CN112841393A - Walnut peptide stable to zinc ions as well as preparation method and application thereof - Google Patents
Walnut peptide stable to zinc ions as well as preparation method and application thereof Download PDFInfo
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- CN112841393A CN112841393A CN202011642742.6A CN202011642742A CN112841393A CN 112841393 A CN112841393 A CN 112841393A CN 202011642742 A CN202011642742 A CN 202011642742A CN 112841393 A CN112841393 A CN 112841393A
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- walnut
- zinc
- enzymolysis
- temperature
- peptide
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- 235000009496 Juglans regia Nutrition 0.000 title claims abstract description 144
- 235000020234 walnut Nutrition 0.000 title claims abstract description 144
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 75
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 title claims abstract description 35
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 240000007049 Juglans regia Species 0.000 title 1
- 241000758789 Juglans Species 0.000 claims abstract description 143
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 73
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 73
- 239000000843 powder Substances 0.000 claims abstract description 49
- 230000001954 sterilising effect Effects 0.000 claims abstract description 30
- 238000001694 spray drying Methods 0.000 claims abstract description 27
- 239000011701 zinc Substances 0.000 claims abstract description 26
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims abstract description 25
- 239000007788 liquid Substances 0.000 claims abstract description 25
- 229910052725 zinc Inorganic materials 0.000 claims abstract description 25
- 238000000034 method Methods 0.000 claims abstract description 19
- 235000013361 beverage Nutrition 0.000 claims abstract description 5
- 235000013365 dairy product Nutrition 0.000 claims abstract description 4
- 239000000243 solution Substances 0.000 claims description 50
- 239000006228 supernatant Substances 0.000 claims description 30
- 239000012460 protein solution Substances 0.000 claims description 27
- 238000001471 micro-filtration Methods 0.000 claims description 26
- 108091005658 Basic proteases Proteins 0.000 claims description 25
- 230000000694 effects Effects 0.000 claims description 19
- 239000002244 precipitate Substances 0.000 claims description 19
- 238000001816 cooling Methods 0.000 claims description 18
- 239000012528 membrane Substances 0.000 claims description 17
- 108090000145 Bacillolysin Proteins 0.000 claims description 16
- 108010059892 Cellulase Proteins 0.000 claims description 16
- 108091005507 Neutral proteases Proteins 0.000 claims description 16
- 102000035092 Neutral proteases Human genes 0.000 claims description 16
- 229940106157 cellulase Drugs 0.000 claims description 16
- 108010038851 tannase Proteins 0.000 claims description 16
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 10
- 238000010438 heat treatment Methods 0.000 claims description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 9
- 238000006243 chemical reaction Methods 0.000 claims description 9
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 9
- 238000010668 complexation reaction Methods 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 108091005804 Peptidases Proteins 0.000 claims description 8
- 239000004365 Protease Substances 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 8
- 229910021641 deionized water Inorganic materials 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 8
- 230000000415 inactivating effect Effects 0.000 claims description 8
- 239000012263 liquid product Substances 0.000 claims description 8
- 238000010008 shearing Methods 0.000 claims description 8
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 claims description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 6
- ZOIORXHNWRGPMV-UHFFFAOYSA-N acetic acid;zinc Chemical compound [Zn].CC(O)=O.CC(O)=O ZOIORXHNWRGPMV-UHFFFAOYSA-N 0.000 claims description 6
- AXCZMVOFGPJBDE-UHFFFAOYSA-L calcium dihydroxide Chemical compound [OH-].[OH-].[Ca+2] AXCZMVOFGPJBDE-UHFFFAOYSA-L 0.000 claims description 6
- WGIWBXUNRXCYRA-UHFFFAOYSA-H trizinc;2-hydroxypropane-1,2,3-tricarboxylate Chemical compound [Zn+2].[Zn+2].[Zn+2].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O WGIWBXUNRXCYRA-UHFFFAOYSA-H 0.000 claims description 6
- 239000004246 zinc acetate Substances 0.000 claims description 6
- 235000013904 zinc acetate Nutrition 0.000 claims description 6
- 239000011746 zinc citrate Substances 0.000 claims description 6
- 235000006076 zinc citrate Nutrition 0.000 claims description 6
- 229940068475 zinc citrate Drugs 0.000 claims description 6
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims description 6
- 229960001763 zinc sulfate Drugs 0.000 claims description 6
- 229910000368 zinc sulfate Inorganic materials 0.000 claims description 6
- 239000002253 acid Substances 0.000 claims description 5
- 239000003513 alkali Substances 0.000 claims description 5
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 5
- 230000035484 reaction time Effects 0.000 claims description 5
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 4
- 239000002131 composite material Substances 0.000 claims description 4
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 4
- 239000011592 zinc chloride Substances 0.000 claims description 4
- 235000005074 zinc chloride Nutrition 0.000 claims description 4
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 3
- WHMDKBIGKVEYHS-IYEMJOQQSA-L Zinc gluconate Chemical compound [Zn+2].OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C([O-])=O WHMDKBIGKVEYHS-IYEMJOQQSA-L 0.000 claims description 3
- CANRESZKMUPMAE-UHFFFAOYSA-L Zinc lactate Chemical compound [Zn+2].CC(O)C([O-])=O.CC(O)C([O-])=O CANRESZKMUPMAE-UHFFFAOYSA-L 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 3
- 239000011670 zinc gluconate Substances 0.000 claims description 3
- 235000011478 zinc gluconate Nutrition 0.000 claims description 3
- 229960000306 zinc gluconate Drugs 0.000 claims description 3
- 239000011576 zinc lactate Substances 0.000 claims description 3
- 235000000193 zinc lactate Nutrition 0.000 claims description 3
- 229940050168 zinc lactate Drugs 0.000 claims description 3
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 claims description 2
- GRYLNZFGIOXLOG-UHFFFAOYSA-N Nitric acid Chemical compound O[N+]([O-])=O GRYLNZFGIOXLOG-UHFFFAOYSA-N 0.000 claims description 2
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 claims description 2
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 claims description 2
- 239000004310 lactic acid Substances 0.000 claims description 2
- 235000014655 lactic acid Nutrition 0.000 claims description 2
- 239000001630 malic acid Substances 0.000 claims description 2
- 235000011090 malic acid Nutrition 0.000 claims description 2
- 229910017604 nitric acid Inorganic materials 0.000 claims description 2
- 239000011975 tartaric acid Substances 0.000 claims description 2
- 235000002906 tartaric acid Nutrition 0.000 claims description 2
- 229960000314 zinc acetate Drugs 0.000 claims description 2
- 229960001939 zinc chloride Drugs 0.000 claims description 2
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims 1
- 235000013305 food Nutrition 0.000 abstract description 9
- 230000008569 process Effects 0.000 abstract description 8
- 238000004659 sterilization and disinfection Methods 0.000 abstract description 8
- 238000005119 centrifugation Methods 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 238000005374 membrane filtration Methods 0.000 abstract 1
- 239000000203 mixture Substances 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 59
- 239000000047 product Substances 0.000 description 23
- 238000011161 development Methods 0.000 description 12
- 238000001556 precipitation Methods 0.000 description 12
- 102000035195 Peptidases Human genes 0.000 description 7
- 239000001913 cellulose Substances 0.000 description 7
- 229920002678 cellulose Polymers 0.000 description 7
- 238000002845 discoloration Methods 0.000 description 7
- 150000008442 polyphenolic compounds Chemical class 0.000 description 7
- 235000013824 polyphenols Nutrition 0.000 description 7
- 239000007787 solid Substances 0.000 description 7
- 102000005158 Subtilisins Human genes 0.000 description 6
- 108010056079 Subtilisins Proteins 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- 230000000593 degrading effect Effects 0.000 description 6
- 238000005189 flocculation Methods 0.000 description 6
- 230000016615 flocculation Effects 0.000 description 6
- 229910052500 inorganic mineral Inorganic materials 0.000 description 6
- 235000010755 mineral Nutrition 0.000 description 6
- 239000011707 mineral Substances 0.000 description 6
- JBQYATWDVHIOAR-UHFFFAOYSA-N tellanylidenegermanium Chemical compound [Te]=[Ge] JBQYATWDVHIOAR-UHFFFAOYSA-N 0.000 description 6
- 102000004196 processed proteins & peptides Human genes 0.000 description 5
- 102000007079 Peptide Fragments Human genes 0.000 description 3
- 108010033276 Peptide Fragments Proteins 0.000 description 3
- 239000013522 chelant Substances 0.000 description 3
- 108010009355 microbial metalloproteinases Proteins 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 235000014571 nuts Nutrition 0.000 description 3
- CMHHMUWAYWTMGS-UHFFFAOYSA-N oxybuprocaine Chemical compound CCCCOC1=CC(C(=O)OCCN(CC)CC)=CC=C1N CMHHMUWAYWTMGS-UHFFFAOYSA-N 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 210000004556 brain Anatomy 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 206010059245 Angiopathy Diseases 0.000 description 1
- 108010059378 Endopeptidases Proteins 0.000 description 1
- 102000005593 Endopeptidases Human genes 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 108010068370 Glutens Proteins 0.000 description 1
- 108010073771 Soybean Proteins Proteins 0.000 description 1
- 235000019498 Walnut oil Nutrition 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 239000002585 base Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000019658 bitter taste Nutrition 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000004438 eyesight Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000005417 food ingredient Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 235000021312 gluten Nutrition 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 235000004280 healthy diet Nutrition 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 208000024348 heart neoplasm Diseases 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 235000021590 normal diet Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000001953 sensory effect Effects 0.000 description 1
- 235000019710 soybean protein Nutrition 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 230000009469 supplementation Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000008170 walnut oil Substances 0.000 description 1
Classifications
-
- 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
- A23J3/14—Vegetable proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G3/00—Sweetmeats; Confectionery; Marzipan; Coated or filled products
- A23G3/34—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof
- A23G3/36—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds
- A23G3/362—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing inorganic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G3/00—Sweetmeats; Confectionery; Marzipan; Coated or filled products
- A23G3/34—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof
- A23G3/36—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds
- A23G3/44—Sweetmeats, confectionery or marzipan; Processes for the preparation thereof characterised by the composition containing organic or inorganic compounds containing peptides or proteins
-
- 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
- A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/385—Concentrates of non-alcoholic beverages
- A23L2/39—Dry compositions
-
- 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
- A23L33/16—Inorganic salts, minerals or trace elements
-
- 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
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- 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)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Inorganic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Mycology (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention discloses a walnut peptide stable to zinc ions, a preparation method and application thereof. The method takes walnut protein powder as a raw material, decomposes the walnut protein into a short peptide mixture by a high-efficiency biological enzymolysis technology, further adds a zinc preparation to complex partial peptide segments, obtains target walnut peptide by centrifugation and membrane filtration methods, and finally prepares a target product by concentration, sterilization and spray drying processes. The walnut peptide can be widely applied to the product fields of formula foods with special medical purposes, oral liquid, dairy products, common beverages and the like.
Description
Technical Field
The invention belongs to the field of food, and particularly relates to a walnut peptide stable to zinc ions, and a preparation method and application thereof.
Background
"daily nuts" has been a deep-hearted healthy diet concept, i.e. proper supplementation of nut foods, outside of the normal diet, daily. The walnut is a very important kind of nut food, and the walnut is usually eaten to prevent and treat heart diseases, cancers and angiopathy, and simultaneously can improve eyesight and strengthen brain. China is a big walnut producing country, germplasm resources reach 380 varieties, most of provinces are cultivated, the main producing areas are concentrated in Yunnan, Sichuan, Xinjiang, Shanxi, Hebei and other provinces, the annual yield can reach 20 million tons at most, and the annual yield is increased year by year.
The walnut oil is an industry which is rapidly developed in recent years, and a large amount of walnut dregs and walnut protein powder are produced every year. The gluten and globulin in the walnut protein have higher hydrophobic amino acid proportion and poorer water solubility, and the functional characteristics of the protein such as emulsibility, gel type, water retention property and the like are poorer than those of the soybean protein isolate, so the application of the walnut protein powder in the food industry is greatly restricted. The modification improves the functional characteristics of the walnut protein, and the walnut protein is hydrolyzed into functional small molecular peptides, which are two deep processing ways for improving the value of the walnut protein.
The applicant of the invention provides a method for industrially producing high-purity walnut peptide (application number: 201711444692.9) by using low-temperature squeezed walnut meal as a raw material, wherein the protein content is more than 90%, the peptide content is more than 85%, the product quality is good, the bitterness is weak, the walnut peptide is easy to match with other food ingredients, the brain strengthening and intelligence improving effects and the immunity enhancing effects are remarkable, and the walnut peptide can be widely applied to various foods and health care products as a high-quality functional base material. In practical application, certain specific application scenes are found, such as oral liquid products, walnut peptides are used together with zinc citrate, the solution is mixed, and precipitates are formed at the bottom of the solution after the solution is kept still for a period of time, so that the sensory quality of the products is seriously influenced. Therefore, there is a need for further improvement of the production process of walnut peptide in order to obtain walnut peptide stable to zinc ions.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a preparation method of walnut peptide stable to zinc ions
The invention also aims to provide the walnut peptide stable to zinc ions, which is obtained by the preparation method.
The invention further aims to provide application of the walnut peptide stable to zinc ions, which can be conveniently applied to products such as formula food, oral liquid, dairy products and common beverages for special medical purposes.
The purpose of the invention is realized by the following technical scheme: a preparation method of walnut peptide stable to zinc ions comprises the following steps:
(1) preparing walnut protein powder into a protein solution with the concentration of 5-20% by mass by using deionized water, and dispersing the walnut protein powder at the shearing speed of 500-5000 r/min;
(2) heating the dispersed walnut protein solution to 90-100 ℃, keeping the temperature for 5-120 min to fully develop the molecular structure of the walnut protein, and then cooling to obtain a walnut protein solution;
(3) adding tannase which is 0.04 to 0.06 percent of the mass of the walnut protein powder and cellulase which is 0.04 to 0.06 percent of the mass of the walnut protein powder into the walnut protein solution for carrying out first-step enzymolysis, degrading polyphenol substances, cellulose and other components in the walnut protein powder, reducing discoloration and being beneficial to further enzymolysis;
(4) adjusting the pH value of the solution to 7.0-9.0 by using alkali liquor, and adding alkaline protease which is 0.5-2.0% of the mass of the walnut protein powder for secondary enzymolysis; after enzymolysis is finished, keeping the temperature at 90-100 ℃ for 5-60 min, passivating the activity of alkaline protease, and then cooling the temperature to the optimal temperature for next enzymolysis;
(5) adjusting the pH value of the system finally obtained in the step (4) to 6.0-7.0 by using acid liquor, and adding neutral protease which is 0.5-2.0% of the mass of the walnut protein powder for third enzymolysis; after enzymolysis is finished, keeping the temperature of 90-100 ℃ for 5-60 min, and inactivating the activity of protease;
(6) carrying out centrifugal separation on the enzymolysis liquid obtained in the step (5), and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain walnut protein peptide powder;
(7) adding a zinc preparation into the supernatant to perform a complex reaction;
(8) further centrifugally separating the feed liquid, and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain zinc-rich walnut protein peptide;
(9) and (4) filtering the supernatant obtained in the step (8) by using a microfiltration membrane, concentrating, sterilizing and spray drying to obtain the walnut peptide stable to zinc ions.
The heating temperature in the step (2) is preferably 90-95 ℃.
The cooling temperature in the step (2) is preferably 60 ℃ or lower; more preferably 30 to 60 ℃.
The dosage of the tannase in the step (3) is preferably 0.05% of the mass of the walnut protein powder.
The dosage of the cellulase in the step (3) is preferably 0.05% of the mass of the walnut protein powder.
The enzymolysis condition of the first step in the step (3) is preferably enzymolysis for 1-3 h at 45-55 ℃; more preferably at 50 ℃ for 2 h.
The alkali liquor in the step (4) is preferably at least one of sodium hydroxide solution, calcium hydroxide solution and potassium hydroxide solution.
The concentration of the alkali liquor in the step (4) is preferably 1-10 mol.L-1。
The alkaline protease in the step (4) is alkaline silk amino acid endopeptidase.
The enzymolysis conditions in the step (4) are as follows: carrying out enzymolysis for 2-20 h at the optimum temperature of the alkaline protease.
The optimum temperature is preferably 60 ℃.
The acid solution in step (5) is preferably at least one of a hydrochloric acid solution, a sulfuric acid solution, a nitric acid solution, a phosphoric acid solution, a citric acid solution, a malic acid solution, a lactic acid solution and a tartaric acid solution.
The concentration of the acid solution is preferably 1-16 mol.L-1。
The enzymolysis conditions in the step (5) are as follows: carrying out enzymolysis for 2-20 h at the optimal temperature of neutral protease.
The optimum temperature is preferably 50 ℃.
The zinc preparation in the step (7) is at least one of zinc gluconate, zinc sulfate, zinc lactate, zinc citrate, zinc chloride and zinc acetate.
The conditions of the complexation reaction described in step (7) are preferably as follows: the addition amount of zinc ions is 0.1-1% of the mass of the walnut protein powder, the reaction temperature is 20-80 ℃, and the reaction time is 10-240 min.
The addition amount of the zinc ions is preferably 0.2-0.4% of the mass of the walnut protein powder.
The conditions for microfiltration described in step (9) are preferably as follows: adopting a microfiltration membrane with the diameter of 0.05-5 mu m, the pressure of 0.05-0.5 MPa and the temperature of 20-60 ℃; more preferably as follows: using a microfiltration membrane with the diameter of 0.05-5 mu m, the pressure of 0.05-0.5 MPa and the temperature of 20-40 ℃.
The concentration degree in the step (9) is preferably concentrated to a concentration of 10-50 Bx;
the conditions for the spray drying described in step (9) are preferably as follows: the air inlet temperature is 140-200 ℃, and the air outlet temperature is 65-95 ℃.
A walnut peptide stable to zinc ions is obtained by the preparation method.
The application of the walnut peptide stable to zinc ions in preparing a zinc-containing composite walnut peptide liquid product.
The form of the liquid product includes but is not limited to oral liquid, dairy products and common beverages.
A zinc-containing compound walnut peptide liquid product contains the above walnut peptide stable to zinc ions and zinc ions.
The mass ratio of the zinc ions to the walnut peptide stabilized by the zinc ions is 1: 100 or less; preferably 1: below 200 a.
Compared with the prior art, the invention has the following advantages and effects:
aiming at some special application scenes of the walnut peptide in the food industry, the invention provides a better solution scheme, namely functional walnut peptide is added, zinc nutrient elements are reinforced, and the product has higher requirements on the appearance stability. The method adds a certain amount of zinc preparation into the prepared walnut peptide feed liquid to perform partial complexation reaction. Some peptide fragments which are easily complexed with zinc ions will preferentially complex to form a precipitate, and this portion of the complex is removed by centrifugation and microfiltration. The precipitate is zinc-rich walnut peptide, and the product obtained by concentration, sterilization and spray drying can be used in products with low clarity requirements, such as solid beverages, tablet candy and the like. In the filtered clear liquid, peptide fragments highly sensitive to zinc ions are few, and the unstable phenomena of turbidity, flocculation, precipitation and the like caused by complex reaction can not occur any more when a zinc preparation with a concentration lower than the last time is added. The clear liquid is further concentrated, sterilized and spray-dried to obtain the product which is the target product of the invention.
The present invention differs from the mineral chelate processing of some protein peptides. Although both involve complexation, they are essentially different in that: (1) the target products of the two are different, and the target product of the mineral chelate of the protein peptide is a precipitate formed by a complexation reaction; the target product of the invention is the clear solution from which the complex product is removed; (2) the two products are different in essence, and the mineral chelate of the protein peptide needs to be added with excessive mineral to ensure that the peptide segment and the mineral are subjected to full complexation reaction. The invention only carries out partial complexation, and the mineral zinc and the walnut peptide are not more than 100: 1, only a few zinc-sensitive peptide fragments are subjected to complexation reaction. The by-product complex precipitate is zinc-rich walnut peptide, the finished product obtained from the clear liquid is essentially walnut peptide, and if high-concentration zinc ions are added again, the complex precipitation reaction still occurs.
Detailed Description
The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.
Example 1
(1) Preparing 50kg of walnut protein powder into 1000kg of protein solution with the concentration of 5% by mass by using deionized water, and dispersing the walnut protein powder at the shearing speed of 500 revolutions per minute;
(2) heating the dispersed walnut protein solution to 90 ℃, keeping the temperature for 120min to fully develop the molecular structure of the walnut protein, and then cooling the walnut protein solution to 60 ℃;
(3) adding 0.025kg of tannase (food-grade tannase FDG-2254 from Xiusheng (Beijing) Biotechnology development Limited) and 0.025kg of cellulase (food-grade solid cellulase FDG-2226 from Xiusheng (Beijing) Biotechnology development Limited) into the walnut protein solution in sequence, and performing enzymolysis for 2h at 50 ℃ to degrade polyphenol substances, cellulose and other components in the walnut protein powder, reduce discoloration and facilitate further enzymolysis;
(4) with a concentration of 1 mol. L-1The pH value of the enzymolysis solution finally obtained in the step (3) is adjusted to 7.0-9.0 by the sodium hydroxide solution, 0.25kg of alkaline protease (Alcalase alkaline protease, Novexin (China) investment Co., Ltd.) is added, the enzymolysis time is 20h, and the enzymolysis temperature is 60 ℃. After enzymolysis is finished, keeping the temperature at 90 ℃ for 60min, inactivating the activity of alkaline protease, and then cooling the temperature to 50 ℃;
(5) with a concentration of 3 mol. L-1The citric acid solution is used for adjusting the pH value of the enzymolysis solution finally obtained in the step (4) to 6.0-7.0, 0.25kg of neutral protease (Neutrase neutral protease, Novestin (China) investment Co., Ltd.) is added, the enzymolysis temperature is 50 ℃, and the enzymolysis time is 20 hours. After enzymolysis is finished, keeping the temperature at 90 ℃ for 60min to passivate the activity of protease;
(6) centrifuging the enzymolysis liquid obtained in the step (5), and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain walnut protein peptide powder;
(7) adding 0.85kg of zinc gluconate into the supernatant at 20 deg.C for 240 min;
(8) further centrifugally separating the feed liquid, and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain 1.3kg of zinc-rich walnut protein peptide;
(9) filtering the supernatant obtained in the step (8) by a microfiltration membrane, wherein the microfiltration process adopts a microfiltration membrane with the diameter of 0.05 mu m, the pressure is 0.05MPa, and the temperature is 20 ℃; then concentrating to 10Bx, sterilizing, and spray drying, wherein the air inlet temperature is 140 ℃, the air outlet temperature is 65 ℃, and the target product is 22.5 kg.
Adding 1g of walnut peptide (target product) prepared in example 1 into 100g of aqueous solution system with the pH value of 6.0-7.0, and adding 5mg of zinc (Zn)2+Calculated, zinc chloride) is sterilized at high temperature of 105 ℃/60min, 121 ℃/30min, 137 ℃/8s and other different strengths, and then is kept in a stable state (namely the turbidity difference is less than 5%) within an observation period of 12 months, and unstable phenomena such as turbidity, flocculation, precipitation and the like do not occur.
Flocculation and precipitation can be directly observed by the naked eye and turbidity can be measured by a spectrophotometer (in Hongypeng, Wukegang, Chaihua. the effect of physicochemical conditions on polypeptide aggregation. food and fermentation industries 2008,34(2): 40-43). At room temperature, a cuvette with a glass thickness of 0.5mm is filled with a solution to be measured, and the absorbance is measured at 660 nm. And comparing the absorbance of the sample to be detected with that of the zinc ion blank control solution, wherein the difference is less than 5%, and judging that the two solutions have no obvious difference. Otherwise, the solution is judged to be mixed.
Example 2
(1) Preparing 200kg of walnut protein powder into 1000kg of protein solution with the concentration of 20% by mass by using deionized water, and dispersing the walnut protein powder at the shearing speed of 500 revolutions per minute;
(2) heating the dispersed walnut protein solution to 95 ℃, keeping for 5min to fully develop the molecular structure of the walnut protein, and then cooling to 60 ℃;
(3) adding 0.1kg of tannase (Xiasang (Beijing) biotechnological development limited, food-grade tannase FDG-2254) and 0.1kg of cellulase (Xiasang (Beijing) biotechnological development limited, food-grade solid cellulase FDG-2226) into the walnut protein solution in sequence, performing enzymolysis for 2 hours at 50 ℃, degrading components such as polyphenols, cellulose and the like in the walnut protein powder, reducing discoloration and facilitating further enzymolysis;
(4) with 10 mol. L-1The pH value of the enzymolysis solution finally obtained in the step (3) is adjusted to 7.0-9.0 by the potassium hydroxide solution, 4kg of alkaline protease (Alcalase alkaline protease, Novexin (China) investment Co., Ltd.) is added, the enzymolysis time is 2h, and the enzymolysis temperature is 60 ℃. After enzymolysis is finished, keeping the temperature at 90 ℃ for 60min, inactivating the activity of alkaline protease, and then cooling the temperature to 50 ℃;
(5) with 16 mol. L-1And (3) regulating the pH value of the enzymolysis solution finally obtained in the step (4) to 6.0-7.0 by using a hydrochloric acid solution, adding 4kg of neutral protease (Neutrase neutral protease, Novestin (China) investment Co., Ltd.), wherein the enzymolysis temperature is 50 ℃ and the enzymolysis time is 2 hours. After enzymolysis is finished, keeping the temperature at 90 ℃ for 60min to passivate the activity of protease;
(6) centrifuging the enzymolysis liquid obtained in the step (5), and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain walnut protein peptide powder;
(7) adding 1.98kg of zinc lactate into the supernatant, wherein the reaction temperature is 80 ℃, and the reaction time is 10 min;
(8) further centrifugally separating the feed liquid, and taking supernatant; 2.1kg of walnut protein peptide rich in zinc is obtained by the precipitation through the processes of concentration, sterilization, spray drying and the like;
(9) filtering the supernatant obtained in the step (8) by a microfiltration membrane, wherein the microfiltration process adopts a microfiltration membrane with the diameter of 5 microns, the pressure is 0.5MPa, and the temperature is 20 ℃; then concentrating to 50Bx, sterilizing, and spray drying, wherein the air inlet temperature is 200 ℃, the air outlet temperature is 90 ℃, and the target product is 41.2 kg.
Adding 1g of walnut peptide (target product) prepared in example 2 into 100g of aqueous solution system with the pH value of 6.0-7.0, and adding 5mg of zinc (Zn)2+Calculated, zinc chloride) is subjected to high-temperature sterilization with different strengths of 105 ℃/60min, 121 ℃/30min, 137 ℃/8s and the like, and then the stable state is kept within the observation period of 12 months, and unstable phenomena such as turbidity, flocculation, precipitation and the like do not occur.
Example 3
(1) Preparing 100kg of walnut protein powder into 1000kg of protein solution with the concentration of 10% by mass by using deionized water, and dispersing the walnut protein powder at the shearing speed of 5000 r/min;
(2) heating the dispersed walnut protein solution to 95 ℃, keeping for 30min to fully develop the molecular structure of the walnut protein, and then cooling to 60 ℃;
(3) adding 0.1kg of tannase (Xiasang (Beijing) biotechnological development limited, food-grade tannase FDG-2254) and 0.1kg of cellulase (Xiasang (Beijing) biotechnological development limited, food-grade solid cellulase FDG-2226) into the walnut protein solution in sequence, performing enzymolysis for 2 hours at 50 ℃, degrading components such as polyphenols, cellulose and the like in the walnut protein powder, reducing discoloration and facilitating further enzymolysis;
(4) with 5 mol. L-1The pH value of the enzymolysis solution finally obtained in the step (3) is adjusted to 7.0-9.0 by the calcium hydroxide solution, 2kg of alkaline protease (Alcalase alkaline protease, Novexin (China) investment Co., Ltd.) is added, the enzymolysis time is 12h, and the enzymolysis temperature is 60 ℃. After enzymolysis is finished, keeping the temperature at 90 ℃ for 60min, inactivating the activity of alkaline protease, and then cooling the temperature to 50 ℃;
(5) with 5 mol. L-1The pH value of the enzymolysis solution finally obtained in the step (4) is adjusted to 6.0-7.0, 2kg of neutral protease (Neaurose neutral protease, Novesin (China) investment Co., Ltd.) is added, the enzymolysis temperature is 50 ℃, and the enzymolysis time is 12 hours. After enzymolysis is finished, keeping the temperature at 90 ℃ for 60min to passivate the activity of protease;
(6) centrifuging the enzymolysis liquid obtained in the step (5), and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain walnut protein peptide powder;
(7) adding 0.94kg of zinc sulfate into the supernatant, wherein the reaction temperature is 60 ℃, and the reaction time is 60 min;
(8) further centrifugally separating the feed liquid, and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain 1.8kg of zinc-rich walnut protein peptide;
(9) filtering the supernatant obtained in the step (8) by a microfiltration membrane, wherein the microfiltration process adopts a microfiltration membrane with the diameter of 0.5 mu m, the pressure is 0.2MPa, and the temperature is 40 ℃; then concentrating to 30Bx, sterilizing, and spray drying, wherein the air inlet temperature is 185 ℃, the air outlet temperature is 95 ℃, and the target product is 45.2 kg.
Adding 1g of the walnut peptide prepared in the embodiment 3 into 100g of an aqueous solution system with the pH value of 6.0-7.0, and further adding 5mg of zinc (Zn)2+Calculated, zinc acetate) is subjected to high-temperature sterilization with different strengths of 105 ℃/60min, 121 ℃/30min, 137 ℃/8s and the like, and then the stable state is kept within the observation period of 12 months, and unstable phenomena such as turbidity, flocculation, precipitation and the like do not occur.
Example 4
Example 4 is substantially identical to the process steps of example 3, except that zinc citrate is used to test the stability of the walnut peptide.
(1) Preparing 100kg of walnut protein powder into 1000kg of protein solution with the concentration of 10% by mass by using deionized water, and dispersing the walnut protein powder at the shearing speed of 5000 r/min;
(2) heating the dispersed walnut protein solution to 95 ℃, keeping for 30min to fully develop the molecular structure of the walnut protein, and then cooling to 60 ℃;
(3) adding 0.1kg of tannase (Xiasang (Beijing) biotechnological development limited, food-grade tannase FDG-2254) and 0.1kg of cellulase (Xiasang (Beijing) biotechnological development limited, food-grade solid cellulase FDG-2226) into the walnut protein solution in sequence, performing enzymolysis for 2 hours at 50 ℃, degrading components such as polyphenols, cellulose and the like in the walnut protein powder, reducing discoloration and facilitating further enzymolysis;
(4) with 5 mol. L-1The pH value of the enzymolysis solution finally obtained in the step (3) is adjusted to 7.0-9.0 by the calcium hydroxide solution, 2kg of alkaline protease (Alcalase alkaline protease, Novexin (China) investment Co., Ltd.) is added, the enzymolysis time is 12h, and the enzymolysis temperature is 60 ℃. After enzymolysis is finished, keeping the temperature at 90 ℃ for 60min, inactivating the activity of alkaline protease, and then cooling the temperature to 50 ℃;
(5) with 5 mol. L-1And (3) adjusting the pH value of the enzymolysis solution finally obtained in the step (4) to 6.0-7.0 by using a phosphoric acid solution, adding 2kg of neutral protease (Neutrase neutral protease, Novestin (China) investment Co., Ltd.), wherein the enzymolysis temperature is 50 ℃ and the enzymolysis time is 12 hours. After the enzymolysis is finishedKeeping the temperature at 90 ℃ for 60min to inactivate the activity of the protease.
(6) Centrifuging the enzymolysis liquid obtained in the step (5), and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain walnut protein peptide powder;
(7) adding 0.94kg zinc sulfate into the supernatant at 60 deg.C for 60 min;
(8) further centrifugally separating the feed liquid, and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain 1.9kg of zinc-rich walnut protein peptide;
(9) filtering the supernatant obtained in the step (8) by a microfiltration membrane, wherein the microfiltration process adopts a microfiltration membrane with the diameter of 0.5 mu m, the pressure is 0.2MPa, and the temperature is 40 ℃; then concentrating, sterilizing and spray drying, wherein the air inlet temperature is 185 ℃, the air outlet temperature is 95 ℃, and the target product is 45.6 kg.
Adding 1g of the walnut peptide prepared in the embodiment 4 into 100g of an aqueous solution system with the pH value of 6.0-7.0, and further adding 5mg of zinc (Zn)2+And calculating zinc citrate), and after high-temperature sterilization with different strengths of 105 ℃/60min, 121 ℃/30min, 137 ℃/8s and the like, the zinc citrate is kept in a stable state within an observation period of 12 months, and unstable phenomena such as turbidity, flocculation, precipitation and the like do not occur.
Comparative example 1
Comparative example 1 compared with example 3, the process steps were completely identical except that step (7) was performed without adding zinc sulfate to perform a complexation reaction, and the centrifugal separation process of step (8) was not performed.
(1) Preparing 100kg of walnut protein powder into 1000kg of protein solution with the concentration of 10% by mass by using deionized water, and dispersing the walnut protein powder at the shearing speed of 5000 r/min;
(2) heating the dispersed walnut protein solution to 95 ℃, keeping for 30min to fully develop the molecular structure of the walnut protein, and then cooling to 60 ℃;
(3) adding 0.1kg of tannase (Xiasang (Beijing) biotechnological development limited, food-grade tannase FDG-2254) and 0.1kg of cellulase (Xiasang (Beijing) biotechnological development limited, food-grade solid cellulase FDG-2226) into the walnut protein solution in sequence, performing enzymolysis for 2 hours at 50 ℃, degrading components such as polyphenols, cellulose and the like in the walnut protein powder, reducing discoloration and facilitating further enzymolysis;
(4) with 5 mol. L-1The pH value of the enzymolysis solution finally obtained in the step (3) is adjusted to 7.0-9.0 by the calcium hydroxide solution, 2kg of alkaline protease (Alcalase alkaline protease, Novexin (China) investment Co., Ltd.) is added, the enzymolysis time is 12h, and the enzymolysis temperature is 60 ℃. After enzymolysis is finished, keeping the temperature at 90 ℃ for 60min, inactivating the activity of alkaline protease, and then cooling the temperature to 50 ℃;
(5) with 5 mol. L-1And (3) adjusting the pH value of the enzymolysis solution finally obtained in the step (4) to 6.0-7.0 by using a phosphoric acid solution, adding 2kg of neutral protease (Neautrase neutral protease, Novesin (China) investment Co., Ltd.), and carrying out enzymolysis at the temperature of 50 ℃ for 12 h. After the enzymolysis is finished, the temperature is kept for 60min at 90 ℃, and the activity of the protease is inactivated.
(6) Centrifuging the enzymolysis liquid obtained in the step (5), and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain walnut protein peptide powder;
(7) filtering the supernatant obtained in the step (6) by a microfiltration membrane, wherein the microfiltration process adopts a microfiltration membrane with the diameter of 0.5 mu m, the pressure is 0.2MPa, and the temperature is 40 ℃; then concentrating, sterilizing and spray drying, wherein the air inlet temperature is 185 ℃, the air outlet temperature is 95 ℃, and the target product is 46.9 kg.
Adding 1g of walnut peptide prepared in the comparative example 1 into 100g of aqueous solution system with the pH value of 6.0-7.0, and then adding 5mg of zinc (Zn)2+Calculated, zinc acetate), the feed liquid is obviously mixed and has precipitation after standing. After high-temperature sterilization with different strengths of 105 ℃/60min, 121 ℃/30min, 137 ℃/8s and the like, the bottom of the sample has obvious precipitation layers within the observation period of 12 months.
Comparative example 2
Comparative example 2 is substantially identical to the process steps of example 3, except that: 15mg of zinc (in Zn) are used2+Calculated, zinc acetate) was used to determine the stability of the walnut peptides.
(1) Preparing 100kg of walnut protein powder into 1000kg of protein solution with the concentration of 10% by mass by using deionized water, and dispersing the walnut protein powder at the shearing speed of 5000 r/min;
(2) heating the dispersed walnut protein solution to 95 ℃, keeping for 30min to fully develop the molecular structure of the walnut protein, and then cooling to 60 ℃;
(3) adding 0.1kg of tannase (Xiasang (Beijing) biotechnological development limited, food-grade tannase FDG-2254) and 0.1kg of cellulase (Xiasang (Beijing) biotechnological development limited, food-grade solid cellulase FDG-2226) into the walnut protein solution in sequence, performing enzymolysis for 2 hours at 50 ℃, degrading components such as polyphenols, cellulose and the like in the walnut protein powder, reducing discoloration and facilitating further enzymolysis;
(4) with 5 mol. L-1The pH value of the enzymolysis solution finally obtained in the step (3) is adjusted to 7.0-9.0 by the calcium hydroxide solution, 2kg of alkaline protease (Alcalase alkaline protease, Novexin (China) investment Co., Ltd.) is added, the enzymolysis time is 12h, and the enzymolysis temperature is 60 ℃. After enzymolysis is finished, keeping the temperature at 90 ℃ for 60min, inactivating the activity of alkaline protease, and then cooling the temperature to 50 ℃;
(5) with 5 mol. L-1And (3) adjusting the pH value of the enzymolysis solution finally obtained in the step (4) to 6.0-7.0 by using a phosphoric acid solution, adding 2kg of neutral protease (Neautrase neutral protease, Novesin (China) investment Co., Ltd.), and carrying out enzymolysis at the temperature of 50 ℃ for 12 h. After the enzymolysis is finished, the temperature is kept for 60min at 90 ℃, and the activity of the protease is inactivated.
(6) Centrifuging the enzymolysis liquid obtained in the step (5), and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain walnut protein peptide powder;
(7) adding 0.18kg of zinc sulfate into the supernatant, wherein the reaction temperature is 60 ℃, and the reaction time is 60 min;
(8) further centrifugally separating the feed liquid, and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain 1.6kg of zinc-rich walnut protein peptide;
(9) filtering the supernatant obtained in the step (8) by a microfiltration membrane, wherein the microfiltration process adopts a microfiltration membrane with the diameter of 0.5 mu m, the pressure is 0.2MPa, and the temperature is 40 ℃; then concentrating, sterilizing and spray drying, wherein the air inlet temperature is 185 ℃, the air outlet temperature is 95 ℃, and the target product is 44.1 kg.
Adding 1g of walnut peptide prepared in comparative example 2 into 100g of aqueous solution system with the pH value of 6.0-7.0, and adding 15mg of zinc (Zn)2+Calculated, zinc acetate), the feed liquid is obviously mixed and has precipitation after standing. After high-temperature sterilization with different strengths of 105 ℃/60min, 121 ℃/30min, 137 ℃/8s and the like, the bottom of the sample has obvious precipitation layers within the observation period of 12 months.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (10)
1. A preparation method of walnut peptide stable to zinc ions is characterized by comprising the following steps:
(1) preparing walnut protein powder into a protein solution with the concentration of 5-20% by mass by using deionized water, and dispersing the walnut protein powder at the shearing speed of 500-5000 r/min;
(2) heating the dispersed walnut protein solution to 90-100 ℃, keeping the temperature for 5-120 min to fully develop the molecular structure of the walnut protein, and then cooling to obtain a walnut protein solution;
(3) adding tannase which is 0.04 to 0.06 percent of the mass of the walnut protein powder and cellulase which is 0.04 to 0.06 percent of the mass of the walnut protein powder into the walnut protein solution for carrying out first-step enzymolysis;
(4) adjusting the pH value of the solution to 7.0-9.0 by using alkali liquor, and adding alkaline protease which is 0.5-2.0% of the mass of the walnut protein powder for secondary enzymolysis; after enzymolysis is finished, keeping the temperature at 90-100 ℃ for 5-60 min, passivating the activity of alkaline protease, and then cooling the temperature to the optimal temperature for next enzymolysis;
(5) adjusting the pH value of the system finally obtained in the step (4) to 6.0-7.0 by using acid liquor, and adding neutral protease which is 0.5-2.0% of the mass of the walnut protein powder for third enzymolysis; after enzymolysis is finished, keeping the temperature of 90-100 ℃ for 5-60 min, and inactivating the activity of protease;
(6) carrying out centrifugal separation on the enzymolysis liquid obtained in the step (5), and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain walnut protein peptide powder;
(7) adding a zinc preparation into the supernatant to perform a complex reaction;
(8) further centrifugally separating the feed liquid, and taking supernatant; concentrating, sterilizing and spray drying the precipitate to obtain zinc-rich walnut protein peptide;
(9) and (4) filtering the supernatant obtained in the step (8) by using a microfiltration membrane, concentrating, sterilizing and spray drying to obtain the walnut peptide stable to zinc ions.
2. The method for preparing walnut peptide stabilized against zinc ions according to claim 1, wherein:
the zinc preparation in the step (7) is at least one of zinc gluconate, zinc sulfate, zinc lactate, zinc citrate, zinc chloride and zinc acetate;
the conditions of the complexation reaction described in step (7) are as follows: the addition amount of zinc ions is 0.1-1% of the mass of the walnut protein powder, the reaction temperature is 20-80 ℃, and the reaction time is 10-240 min;
the microfiltration conditions described in step (9) are as follows: a microfiltration membrane with the diameter of 0.05-5 mu m is adopted, the pressure is 0.05-0.5 MPa, and the temperature is 20-60 ℃.
3. The method for preparing walnut peptide stabilized against zinc ions according to claim 1, wherein:
the using amount of the tannase in the step (3) is equivalent to 0.05% of the mass of the walnut protein powder;
the dosage of the cellulase in the step (3) is equivalent to 0.05 percent of the mass of the walnut protein powder;
the enzymolysis condition of the first step in the step (3) is enzymolysis for 1-3 h at 45-55 ℃;
the enzymolysis conditions in the step (4) are as follows: carrying out enzymolysis for 2-20 h at the optimum temperature of alkaline protease;
the enzymolysis conditions in the step (5) are as follows: carrying out enzymolysis for 2-20 h at the optimal temperature of neutral protease.
4. The method for preparing walnut peptide stabilized against zinc ions according to claim 1, wherein:
the alkali liquor in the step (4) is at least one of a sodium hydroxide solution, a calcium hydroxide solution and a potassium hydroxide solution;
the acid solution in the step (5) is at least one of hydrochloric acid solution, sulfuric acid solution, nitric acid solution, phosphoric acid solution, citric acid solution, malic acid solution, lactic acid solution and tartaric acid solution.
5. The method for preparing walnut peptide stabilized against zinc ions according to claim 1, wherein:
the heating temperature in the step (2) is 90-95 ℃;
the cooling temperature in the step (2) is below 60 ℃;
the concentration degree in the step (9) is to be concentrated to a concentration of 10-50 Bx;
the spray drying conditions described in step (9) are as follows: the air inlet temperature is 140-200 ℃, and the air outlet temperature is 65-95 ℃.
6. A walnut peptide stable to zinc ions, which is characterized in that: the preparation method of any one of claims 1 to 5.
7. Use of a walnut peptide stabilized with zinc ions according to claim 6 for the preparation of a zinc-containing composite walnut peptide liquid product.
8. Use according to claim 7, characterized in that: the liquid product is in the form of oral liquid, dairy products or common beverages.
9. A zinc-containing composite walnut peptide liquid product is characterized in that: comprises the walnut peptide stabilized against zinc ions according to claim 6 and zinc ions.
10. The zinc-containing composite walnut peptide liquid product of claim 9, wherein: the mass ratio of the zinc ions to the walnut peptide stabilized by the zinc ions is 1: 100 or less.
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| CN101731629A (en) * | 2009-12-29 | 2010-06-16 | 中国农业大学 | Preparation method of peptide-calcium composite |
| CN108085356A (en) * | 2017-12-27 | 2018-05-29 | 广州合诚实业有限公司 | Using cold pressing walnut dregs as the method for primary industry metaplasia production of high purity walnut peptide |
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| CN101731629A (en) * | 2009-12-29 | 2010-06-16 | 中国农业大学 | Preparation method of peptide-calcium composite |
| CN108085356A (en) * | 2017-12-27 | 2018-05-29 | 广州合诚实业有限公司 | Using cold pressing walnut dregs as the method for primary industry metaplasia production of high purity walnut peptide |
| CN108295240A (en) * | 2018-04-08 | 2018-07-20 | 黑龙江军门现代农业发展有限公司 | A kind of ginseng oral liquid of invigorating qi for tranquilization and preparation method thereof |
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