CN-121652971-B - Salt-containing probiotic composition and preparation method thereof

CN121652971BCN 121652971 BCN121652971 BCN 121652971BCN-121652971-B

Abstract

The invention relates to the technical field of microbial preparations, in particular to a salt-containing probiotic composition and a preparation method thereof, wherein the salt-containing probiotic composition comprises refined dyers woad leaf salt, modified carboxymethyl chitosan, lactic acid-glucolactone compound acidulant, sodium alginate, calcium chloride, lactobacillus plantarum freeze-dried bacterial powder and trehalose; the hydrogen peroxide is used to oxidize Fe 2+ to Fe 3+ in the dyer's salt, and the citric acid chelates Fe 3+ and Cu 2+ . Tannic acid replaces Fe 3+ , and a coordination reaction occurs, so that a metal-polyphenol network is formed on the surface of the probiotics, and physical barrier protection is provided. The lactic acid-glucolactone compound acidulant gradually reduces the pH value, and tannic acid orderly captures Fe 3+ and MPNs to uniformly grow. The sodium alginate-Ca 2+ gel network and the modified carboxymethyl chitosan form a composite gel network, and the double-layer protection of the sodium alginate-Ca 2+ gel network and the MPNs network obviously improves the survival rate and the functional activity of probiotics.

Inventors

YUAN SU
CAO PING
FANG YONG
ZHE BO

Assignees

内蒙古蒙盐科技有限责任公司

Dates

Publication Date: 20260505
Application Date: 20260204

Claims (4)

1. The salt-containing probiotic composition is characterized by comprising, by weight, 2-5 parts of refined dyers woad salt, 8-15 parts of modified carboxymethyl chitosan, 1.5-3 parts of lactic acid-glucolactone compound acidulant, 1.5-3 parts of sodium alginate, 0.5-1.5 parts of calcium chloride, 5-15 parts of lactobacillus plantarum freeze-dried bacterial powder and 2-5 parts of trehalose; The refined dyer salt is treated by hydrogen peroxide to oxidize Fe 2+ into Fe 3+ , citric acid is combined with free Fe 3+ and Cu 2+ , and finally stearic acid is used for hydrophobic passivation treatment; The modified carboxymethyl chitosan is modified by tannic acid and gallic acid together; the refined dyers woad salt and the modified carboxymethyl chitosan are bridged by Fe 3+ to form a metal-polyphenol network, and the metal-polyphenol network and the sodium alginate-calcium chloride gel network form a double-layer protection structure together; the lactic acid-glucolactone compound acidulant is used for controlling the release rate of Fe 3+ so as to form a uniform MPNs network; the preparation method of the refined dyers woad salt comprises the following steps: s11, loading the coarse powder of the dyers woad salt into a ball milling tank, adding absolute ethyl alcohol and grinding balls, and grinding and crushing to obtain dyers woad salt slurry; s12, adding hydrogen peroxide into absolute ethyl alcohol, slowly dropwise adding the obtained solution into the dyer woad salt slurry, and magnetically stirring to obtain a first compound; s13, dissolving citric acid in absolute ethyl alcohol, slowly dropwise adding the obtained citric acid solution into the first compound, and continuously stirring to obtain a second compound; s14, dissolving stearic acid in absolute ethyl alcohol, adding the obtained stearic acid solution into a second compound, continuously stirring, concentrating the obtained slurry by vacuum rotary evaporation, vacuum drying, grinding and dispersing to obtain refined dyers woad leaf salt; The preparation method of the modified carboxymethyl chitosan comprises the following steps: S21, dissolving carboxymethyl chitosan in deionized water, and regulating the pH value by using HCl to obtain carboxymethyl chitosan solution; S22, dissolving gallic acid in deionized water, adding EDC, HCl and NHS, stirring and activating to obtain a gallic acid solution, slowly dripping the gallic acid solution into carboxymethyl chitosan solution, regulating pH with NaOH, stirring and reacting to obtain a reaction solution, dialyzing and purifying to obtain gallic acid-carboxymethyl chitosan; S23, dissolving tannic acid in deionized water, regulating the pH value by using NaOH, adding the obtained tannic acid solution into gallic acid-carboxymethyl chitosan, stirring for reaction, adding the obtained reaction solution into absolute ethyl alcohol for precipitation, centrifuging, collecting the precipitate, washing with absolute ethyl alcohol, re-dissolving in deionized water, and freeze-drying to obtain modified carboxymethyl chitosan; The mass ratio of the carboxymethyl chitosan to the gallic acid to the tannic acid is 1:0.3-0.6:0.5-1.5.
2. The salt-containing probiotic composition according to claim 1, wherein the mass ratio of the dyer woad salt coarse powder to the citric acid is 100:5-8.
3. A salt-containing probiotic composition according to claim 1, characterized in that the method for preparing the lactic acid-gluconolactone complex acidulant comprises the steps of: s31, dissolving sodium lactate in deionized water, adding lactic acid, uniformly stirring, and regulating the pH value by using NaOH to obtain a lactic acid-sodium lactate buffer solution; S32, dissolving the glucolactone in deionized water, adding the deionized water into the lactic acid-sodium lactate buffer solution, and uniformly stirring to obtain the lactic acid-glucolactone compound acidifier.
4. A method for preparing a salt-containing probiotic composition, applied to preparing a salt-containing probiotic composition according to any one of claims 1 to 3, characterized in that the method comprises the following steps: S1, dispersing lactobacillus plantarum freeze-dried powder into PBS buffer solution to obtain probiotic suspension; S2, dispersing refined dyers woad salt and modified carboxymethyl chitosan in deionized water respectively, sequentially adding the deionized water into a probiotic suspension, and uniformly mixing to obtain a first mixture; S3, slowly adding the lactic acid-glucolactone compound acidulant into the first mixture, uniformly stirring, and standing for reaction to obtain a second mixture; S4, adding the sodium alginate solution into the second mixture, uniformly stirring, dropwise adding the obtained mixed solution into the CaCl 2 solution, carrying out curing reaction, washing the obtained product with deionized water, adding trehalose, and freeze-drying to obtain the salt-containing probiotic composition.

Description

Salt-containing probiotic composition and preparation method thereof Technical Field The invention belongs to the technical field of microbial preparations, and particularly relates to a salt-containing probiotic composition and a preparation method thereof. Background Probiotics are active microorganisms beneficial to hosts and have been widely used in the fields of foods, health products and medicines. The dyers woad leaf salt is taken as a natural mineral salt, is rich in various microelements, has unique medicinal value and health care function, can be taken as a probiotic reinforcing agent, and can promote the growth of microorganisms and enhance the stability of cell membranes. However, the traditional dyers 'salt has larger particle size, is difficult to form effective synergistic effect with probiotics on a microscopic level, hydrogen peroxide may be generated by metabolism of the probiotics, and transition metal ions (such as Fe 2+、Cu2+ and the like) contained in the dyers' salt undergo Fenton reaction in a free state to generate active oxygen, so that oxidation damage is caused to cell membranes of the probiotics, and the survival rate of the probiotics is reduced sharply. Disclosure of Invention (1) Technical problem to be solved The invention aims to provide a salt-containing probiotic composition and a preparation method thereof, which aim to solve the problem that the Fenton reaction of transition metal ions in dyers woad salt damages the cell membrane of the probiotic. (2) Technical proposal In order to achieve the aim, on the one hand, the invention provides a salt-containing probiotic composition, which comprises, by weight, 2-5 parts of refined dyers woad salt, 8-15 parts of modified carboxymethyl chitosan, 1.5-3 parts of lactic acid-glucolactone compound acidulant, 1.5-3 parts of sodium alginate, 0.5-1.5 parts of calcium chloride, 5-15 parts of lactobacillus plantarum freeze-dried bacterial powder and 2-5 parts of trehalose; The refined dyer salt is treated by hydrogen peroxide to oxidize Fe 2+ into Fe 3+, citric acid is combined with free Fe 3+ and Cu 2+, and finally stearic acid is used for hydrophobic passivation treatment; the modified carboxymethyl chitosan is modified by tannic acid and gallic acid together. Further, the preparation method of the refined dyers woad salt comprises the following steps: s11, loading the coarse powder of the dyers woad salt into a ball milling tank, adding absolute ethyl alcohol and grinding balls, and grinding and crushing to obtain dyers woad salt slurry; s12, adding hydrogen peroxide into absolute ethyl alcohol, slowly dropwise adding the obtained solution into the dyer woad salt slurry, and magnetically stirring to obtain a first compound; s13, dissolving citric acid in absolute ethyl alcohol, slowly dropwise adding the obtained citric acid solution into the first compound, and continuously stirring to obtain a second compound; S14, dissolving stearic acid in absolute ethyl alcohol, adding the obtained stearic acid solution into the second compound, continuously stirring, concentrating the obtained slurry by vacuum rotary evaporation, vacuum drying, grinding and dispersing to obtain the refined dyers woad leaf salt. Further, the mass ratio of the dyer woad salt coarse powder to the citric acid is 100:5-8. Further, the preparation method of the modified carboxymethyl chitosan comprises the following steps: S21, dissolving carboxymethyl chitosan in deionized water, and regulating the pH value by using HCl to obtain carboxymethyl chitosan solution; S22, dissolving gallic acid in deionized water, adding EDC, HCl and NHS, stirring and activating to obtain a gallic acid solution, slowly dripping the gallic acid solution into carboxymethyl chitosan solution, regulating pH with NaOH, stirring and reacting to obtain a reaction solution, dialyzing and purifying to obtain gallic acid-carboxymethyl chitosan; s23, dissolving tannic acid in deionized water, regulating the pH value by using NaOH, adding the obtained tannic acid solution into gallic acid-carboxymethyl chitosan, stirring for reaction, adding the obtained reaction solution into absolute ethyl alcohol for precipitation, centrifuging, collecting the precipitate, washing with absolute ethyl alcohol, redissolving with deionized water, and freeze-drying to obtain the modified carboxymethyl chitosan. Further, the mass ratio of the carboxymethyl chitosan to the gallic acid to the tannic acid is 1:0.3-0.6:0.5-1.5. Further, the preparation method of the lactic acid-glucolactone compound acidulant comprises the following steps: s31, dissolving sodium lactate in deionized water, adding lactic acid, uniformly stirring, and regulating the pH value by using NaOH to obtain a lactic acid-sodium lactate buffer solution; S32, dissolving the glucolactone in deionized water, adding the deionized water into the lactic acid-sodium lactate buffer solution, and uniformly stirring to obtain the lactic acid-glucolactone compo