enzymolysis liquid;
(2) high molecular weight protein is dispeled in ultrafiltration; The enzymolysis residue is dispeled in the enzymolysis liquid filtration, and crossing molecular weight is the milipore filter of 50KDa, gets through liquid;
(3) ion exchange resin: will pass through 717 strong basic type anion-exchange resins through liquid, be eluant, eluent with the pure water, collects eluent;
(4) Maillard reaction: it is 20% (w/w) that eluent is concentrated into solid content, adds 2.1% (w/w) wood sugar, and 110 ℃ of reaction 60min get maillard reaction product;
(5) concentrated, spray-drying, get anti-oxidation peptide: vacuum concentrates maillard reaction product, and spray-drying gets anti-oxidation peptide product 2.
By table 1 as seen, the ABTS radical scavenging activity of the prepared anti-oxidation peptide of the present invention is far above synthetized oxidation preventive agent BHA and BHT and natural V
CAnd V
E
By table 2 as seen, the prepared anti-oxidation peptide of the present invention is added into Guangdong style sausage with 0.05% (w/w) addition and can obviously suppresses peroxide value rising in the Guangdong style sausage storage process, and effect is significantly better than BHA.
Embodiment 3
(1) control enzymolysis; With the dregs of beans is raw material, adds water and makes protein content reach 9.5%, 90 ℃ of sterilization 20min, is cooled to 50 ℃, and adding Protamex enzymolysis to degree of hydrolysis is 13.8%, 85 ℃ of enzyme 20min that goes out, and gets enzymolysis liquid;
(2) high molecular weight protein is dispeled in ultrafiltration; The enzymolysis residue is dispeled in the enzymolysis liquid filtration, and crossing molecular weight is the milipore filter of 10KDa, gets through liquid;
(3) ion exchange resin: will pass through 717 strong basic type anion-exchange resins through liquid, be eluant, eluent with the pure water, collects eluent;
(4) Maillard reaction: it is 20% (w/w) that eluent is concentrated into solid content, adds 3% (w/w) wood sugar, and 121 ℃ of reaction 15min get maillard reaction product;
(5) concentrated, spray-drying, get anti-oxidation peptide: vacuum concentrates maillard reaction product, and spray-drying gets the anti-oxidation peptide product.
Embodiment 4
(1) control enzymolysis; With the dregs of beans is raw material, adds water and makes protein content reach 9.8%, 100 ℃ of sterilization 20min, is cooled to 55 ℃, and adding Alcalase enzymolysis to degree of hydrolysis is 14.3%, 85 ℃ of enzyme 20min that goes out, and gets enzymolysis liquid;
(2) high molecular weight protein is dispeled in ultrafiltration; The enzymolysis residue is dispeled in the enzymolysis liquid filtration, and crossing molecular weight is the milipore filter of 10KDa, gets through liquid;
(3) ion exchange resin: will pass through 717 strong basic type anion-exchange resins through liquid, be eluant, eluent with the pure water, collects eluent;
(4) Maillard reaction: it is 23% (w/w) that eluent is concentrated into solid content, adds 3% (w/w) glucose, and 121 ℃ of reaction 10min get maillard reaction product;
(5) concentrated, spray-drying, get anti-oxidation peptide: vacuum concentrates maillard reaction product, and spray-drying gets the anti-oxidation peptide product.
Table 1
Table 2