CN112136874B - Modified protocatechuic acid liposome for storage and preservation of shrimps and crabs as well as preparation method and application of modified protocatechuic acid liposome - Google Patents
Modified protocatechuic acid liposome for storage and preservation of shrimps and crabs as well as preparation method and application of modified protocatechuic acid liposome Download PDFInfo
- Publication number
- CN112136874B CN112136874B CN202010903406.6A CN202010903406A CN112136874B CN 112136874 B CN112136874 B CN 112136874B CN 202010903406 A CN202010903406 A CN 202010903406A CN 112136874 B CN112136874 B CN 112136874B
- Authority
- CN
- China
- Prior art keywords
- bacterial cellulose
- protocatechuic acid
- modified
- liposome
- shrimps
- 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
- YQUVCSBJEUQKSH-UHFFFAOYSA-N 3,4-dihydroxybenzoic acid Chemical class OC(=O)C1=CC=C(O)C(O)=C1 YQUVCSBJEUQKSH-UHFFFAOYSA-N 0.000 title claims abstract description 160
- 239000002502 liposome Substances 0.000 title claims abstract description 103
- 241000238557 Decapoda Species 0.000 title claims abstract description 79
- 238000003860 storage Methods 0.000 title claims abstract description 28
- 238000004321 preservation Methods 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 title claims abstract description 10
- 229920002749 Bacterial cellulose Polymers 0.000 claims abstract description 137
- 239000005016 bacterial cellulose Substances 0.000 claims abstract description 137
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 claims abstract description 56
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 56
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 claims abstract description 32
- 229940083466 soybean lecithin Drugs 0.000 claims abstract description 32
- PYMYPHUHKUWMLA-UHFFFAOYSA-N 2,3,4,5-tetrahydroxypentanal Chemical compound OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims abstract description 31
- 229940040387 citrus pectin Drugs 0.000 claims abstract description 31
- 239000009194 citrus pectin Substances 0.000 claims abstract description 31
- 229930003427 Vitamin E Natural products 0.000 claims abstract description 28
- 235000012000 cholesterol Nutrition 0.000 claims abstract description 28
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 claims abstract description 28
- 235000019165 vitamin E Nutrition 0.000 claims abstract description 28
- 229940046009 vitamin E Drugs 0.000 claims abstract description 28
- 239000011709 vitamin E Substances 0.000 claims abstract description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000002131 composite material Substances 0.000 claims abstract description 27
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 63
- 239000001963 growth medium Substances 0.000 claims description 50
- 239000012528 membrane Substances 0.000 claims description 41
- 238000000855 fermentation Methods 0.000 claims description 37
- 230000004151 fermentation Effects 0.000 claims description 37
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 36
- 239000000243 solution Substances 0.000 claims description 34
- 230000001580 bacterial effect Effects 0.000 claims description 29
- 238000005406 washing Methods 0.000 claims description 28
- 238000002791 soaking Methods 0.000 claims description 26
- 239000007788 liquid Substances 0.000 claims description 24
- FYSNRJHAOHDILO-UHFFFAOYSA-N thionyl chloride Chemical compound ClS(Cl)=O FYSNRJHAOHDILO-UHFFFAOYSA-N 0.000 claims description 24
- 150000004676 glycans Chemical class 0.000 claims description 22
- 239000008055 phosphate buffer solution Substances 0.000 claims description 22
- 229920001282 polysaccharide Polymers 0.000 claims description 22
- 239000005017 polysaccharide Substances 0.000 claims description 22
- 238000011218 seed culture Methods 0.000 claims description 22
- 239000001888 Peptone Substances 0.000 claims description 21
- 108010080698 Peptones Proteins 0.000 claims description 21
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 21
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 21
- 238000012258 culturing Methods 0.000 claims description 21
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 21
- 235000019319 peptone Nutrition 0.000 claims description 21
- 239000000843 powder Substances 0.000 claims description 21
- 230000008014 freezing Effects 0.000 claims description 20
- 238000007710 freezing Methods 0.000 claims description 20
- 150000002632 lipids Chemical class 0.000 claims description 20
- 239000000725 suspension Substances 0.000 claims description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 19
- 239000002609 medium Substances 0.000 claims description 17
- 238000003756 stirring Methods 0.000 claims description 17
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 16
- 229920000053 polysorbate 80 Polymers 0.000 claims description 16
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 15
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 14
- 229930006000 Sucrose Natural products 0.000 claims description 14
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 14
- 239000005720 sucrose Substances 0.000 claims description 14
- 238000001914 filtration Methods 0.000 claims description 11
- 238000009210 therapy by ultrasound Methods 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 9
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical compound NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 claims description 8
- 238000002390 rotary evaporation Methods 0.000 claims description 8
- 244000235858 Acetobacter xylinum Species 0.000 claims description 7
- 235000002837 Acetobacter xylinum Nutrition 0.000 claims description 7
- 229920001817 Agar Polymers 0.000 claims description 7
- 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 claims description 7
- 230000004913 activation Effects 0.000 claims description 7
- 239000008272 agar Substances 0.000 claims description 7
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- 238000011081 inoculation Methods 0.000 claims description 7
- 230000007935 neutral effect Effects 0.000 claims description 7
- 238000010992 reflux Methods 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 239000008363 phosphate buffer Substances 0.000 claims description 3
- 230000006870 function Effects 0.000 abstract description 9
- 238000000034 method Methods 0.000 description 27
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 20
- 230000000052 comparative effect Effects 0.000 description 15
- 230000001954 sterilising effect Effects 0.000 description 15
- 239000001814 pectin Substances 0.000 description 13
- 235000010987 pectin Nutrition 0.000 description 13
- 229920001277 pectin Polymers 0.000 description 13
- 230000000694 effects Effects 0.000 description 11
- 230000008569 process Effects 0.000 description 11
- 229910052757 nitrogen Inorganic materials 0.000 description 10
- 206010016807 Fluid retention Diseases 0.000 description 9
- 241000121184 Monodon Species 0.000 description 8
- 241000238552 Penaeus monodon Species 0.000 description 8
- 239000000126 substance Substances 0.000 description 7
- 238000007865 diluting Methods 0.000 description 6
- 241000283070 Equus zebra Species 0.000 description 5
- 238000004132 cross linking Methods 0.000 description 5
- 230000003647 oxidation Effects 0.000 description 5
- 238000007254 oxidation reaction Methods 0.000 description 5
- 230000008859 change Effects 0.000 description 4
- 239000013078 crystal Substances 0.000 description 4
- 239000000835 fiber Substances 0.000 description 4
- 229910052739 hydrogen Inorganic materials 0.000 description 4
- 239000001257 hydrogen Substances 0.000 description 4
- 230000009471 action Effects 0.000 description 3
- 230000003064 anti-oxidating effect Effects 0.000 description 3
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 3
- 239000011162 core material Substances 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- WTEVQBCEXWBHNA-UHFFFAOYSA-N Citral Natural products CC(C)=CCCC(C)=CC=O WTEVQBCEXWBHNA-UHFFFAOYSA-N 0.000 description 2
- 229920001410 Microfiber Polymers 0.000 description 2
- 241001585141 Rimapenaeus pacificus Species 0.000 description 2
- 125000006297 carbonyl amino group Chemical group [H]N([*:2])C([*:1])=O 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 229940043350 citral Drugs 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000006866 deterioration Effects 0.000 description 2
- 238000005538 encapsulation Methods 0.000 description 2
- 238000001704 evaporation Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- WTEVQBCEXWBHNA-JXMROGBWSA-N geranial Chemical compound CC(C)=CCC\C(C)=C\C=O WTEVQBCEXWBHNA-JXMROGBWSA-N 0.000 description 2
- 239000003658 microfiber Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- PFTAWBLQPZVEMU-DZGCQCFKSA-N (+)-catechin Chemical compound C1([C@H]2OC3=CC(O)=CC(O)=C3C[C@@H]2O)=CC=C(O)C(O)=C1 PFTAWBLQPZVEMU-DZGCQCFKSA-N 0.000 description 1
- RVBUGGBMJDPOST-UHFFFAOYSA-N 2-thiobarbituric acid Chemical compound O=C1CC(=O)NC(=S)N1 RVBUGGBMJDPOST-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000907635 Farfantepenaeus duorarum Species 0.000 description 1
- 235000005206 Hibiscus Nutrition 0.000 description 1
- 235000007185 Hibiscus lunariifolius Nutrition 0.000 description 1
- 244000284380 Hibiscus rosa sinensis Species 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- OWIKHYCFFJSOEH-UHFFFAOYSA-N Isocyanic acid Chemical compound N=C=O OWIKHYCFFJSOEH-UHFFFAOYSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- 241000533950 Leucojum Species 0.000 description 1
- 239000000232 Lipid Bilayer Substances 0.000 description 1
- 241000304195 Salvia miltiorrhiza Species 0.000 description 1
- 235000011135 Salvia miltiorrhiza Nutrition 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 238000005576 amination reaction Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 239000008346 aqueous phase Substances 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- ADRVNXBAWSRFAJ-UHFFFAOYSA-N catechin Natural products OC1Cc2cc(O)cc(O)c2OC1c3ccc(O)c(O)c3 ADRVNXBAWSRFAJ-UHFFFAOYSA-N 0.000 description 1
- 235000005487 catechin Nutrition 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229950001002 cianidanol Drugs 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 235000012055 fruits and vegetables Nutrition 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 150000007965 phenolic acids Chemical class 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 210000002706 plastid Anatomy 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000001338 self-assembly Methods 0.000 description 1
- 238000013268 sustained release Methods 0.000 description 1
- 239000012730 sustained-release form Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/06—Freezing; Subsequent thawing; Cooling
- A23B4/08—Freezing; Subsequent thawing; Cooling with addition of chemicals or treatment with chemicals before or during cooling, e.g. in the form of an ice coating or frozen block
- A23B4/09—Freezing; Subsequent thawing; Cooling with addition of chemicals or treatment with chemicals before or during cooling, e.g. in the form of an ice coating or frozen block with direct contact between the food and the chemical, e.g. liquid N2, at cryogenic temperature
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23B—PRESERVING, e.g. BY CANNING, MEAT, FISH, EGGS, FRUIT, VEGETABLES, EDIBLE SEEDS; CHEMICAL RIPENING OF FRUIT OR VEGETABLES; THE PRESERVED, RIPENED, OR CANNED PRODUCTS
- A23B4/00—General methods for preserving meat, sausages, fish or fish products
- A23B4/12—Preserving with acids; Acid fermentation
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
-
- 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
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to the technical field of aquatic product processing and preservation, and discloses a modified protocatechuic acid liposome for shrimp and crab storage and preservation, and a preparation method and application thereof. The modified protocatechuic acid liposome comprises the following three-layer structure from inside to outside: protocatechuic acid core, soybean lecithin/cholesterol/vitamin E composite layer, and citrus pectin/aminated bacterial cellulose composite layer. According to the modified protocatechuic acid liposome, the citrus pectin/aminated bacterial cellulose composite layer is coated outside the protocatechuic acid liposome, so that the viscosity of the liposome can be improved, the liposome has higher stability, and the functions of slowly releasing protocatechuic acid and retaining water can be better exerted.
Description
Technical Field
The invention relates to the technical field of aquatic product processing and preservation, in particular to a modified protocatechuic acid liposome for shrimp and crab storage and preservation and a preparation method and application thereof.
Background
The shrimp and crab meat is fresh and tender, delicious in taste and rich in nutrition, and is deeply favored by people. Compared with other aquatic products, the shrimp and crab feed has higher protein and water content and a large amount of high-activity enzymes and tyrosine in the bodies, so the shrimp and crab feed is easy to decay and deteriorate and is easy to be oxidized to generate blackening in the processes of fishing, transporting, processing and storing, and the commercial value of the shrimps and crabs is reduced. Along with the improvement of living standard of people, the requirement on freshness of people is gradually improved, so that the method for keeping freshness of the fresh vegetables is of great significance for finding a green, safe and efficient preservation method.
At present, the common preservation methods for shrimps and crabs include preservation by adopting a biological preservative solution, low-temperature preservation, modified atmosphere preservation and the like. Low-temperature freezing is a common preservation method for aquatic products, and the frozen aquatic products in China account for about 60 percent of the total quantity of the aquatic products. However, the growth of ice crystals in freezing and frozen storage can cause the change of the organization structure of the product, and the phenomena of freezing damage and juice loss after thawing are serious, thereby causing serious influence on the appearance quality and the market value of the product.
Protocatechuic acid is a phenolic acid substance naturally existing in a plurality of vegetables and fruits, is an effective active ingredient in a plurality of traditional Chinese medicines (such as salvia miltiorrhiza, hibiscus, and the like), and has been applied to the fields of medicine, health care, and the like due to the physiological functions of antioxidation, anti-inflammation, bacteriostasis, and the like. Its low water solubility and chemical instability greatly limit its application in the food industry.
The liposome is a micro vesicle with a double-layer membrane structure formed by self-assembly of an amphiphilic lipid bilayer coated with certain components, and the aqueous phase core and the bilayer can be coated with hydrophilic and lipophilic core materials to achieve the purposes of protecting, slowly releasing, targeting and the like on an embedded object, thereby being one of international leading-edge technologies in the field of biomedical food and chemical industry at present. Chinese patent publication No. CN110754522A discloses a method for preparing a citral liposome fruit and vegetable antibacterial antistaling agent, which reduces the volatility and chemical instability of citral by liposome encapsulation, but the liposome encapsulation also has the following defects: plastids suffer from physicochemical instability (e.g., aggregation, rupture, fusion, etc.) resulting in particle size variation and core leakage.
Disclosure of Invention
In order to solve the technical problems, the invention provides a modified protocatechuic acid liposome for storage and preservation of shrimps and crabs, and a preparation method and application thereof. The modified protocatechuic acid liposome is coated with a citrus pectin/aminated bacterial cellulose composite layer, so that the viscosity of the liposome can be improved, the liposome has higher stability, and the functions of slowly releasing protocatechuic acid and retaining water can be better exerted.
The specific technical scheme of the invention is as follows:
a modified protocatechuic acid liposome for storage and preservation of shrimps and crabs comprises the following three-layer structure from inside to outside: protocatechuic acid core, soybean lecithin/cholesterol/vitamin E composite layer, and citrus pectin/aminated bacterial cellulose composite layer.
According to the invention, the soybean lecithin/cholesterol/vitamin E composite layer is coated outside the protocatechuic acid to form the liposome, so that the chemical stability of the protocatechuic acid can be improved, the protocatechuic acid is slowly released in the storage process of the shrimps and crabs, the long-acting bacteriostasis and antioxidation can be realized, and the bacterial breeding and the oxidative blackening in the shrimps and crabs can be inhibited; meanwhile, the liposome covers the surfaces of the shrimps and crabs to form a film, so that the effects of water retention and freezing resistance can be achieved, the generation and growth of ice crystals in the freezing storage process are inhibited, and the mechanical damage of the frozen ice crystals to cell membranes of the shrimps and crabs is reduced.
The liposome has the problem of unstable physical and chemical properties, and is easy to generate particle size change and excessive leakage of protocatechuic acid in the process of frozen storage. In order to solve the problem, the invention coats a citrus pectin/aminated bacterial cellulose composite layer outside the liposome, can change the electrostatic action and the steric hindrance action of a liposome system, improve the viscosity of the system, be conductive to improving the stability of the liposome and enhance the water retention action of the liposome system. The purpose of adopting the aminated bacterial cellulose is as follows: the pectin molecules have negative charges, molecular chains tend to repel each other, so that the improvement effect on the viscosity of a liposome system is weak, the bacterial cellulose has a typical microfiber network characteristic in structure, and the crosslinking degree of a pectin layer can be improved after the bacterial cellulose is crosslinked with the pectin molecular chains through hydrogen bonds, so that the viscosity of the liposome system is effectively improved, the liposome system has higher stability, and the functions of slowly releasing protocatechuic acid and retaining water can be better exerted; the bacterial cellulose is aminated, so that the bacterial cellulose and pectin can form more stable covalent connection through the carbonylamino reaction between amino and carbonyl besides a hydrogen bond, and a cross-linked network structure in a citrus pectin/aminated bacterial cellulose composite layer is more stable in a frozen storage process, so that the stability of the liposome can be further improved, and the liposome can play a better water retention role.
Preferably, the preparation method of the aminated bacterial cellulose comprises the following steps: dispersing bacterial cellulose into dimethylformamide, dropwise adding thionyl chloride, carrying out reflux reaction on the bacterial cellulose and the thionyl chloride for 4-5 h at the temperature of 70-80 ℃, filtering, and washing to obtain chlorinated bacterial cellulose, wherein the mass ratio of the bacterial cellulose to the thionyl chloride is 1.5-2.0; re-dispersing the chlorinated bacterial cellulose into dimethyl formamide, adding ethylenediamine, reacting at 75-85 ℃ for 3-4 h, filtering, and washing with anhydrous acetone and water in sequence to obtain the aminated bacterial cellulose, wherein the mass-volume ratio of the chlorinated bacterial cellulose to the ethylenediamine is 1.
Preferably, the preparation method of the bacterial cellulose comprises the following steps:
(i) And (3) activation: inoculating acetobacter xylinum to a slant culture medium, and culturing for 48-60 hours in an incubator at 28-30 ℃ to obtain an activated strain;
(ii) Seed culture: selecting 2-3 ring activated strains to inoculate into 45-55 mL seed culture medium, and culturing for 24-30 h at 28-30 ℃ in a shaking incubator of 120-150 r/min to obtain seed bacterial liquid;
(iii) Fermentation culture: inoculating the seed bacterial liquid into a fermentation culture medium according to the inoculation amount of 5-10 wt%, and fully shaking to separate out the bacterial strains and fully disperse the bacterial strains into the fermentation culture medium; then culturing the mixture for 6 to 8 days in a shaking incubator at the temperature of between 28 and 30 ℃ and at the speed of between 140 and 170r/min, and forming a bacterial cellulose membrane on the surface of a fermentation medium;
(iv) Taking out the bacterial cellulose membrane, washing the bacterial cellulose membrane with water until no floccule is left, soaking the bacterial cellulose membrane in 0.1-0.15 mol/L sodium hydroxide solution for 30-40 min at the temperature of 75-85 ℃, and then washing the bacterial cellulose membrane to be neutral to obtain the bacterial cellulose.
The bacterial cellulose obtained by the method is in a snowflake shape, a plurality of fine villus fibers are attached to the periphery of each main fiber, and the high-branch structure is favorable for forming a cross-linked network structure among pectin molecular chains, so that the viscosity of a liposome system is improved, the liposome is more stable in the freezing and storing process, and the functions of slowly releasing protocatechuic acid and retaining water are favorably realized.
Preferably, the slant culture medium comprises the following components: 20g/L of glucose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate, 1g/L of citric acid, 20g/L of agar and 0.5g/L of calcium carbonate.
Preferably, the seed culture medium comprises the following components: 20g/L of sucrose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate and 1g/L of citric acid.
Preferably, the fermentation medium comprises the following components: 20g/L of sucrose, 8g/L of peptone, 10.55g/L of yeast powder, 2.7g/L of disodium hydrogen phosphate and 1g/L of citric acid.
Preferably, the pH of the slant medium, the seed medium, and the fermentation medium is 6.0 to 6.5.
A method for preparing the modified protocatechuic acid liposome comprises the following steps:
(1) Adding soybean lecithin, cholesterol, tween-80 and vitamin E into absolute ethyl alcohol, stirring and dissolving completely to form a uniform film;
(2) Carrying out reduced pressure rotary evaporation to remove the absolute ethyl alcohol to form a film; adding a phosphate buffer solution containing protocatechuic acid to perform constant volume membrane washing, and performing ultrasonic treatment to obtain a crude lipid suspension;
(3) Dissolving citrus pectin and aminated bacterial cellulose into a phosphate buffer solution to obtain a polysaccharide solution;
(4) And dropwise adding the crude lipid suspension into the polysaccharide solution, stirring and reacting at 60-70 ℃ for 30-40 min, and continuously standing at the temperature for 50-60 min to obtain the modified protocatechuic acid liposome.
In the step (4), after the crude lipid suspension is mixed with the polysaccharide solution, carbonyl-ammonia reaction is carried out between carbonyl in pectin and amino in the aminated bacterial cellulose at the temperature of 60-70 ℃, covalent crosslinking is formed between pectin molecular chains and cellulose molecular chains, and a crosslinked net structure in the citrus pectin/aminated bacterial cellulose composite layer is more stable, so that the liposome can better play roles of water retention and slow release.
Preferably, in the step (1), the mass ratio of the soybean lecithin to the cholesterol to the tween-80 to the vitamin E is 1.0-4.5.
Preferably, in the protocatechuic acid-containing phosphate buffer solution in the step (2), the mass concentration of the protocatechuic acid is 100-300 mg/L; and (3) the mass-to-volume ratio of the soybean lecithin in the step (1) to the mixed solution with constant volume in the step (2) is 1.0-4.5g.
Preferably, in the step (1), the mass volume ratio of the soybean lecithin to the absolute ethyl alcohol is 1.0-4.5g.
Preferably, in the step (2), the temperature of the reduced pressure rotary evaporation is 40 to 60 ℃.
Preferably, in the step (2), the phosphate buffer has a concentration of 0.04 to 0.06mol/L and a pH of 6.6 to 7.0.
Preferably, in the step (2), the time of the ultrasonic treatment is 15 to 30min.
Preferably, in the step (3), the phosphate buffer has a concentration of 0.04 to 0.06mol/L and a pH of 7.1 to 7.3.
Preferably, in the step (3), the mass fraction of the citrus pectin and the mass fraction of the bacterial cellulose in the polysaccharide solution are respectively 0.1 to 0.4% and 0.01 to 0.04%.
Preferably, in the step (4), the volume ratio of the crude lipid suspension to the polysaccharide solution is 1.0-1.3.
The application of the modified protocatechuic acid liposome in the storage and the fresh keeping of shrimps and crabs comprises the following steps: soaking the shrimps and crabs in the modified protocatechuic acid liposome at the temperature of 8-10 ℃ for 1-2 h, taking out, freezing and storing at the temperature of-18-30 ℃.
Preferably, the specific process of freezing is as follows: and (3) immersing the soaked shrimps and crabs into liquid nitrogen for freezing, so that the central temperature of the shrimps and crabs is reduced to-18 to-30 ℃ within 15-24 min.
Compared with the prior art, the invention has the following advantages:
(1) When the modified protocatechuic acid liposome is applied to the freshness preservation of shrimps and crabs, the protocatechuic acid liposome can slowly release protocatechuic acid, can inhibit the putrefaction deterioration and oxidation blackening of the shrimps and crabs for a long time, has the functions of water retention and freezing resistance, and can prevent cell membranes of the shrimps and crabs from being damaged by ice crystals;
(2) According to the invention, pectin is modified on the surface of the protocatechuic acid liposome, and a fiber mesh structure is formed by bacterial cellulose and pectin, so that the viscosity of the liposome can be improved, the liposome has higher stability, and the functions of slowly releasing protocatechuic acid and retaining water can be better exerted;
(3) The bacterial cellulose adopted by the invention has a snowflake-shaped highly branched structure, so that the crosslinking degree of a citrus pectin/aminated bacterial cellulose composite layer can be improved, the viscosity of the liposome can be further improved, and the functions of slowly releasing protocatechuic acid and retaining water can be favorably exerted;
(4) According to the invention, bacterial cellulose is subjected to amination modification, so that covalent crosslinking is formed between the bacterial cellulose and pectin, a fiber network has higher stability in a frozen storage process, and the functions of slowly releasing protocatechuic acid and retaining water of liposome are facilitated.
Detailed Description
The present invention will be further described with reference to the following examples.
General examples
Bacterial cellulose is prepared by the following steps:
(i) The culture medium is prepared according to the following formula:
slant culture medium: 20g/L of glucose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate, 1g/L of citric acid, 20g/L of agar and 0.5g/L of calcium carbonate; the pH value is 6.0-6.5; sterilizing at 121 deg.C and 0.1MPa for 30min;
seed culture medium: 20g/L of sucrose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH value is 6.0-6.5; sterilizing at 121 deg.C under 0.1MPa for 30min;
the fermentation medium comprises the following components: 20g/L of sucrose, 8g/L of peptone, 10.55g/L of yeast powder, 2.7g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH value is 6.0-6.5; sterilizing at 121 deg.C and 0.1MPa for 30min;
(ii) And (3) activation: inoculating acetobacter xylinum to a slant culture medium, and culturing for 48-60 h in an incubator at 28-30 ℃ to obtain an activated strain;
(iii) Seed culture: selecting 2-3 rings of activated strains, inoculating the strains into 45-55 mL of seed culture medium, and culturing for 24-30 h at 28-30 ℃ in a shaking table incubator at 120-150 r/min to obtain seed bacterial liquid;
(iv) Fermentation culture: inoculating the seed bacterial liquid into a fermentation culture medium according to the inoculation amount of 5-10 wt%, and fully shaking to separate out the bacterial strain and fully disperse the bacterial strain into the fermentation culture medium; then culturing the mixture for 6 to 8 days in a shaking incubator at the temperature of between 28 and 30 ℃ and at the speed of between 140 and 170r/min, and forming a bacterial cellulose membrane on the surface of a fermentation medium;
(v) Taking out the bacterial cellulose membrane, washing the bacterial cellulose membrane with water until no floccule exists, soaking the bacterial cellulose membrane in 0.1-0.15 mol/L sodium hydroxide solution for 30-40 min at the temperature of 75-85 ℃, and then washing the bacterial cellulose membrane to be neutral to obtain the bacterial cellulose.
Using the obtained bacterial cellulose, preparing aminated bacterial cellulose by the following steps:
(I) Dispersing bacterial cellulose into dimethyl formamide, dropwise adding thionyl chloride, carrying out reflux reaction on the bacterial cellulose and the thionyl chloride for 4-5 h at 70-80 ℃, filtering, and washing to obtain chlorinated bacterial cellulose, wherein the mass ratio of the bacterial cellulose to the thionyl chloride is 1.5-2.0;
(II) dispersing the chlorinated bacterial cellulose into dimethyl formamide again, adding ethylenediamine, reacting for 3-4 h at 75-85 ℃ with the mass-volume ratio of 1.
Preparing a modified protocatechuic acid liposome by using the obtained aminated bacterial cellulose through the following steps:
(1) Adding soybean lecithin, cholesterol, tween-80 and vitamin E into absolute ethyl alcohol, stirring and dissolving completely to form a uniform film;
the mass ratio of the soybean lecithin to the cholesterol to the Tween-80 to the vitamin E is (1.0-4.5g);
(2) Carrying out reduced pressure rotary evaporation at 40-60 ℃ to remove absolute ethyl alcohol to form a film; adding a phosphate buffer solution containing protocatechuic acid to perform constant volume membrane washing, and performing ultrasonic treatment for 15-30 min to obtain a crude lipid suspension;
the concentration of the phosphate buffer solution is 0.04-0.06 mol/L, and the pH value is 6.6-7.0;
in the phosphate buffer solution containing protocatechuic acid, the mass concentration of protocatechuic acid is 100-300 mg/L;
the mass-to-volume ratio of the soybean lecithin in the step (1) to the mixed solution with constant volume in the step (2) is 1.0-4.5g;
(3) Dissolving citrus pectin and aminated bacterial cellulose into a phosphate buffer solution to obtain a polysaccharide solution;
the concentration of the phosphate buffer solution is 0.04-0.06 mol/L, and the pH value is 7.1-7.3;
in the polysaccharide solution, the mass fraction of the citrus pectin is 0.1-0.4%, and the mass fraction of the bacterial cellulose is 0.01-0.04%;
(4) Dropwise adding the crude lipid suspension into the polysaccharide solution, stirring and reacting at 60-70 ℃ for 30-40 min, and then continuously standing at the temperature for 50-60 min to obtain modified protocatechuic acid liposome;
the volume ratio of the crude lipid suspension to the polysaccharide solution is 1.0-1.3.
The obtained modified protocatechuic acid liposome comprises the following three-layer structure from inside to outside: protocatechuic acid core, soybean lecithin/cholesterol/vitamin E composite layer, and citrus pectin/aminated bacterial cellulose composite layer.
The shrimp and crab storage and preservation are carried out by using the obtained modified protocatechuic acid liposome, and the method comprises the following steps: soaking the shrimps and the crabs in the modified protocatechuic acid liposome at the temperature of between 8 and 10 ℃ for 1 to 2 hours, then taking out the shrimps and the crabs, soaking the shrimps and the crabs in liquid nitrogen for freezing to ensure that the central temperature of the shrimps and the crabs is reduced to between 18 below zero and 30 below zero in 15 to 24 minutes, and then storing the shrimps and the crabs at the temperature of between 18 below zero and 30 below zero.
Example 1
Bacterial cellulose is prepared by the following steps:
(i) The culture medium is prepared according to the following formula:
slant culture medium: 20g/L of glucose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate, 1g/L of citric acid, 20g/L of agar and 0.5g/L of calcium carbonate; the pH was 6.2; sterilizing at 121 deg.C under 0.1MPa for 30min;
seed culture medium: 20g/L of sucrose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH was 6.2; sterilizing at 121 deg.C under 0.1MPa for 30min;
the fermentation medium comprises the following components: 20g/L of sucrose, 8g/L of peptone, 10.55g/L of yeast powder, 2.7g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH was 6.2; sterilizing at 121 deg.C under 0.1MPa for 30min;
(ii) Activation: inoculating acetobacter xylinum to a slant culture medium, and culturing in an incubator at 29 ℃ for 54h to obtain an activated strain;
(iii) Seed culture: selecting 3-ring activated strains, inoculating the strains into 50mL of seed culture medium, and culturing for 27h at 29 ℃ in a shaking incubator at 140r/min to obtain seed bacterial liquid;
(iv) Fermentation culture: inoculating the seed bacterial liquid into a fermentation culture medium according to the inoculation amount of 8wt%, and fully shaking to separate out the bacterial strain and fully disperse the bacterial strain into the fermentation culture medium; then culturing for 7d in a shaking incubator at 29 ℃ and 160r/min, and forming a bacterial cellulose membrane on the surface of a fermentation medium;
(v) Taking out the bacterial cellulose membrane, washing the bacterial cellulose membrane with water until no floccule is washed, soaking the bacterial cellulose membrane in 0.13mol/L sodium hydroxide solution at the temperature of 80 ℃ for 35min, and then washing the bacterial cellulose membrane to be neutral to obtain the bacterial cellulose.
Using the obtained bacterial cellulose, preparing aminated bacterial cellulose by the following steps:
(I) Dispersing 5g of bacterial cellulose into 100mL of dimethylformamide, dropwise adding 7.5g of thionyl chloride, carrying out reflux reaction at 75 ℃ for 4.5h, filtering, and washing with water to obtain chlorinated bacterial cellulose;
(II) re-dispersing 5g of chlorinated bacterial cellulose into 100mL of dimethylformamide, adding 45g of ethylenediamine, reacting at 80 ℃ for 3.5h, filtering, and washing with anhydrous acetone and water in sequence to obtain the aminated bacterial cellulose.
Preparing a modified protocatechuic acid liposome by using the obtained aminated bacterial cellulose through the following steps:
(1) Adding 3g of soybean lecithin, 1g of cholesterol, 1g of tween-80 and 0.2g of vitamin E into 60mL of absolute ethyl alcohol, and stirring to completely dissolve the soybean lecithin, the cholesterol, the tween-80 and the vitamin E to uniformly form a film;
(2) Rotary evaporating at 50 deg.C under reduced pressure to remove anhydrous ethanol to form film; adding 0.05mol/L phosphate buffer solution containing 200mg/L protocatechuic acid and having pH of 6.8, diluting to 100mL, and performing ultrasonic treatment for 30min to obtain crude lipid suspension;
(3) Dissolving 0.22g of citrus pectin and 0.022g of aminated bacterial cellulose into 110mL of 0.05mol/L phosphate buffer solution with pH of 7.2 to obtain a polysaccharide solution;
(4) And (3) dropwise adding 100mL of the crude lipid suspension into 110mL of the polysaccharide solution, stirring and reacting at 65 ℃ for 35min, and continuously standing at the temperature for 55min to obtain the modified protocatechuic acid liposome.
The obtained modified protocatechuic acid liposome comprises the following three-layer structure from inside to outside: protocatechuic acid core, soybean lecithin/cholesterol/vitamin E composite layer, and citrus pectin/aminated bacterial cellulose composite layer.
The method for storing and preserving the prawns in the zebra by using the obtained modified protocatechuic acid liposome comprises the following steps: soaking the Penaeus monodon in modified protocatechuic acid liposome at 9 deg.C for 2 hr, taking out, soaking in liquid nitrogen, freezing to reduce the central temperature of Penaeus monodon to-20 deg.C within 20min, and storing at-20 deg.C.
Example 2
Bacterial cellulose is prepared by the following steps:
(i) The culture medium is prepared according to the following formula:
slant culture medium: 20g/L of glucose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate, 1g/L of citric acid, 20g/L of agar and 0.5g/L of calcium carbonate; the pH was 6.0; sterilizing at 121 deg.C and 0.1MPa for 30min;
seed culture medium: 20g/L of sucrose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH was 6.0; sterilizing at 121 deg.C under 0.1MPa for 30min;
the fermentation medium comprises the following components: 20g/L of sucrose, 8g/L of peptone, 10.55g/L of yeast powder, 2.7g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH was 6.0; sterilizing at 121 deg.C and 0.1MPa for 30min;
(ii) Activation: inoculating acetobacter xylinum to a slant culture medium, and culturing in an incubator at 28 ℃ for 48 hours to obtain an activated strain;
(iii) Seed culture: selecting 3-ring activated strains, inoculating the strains into 455mL of seed culture medium, and culturing for 24 hours in a shaking incubator at 28 ℃ and 120r/min to obtain seed bacterial liquid;
(iv) Fermentation culture: inoculating the seed bacterial liquid into a fermentation culture medium according to the inoculation amount of 10wt%, and fully shaking to separate out the bacterial strain and fully disperse the bacterial strain into the fermentation culture medium; then culturing for 6 days in a shaking incubator at 28 ℃ and 140r/min, and forming a bacterial cellulose membrane on the surface of a fermentation medium;
(v) Taking out the bacterial cellulose membrane, washing the bacterial cellulose membrane by water until no floccule is washed, soaking the bacterial cellulose membrane in 0.1mol/L sodium hydroxide solution for 40min at 75 ℃, and then washing the bacterial cellulose membrane to be neutral to obtain the bacterial cellulose.
Using the obtained bacterial cellulose to prepare the aminated bacterial cellulose by the following steps:
(I) Dispersing 5g of bacterial cellulose into 100mL of dimethylformamide, dropwise adding 8.5g of thionyl chloride, carrying out reflux reaction at 70 ℃ for 5 hours, filtering, and washing with water to obtain chlorinated bacterial cellulose;
(II) dispersing 5g of chlorinated bacterial cellulose into 100mL of dimethylformamide again, adding 42.5g of ethylenediamine, reacting for 4h at 75 ℃, filtering, and washing with anhydrous acetone and water in sequence to obtain the aminated bacterial cellulose.
Preparing a modified protocatechuic acid liposome by using the obtained aminated bacterial cellulose through the following steps:
(1) Adding 1.0g of soybean lecithin, 0.5g of cholesterol, 0.5g of tween-80 and 0.1g of vitamin E into 60mL of absolute ethyl alcohol, and stirring for completely dissolving to uniformly form a film;
(2) Performing rotary evaporation at 40 deg.C under reduced pressure to remove anhydrous ethanol to form film; adding 0.04mol/L phosphate buffer solution containing 100mg/L protocatechuic acid and having pH of 6.6, diluting to 100mL, and performing ultrasonic treatment for 15min to obtain crude lipid suspension;
(3) Dissolving 0.13g of citrus pectin and 0.013g of aminated bacterial cellulose in 130mL of 0.04mol/L phosphate buffer solution with pH of 7.1 to obtain a polysaccharide solution;
(4) And (3) dropwise adding 100mL of the crude lipid suspension into 130mL of the polysaccharide solution, stirring and reacting at 60 ℃ for 40min, and continuously standing at the temperature for 50min to obtain the modified protocatechuic acid liposome.
The obtained modified protocatechuic acid liposome comprises the following three-layer structure from inside to outside: protocatechuic acid core, soybean lecithin/cholesterol/vitamin E composite layer, and citrus pectin/aminated bacterial cellulose composite layer.
The method for storing and preserving the prawns in the zebra by using the obtained modified protocatechuic acid liposome comprises the following steps: soaking the Penaeus monodon in modified protocatechuic acid liposome at 10 deg.C for 1.5h, taking out, soaking in liquid nitrogen, freezing to reduce the central temperature of Penaeus monodon to-30 deg.C in 24min, and storing at-30 deg.C.
Example 3
Bacterial cellulose is prepared by the following steps:
(i) The culture medium is prepared according to the following formula:
slant culture medium: 20g/L of glucose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate, 1g/L of citric acid, 20g/L of agar and 0.5g/L of calcium carbonate; the pH was 6.5; sterilizing at 121 deg.C and 0.1MPa for 30min;
seed culture medium: 20g/L of sucrose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH was 6.5; sterilizing at 121 deg.C under 0.1MPa for 30min;
the fermentation medium comprises the following components: 20g/L of sucrose, 8g/L of peptone, 10.55g/L of yeast powder, 2.7g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH was 6.5; sterilizing at 121 deg.C and 0.1MPa for 30min;
(ii) Activation: inoculating acetobacter xylinum to a slant culture medium, and culturing in an incubator at 30 ℃ for 60 hours to obtain an activated strain;
(iii) Seed culture: selecting 2-ring activated strains, inoculating the strains into 55mL of seed culture medium, and culturing for 30h in a shaking table incubator at 30 ℃ and 150r/min to obtain seed bacterial liquid;
(iv) Fermentation culture: inoculating the seed bacterial liquid into a fermentation culture medium according to the inoculation amount of 5wt%, and fully shaking to separate out the bacterial strains and fully disperse the bacterial strains into the fermentation culture medium; then culturing for 8 days in a shaking incubator at 30 ℃ and 170r/min, and forming a bacterial cellulose membrane on the surface of a fermentation medium;
(v) Taking out the bacterial cellulose membrane, washing the bacterial cellulose membrane with water until no floccule is washed, soaking the bacterial cellulose membrane in 0.15mol/L sodium hydroxide solution for 30min at the temperature of 85 ℃, and then washing the bacterial cellulose membrane to be neutral to obtain the bacterial cellulose.
Using the obtained bacterial cellulose, preparing aminated bacterial cellulose by the following steps:
(I) Dispersing 5g of bacterial cellulose into 100mL of dimethylformamide, dropwise adding 10g of thionyl chloride, carrying out reflux reaction at 80 ℃ for 4 hours, filtering, and washing with water to obtain chlorinated bacterial cellulose;
(II) dispersing 5g of chlorinated bacterial cellulose into 100mL of dimethylformamide again, adding 47.5g of ethylenediamine, reacting at 85 ℃ for 3 hours, filtering, and washing with anhydrous acetone and water in sequence to obtain the aminated bacterial cellulose.
Preparing a modified protocatechuic acid liposome by using the obtained aminated bacterial cellulose through the following steps:
(1) Adding 4.5g of soybean lecithin, 1.5g of cholesterol, 2.0g of Tween-80 and 0.05g of vitamin E into 60mL of absolute ethyl alcohol, and stirring for completely dissolving to uniformly form a film;
(2) Rotary evaporating at 60 deg.C under reduced pressure to remove anhydrous ethanol to form film; adding 0.06mol/L phosphate buffer solution containing 300mg/L protocatechuic acid and having pH of 7.0, diluting to 100mL, and performing ultrasonic treatment for 25min to obtain crude lipid suspension;
(3) Dissolving 0.4g of citrus pectin and 0.04g of aminated bacterial cellulose into 100mL of 0.06mol/L phosphate buffer solution with pH of 7.3 to obtain polysaccharide solution;
(4) And (3) dropwise adding 100mL of the crude lipid suspension into 100mL of the polysaccharide solution, stirring and reacting at 70 ℃ for 30min, and continuously standing at the temperature for 60min to obtain the modified protocatechuic acid liposome.
The obtained modified protocatechuic acid liposome comprises the following three-layer structure from inside to outside: protocatechuic acid core, soybean lecithin/cholesterol/vitamin E composite layer, and citrus pectin/aminated bacterial cellulose composite layer.
The method for storing and preserving the prawns in the zebra by using the obtained modified protocatechuic acid liposome comprises the following steps: soaking the Mylopsis monodon in modified protocatechuic acid liposome at 8 deg.C for 1 hr, taking out, soaking in liquid nitrogen, freezing to reduce the central temperature of Mylopsis monodon to-18 deg.C within 15min, and storing at-18 deg.C.
Comparative example 1
Soaking the Mylopsis monodon in liquid nitrogen for freezing to reduce the central temperature of Mylopsis monodon to-20 deg.C within 20min, and storing at-20 deg.C.
Comparative example 2
The method for storing and preserving the penaeus monodon by using the protocatechuic acid solution comprises the following steps: soaking the Mylopsis prawns in 0.05mol/L phosphate buffer solution with pH 7 and containing 100g/mL protocatechuic acid at 9 deg.C for 2h, taking out, soaking in liquid nitrogen, freezing to reduce the central temperature of Mylopsis prawns to-20 deg.C within 20min, and storing at-20 deg.C.
Comparative example 3
Protocatechuic acid liposomes were prepared by the following steps:
(1) Adding 3g of soybean lecithin, 1g of cholesterol, 1g of tween-80 and 0.2g of vitamin E into 60mL of absolute ethyl alcohol, and stirring to completely dissolve the soybean lecithin, the cholesterol, the tween-80 and the vitamin E to uniformly form a film;
(2) Performing rotary evaporation at 50 deg.C under reduced pressure to remove anhydrous ethanol to form film; adding 0.05mol/L phosphate buffer solution containing 200mg/L protocatechuic acid and having pH of 6.8, diluting to 100mL membrane, and performing ultrasonic treatment for 30min to obtain protocatechuic acid liposome.
The obtained protocatechuic acid liposome comprises the following three-layer structure from inside to outside: protocatechuic acid core, soybean lecithin/cholesterol/vitamin E composite layer.
The obtained protocatechuic acid liposome is used for the storage and the fresh keeping of the prawns, and the method comprises the following steps: soaking the Penaeus monodon in protocatechuic acid liposome at 9 deg.C for 2 hr, taking out, soaking in liquid nitrogen, freezing to reduce the central temperature of Penaeus monodon to-20 deg.C within 20min, and storing at-20 deg.C.
Comparative example 4
Modified protocatechuic acid liposomes were prepared by the following steps:
(1) Adding 3g of soybean lecithin, 1g of cholesterol, 1g of tween-80 and 0.2g of vitamin E into 60mL of absolute ethyl alcohol, and stirring to completely dissolve the soybean lecithin, the cholesterol, the tween-80 and the vitamin E to uniformly form a film;
(2) Performing rotary evaporation at 50 deg.C under reduced pressure to remove anhydrous ethanol to form film; adding 0.05mol/L phosphate buffer solution containing 200mg/L protocatechuic acid and having pH of 6.8, diluting to 100mL, and performing ultrasonic treatment for 30min to obtain crude lipid suspension;
(3) Dissolving 0.22g of citrus pectin in 110mL0.05mol/L phosphate buffer solution with pH of 7.2 to obtain polysaccharide solution;
(4) And (3) dropwise adding 100mL of the crude lipid suspension into 110mL of the polysaccharide solution, stirring and reacting at 65 ℃ for 35min, and continuously standing at the temperature for 55min to obtain the modified protocatechuic acid liposome.
The obtained modified protocatechuic acid liposome comprises the following three-layer structure from inside to outside: protocatechuic acid core, soybean lecithin/cholesterol/vitamin E composite layer, and citrus pectin layer.
The method for storing and preserving the prawns in the zebra by using the obtained modified protocatechuic acid liposome comprises the following steps: soaking the Mylopsis monodon in modified protocatechuic acid liposome at 9 deg.C for 2 hr, taking out, soaking in liquid nitrogen, freezing to reduce the central temperature of Mylopsis monodon to-20 deg.C within 20min, and storing at-20 deg.C.
Comparative example 5
Bacterial cellulose is prepared by the following steps:
(i) The culture medium is prepared according to the following formula:
slant culture medium: 20g/L of glucose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate, 1g/L of citric acid, 20g/L of agar and 0.5g/L of calcium carbonate; the pH was 6.2; sterilizing at 121 deg.C under 0.1MPa for 30min;
seed culture medium: 20g/L of sucrose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH was 6.2; sterilizing at 121 deg.C under 0.1MPa for 30min;
the fermentation medium comprises the following components: 20g/L of sucrose, 8g/L of peptone, 10.55g/L of yeast powder, 2.7g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH was 6.2; sterilizing at 121 deg.C and 0.1MPa for 30min;
(ii) And (3) activation: inoculating acetobacter xylinum to a slant culture medium, and culturing in an incubator at 29 ℃ for 54h to obtain an activated strain;
(iii) Seed culture: selecting 3-ring activated strains, inoculating the strains into 50mL of seed culture medium, and culturing for 27h at 29 ℃ in a shaking incubator at 140r/min to obtain seed bacterial liquid;
(iv) Fermentation culture: inoculating the seed bacterial liquid into a fermentation culture medium according to the inoculation amount of 8wt%, and fully shaking to separate out the bacterial strains and fully disperse the bacterial strains into the fermentation culture medium; then culturing for 7d in a shaking incubator at 29 ℃ and 160r/min, and forming a bacterial cellulose membrane on the surface of a fermentation medium;
(v) Taking out the bacterial cellulose membrane, washing the bacterial cellulose membrane with water until no floccule is washed, soaking the bacterial cellulose membrane in 0.13mol/L sodium hydroxide solution at the temperature of 80 ℃ for 35min, and then washing the bacterial cellulose membrane to be neutral to obtain the bacterial cellulose.
Preparing the modified protocatechuic acid liposome by using the obtained bacterial cellulose through the following steps:
(1) Adding 3g of soybean lecithin, 1g of cholesterol, 1g of tween-80 and 0.2g of vitamin E into 60mL of absolute ethyl alcohol, and stirring to completely dissolve the soybean lecithin, the cholesterol, the tween-80 and the vitamin E to form a film uniformly;
(2) Performing rotary evaporation at 50 deg.C under reduced pressure to remove anhydrous ethanol to form film; adding 0.05mol/L phosphate buffer solution containing 200mg/L protocatechuic acid and having pH of 6.8, diluting to 100mL, and performing ultrasonic treatment for 30min to obtain crude lipid suspension;
(3) Dissolving 0.22g of citrus pectin and 0.022g of bacterial cellulose in 110mL of 0.05mol/L phosphate buffer solution with the pH value of 7.2 to obtain a polysaccharide solution;
(4) And (3) dropwise adding 100mL of the crude lipid suspension into 110mL of the polysaccharide solution, stirring and reacting at 65 ℃ for 35min, and continuously standing at the temperature for 55min to obtain the modified protocatechuic acid liposome.
The obtained modified protocatechuic acid liposome comprises the following three-layer structure from inside to outside: protocatechuic acid core, soybean lecithin/cholesterol/vitamin E composite layer, and citrus pectin/bacterial cellulose composite layer.
The method for storing and preserving the prawns in the zebra by using the obtained modified protocatechuic acid liposome comprises the following steps: soaking the Mylopsis monodon in modified protocatechuic acid liposome at 9 deg.C for 2 hr, taking out, soaking in liquid nitrogen, freezing to reduce the central temperature of Mylopsis monodon to-20 deg.C within 20min, and storing at-20 deg.C.
Test example
Samples were periodically taken during the storage of the penaeus monodon of examples 1 to 3 and comparative examples 1 to 5 to test the thiobarbituric acid value (TBARS) and the Water Holding Capacity (WHC), and the results are shown in Table 1. The TBARS value is an important index for judging the fat oxidation of aquatic products, and ketones and aldehydes generated by the fat oxidation and hydrolysis are one of important reasons for causing the quality deterioration of the aquatic products. The WHC is the capability of water retention of the muscle tissue of an aquatic product when the muscle tissue is subjected to external force, has a certain effect on the nutrition and flavor of the aquatic product, and finally influences the eating mouthfeel such as elasticity, chewiness and the like of consumers in the eating process.
TABLE 1
Comparative example 2 on the basis of comparative example 1, the protocatechuic acid solution is adopted to pretreat the zebra shrimps, and then the zebra shrimps are stored at low temperature, so that TBARS is reduced to a certain extent, and WHC is increased to a certain extent, which shows that the protocatechuic acid solution has the effects of slowing down fat oxidation and water retention of aquatic products.
Comparative example 3 on the basis of comparative example 2, protocatechuic acid is made into liposome, TBARS is obviously reduced, and WHC is obviously increased, because a soybean lecithin/cholesterol/vitamin E composite layer is coated outside the protocatechuic acid, the chemical stability of the protocatechuic acid can be improved, and the protocatechuic acid is slowly released in the process of shrimp and crab storage, so that long-acting oxidation resistance is realized, and meanwhile, a liposome system covers the surface of the spotted shrimp, and a good water retention effect can be provided.
Comparative example 4 on the basis of comparative example 3, the citrus pectin layer is coated outside the liposome, the TBARS is reduced, and the WHC is increased, because the citrus pectin layer can change the electrostatic effect and the steric hindrance effect of the liposome system, and the viscosity of the liposome is improved, so that the stability of the liposome is improved, the catechin is slowly released in the storage process to provide the antioxidation effect, and meanwhile, the water retention effect of the liposome system can be enhanced due to the improvement of the viscosity.
Comparative example 5 on the basis of comparative example 4, bacterial cellulose is doped into the citrus pectin layer to form a citrus pectin/bacterial cellulose composite layer, the TBARS is reduced, and the WHC is increased, because the bacterial cellulose has a typical microfiber network characteristic in structure, and after being crosslinked with a pectin molecular chain through a hydrogen bond, the crosslinking degree of the pectin layer can be improved, so that the viscosity of a liposome system is effectively improved, the liposome system has higher stability, and the functions of slowly releasing protocatechuic acid and retaining water can be better exerted.
Example 1 based on comparative example 5, bacterial cellulose was aminated, TBARS was decreased, and WHC was increased because, in addition to hydrogen bonding, aminated bacterial cellulose and pectin also formed more stable covalent bonding through the carbonylamino reaction between amino and carbonyl groups, and the cross-linked network structure in the citrus pectin/aminated bacterial cellulose complex layer was made more stable during frozen storage, thereby further improving the stability of liposomes and allowing them to exert the effects of sustained release of protocatechuic acid and retention of water better.
The raw materials and equipment used in the invention are common raw materials and equipment in the field if not specified; the methods used in the present invention are conventional in the art unless otherwise specified.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, alterations and equivalents of the above embodiments according to the technical spirit of the present invention are still within the protection scope of the technical solution of the present invention.
Claims (9)
1. A modified protocatechuic acid liposome for shrimp and crab storage and preservation is characterized by comprising the following three-layer structure from inside to outside: protocatechuic acid core, soybean lecithin/cholesterol/vitamin E composite layer, citrus pectin/aminated bacterial cellulose composite layer; the preparation method of the modified protocatechuic acid liposome comprises the following steps:
(1) Adding soybean lecithin, cholesterol, tween-80 and vitamin E into absolute ethyl alcohol, stirring and dissolving completely to form a uniform film;
(2) Carrying out reduced pressure rotary evaporation to remove the absolute ethyl alcohol to form a film; adding a phosphate buffer solution containing protocatechuic acid to perform constant volume membrane washing, and performing ultrasonic treatment to obtain a crude lipid suspension;
(3) Dissolving citrus pectin and aminated bacterial cellulose into a phosphate buffer solution to obtain a polysaccharide solution;
(4) And dropwise adding the crude lipid suspension into the polysaccharide solution, stirring and reacting at 60-70 ℃ for 30-40min, and continuously standing at the temperature for 50-60min to obtain the modified protocatechuic acid liposome.
2. The modified protocatechuic acid liposome for the storage and the preservation of shrimps and crabs as claimed in claim 1, wherein the preparation method of the aminated bacterial cellulose is as follows: dispersing bacterial cellulose into dimethylformamide, dropwise adding thionyl chloride, carrying out reflux reaction on the bacterial cellulose and the thionyl chloride for 4 to 5 hours at the temperature of 70 to 80 ℃, filtering, and washing to obtain chlorinated bacterial cellulose, wherein the mass of the bacterial cellulose and the thionyl chloride is 1.5 to 2.0; re-dispersing the chlorinated bacterial cellulose into dimethylformamide, adding ethylenediamine, reacting at 75-85 ℃ for 3-4 h, filtering, and washing with anhydrous acetone and water in sequence to obtain the aminated bacterial cellulose, wherein the mass volume ratio of the chlorinated bacterial cellulose to the ethylenediamine is 1.5-9.5.
3. The modified protocatechuic acid liposome for the storage and the preservation of shrimps and crabs as claimed in claim 2, wherein the preparation method of the bacterial cellulose is as follows:
(i) Activation: inoculating acetobacter xylinum to a slant culture medium, and culturing in an incubator at 28-30 ℃ for 48-60h to obtain an activated strain;
(ii) Seed culture: selecting 2 to 3 ring activated strains, inoculating the strains into 45 to 55mL seed culture medium, and culturing for 24 to 30h in a shaking incubator at 28 to 30 ℃ and 120 to 150r/min to obtain seed bacterial liquid;
(iii) Fermentation culture: inoculating the seed bacterial liquid into a fermentation culture medium according to the inoculation amount of 5-10wt%, and fully shaking to separate out the bacterial strain and fully disperse the bacterial strain into the fermentation culture medium; then culturing the mixture for 6 to 8 days in a shaking incubator at the temperature of 28 to 30 ℃ and at the speed of 140 to 170r/min, and forming a bacterial cellulose membrane on the surface of a fermentation medium;
(iv) Taking out the bacterial cellulose membrane, washing the bacterial cellulose membrane with water until no floccule exists, soaking the bacterial cellulose membrane in a sodium hydroxide solution of 0.1 to 0.15mol/L at 75 to 85 ℃ for 30 to 40min, and then washing the bacterial cellulose membrane with water until the bacterial cellulose membrane is neutral to obtain the bacterial cellulose.
4. The modified protocatechuic acid liposome for shrimp and crab storage and preservation according to claim 3, wherein the slant culture medium comprises the following components: 20g/L of glucose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate, 1g/L of citric acid, 20g/L of agar and 0.5g/L of calcium carbonate; the seed culture medium comprises the following components: 20g/L of sucrose, 5g/L of peptone, 5g/L of yeast powder, 2g/L of disodium hydrogen phosphate and 1g/L of citric acid; the fermentation medium comprises the following components: 20g/L of sucrose, 8g/L of peptone, 10.55g/L of yeast powder, 2.7g/L of disodium hydrogen phosphate and 1g/L of citric acid; the pH value of the slant culture medium, the seed culture medium and the fermentation culture medium is 6.0 to 6.5.
5. The modified protocatechuic acid liposome for the storage and preservation of shrimps and crabs as claimed in claim 1, wherein in the step (1), the mass ratio of the soybean lecithin to the cholesterol to the tween-80 to the vitamin E is 1.0 to 4.5 to 1.5.
6. The modified protocatechuic acid liposome for the storage and the preservation of shrimps and crabs as claimed in claim 1, wherein the mass concentration of protocatechuic acid in the protocatechuic acid-containing phosphate buffer in the step (2) is 100 to 300mg/L; the mass-to-volume ratio of the soybean lecithin in the step (1) to the mixed solution subjected to volume metering in the step (2) is 1.0-4.5 g.
7. The modified protocatechuic acid liposome for the storage and preservation of shrimps and crabs as claimed in claim 1, wherein in the step (3), the mass fraction of citrus pectin is 0.1 to 0.4%, and the mass fraction of bacterial cellulose is 0.01 to 0.04%.
8. The modified protocatechuic acid liposome for the storage and the preservation of shrimps and crabs as claimed in claim 1, wherein in the step (4), the volume ratio of the crude lipid suspension to the polysaccharide solution is 1.0 to 1.3.
9. The use of the modified protocatechuic acid liposome as defined in any one of claims 1 to 8 for storage and preservation of shrimps and crabs, comprising the steps of: soaking the shrimps and the crabs in the modified protocatechuic acid liposome at the temperature of 8 to 10 ℃ for 1 to 2h, taking out the shrimps and the crabs, freezing and storing at the temperature of minus 18 to minus 30 ℃.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010903406.6A CN112136874B (en) | 2020-09-01 | 2020-09-01 | Modified protocatechuic acid liposome for storage and preservation of shrimps and crabs as well as preparation method and application of modified protocatechuic acid liposome |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010903406.6A CN112136874B (en) | 2020-09-01 | 2020-09-01 | Modified protocatechuic acid liposome for storage and preservation of shrimps and crabs as well as preparation method and application of modified protocatechuic acid liposome |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112136874A CN112136874A (en) | 2020-12-29 |
| CN112136874B true CN112136874B (en) | 2022-11-01 |
Family
ID=73890504
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010903406.6A Active CN112136874B (en) | 2020-09-01 | 2020-09-01 | Modified protocatechuic acid liposome for storage and preservation of shrimps and crabs as well as preparation method and application of modified protocatechuic acid liposome |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112136874B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113412857A (en) * | 2021-06-28 | 2021-09-21 | 北京市农林科学院 | Protocatechuic acid ester-containing fruit and vegetable sterilization and fresh-keeping agent |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102276736A (en) * | 2011-07-10 | 2011-12-14 | 东华大学 | Selective amination method of bacterial cellulose |
| CN102389150A (en) * | 2011-10-09 | 2012-03-28 | 中国计量学院 | Method for preparing liposome enveloped by catechin |
| CN103610642A (en) * | 2013-12-10 | 2014-03-05 | 中国计量学院 | Lipidosome encapsulating epigallocatechin gallate and preparation method thereof |
| CN106633127A (en) * | 2016-10-18 | 2017-05-10 | 东北农业大学 | Application of bacterial cellulose-tea polyphenol composite film in food fresh keeping |
| CN108685711A (en) * | 2018-08-04 | 2018-10-23 | 东莞市联洲知识产权运营管理有限公司 | A kind of bacterial cellulose facial mask and preparation method thereof that the lipid of load essential oil is modifies |
| CN108713585A (en) * | 2017-11-21 | 2018-10-30 | 南京农业大学 | A kind of fresh-water fishes composite preservative and the preparation method and application thereof |
| CN108822323A (en) * | 2018-04-10 | 2018-11-16 | 上海应用技术大学 | A kind of preparation method with the bacteria cellulose film of moisture-keeping bacterium-restraining |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108385387A (en) * | 2018-02-11 | 2018-08-10 | 芦山华美包纱有限公司 | A kind of antimicrobial fibre cellulose fiber and production method and its application |
-
2020
- 2020-09-01 CN CN202010903406.6A patent/CN112136874B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102276736A (en) * | 2011-07-10 | 2011-12-14 | 东华大学 | Selective amination method of bacterial cellulose |
| CN102389150A (en) * | 2011-10-09 | 2012-03-28 | 中国计量学院 | Method for preparing liposome enveloped by catechin |
| CN103610642A (en) * | 2013-12-10 | 2014-03-05 | 中国计量学院 | Lipidosome encapsulating epigallocatechin gallate and preparation method thereof |
| CN106633127A (en) * | 2016-10-18 | 2017-05-10 | 东北农业大学 | Application of bacterial cellulose-tea polyphenol composite film in food fresh keeping |
| CN108713585A (en) * | 2017-11-21 | 2018-10-30 | 南京农业大学 | A kind of fresh-water fishes composite preservative and the preparation method and application thereof |
| CN108822323A (en) * | 2018-04-10 | 2018-11-16 | 上海应用技术大学 | A kind of preparation method with the bacteria cellulose film of moisture-keeping bacterium-restraining |
| CN108685711A (en) * | 2018-08-04 | 2018-10-23 | 东莞市联洲知识产权运营管理有限公司 | A kind of bacterial cellulose facial mask and preparation method thereof that the lipid of load essential oil is modifies |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112136874A (en) | 2020-12-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11612169B2 (en) | Preparation method for water-retaining nanoagent and process of treatment crustacean product by water-retaining nanoagent prepared by thereof | |
| CN102940032B (en) | Composite biological coating agent and method for using the same in fresh keeping of blueberry | |
| CN104322650B (en) | A kind of large yellow croaker coating preservation method | |
| CN103929982A (en) | Natural garden stuff enzyme beverage and preparation method therefor | |
| CN112136874B (en) | Modified protocatechuic acid liposome for storage and preservation of shrimps and crabs as well as preparation method and application of modified protocatechuic acid liposome | |
| CN108835223A (en) | A kind of complex biological preservative and its preservation method of pagrosomus fish | |
| JP2024505861A (en) | Oligogalacturonic acid polysaccharide, complex and its preparation method and use thereof | |
| CN110235935B (en) | Fresh bamboo shoot preservative and preparation method and application thereof | |
| CN114231381A (en) | Preparation method of mixed-strain compound fermented roxburgh rose fruit vinegar | |
| CN112088882B (en) | Antibacterial nanocellulose microcapsule and preparation method thereof | |
| CN111449130A (en) | Green preservative for fresh-cut Nanguo pear | |
| CN111449181A (en) | Method for preparing fermentation type composite fruit and vegetable beverage through ultrahigh pressure auxiliary enzyme treatment | |
| CN101438735A (en) | Film-coating fresh-keeping method for American red globe grape fruit ear | |
| US20230043870A1 (en) | Liquid nitrogen quick-freezing preservation method for fresh bamboo shoots | |
| CN111513227A (en) | Processing method of dragon fruit instant powder | |
| CN107691638A (en) | A kind of store method of blue or green jujube | |
| CN112390900A (en) | Method for extracting chitosan from snow crab shells | |
| CN112494347A (en) | Skin care water emulsion and preparation method thereof | |
| Pang et al. | Extraction, Separation and Purification of Seaweed Polysaccharide and its Application in Food Industry | |
| CN115005268B (en) | Composite antibacterial strawberry preservative solution and preparation method thereof | |
| CN115989828B (en) | Method for preventing and treating postharvest diseases of tomatoes by combining chitosan oligosaccharide with pichia caribbica and storing and preserving method | |
| KR19980020653A (en) | Manufacturing method of health supplement food containing calcium hydroxide as main ingredient | |
| CN107495187A (en) | A kind of method of waterishlogged Stichopus japonicus | |
| CN111671025B (en) | Sugar electrolyte beverage for relieving alcoholism and preparation method thereof | |
| CN117050586B (en) | Coating agent for preserving and fresh-keeping of picked fruits, preparation method and application |
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 |