NL2026059B1 - Sodium alginate chitosan microcapsule containing polylisine and nisin and preparation method therefor - Google Patents
Sodium alginate chitosan microcapsule containing polylisine and nisin and preparation method therefor Download PDFInfo
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- NL2026059B1 NL2026059B1 NL2026059A NL2026059A NL2026059B1 NL 2026059 B1 NL2026059 B1 NL 2026059B1 NL 2026059 A NL2026059 A NL 2026059A NL 2026059 A NL2026059 A NL 2026059A NL 2026059 B1 NL2026059 B1 NL 2026059B1
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- solution
- nisin
- sodium alginate
- chitosan
- polylysine
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- NVNLLIYOARQCIX-MSHCCFNRSA-N Nisin Chemical compound N1C(=O)[C@@H](CC(C)C)NC(=O)C(=C)NC(=O)[C@@H]([C@H](C)CC)NC(=O)[C@@H](NC(=O)C(=C/C)/NC(=O)[C@H](N)[C@H](C)CC)CSC[C@@H]1C(=O)N[C@@H]1C(=O)N2CCC[C@@H]2C(=O)NCC(=O)N[C@@H](C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(NCC(=O)N[C@H](C)C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCSC)C(=O)NCC(=O)N[C@H](CS[C@@H]2C)C(=O)N[C@H](CC(N)=O)C(=O)N[C@H](CCSC)C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(N[C@H](C)C(=O)N[C@@H]3C(=O)N[C@@H](C(N[C@H](CC=4NC=NC=4)C(=O)N[C@H](CS[C@@H]3C)C(=O)N[C@H](CO)C(=O)N[C@H]([C@H](C)CC)C(=O)N[C@H](CC=3NC=NC=3)C(=O)N[C@H](C(C)C)C(=O)NC(=C)C(=O)N[C@H](CCCCN)C(O)=O)=O)CS[C@@H]2C)=O)=O)CS[C@@H]1C NVNLLIYOARQCIX-MSHCCFNRSA-N 0.000 title claims abstract description 54
- 108010053775 Nisin Proteins 0.000 title claims abstract description 54
- 239000004309 nisin Substances 0.000 title claims abstract description 54
- 235000010297 nisin Nutrition 0.000 title claims abstract description 54
- 239000000661 sodium alginate Substances 0.000 title claims abstract description 54
- 229940005550 sodium alginate Drugs 0.000 title claims abstract description 54
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 title claims abstract description 50
- 235000010413 sodium alginate Nutrition 0.000 title claims abstract description 50
- 229920001661 Chitosan Polymers 0.000 title claims abstract description 41
- 239000003094 microcapsule Substances 0.000 title claims abstract description 41
- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 239000000243 solution Substances 0.000 claims abstract description 48
- 229920000656 polylysine Polymers 0.000 claims abstract description 47
- 108010039918 Polylysine Proteins 0.000 claims abstract description 46
- 239000011248 coating agent Substances 0.000 claims abstract description 18
- 238000000576 coating method Methods 0.000 claims abstract description 18
- 235000010410 calcium alginate Nutrition 0.000 claims abstract description 13
- 239000000648 calcium alginate Substances 0.000 claims abstract description 13
- 229960002681 calcium alginate Drugs 0.000 claims abstract description 13
- OKHHGHGGPDJQHR-YMOPUZKJSA-L calcium;(2s,3s,4s,5s,6r)-6-[(2r,3s,4r,5s,6r)-2-carboxy-6-[(2r,3s,4r,5s,6r)-2-carboxylato-4,5,6-trihydroxyoxan-3-yl]oxy-4,5-dihydroxyoxan-3-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylate Chemical compound [Ca+2].O[C@@H]1[C@H](O)[C@H](O)O[C@@H](C([O-])=O)[C@H]1O[C@H]1[C@@H](O)[C@@H](O)[C@H](O[C@H]2[C@H]([C@@H](O)[C@H](O)[C@H](O2)C([O-])=O)O)[C@H](C(O)=O)O1 OKHHGHGGPDJQHR-YMOPUZKJSA-L 0.000 claims abstract description 13
- 239000011259 mixed solution Substances 0.000 claims abstract description 13
- 238000002156 mixing Methods 0.000 claims abstract description 13
- 238000000034 method Methods 0.000 claims abstract description 12
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims abstract description 9
- 239000002775 capsule Substances 0.000 claims abstract description 9
- 239000001509 sodium citrate Substances 0.000 claims abstract description 9
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 claims abstract description 9
- 239000002131 composite material Substances 0.000 claims abstract description 6
- 238000005406 washing Methods 0.000 claims description 24
- 239000007864 aqueous solution Substances 0.000 claims description 18
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 12
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 12
- 229920000053 polysorbate 80 Polymers 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- 239000000839 emulsion Substances 0.000 claims description 8
- 239000011780 sodium chloride Substances 0.000 claims description 8
- 239000006228 supernatant Substances 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 5
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims description 4
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 238000001879 gelation Methods 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 4
- 239000004005 microsphere Substances 0.000 claims description 4
- 239000011734 sodium Substances 0.000 claims description 4
- 229910052708 sodium Inorganic materials 0.000 claims description 4
- 238000001291 vacuum drying Methods 0.000 claims description 4
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 2
- 230000014759 maintenance of location Effects 0.000 claims 1
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 8
- 235000013305 food Nutrition 0.000 abstract description 7
- 238000005516 engineering process Methods 0.000 abstract description 3
- 230000002335 preservative effect Effects 0.000 abstract description 3
- 102000004190 Enzymes Human genes 0.000 abstract description 2
- 108090000790 Enzymes Proteins 0.000 abstract description 2
- 238000000855 fermentation Methods 0.000 abstract description 2
- 230000004151 fermentation Effects 0.000 abstract description 2
- 235000021107 fermented food Nutrition 0.000 abstract description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 235000019249 food preservative Nutrition 0.000 description 2
- 239000005452 food preservative Substances 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- CFYIUBWVKZQDOG-UHFFFAOYSA-N 4-[[2-[[2-[[1-(4-nitroanilino)-1-oxo-3-phenylpropan-2-yl]amino]-2-oxoethyl]amino]-2-oxoethyl]amino]-4-oxobutanoic acid Chemical compound C=1C=C([N+]([O-])=O)C=CC=1NC(=O)C(NC(=O)CNC(=O)CNC(=O)CCC(=O)O)CC1=CC=CC=C1 CFYIUBWVKZQDOG-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 239000000022 bacteriostatic agent Substances 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 235000015895 biscuits Nutrition 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 235000019688 fish Nutrition 0.000 description 1
- 229920001519 homopolymer Polymers 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 125000003588 lysine group Chemical group [H]N([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 235000015067 sauces Nutrition 0.000 description 1
- 235000014102 seafood Nutrition 0.000 description 1
- 235000013555 soy sauce Nutrition 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Abstract
A sodium alginate chitosan microcapsule containing polylysine and Nisin and a preparation method therefor belong to the field of food engineering technology. The method has steps of: 5 mixing Nisin with sodium alginate, dripping a calcium chloride solution to make into a calcium alginate microcapsule, then adding a mixed solution of polylysine and chitosan for secondary coating, and then coating in a sodium alginate solution again, and liquefying a capsule core with a sodium citrate solution, so as to prepare the sodium alginate chitosan microcapsules containing polylysine and Nisin. Compared to free and single polylysine, Nisin and chitosan, 10 the microencapsulated composite antibacterial products containing polylysine, Nisin and chitosan has increased resistance to pH, temperature and enzymes, has increased preservative effect, and may be processed with food without affecting the fermentation process of fermented food.
Description
1 001618P-NL
[0001] The disclosure belongs to the field of food engineering technology, in particular, to a sodium alginate chitosan microcapsule containing polylysine and Nisin and a preparation method therefor.
[0002] Polylysine is a homopolymer containing 25-30 lysine residues, which has good bactericidal ability and thermal stability. It is currently approved for use as a preservative in foods, and is widely used in the preservation of instant rice, wet cooked noodles, cooked vegetables, seafood, sauces, soy sauce, fish fillets and biscuits. It can be broken down into lysine in the human body, which can be completely digested and absorbed by the human body. Further, lysine is one of the eight amino acids necessary for the human body to allow fortified use in foods. At the same time, it has been found that the combined use of polylysine and other natural bacteriostatic agents has a significant synergistic effect and can improve its bacteriostatic ability.
[0003] Nisin can inhibit most Gram-positive bacteria and has a strong inhibitory effect on Bacillus spores. Therefore, it 1s widely used as a food preservative in the food industry. After being consumed, it is quickly hydrolyzed into amino acids under the physiological pH conditions of the human body and the action of a-chymotrypsin, without changing the normal flora in the human intestine and producing resistance problems like other antibiotics, even without having cross-resistance with other antibiotics. So, it is a natural food preservative with high efficiency, non-toxicity, safety and no side effects.
[0004] In view of the problems in the prior art, the disclosure aims at designing a technical solution in which a sodium alginate chitosan microcapsule containing polylysine and Nisin and a preparation method therefor are provided.
[0005] The method for preparing a sodium alginate chitosan microcapsule containing
2 001618P-NL polylysine and Nisin includes process steps of: mixing Nisin with sodium alginate, dripping a calcium chloride solution to make into a calcium alginate microcapsule, then adding a mixed solution of polylysine and chitosan for secondary coating, and then coating in a sodium alginate solution again and liquefying a capsule core with a sodium citrate solution, so as to prepare the sodium alginate chitosan microcapsule containing polylysine and Nisin.
[0006] The method for preparing the sodium alginate chitosan microcapsule containing polylysine and Nisin specifically includes process steps of:
[0007] preparation of sodium alginate solution: preparing a 0.8 to 2 wt% sodium alginate aqueous solution;
[0008] preparation of Nisin sodium alginate solution: adding sodium alginate and Nisin to the sodium alginate aqueous solution in a mass ratio of 1: 5 to 1:20, mixing well and filtering;
[0009] formation of microcapsule: dripping the mixed solution in step (2) into a 1.0 to 2.0% (w/v) calcium chloride solution containing 0.1% (v/v) Tween 80 by an electrostatic droplet generator for gelation reaction with a temperature of 20 to 50°C and a standing time of to 100min, to form a Nisin calcium alginate microsphere emulsion;
[0010] washing and drying: then washing the microcapsules with a distilled aqueous solution containing 0.5 to 1.5wt% Tween-80 to collect retentates for vacuum drying or vacuum freeze drying for future use after removing un-coated Nisin and wall materials;
[0011] preparation of polylysine solution: preparing a 0.2 to 2.0 wt% polylysine 20 aqueous solution;
[0012] preparation of chitosan emulsion: dissolving chitosan to prepare a 0.2 to 2.0wt% chitosan with a 0.5 to 2% (v/v) acetic acid solution;
[0013] preparation of the mixed polylysine and chitosan solution: mixing the solution in the step (5) and the solution in the step (6) at a ratio of 1:1 to 1:10;
[0014] secondary coating: putting 1g dry Nisin calcium alginate microcapsule prepared in the step (4) into the mixed solution of 30 to 70mL in the step (7) for shaking for 10 to 100min to remove a supernatant, and washing the microcapsules 3 times with a distilled aqueous solution containing 0.5 to 1.5 wt% Tween-80;
[0015] (9) 3-times coating: putting the sodium alginate solution in the step (1) into the step (8) for shaking for 10 to 100min to remove a supernatant, and washing with saline;
3 001618P-NL
[0016] washing and drying: finally liquefying the capsule core with a 1.2 to 2.0wt% sodium citrate solution for 5 to 60min, and washing with saline, to obtain a sodium alginate- polylysine/chitosan-sodium alginate composite membrane microcapsule encapsulated with Nisin.
[0017] The sodium alginate chitosan microcapsule containing polylysine and Nisin is prepared by steps of: mixing Nisin with sodium alginate, dripping a calcium chloride solution to make into a calcium alginate microcapsule, then adding a mixed solution of polylysine and chitosan for secondary coating, and then coating in a sodium alginate solution again, and liquetying a capsule core with a sodium citrate solution, so as to prepare the sodium alginate chitosan microcapsules containing polylysine and Nisin.
[0018] Compared to free and single polylysine, Nisin and chitosan, the microencapsulated composite antibacterial products containing polylysine, Nisin and chitosan has increased resistance to pH, temperature and enzymes, has increased preservative effect, and may be processed with food without affecting the fermentation process of fermented food; meanwhile, the high temperature and high pressure during processing may also make Nisin and polylysine release rapidly, thereby removing the effect of coated wall materials on the antibacterial effect of polylysine and Nisin.
[0019] The present invention is further illustrated by the following description in conjunction with the embodiments.
[0020] Embodiment 1:
[0021] (1) preparation of sodium alginate solution: preparing a 2.0g sodium alginate dissolved in 250ml distilled water;
[0022] (2) preparation of Nisin sodium alginate solution: adding sodium alginate and Nisin to the sodium alginate aqueous solution in a mass ratio of 1: 5, mixing well and filtering;
[0023] (3) formation of microcapsule: dripping the mixed solution in step (2) into a
1.0% (w/v) calcium chloride solution containing 250uL Tween 80 by an electrostatic droplet generator for gelation reaction with a temperature of 20°C and a standing time of 20min, to
4 001618P-NL form a Nisin calcium alginate microsphere emulsion;
[0024] (4) washing and drying: then washing the microcapsules with a distilled aqueous solution containing 0.5wt% Tween-80 to collect retentates for vacuum drying or vacuum freeze drying for future use after removing un-coated Nisin and wall materials;
[0025] (5) preparation of polylysine solution: preparing a 0.2wt% polylysine aqueous solution;
[0026] (6) preparation of chitosan emulsion: dissolving chitosan to prepare a 0.2wt% chitosan with a 0.5% (v/v) acetic acid solution;
[0027] (7) preparation of the mixed polylysine and chitosan solution: mixing the solution in the step (5) and the solution in the step (6) at a ratio of 1:1;
[0028] (8) secondary coating: putting lg dry Nisin calcium alginate microcapsule prepared in the steps (3) and (4) into the mixed solution of 30 mL in the step (7) for shaking for 10min to remove a supernatant, and washing the microcapsules 3 times with a distilled aqueous solution containing 0.5 wt% Tween-80 (collecting retentates);
[0029] (9) 3-times coating: putting the sodium alginate solution in the step (1) into the step (8) for shaking for 10min to remove a supernatant, and washing with saline;
[0030] (10) washing and drying: finally liquefying the capsule core with a 1.2wt% sodium citrate solution for 5min, and washing with saline, to obtain a sodium alginate- polylysine/chitosan-sodium alginate composite membrane microcapsule encapsulated with Nisin;
[0031] (11) detection and application results: the formula results in 70% coating rate for Nisin and 60% for polylysine. After 15min of high-temperature steam sterilization, the titer is measured by the paper diffusion method; compared with free Nisin, its antibacterial activity is
1.2 times higher, and compared with free polylysine, its antibacterial activity is 1.1 times higher. It may be added with the flour at 0.05% to make Cantonese-style moon cakes, the shelf life may be extended for 30 days without affecting the making process of the moon cake.
[0032] Embodiment 2:
[0033] (1) preparation of sodium alginate solution: preparing a 1 wt% sodium alginate aqueous solution;
[0034] (2) preparation of Nisin sodium alginate solution: adding sodium alginate and
001618P-NL Nisin to the sodium alginate aqueous solution in a mass ratio of 1: 10, mixing well and filtering;
[0035] (3) formation of microcapsule: dripping the mixed solution in step (2) into a
1.5% (w/v) calcium chloride solution containing 0.1% (v/v) Tween 80 by an electrostatic droplet generator for gelation reaction with a temperature of 30°C and a standing time of 40 5 min, to form a Nisin calcium alginate microsphere emulsion;
[0036] (4) washing and drying: then washing the microcapsules with a distilled aqueous solution containing 1.0wt% Tween-80 to collect retentates for vacuum drying or vacuum freeze drying for future use after removing un-coated Nisin and wall materials;
[0037] (5) preparation of polylysine solution: preparing a 1.0 wt% polylysine aqueous solution;
[0038] (6) preparation of chitosan emulsion: dissolving chitosan to prepare a 1.0wt% chitosan with a 1% (v/v) acetic acid solution;
[0039] (7) preparation of the mixed polylysine and chitosan solution: mixing the solution in the step (5) and the solution in the step (6) at a ratio of 1:5;
[0040] (8) secondary coating: putting lg dry Nisin calcium alginate microcapsule prepared in the steps (3) and (4) into the mixed solution of 50 mL in the step (7) for shaking for 60 min to remove a supernatant, and washing the microcapsules 3 times with a distilled aqueous solution containing 1.0 wt% Tween-80 (collecting retentates);
[0041] (9) 3-times coating: putting the sodium alginate solution in the step (1) into the step (8) for shaking for 40 min to remove a supernatant, and washing with saline;
[0042] (10) washing and drying: finally liquefying the capsule core with a 1.6 wt% sodium citrate solution for 20 min, and washing with saline, to obtain a sodium alginate- polylysine/chitosan-sodium alginate composite membrane microcapsule encapsulated with Nisin;
[0043] (11) detection and application results: the formula results in 80% coating rate for Nisin and 70% for polylysine. After 15min of high-temperature steam sterilization, the titer is measured by the paper diffusion method; compared with free Nisin, its antibacterial activity is
1.8 times higher, and compared with free polylysine, its antibacterial activity is 1.5 times higher. It may be added with the flour at 0.03% to make Cantonese-style moon cakes, the shelf life may be extended for 20 days without affecting the making process of the moon cake.
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| NL2026059A NL2026059B1 (en) | 2020-07-15 | 2020-07-15 | Sodium alginate chitosan microcapsule containing polylisine and nisin and preparation method therefor |
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| NL2026059B1 true NL2026059B1 (en) | 2021-05-17 |
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