CN115381095B - Method for improving load and stability of fat-soluble active factor - Google Patents
Method for improving load and stability of fat-soluble active factor Download PDFInfo
- Publication number
- CN115381095B CN115381095B CN202211046117.4A CN202211046117A CN115381095B CN 115381095 B CN115381095 B CN 115381095B CN 202211046117 A CN202211046117 A CN 202211046117A CN 115381095 B CN115381095 B CN 115381095B
- Authority
- CN
- China
- Prior art keywords
- fat
- stability
- protein
- soluble active
- particles
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 22
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 75
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 75
- 230000013595 glycosylation Effects 0.000 claims abstract description 24
- 238000006206 glycosylation reaction Methods 0.000 claims abstract description 24
- 230000004048 modification Effects 0.000 claims abstract description 22
- 238000012986 modification Methods 0.000 claims abstract description 22
- 239000013543 active substance Substances 0.000 claims abstract description 5
- 238000005516 engineering process Methods 0.000 claims abstract description 4
- 239000002245 particle Substances 0.000 claims description 40
- 239000000243 solution Substances 0.000 claims description 23
- 239000007788 liquid Substances 0.000 claims description 22
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 22
- 239000006185 dispersion Substances 0.000 claims description 19
- 238000003756 stirring Methods 0.000 claims description 17
- 239000011162 core material Substances 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- 239000000084 colloidal system Substances 0.000 claims description 11
- 239000000463 material Substances 0.000 claims description 11
- 239000008055 phosphate buffer solution Substances 0.000 claims description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- 108090000790 Enzymes Proteins 0.000 claims description 8
- 102000004190 Enzymes Human genes 0.000 claims description 8
- 230000001965 increasing effect Effects 0.000 claims description 8
- 229920002307 Dextran Polymers 0.000 claims description 6
- 102000004142 Trypsin Human genes 0.000 claims description 6
- 108090000631 Trypsin Proteins 0.000 claims description 6
- 239000012588 trypsin Substances 0.000 claims description 6
- 238000002604 ultrasonography Methods 0.000 claims description 6
- 239000002131 composite material Substances 0.000 claims description 5
- 238000000502 dialysis Methods 0.000 claims description 4
- 230000009144 enzymatic modification Effects 0.000 claims description 4
- 238000000108 ultra-filtration Methods 0.000 claims description 3
- 239000004365 Protease Substances 0.000 claims description 2
- 230000007613 environmental effect Effects 0.000 claims description 2
- 238000000265 homogenisation Methods 0.000 claims description 2
- 230000003993 interaction Effects 0.000 claims description 2
- 230000004044 response Effects 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims 2
- 108091005804 Peptidases Proteins 0.000 claims 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims 1
- 239000012634 fragment Substances 0.000 claims 1
- 239000000693 micelle Substances 0.000 claims 1
- 238000002360 preparation method Methods 0.000 claims 1
- 230000002209 hydrophobic effect Effects 0.000 abstract description 12
- 235000013305 food Nutrition 0.000 abstract description 11
- 238000012545 processing Methods 0.000 abstract description 7
- 102000008934 Muscle Proteins Human genes 0.000 abstract description 2
- 108010074084 Muscle Proteins Proteins 0.000 abstract description 2
- 235000019629 palatability Nutrition 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 230000002797 proteolythic effect Effects 0.000 abstract description 2
- 239000004480 active ingredient Substances 0.000 abstract 1
- 230000003078 antioxidant effect Effects 0.000 abstract 1
- 238000011156 evaluation Methods 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 63
- REFJWTPEDVJJIY-UHFFFAOYSA-N Quercetin Chemical compound C=1C(O)=CC(O)=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 REFJWTPEDVJJIY-UHFFFAOYSA-N 0.000 description 20
- 239000000047 product Substances 0.000 description 15
- 229960001285 quercetin Drugs 0.000 description 11
- ZVOLCUVKHLEPEV-UHFFFAOYSA-N Quercetagetin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=C(O)C(O)=C(O)C=C2O1 ZVOLCUVKHLEPEV-UHFFFAOYSA-N 0.000 description 10
- HWTZYBCRDDUBJY-UHFFFAOYSA-N Rhynchosin Natural products C1=C(O)C(O)=CC=C1C1=C(O)C(=O)C2=CC(O)=C(O)C=C2O1 HWTZYBCRDDUBJY-UHFFFAOYSA-N 0.000 description 10
- MWDZOUNAPSSOEL-UHFFFAOYSA-N kaempferol Natural products OC1=C(C(=O)c2cc(O)cc(O)c2O1)c3ccc(O)cc3 MWDZOUNAPSSOEL-UHFFFAOYSA-N 0.000 description 10
- 235000005875 quercetin Nutrition 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 238000001816 cooling Methods 0.000 description 8
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 description 8
- 239000003094 microcapsule Substances 0.000 description 8
- 239000000843 powder Substances 0.000 description 8
- 229940088598 enzyme Drugs 0.000 description 7
- 230000014759 maintenance of location Effects 0.000 description 7
- 239000001168 astaxanthin Substances 0.000 description 6
- 229940022405 astaxanthin Drugs 0.000 description 6
- JEBFVOLFMLUKLF-IFPLVEIFSA-N Astaxanthin Natural products CC(=C/C=C/C(=C/C=C/C1=C(C)C(=O)C(O)CC1(C)C)/C)C=CC=C(/C)C=CC=C(/C)C=CC2=C(C)C(=O)C(O)CC2(C)C JEBFVOLFMLUKLF-IFPLVEIFSA-N 0.000 description 5
- 235000013793 astaxanthin Nutrition 0.000 description 5
- MQZIGYBFDRPAKN-ZWAPEEGVSA-N astaxanthin Chemical compound C([C@H](O)C(=O)C=1C)C(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)C(=O)[C@@H](O)CC1(C)C MQZIGYBFDRPAKN-ZWAPEEGVSA-N 0.000 description 5
- 239000004148 curcumin Substances 0.000 description 5
- 229940109262 curcumin Drugs 0.000 description 5
- QRYRORQUOLYVBU-VBKZILBWSA-N Carnosic acid Natural products CC([C@@H]1CC2)(C)CCC[C@]1(C(O)=O)C1=C2C=C(C(C)C)C(O)=C1O QRYRORQUOLYVBU-VBKZILBWSA-N 0.000 description 4
- 108010087806 Carnosine Proteins 0.000 description 4
- CQOVPNPJLQNMDC-UHFFFAOYSA-N N-beta-alanyl-L-histidine Natural products NCCC(=O)NC(C(O)=O)CC1=CN=CN1 CQOVPNPJLQNMDC-UHFFFAOYSA-N 0.000 description 4
- 238000009835 boiling Methods 0.000 description 4
- 229940044199 carnosine Drugs 0.000 description 4
- CQOVPNPJLQNMDC-ZETCQYMHSA-N carnosine Chemical compound [NH3+]CCC(=O)N[C@H](C([O-])=O)CC1=CNC=N1 CQOVPNPJLQNMDC-ZETCQYMHSA-N 0.000 description 4
- 235000012754 curcumin Nutrition 0.000 description 4
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 description 4
- 150000004676 glycans Chemical class 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 239000000413 hydrolysate Substances 0.000 description 4
- 239000005457 ice water Substances 0.000 description 4
- 230000000415 inactivating effect Effects 0.000 description 4
- 239000008363 phosphate buffer Substances 0.000 description 4
- 229920001282 polysaccharide Polymers 0.000 description 4
- 239000005017 polysaccharide Substances 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- 238000009777 vacuum freeze-drying Methods 0.000 description 4
- 108010073771 Soybean Proteins Proteins 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 238000005286 illumination Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000000975 bioactive effect Effects 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- 235000013376 functional food Nutrition 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 102000035118 modified proteins Human genes 0.000 description 2
- 108091005573 modified proteins Proteins 0.000 description 2
- 239000003921 oil Substances 0.000 description 2
- 230000001590 oxidative effect Effects 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 230000009145 protein modification Effects 0.000 description 2
- 238000001338 self-assembly Methods 0.000 description 2
- 235000019710 soybean protein Nutrition 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 108010070551 Meat Proteins Proteins 0.000 description 1
- 102000014171 Milk Proteins Human genes 0.000 description 1
- 108010011756 Milk Proteins Proteins 0.000 description 1
- 108090000526 Papain Proteins 0.000 description 1
- 108010064851 Plant Proteins Proteins 0.000 description 1
- 229930182558 Sterol Natural products 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000006315 carbonylation Effects 0.000 description 1
- 238000005810 carbonylation reaction Methods 0.000 description 1
- 150000001746 carotenes Chemical class 0.000 description 1
- 235000005473 carotenes Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 235000021245 dietary protein Nutrition 0.000 description 1
- 102000038379 digestive enzymes Human genes 0.000 description 1
- 108091007734 digestive enzymes Proteins 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 235000013622 meat product Nutrition 0.000 description 1
- 235000021239 milk protein Nutrition 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 229940055729 papain Drugs 0.000 description 1
- 235000019834 papain Nutrition 0.000 description 1
- 235000021118 plant-derived protein Nutrition 0.000 description 1
- 150000008442 polyphenolic compounds Chemical class 0.000 description 1
- 235000013824 polyphenols Nutrition 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229940001941 soy protein Drugs 0.000 description 1
- 150000003432 sterols Chemical class 0.000 description 1
- 235000003702 sterols Nutrition 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000009469 supplementation Effects 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Biochemistry (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Botany (AREA)
- Zoology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention discloses a method for improving the load capacity and stability of a fat-soluble active factor, which belongs to the technical field of food processing and comprises enzymolysis of myofibrillar protein, glycosylation modification of an enzymolysis product, embedding of a fat-soluble active ingredient, stability evaluation and the like. According to the invention, the amphipathy of the protein is improved through enzymolysis, the stability and the antioxidant activity of a proteolytic product are further improved through glycosylation modification, the application of the hydrophobic protein as a carrier in the efficient entrapment of the fat-soluble functional factors is realized by combining the advantages of the two technologies, the utilization way of the muscle protein is widened, and the problems of poor stability, palatability and bioavailability of the fat-soluble active substances in the food processing process are solved.
Description
Technical Field
The invention relates to the technical field of food processing, in particular to a method for improving the loading capacity and stability of a fat-soluble active factor.
Background
The development of functional foods requires the enhancement or supplementation of bioactive factors such as carotenes, astaxanthin, polyphenols, sterols, vitamins, functional oils, active peptides, etc. in foods. However, many active factors have problems of poor solubility, poor palatability, low bioavailability, easy degradation during processing or storage, poor stability in the gastrointestinal tract environment, and the like due to the influence of factors such as chemical structures and the like. How to improve the water solubility of these bioactive factors, and to improve the stability and bioavailability thereof, is always a difficulty in food processing and is also a hot spot problem in research in the field of food science.
The food protein is widely used for constructing a food colloid delivery system because of natural amphipathy and colloid self-assembly, is recognized as an excellent carrier for protecting and delivering functional factors, and aims to solve the problems of weak solubility, low bioavailability, poor processing or storage stability and the like of active factors. Currently, proteins for industrial application are mainly concentrated on plant proteins and milk proteins which are more water-soluble, while the use of proteins which are more hydrophobic is extremely limited. Compared with water-soluble proteins, the hydrophobic proteins have more abundant hydrophobic sites, can realize embedding and conveying of fat-soluble substances by means of hydrophobic interaction, and have potential application prospects. However, the poor water solubility makes it difficult to disperse uniformly in the system, limiting its wide application.
Myofibrillar proteins are important structural proteins in muscle, accounting for 50-55% of the total amount of muscle proteins, and play an important role in maintaining the water-holding capacity, gel property, emulsifying property and other functions of meat products. However, its solubility in aqueous solutions is not high and its potential as an emulsifier alone is limited, limiting its use in certain foods. Therefore, in order to widen the application range of myofibrillar proteins, a widening idea is needed, and a proper technology is sought for modifying the myofibrillar proteins. Currently, many methods for protein modification mainly include physical modification, chemical modification, enzymatic modification, and the like. The protein is modified by the polysaccharide, so that aggregation of the protein can be effectively inhibited by means of steric hindrance effect of polysaccharide substances, and the solubility, stability and colloid stability of the protein are improved. For example, chinese patent application CN201310697737.9 proposes glycosylation modification of isolated soy protein with glucose, resulting in a 4.38-fold improvement in protein solubility over prior modification, but the improvement in myofibrillar protein solubility by this method is still very limited. In addition, myofibrillar proteins have poor oxidative stability, and are subject to structural changes (increased carbonylation, increased conversion of thiol groups to disulfide bonds, etc.) under oxidative conditions, and the amphiphilicity, colloidal properties, etc. of oxidized proteins are changed. Thus, for less water-soluble myofibrillar proteins, there is a need to simultaneously increase the solubility, stability and colloidal properties of the proteins by suitable molecular modification techniques.
Enzymatic modification is a common way of protein modification. Huang Jianzhao and the like modify SPI by papain, and the solubility of SPI is obviously improved; the Radha et al enzymatically modify soybean protein to find that the solubility of the protein is improved after enzymolysis and that other functions of the protein are not affected. The Chinese patent CN201510013536.1 utilizes the ultrahigh pressure homogenization combined enzyme method to modify the soybean protein isolate, and also discovers that the solubility, the water retention and the oil retention of the protein are improved. However, there are currently fewer reports of enzymatic modification of meat proteins. The hydrophobic protein is modified by combining the advantages of larger steric hindrance and enzymolysis of polysaccharide in improving the amphipathy of the protein, and the application of the hydrophobic protein serving as a carrier of a hydrophobic active substance in a food colloid conveying system is expected to be realized.
However, the protein used for constructing the food colloid delivery system is mainly concentrated on the protein with stronger water solubility, the successful case of the hydrophobic protein is rare, and no case of improving the solubility and colloid property of the hydrophobic protein by combining enzymolysis and glycosylation modification is disclosed at home and abroad.
Disclosure of Invention
The present invention aims to solve the above problems by providing a method for improving the load and stability of a fat-soluble active factor.
Methods for increasing the loading and stability of fat-soluble active factors:
firstly, carrying out limited enzymolysis on protein by gastrointestinal digestive enzyme to expose hydrophilic groups in the molecular structure of the protein, and improving the amphipathy and self-assembly characteristics of fragmented protein; then carrying out glycosylation treatment on the proteolytic products by utilizing polysaccharide to obtain modified protein recombinant particles; and then adding fat-soluble active factors, inducing the entrapment of the modified protein recombinant particles and the hydrophobic ligand by utilizing the environmental response characteristic, and obtaining the compound entrapped with the fat-soluble active factors after freeze drying.
The method comprises the following specific steps:
s1, enzymolysis modification of myofibrillar protein: dissolving and dispersing myofibrillar protein in 20mM phosphate buffer (containing 0.6mol/L NaCl, pH 7.4) to make the protein concentration 20mg/mL, taking 50mL, adjusting pH value to 7.0, adding 0.1g trypsin (1500U/mg), continuously stirring at 37 ℃ for enzymolysis for 2h, inactivating enzyme by boiling water after enzymolysis for 5min, cooling by ice water, centrifuging the hydrolysate at 10000rpm for 20min after cooling, and removing precipitate;
s2, glycosylation modification of a myofibrillar enzymolysis product: placing the enzymolysis liquid in a magnetic stirrer (300 rpm), slowly adding D-dextran powder until the addition amount reaches 20mg/mL, adjusting the pH of the solution to 7.0, continuously and fully stirring and reacting for 4 hours, performing vacuum freeze drying on the obtained solution, and reacting the obtained powder in a saturated KBr solution with the temperature of 60 ℃ and the Relative Humidity (RH) for 24 hours to obtain a myofibrillar protein peptide glycosylation product;
s3, preparing D-dextran-myofibrillar protein peptide nanometer self-assembled particles: stirring the peptide glycosylation product obtained in the step (2) in a water bath at 25 ℃ for 3 hours, redispersing the peptide glycosylation product in a 20mM phosphate buffer solution until the concentration of particles is 2mg/mL, transferring the peptide glycosylation product to 4 ℃ and standing overnight, and completely expanding the particles;
s4, preparing glycosylated myofibrillar protein peptide-fat-soluble factor composite colloid particles: dispersing the above-mentioned carnosine glycosylated particles in phosphate buffer solution with pH7.4 by utilizing ultrasonic waves (250W, 10 min) to obtain 2mg/mL wall material dispersion liquid; respectively dispersing active factors in absolute ethyl alcohol by utilizing ultrasonic waves (250W, 10 min) to obtain core material dispersion liquid with the active factor content of 10mg/mL, taking out 0.5mL of core material dispersion liquid, dropwise adding the core material dispersion liquid into 50mL-100mL of wall material dispersion liquid in a magnetic stirrer environment (300 rpm), stirring for 10min, and continuing ultrasonic waves of the solution at 250W for 10min to obtain glycosylated myofibrillar peptide nano-assembly particles embedded with fat-soluble factors.
The invention has the beneficial effects that:
compared with unmodified myofibrillar proteins, single enzymatic modified myofibrillar proteins, single glycosylation modified myofibrillar proteins and the like, the recombinant myofibrillar protein particles after combined enzymatic and glycosylation modification have higher solubility, and the stability of entrapped astaxanthin, curcumin, quercetin and other fat-soluble components is better, so that the system still presents a uniform state in the whole storage process. The embedding rate of the prepared embedded particles is higher than 80%. The modification of myofibrillar protein by the technology not only improves the solubility and stability of protein, but also obviously improves the embedding rate and stability of fat-soluble active factors, thereby being beneficial to realizing the application of the hydrophobic protein as a carrier of a food colloid conveying system in the processing of functional foods.
Drawings
FIG. 1 shows the change in solubility before and after MP modification;
FIG. 2 shows the entrapment rate of active factors in microcapsules;
FIG. 3 shows the stability of active factors in microcapsules under different light environments;
figure 4 shows the stability of the active factors in the microcapsules under different heating environments.
Detailed Description
The technical scheme of the present invention will be described in further detail with reference to the accompanying drawings in the embodiments of the present invention, but the present invention is not limited to the following embodiments. All other embodiments, which are derived from the embodiments of the invention without creative efforts of a person skilled in the art, belong to the protection scope of the present invention.
Example 1 method for increasing load and stability of astaxanthin using myofibrillar proteins:
s1, enzymolysis modification of myofibrillar protein: dissolving and dispersing myofibrillar protein in 20mM phosphate buffer (containing 0.6mol/L NaCl, pH 7.4) to make the protein concentration 20mg/mL, taking 50mL, adjusting pH value to 7.0, adding 0.1g trypsin (1500U/mg), continuously stirring at 37 ℃ for enzymolysis for 2h, inactivating enzyme by boiling water after enzymolysis for 5min, cooling by ice water, centrifuging the hydrolysate at 10000rpm for 20min after cooling, and removing precipitate;
s2, glycosylation modification of a myofibrillar enzymolysis product: placing the enzymolysis liquid in a magnetic stirrer (300 rpm), slowly adding D-dextran powder until the addition amount reaches 10mg/mL-20mg/mL, adjusting the pH of the solution to 7.0, continuously stirring and reacting for 4 hours, performing vacuum freeze drying on the obtained solution, and reacting the obtained powder in a saturated KBr solution with the temperature of 60 ℃ and the Relative Humidity (RH) for 24 hours to obtain a myofibrillar protein peptide glycosylation product, wherein the solubility is reached;
s3, preparing D-dextran-myofibrillar protein peptide nanometer self-assembled particles: redispersing the peptide glycosylation product obtained in the step (2) in a 20mM phosphate buffer solution by using a homogenizer (3000 rpm,30 s) until the particle concentration is 2mg/mL, stirring the solution in a water bath at 25 ℃ for 3 hours, transferring to 4 ℃ and standing for 8 hours to allow the particles to fully expand;
s4, preparing glycosylated myofibrillar protein peptide-astaxanthin composite colloid particles: dispersing the above-mentioned carnosine glycosylated particles in phosphate buffer solution with pH7.4 by utilizing ultrasonic waves (250W, 10 min) to obtain 2mg/mL wall material dispersion liquid; dispersing astaxanthin in absolute ethyl alcohol by utilizing ultrasonic waves (250W, 10 min) to obtain core material dispersion liquid with the active factor content of 10mg/mL, taking out 0.5mL of alcohol solution, dropwise adding the core material dispersion liquid into 50mL-100mL of wall material dispersion liquid in a magnetic stirrer environment (300 rpm), stirring for 10min, continuing ultrasonic waves for 10min at 250W to obtain glycosylated myofibrillar peptide nano-assembly particles embedded with fat-soluble factors, wherein the embedding rate of astaxanthin in the microcapsules reaches 85.40%, the retention rate of quercetin after 48h illumination reaches 64.28%, and the retention rate of quercetin is 57.28% after heating for 30min at 90 ℃.
Example 2 method for increasing the load and stability of curcumin using myofibrillar proteins:
s1, enzymolysis modification of myofibrillar protein: dissolving and dispersing myofibrillar protein in 20mM phosphate buffer (containing 0.6mol/L NaCl, pH 7.4) to make the protein concentration 20mg/mL, taking 50mL, adjusting pH value to 7.0, adding 0.01g trypsin, continuously stirring and hydrolyzing at 37 ℃ for 2h, inactivating enzyme by boiling water for 15min after enzymolysis, cooling by ice water after enzyme inactivation, centrifuging the hydrolysate at 10000rpm for 20min after cooling, removing large particle matters by using an ultrafiltration membrane of 10kDa, and dialyzing overnight by using a 100Da dialysis bag in a refrigerator at 4 ℃;
s2, glycosylation modification of a myofibrillar enzymolysis product: placing the enzymolysis solution in a magnetic stirrer (300 rpm), slowly adding D-dextran powder until the addition amount reaches 20mg/mL, adjusting the pH of the solution to 7.0, continuously stirring and reacting for 4 hours, performing vacuum freeze drying on the obtained solution, and reacting the obtained powder in a saturated KBr solution with the temperature of 60 ℃ and the Relative Humidity (RH) for 24 hours to obtain a myofibrillar protein peptide glycosylation product;
s3, preparing D-dextran-myofibrillar protein peptide nanometer self-assembled particles: redispersing the peptide glycosylation product obtained in the step (2) in a 20mM phosphate buffer solution by using a homogenizer (3000 rpm,30 s) until the particle concentration is 2mg/mL, stirring the solution in a water bath at 25 ℃ for 3 hours, transferring to 4 ℃ and standing for 8 hours to allow the particles to fully expand;
s4, preparing glycosylated myofibrillar protein peptide-curcumin composite colloid particles: dispersing the above-mentioned carnosine glycosylated particles in phosphate buffer solution with pH7.4 by utilizing ultrasonic waves (250W, 10 min) to obtain 2mg/mL wall material dispersion liquid; dispersing curcumin in absolute ethyl alcohol by utilizing ultrasonic waves (250W, 10 min) to obtain core material dispersion liquid with the active factor content of 10mg/mL, taking out 0.5mL of alcohol solution, dropwise adding the core material dispersion liquid into 50mL-100mL of wall material dispersion liquid in a magnetic stirrer environment (300 rpm), stirring for 10min, continuing ultrasonic waves for 10min at 250W to obtain glycosylated myofibrillar peptide nano-assembly particles embedded with fat-soluble factors, wherein the embedding rate of curcumin in the microcapsule reaches 88.6%, the quercetin retention rate of the microcapsule particles reaches 72.19% after 48h illumination, and the quercetin retention rate is 65.19% after heating for 30min at 90 ℃.
Example 3 method of increasing the load and stability of quercetin using myofibrillar proteins:
s1, enzymolysis modification of myofibrillar protein: the myofibrillar protein is dissolved and dispersed in 20mM phosphate buffer (containing 0.6mol/L NaCl, pH 7.4) to make the protein concentration 20mg/mL, 50mL is taken, the pH value is regulated to 7.0, 0.01g trypsin is added, continuous stirring and enzymolysis are carried out for 2h at 37 ℃, boiling water is used for inactivating enzyme for 15min after enzymolysis, ice water is used for cooling after enzyme inactivation, hydrolysate is centrifuged at 10000rpm for 20min after cooling, then large particle matters are removed by an ultrafiltration membrane of 10kDa, and then dialysis is carried out in a 100Da dialysis bag in a refrigerator at 4 ℃ for overnight.
S2, glycosylation modification of a myofibrillar enzymolysis product: placing the enzymolysis liquid in a magnetic stirrer (300 rpm), slowly adding D-dextran powder until the addition amount reaches 10mg/mL-20mg/mL, adjusting the pH of the solution to 7.0, continuously stirring and reacting for 4 hours, performing vacuum freeze drying on the obtained solution, and reacting the obtained powder in a saturated KBr solution with the temperature of 60 ℃ and the Relative Humidity (RH) for 24 hours to obtain a myofibrillar protein peptide glycosylation product;
s3, preparing D-dextran-myofibrillar protein peptide nanometer self-assembled particles: redispersing the peptide glycosylation product obtained in the step (2) in a 20mM phosphate buffer solution by using a homogenizer (3000 rpm,30 s) until the particle concentration is 2mg/mL, stirring the solution in a water bath at 25 ℃ for 3 hours, transferring to 4 ℃ and standing for 8 hours to allow the particles to fully expand;
s4, preparing glycosylated myofibrillar protein peptide-quercetin composite colloidal particles: dispersing the above-mentioned carnosine glycosylated particles in phosphate buffer solution with pH7.4 by utilizing ultrasonic waves (250W, 10 min) to obtain 2mg/mL wall material dispersion liquid; dispersing quercetin in absolute ethyl alcohol by utilizing ultrasound (250W, 10 min) to obtain core material dispersion liquid with the active factor content of 10mg/mL, taking out 0.5mL of alcohol solution, dropwise adding the core material dispersion liquid into 50mL-100mL of wall material dispersion liquid in a magnetic stirrer environment (300 rpm), stirring for 10min, continuing to carry out ultrasound on the solution at 250W for 10min to obtain glycosylated myofibrillar peptide nano-assembly particles embedded with fat-soluble factors, wherein the embedding rate of quercetin in the microcapsule reaches 90.7%, the retention rate of quercetin after 48h illumination of the microcapsule reaches 75.29%, and the retention rate of quercetin after 30min heating at 90 ℃ is 64.29%.
The foregoing examples are merely illustrative of the technical concept and features of the present invention, but the embodiments of the present invention are not limited to the foregoing examples, which are intended to enable those skilled in the art to understand the present invention and implement the same, and thus are not intended to limit the scope of the present invention. Any other changes, modifications, substitutions, combinations, and simplifications that do not depart from the spirit and principles of the invention are intended to be equivalent substitutes for those that do not depart from the spirit and principles of the invention.
Claims (6)
1. A method for increasing the loading and stability of a fat-soluble active agent comprising the steps of:
s1, enzymolysis modification of myofibrillar protein: carrying out limited enzymolysis on myofibrillar protein by trypsin, and separating the enzymolyzed fragmented protein by centrifugation;
s2, glycosylation modification of a myofibrillar enzymolysis product: carrying out glycosylation modification on the enzymolysis-carried myofibrillar protein peptide by using D-dextran through a dry-heat Maillard technology to obtain D-dextran-myofibrillar protein enzymolysis product conjugate particles;
s3, preparing D-dextran-myofibrillar protein peptide nanometer self-assembled particles: redispersing the peptide glycosylation product obtained in the step S2 in a 20mM phosphate buffer solution by using a homogenizer until the concentration of particles is 2mg/mL, stirring the solution in a water bath at 25 ℃ for 3 hours under the homogenization condition of 3000rpm and 30S, transferring to 4 ℃ and standing for 8 hours, and completely expanding the particles to form an assembled micelle;
s4, preparing glycosylated myofibrillar protein peptide-fat-soluble factor composite colloid particles: the self-assembled particles are dissolved in phosphate buffer solution to obtain wall material dispersion liquid by means of ultrasound, the fat-soluble active factors are dissolved in ethanol solution to obtain core material dispersion liquid, and the interaction between the assembled particles and the fat-soluble active factors is induced by using environmental response characteristics and ultrasound to realize the entrapment of the fat-soluble active factors.
2. The method for improving the loading and stability of a fat-soluble active factor according to claim 1, wherein trypsin is used for enzymatic modification of myofibrillar proteins in step S1, and the amount of enzyme added is 0.1g/g protein.
3. The method for improving the loading and stability of the fat-soluble active factors according to claim 1, wherein the protease in the step S1 is followed by centrifugation, ultrafiltration and dialysis to remove non-enzymatically hydrolyzed particles, thereby obtaining an enzymatically hydrolyzed fragment with better amphipathy.
4. The method for increasing the loading and stability of a fat-soluble active agent according to claim 1, wherein the D-dextran has a molecular weight of 40000 in step S2.
5. The method for improving the loading capacity and stability of the fat-soluble active factors according to claim 1, wherein in the preparation process of the active factor-loaded colloidal particles in step S4, the wall material and the core material are fully dispersed under the assistance of ultrasound, and the ultrasound condition is 250w for 10min.
6. The method for improving the loading and stability of a fat-soluble active agent according to claim 1, wherein the mass ratio of the wall material to the core material in step S4 is 20:1-40:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211046117.4A CN115381095B (en) | 2022-08-30 | 2022-08-30 | Method for improving load and stability of fat-soluble active factor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211046117.4A CN115381095B (en) | 2022-08-30 | 2022-08-30 | Method for improving load and stability of fat-soluble active factor |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN115381095A CN115381095A (en) | 2022-11-25 |
| CN115381095B true CN115381095B (en) | 2023-04-28 |
Family
ID=84123371
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211046117.4A Active CN115381095B (en) | 2022-08-30 | 2022-08-30 | Method for improving load and stability of fat-soluble active factor |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN115381095B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102793138A (en) * | 2012-08-10 | 2012-11-28 | 广东兴亿海洋生物工程有限公司 | Shellfish flavor peptide and preparation method thereof |
| CN111480800A (en) * | 2019-01-27 | 2020-08-04 | 西北农林科技大学 | Method for improving flavor of meat product |
| CN114081096A (en) * | 2021-10-18 | 2022-02-25 | 广东省农业科学院蚕业与农产品加工研究所 | Preparation method of milk protein loaded astaxanthin emulsion combined with polyphenol and glycosylation modification |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080241320A1 (en) * | 2007-03-30 | 2008-10-02 | Dsm Ip Assets B.V. | Protective hydrocolloid for active ingredients |
-
2022
- 2022-08-30 CN CN202211046117.4A patent/CN115381095B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102793138A (en) * | 2012-08-10 | 2012-11-28 | 广东兴亿海洋生物工程有限公司 | Shellfish flavor peptide and preparation method thereof |
| CN111480800A (en) * | 2019-01-27 | 2020-08-04 | 西北农林科技大学 | Method for improving flavor of meat product |
| CN114081096A (en) * | 2021-10-18 | 2022-02-25 | 广东省农业科学院蚕业与农产品加工研究所 | Preparation method of milk protein loaded astaxanthin emulsion combined with polyphenol and glycosylation modification |
Non-Patent Citations (2)
| Title |
|---|
| 杨宇鸿 ; 董士远 ; 靳卫亚 ; 毛振杰 ; 苏明月 ; 岳敏 ; .草鱼肌原纤维蛋白-葡萄糖糖基化产物的理化特性及乳化特性研究.食品工业科技.2018,(18),77-82，142. * |
| 荣婧 ; 仇超颖 ; 胡晓 ; 杨贤庆 ; 李来好 ; .鸢乌贼肌原纤维蛋白糖基化产物功能特性研究.南方水产科学.2018,(01),68-76. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN115381095A (en) | 2022-11-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Mao et al. | Development of soy protein isolate-carrageenan conjugates through Maillard reaction for the microencapsulation of Bifidobacterium longum | |
| CN110089753A (en) | A kind of preparation method for the nanoparticle improving curcumin bioavailability | |
| WO2021098492A1 (en) | Soybean polypeptide-based nanoparticle with high-load curcumin, and ph-driven preparation method therefor and use thereof | |
| US20010016220A1 (en) | Encapsulated multifunctional biologically active food component, process for its production and its use | |
| CN105639651A (en) | Preparation method and application of polyphenol-protein/polypeptide-carbohydrate covalent complexes | |
| CN101058649A (en) | Stable nano gel with core-shell structure, preparation method and application thereof | |
| CN104605228B (en) | A kind of EGCG chitosans/beta lactoglobulin composite nano-granule and preparation method thereof | |
| CN108850790A (en) | A kind of monascorubin microcapsules and preparation method thereof | |
| Chang et al. | Protein particle-based vehicles for encapsulation and delivery of nutrients: Fabrication, digestion, and release properties | |
| CN110393295A (en) | A kind of curcumin composite nanometer particle solution and preparation method thereof with anti-oxidation function | |
| CN110679953A (en) | Preparation method of nano liposome embedded with egg white source active peptide | |
| CN112273654A (en) | Method for preparing soybean protein enzymolysis aggregate embedded curcumin nanoparticles by pH driving method and application thereof | |
| CN114081096A (en) | Preparation method of milk protein loaded astaxanthin emulsion combined with polyphenol and glycosylation modification | |
| Fu et al. | Modification of soy protein isolate by Maillard reaction and its application in microencapsulation of Limosilactobacillus reuteri | |
| Yang et al. | Stability and bioavailability of protein matrix‐encapsulated astaxanthin ester microcapsules | |
| CN108096188B (en) | Load dewatering medicament and the oil-in-water composite Nano lotion of nutrients and preparation method thereof | |
| Ngwuluka et al. | Natural polymers in micro-and nanoencapsulation for therapeutic and diagnostic applications: part II-polysaccharides and proteins | |
| Jiao et al. | Improvement in entrapment efficiency and in vitro digestion stability of lutein by zein nanocarriers with pepsin hydrolysis | |
| CN115381095B (en) | Method for improving load and stability of fat-soluble active factor | |
| Yang et al. | Proteins from leguminous plants: from structure, property to the function in encapsulation/binding and delivery of bioactive compounds | |
| Tang | Nanostructures of soy proteins for encapsulation of food bioactive ingredients | |
| CN101455645A (en) | Preparation method of medicine microspheres using plant soy protein as carrier | |
| Fan et al. | Soy protein isolate (SPI)-hemin complex nanoparticles as a novel water-soluble iron-fortifier: Fabrication, formation mechanism and in vitro bioavailability | |
| JPWO2005025609A1 (en) | Lactoferrin material composition | |
| CN102010513A (en) | Stable polysaccharide modified gelatin nano particle and preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |