CN113017047A - Preparation method of pickled vegetables - Google Patents

Preparation method of pickled vegetables Download PDF

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Publication number
CN113017047A
CN113017047A CN202110286596.6A CN202110286596A CN113017047A CN 113017047 A CN113017047 A CN 113017047A CN 202110286596 A CN202110286596 A CN 202110286596A CN 113017047 A CN113017047 A CN 113017047A
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China
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solution
vegetables
probiotic
modified chitosan
oligosaccharide
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邵胜荣
张铁成
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Xingtanju Hangzhou Network Technology Co ltd
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Xingtanju Hangzhou Network Technology Co ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L19/00—Products from fruits or vegetables; Preparation or treatment thereof
    • A23L19/20—Products from fruits or vegetables; Preparation or treatment thereof by pickling, e.g. sauerkraut or pickles
    • 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
    • A23B7/00—Preservation or chemical ripening of fruit or vegetables
    • A23B7/14—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10
    • A23B7/153—Preserving or ripening with chemicals not covered by groups A23B7/08 or A23B7/10 in the form of liquids or solids
    • A23B7/154—Organic compounds; Microorganisms; Enzymes
    • A23B7/155—Microorganisms; Enzymes; Antibiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
    • A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L5/00—Preparation or treatment of foods or foodstuffs, in general; Food or foodstuffs obtained thereby; Materials therefor
    • A23L5/20—Removal of unwanted matter, e.g. deodorisation or detoxification
    • A23L5/23—Removal of unwanted matter, e.g. deodorisation or detoxification by extraction with solvents
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
    • A23P10/00—Shaping or working of foodstuffs characterised by the products
    • A23P10/30—Encapsulation of particles, e.g. foodstuff additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/113—Acidophilus
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/125—Casei
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/513—Adolescentes
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/517—Bifidum
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/529—Infantis
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/531—Lactis
    • 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
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/51—Bifidobacterium
    • A23V2400/533—Longum
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation

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Abstract

The invention discloses a preparation method of pickled vegetables, which comprises the following steps: (1) slitting; (2) pickling: mixing the dried vegetables with salt, and sealing and pickling; (3) desalting and dehydrating: placing the pickled vegetables in clear water, ultrasonically cleaning, then carrying out flowing cleaning, squeezing to remove salt water, taking out and airing; (4) preparing materials: adding seasonings into the aired vegetables, mixing and stirring the seasonings, sterilizing, and then adding probiotic powder and uniformly mixing; (5) subpackaging: in N2And CO2Under the protection of mixed gasAnd (5) packaging to obtain the pickled vegetables. The method has the advantages that the vegetables are desalted after being salted by the salt, and the desalting process is controlled, so that the pickled vegetables are guaranteed to have good taste, the salt content of the pickled vegetables is reduced, and the influence on human health caused by overhigh salt content is avoided; meanwhile, the probiotic powder is added into the pickled vegetables, so that the pickled vegetables have health-care effects of maintaining intestinal health, promoting nutrient absorption, regulating immune system and the like.

Description

Preparation method of pickled vegetables
Technical Field
The invention relates to the field of food, in particular to a preparation method of pickled vegetables.
Background
The pickled vegetables are typical representatives and important components of traditional foods in China, and are vegetable products obtained by pickling or products pickled by raw materials such as sauce, salt, sugar, vinegar and the like. The pickled vegetables have unique flavors of fresh, sweet, crisp and tender, or salty, fresh, spicy and the like, have certain nutritional value, are deeply favored by the masses and become indispensable seasoning non-staple food in daily life of people.
The traditional preparation method of pickles generally comprises the steps of cleaning vegetables, slicing, pickling with high salt content, dehydrating and the like, for example, the Chinese patent document discloses 'pickled dried turnip', the publication number of which is CN105876702A, and the preparation method comprises the following steps: firstly removing pedicles of fresh radishes, cleaning, cutting into slices, and pickling with salt for 12-18 hours according to the mass ratio of the fresh radishes to the salt of 20: 1; then, airing the pickled radishes for 18-24 hours; adding salt according to the mass ratio of the fresh radish to the salt of 20:1, adding five-spice powder according to the mass ratio of the fresh radish to the five-spice powder of 20:1, mixing and kneading for 20-30 minutes; and finally, airing for 6-10 hours to obtain the pickled dried turnips.
However, the traditional pickled vegetables with high salt content are too high in salt content, and diseases such as hypertension can be caused after long-term eating, so that the traditional pickled vegetables with high salt content are not beneficial to human health. With the continuous improvement of the living standard of people, the quality requirement of people on the pickled vegetables is higher and higher, and in order to meet the requirements of consumers, the development of the pickled vegetables which are fresh, fragrant, crisp and tender, unique in flavor and have the nutrition and health care effects has important value.
Disclosure of Invention
The invention provides a preparation method of pickled vegetables, aiming at overcoming the problems that the pickled vegetables in the prior art have too high salt content, do not have other nutrition health-care effects and are not beneficial to human health after being eaten for a long time, and the salt content of the pickled vegetables is reduced while the pickled vegetables have good taste by controlling a desalting process; meanwhile, probiotics are added into the pickled vegetables, so that the pickled vegetables have the effects of maintaining intestinal health and promoting nutrient absorption, and are beneficial to human health.
In order to achieve the purpose, the invention adopts the following technical scheme:
a preparation method of pickles comprises the following steps:
(1) slitting: cleaning fresh vegetables, cutting and airing;
(2) pickling: mixing the dried vegetables with salt, and sealing and pickling;
(3) desalting and dehydrating: placing the pickled vegetables in clear water, ultrasonically cleaning for 20-30 min, then carrying out flowing cleaning for 10-20 min, squeezing out salt water, taking out and airing;
(4) preparing materials: adding seasonings into the aired vegetables, mixing and stirring the seasonings, sterilizing, and then adding probiotic powder and uniformly mixing;
(5) subpackaging: in N2And CO2And (5) subpackaging under the protection of mixed gas to obtain the pickled vegetables.
The invention carries out desalination treatment on the vegetables after being salted by salt, and reduces the salt content of the pickles while ensuring the pickles to have good taste by controlling the desalination process, thereby avoiding the influence on human health caused by overhigh salt content. Meanwhile, probiotic powder is added into the pickled vegetables, so that the pickled vegetables have health-care effects of maintaining intestinal health, promoting nutrient absorption, regulating immune system and the like, and the pickled vegetables are fresh, fragrant, crisp and tender, have unique flavor and have the nutrition and health-care effects.
The invention sterilizes the pickles before adding the probiotic powder, and adds the probiotic powder and then adds the probiotic powder into the pickles2And CO2The mixed gas is used for sub-packaging under protection, and the probiotics activity is kept, so that the breeding of microorganisms in the process of storing the pickled vegetables can be reduced, and the storage time of the pickled vegetables is effectively prolonged.
Preferably, the vegetables in step (1) are selected from one or more of radish, cucumber, mustard, asparagus lettuce and turnip cabbage; and the airing time is 20-30 h.
More preferably, the vegetable in step (1) is radish or cucumber.
Preferably, the mass ratio of the vegetables to the salt in the step (2) is 15-20: 1; the pickling time is 3-12 months.
Preferably, the airing time in the step (3) is 18-24 h.
Preferably, the seasoning in the step (4) comprises the following components in parts by weight: 5-10 parts of soy sauce, 5-10 parts of vinegar, 3-5 parts of chili powder, 3-5 parts of pepper powder, 3-5 parts of white granulated sugar, 3-5 parts of monosodium glutamate, 2-3 parts of sesame oil and 2-5 parts of vegetable oil; the mass ratio of the added seasonings to the vegetables is 1: 80-100.
Preferably, the pasteurization method in the step (4) adopts pasteurization, the sterilization temperature is 80-85 ℃, and the sterilization time is 8-10 min.
Preferably, the preparation method of the probiotic powder in the step (4) comprises the following steps:
A) adding chitosan into a mixed solvent of water and ethanol in a volume ratio of 1: 1-2, then adding epoxy chloropropane, stirring and reacting for 3-4 h at 70-75 ℃, filtering, washing and drying a product to obtain epoxy chloropropane modified chitosan; wherein the adding proportion of the chitosan, the mixed solvent and the epichlorohydrin is 1 g: 40-60 mL: 12-13 mL;
B) adding L-aspartic acid and glycine into water for dissolving, adding sodium carbonate to adjust the pH value of the solution to 10.5-11.5, then adding epoxy chloropropane modified chitosan, and reacting for 12-18 h at 75-85 ℃; then adding sodium carbonate to adjust the pH value of the solution to 12-13, continuously reacting for 6-8 h at 75-85 ℃, filtering, washing and drying the product to obtain amino acid modified chitosan, wherein the mass ratio of L-aspartic acid to glycine to epichlorohydrin modified chitosan is 4-5: 1-2: 1;
C) dissolving amino acid modified chitosan in an acetic acid-sodium acetate buffer solution with the pH value of 5.5-6.5 to obtain a modified chitosan solution;
D) mixing the modified chitosan solution with a gelatin solution, a probiotic suspension, an oligosaccharide solution and an emulsifier at 35-45 ℃, and uniformly stirring to obtain an emulsion, wherein the mass ratio of the modified chitosan to the gelatin to the probiotic suspension to the oligosaccharide to the emulsifier is 6-8: 8-10: 1: 0.1-0.5: 0.1-0.3, and the mass concentration of the modified chitosan is 6-8%; adjusting the pH of the emulsion to 4.0-4.2 by using an acetic acid solution, and stirring and reacting at a constant temperature for 20-30 min;
E) and cooling the reacted mixed solution to 4-8 ℃, adding glutaraldehyde with the volume ratio of 1: 8-10 to the emulsion, continuing to stir for reaction for 20-30 min, adjusting the pH of the system to 8-9 by using a sodium hydroxide solution, continuing to stir for 15-20 min, and filtering, washing and drying the product to obtain the probiotic powder.
The number of live bacteria is greatly reduced because probiotics are easily influenced by factors such as salt, other seasoning components, microorganisms, storage temperature and the like in the processes of storage, transportation, sale and consumption of the pickled vegetables; and most of the probiotics are inactivated under the strong acid condition of the stomach after eating, so that the number of the viable bacteria which can effectively enter and be planted in the intestinal tract is far less than the intake amount, and the health care effect of the probiotics is difficult to be really played. Therefore, the invention wraps the probiotics and the oligosaccharide in the wall material consisting of the gelatin and the amino acid modified chitosan to prepare the microcapsule powder, and the influence of external environmental factors on the activity of the probiotics in the storage process can be reduced by utilizing the protection effect of the wall material; meanwhile, the microcapsule wall material in the invention can not be dissolved in gastric acid, but can be quickly disintegrated in intestinal tracts, so that the problems of short storage period, poor gastric acid resistance and the like of probiotics are effectively solved, and the health effect of the probiotics can be effectively exerted when the microcapsule wall material is added into pickled vegetables.
The probiotic microcapsule powder is prepared by the steps A) and B) firstly, and L-aspartic acid and glycine are modified on chitosan through epoxy chloropropane to obtain amino acid modified chitosan. Because chitosan contains a large amount of amino groups, the amino groups are easy to protonate to form the chitosan with-NH in strongly acidic gastric juice (pH 1-3)3 +The polycation has good solubility, and amino groups in an intestinal tract (pH is 6-7) with the pH close to neutral are deprotonated, so that the solubility is reduced, therefore, chitosan is directly used as a wall material to wrap probiotics, and the wall material is easy to damage in gastric juice, so that the probiotics are inactivated and cannot effectively enter the intestinal tract to play a role. Therefore, the invention modifies L-aspartic acid and glycine on chitosan, introduces carboxyl into chitosan molecular chain, neutralizes cation formed by protonation of amino group by anion formed by dissociation of carboxyl, and makes net charge of amino acid modified chitosan in gastric acid environment close to zero, thereby reducing solubility; the amino group is deprotonated at higher pH in the intestinal tract, the amino acid modified chitosan is negatively charged under the dissociation action of carboxyl group, and the solubility is increased, so that the wall material cannot be dissolved in the stomach, and can be disintegrated after entering the intestinal tractAnd is beneficial to the probiotics in the core material to play a role in the intestinal tract. The invention controls the grafting amount of L-aspartic acid and glycine with different isoelectric points by adjusting the adding amount of L-aspartic acid and glycine in the step B) and the pH value of the reaction, so that the probiotic microcapsule has higher embedding rate, the wall material is not damaged in gastric acid for 2h, and the probiotic can be rapidly disintegrated and released after entering the small intestine, thereby exerting the health care effect of the probiotic.
And then, by steps D) and E), glutaraldehyde is used as a cross-linking agent, complex coacervation of amino acid modified chitosan and gelatin is utilized to form a wall material, and the probiotics and oligosaccharide are wrapped to obtain the probiotic microcapsule. Gelatin has the characteristic of amphoteric polyelectrolyte, the isoelectric point of gelatin is pH 5.0, gelatin and amino acid modified chitosan are negatively charged in neutral emulsion with pH more than 5, no reaction occurs, when the pH of the emulsion is adjusted to 4.0-4.2, gelatin is positively charged, amino acid modified chitosan is still negatively charged, due to the electrostatic adsorption effect, the gelatin and the amino acid modified chitosan are rapidly condensed, and a wall material is formed under the action of a cross-linking agent to wrap probiotics and oligosaccharide to form microcapsules.
In the probiotic microcapsule powder prepared by the invention, gelatin, chitosan and amino acid in the wall material are edible natural biological materials and are harmless to human bodies; the core material is embedded with probiotics and oligosaccharide which is an effective proliferation factor of the probiotics, can assist in improving the storage stability of the probiotics and the exertion of the effect of the probiotics in intestinal tracts, and is beneficial to improving the nutrition and health care effects of the pickled vegetables.
Preferably, the culture method of the probiotic bacterial suspension in the step D) comprises the following steps: inoculating the activated probiotics into an MRS culture medium, and carrying out anaerobic culture at 35-40 ℃ to obtain a seed solution; after the multi-generation seed liquid amplification culture, inoculating the seed liquid in a fermentation tank, and carrying out anaerobic culture in an MRS culture medium at 35-40 ℃ to the late logarithmic phase to obtain probiotic fermentation liquid; and centrifuging the fermentation liquor, collecting bacterial sludge, and washing with a phosphate buffer solution with the pH value of 6.5-7.0 to obtain the probiotic bacterial suspension.
Preferably, the mass ratio of the probiotic powder to the vegetables added in the step (4) is 1: 200-300; the probiotic is selected from one or more of Bifidobacterium lactis, Bifidobacterium adolescentis, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus casei and Bacillus coagulans.
More preferably, the probiotic is bacillus coagulans.
Preferably, the oligosaccharide in step D) is selected from one or more of fructo-oligosaccharide, xylo-oligosaccharide, galacto-oligosaccharide, malto-oligosaccharide, stachyose, raffinose, soybean oligosaccharide and isomalto-oligosaccharide.
Preferably, N in step (5)2And CO2The volume ratio of (A) to (B) is 1: 0.1-0.3.
Therefore, the invention has the following beneficial effects:
(1) the vegetables are desalted after being salted, and the desalting process is controlled, so that the pickled vegetables have good taste, the salt content of the pickled vegetables is reduced, and the influence on human health caused by overhigh salt content is avoided;
(2) sterilizing the pickles before adding the probiotic powder, and adding the probiotic powder after N2And CO2The mixed gas is used for sub-packaging under the protection of the mixed gas, so that the growth of microorganisms in the process of storing the pickled vegetables can be reduced while the activity of probiotics is maintained, and the storage time of the pickled vegetables is prolonged;
(3) the probiotics and the oligosaccharide are wrapped in the wall material consisting of gelatin and amino acid modified chitosan to prepare the probiotic powder, and the influence of external environmental factors on the activity of the probiotics in the storage process can be reduced by utilizing the protective effect of the wall material; meanwhile, the wall material can not be dissolved in gastric acid, but can be quickly disintegrated in intestinal tracts, so that the problems of short storage period, poor gastric acid resistance and the like of probiotics are effectively solved, and the health effect of the probiotics can be effectively exerted when the wall material is added into pickled vegetables.
Detailed Description
The invention is further described with reference to specific embodiments.
In the present invention, all the equipment and materials are commercially available or commonly used in the art, and the methods in the following examples are conventional in the art unless otherwise specified.
Example 1:
a preparation method of sauce pickled radish comprises the following steps:
(1) slitting: cleaning fresh radish, cutting into strips, and airing at normal temperature for 24 h;
(2) pickling: mixing the dried radish and salt according to the mass ratio of 18:1, and sealing and pickling for 9 months;
(3) desalting and dehydrating: ultrasonically cleaning pickled radish in clear water for 25min, then cleaning for 15min in a flowing manner, squeezing to remove salt water, taking out, and airing for 20 h;
(4) preparing materials: adding seasonings into the aired radish, wherein the seasonings comprise the following components in parts by weight: 8 parts of soy sauce, 8 parts of vinegar, 4 parts of chilli powder, 4 parts of pepper powder, 4 parts of white granulated sugar, 4 parts of monosodium glutamate, 2.5 parts of sesame oil and 4 parts of peanut oil; the mass ratio of the added seasoning to the radish is 1: 90; mixing and stirring materials, and performing pasteurization at 83 deg.C for 9 min; then adding probiotic powder with the mass ratio of 1:250 to the radish and uniformly mixing;
(5) subpackaging: n in a volume ratio of 1:0.22And CO2Subpackaging under the protection of mixed gas to obtain the sauce pickled radish, and refrigerating and storing at the temperature below 0 ℃.
The preparation method of the probiotic powder comprises the following steps:
A) adding chitosan into a mixed solvent of water and ethanol in a volume ratio of 1:1.5, then adding epoxy chloropropane, stirring and reacting for 3.5h at 72 ℃, filtering, washing and drying a product to obtain epoxy chloropropane modified chitosan; wherein the adding proportion of the chitosan, the mixed solvent and the epichlorohydrin is 1 g: 50mL of: 12.5 mL;
B) adding L-aspartic acid and glycine into water for dissolving, adding sodium carbonate to adjust the pH of the solution to 11.1, then adding epichlorohydrin modified chitosan, and reacting for 16h at 80 ℃; then adding sodium carbonate to adjust the pH value of the solution to 12.5, continuously reacting for 7 hours at the temperature of 80 ℃, filtering, washing and drying the product to obtain amino acid modified chitosan, wherein the mass ratio of the L-aspartic acid to the glycine to the epoxy chloropropane modified chitosan is 4.5:1.5: 1;
C) dissolving amino acid modified chitosan in acetic acid-sodium acetate buffer solution with pH of 6.0 to obtain modified chitosan solution;
D) activating bacillus coagulans, inoculating the activated bacillus coagulans into an MRS culture medium, and performing anaerobic culture at 37 ℃ to obtain a seed solution; after the multi-generation seed liquid amplification culture, inoculating the seed liquid in a fermentation tank, and carrying out anaerobic culture in an MRS culture medium at 37 ℃ to the late logarithmic phase to obtain probiotic fermentation liquid; centrifuging the fermentation liquor at 4000r/min and 4 ℃ for 10min, collecting bacterial sludge, and washing out the bacterial sludge by using a phosphate buffer solution with the pH value of 6.8 to obtain a probiotic bacterial suspension;
E) mixing the modified chitosan solution with a gelatin solution, a probiotic bacterial suspension, xylo-oligosaccharide, fructo-oligosaccharide solution and an emulsifier Tween 80 at 40 ℃, and uniformly stirring to obtain an emulsion, wherein the mass ratio of the modified chitosan to the gelatin to the probiotic bacterial suspension to the xylo-oligosaccharide to the fructo-oligosaccharide to the Tween 80 is 7:9:1:0.1:0.2:0.2, and the mass concentration of the modified chitosan is 7%; adjusting the pH of the emulsion to 4.1 by using an acetic acid solution, and stirring at constant temperature for reaction for 25 min;
F) and cooling the reacted mixed solution to 5 ℃, adding glutaraldehyde with the volume ratio of 1:9 to the emulsion, continuing to stir for reaction for 25min, adjusting the pH of the system to 8.5 by using a sodium hydroxide solution, continuing to stir for 18min, and filtering, washing and drying the product to obtain the probiotic powder.
Example 2:
a preparation method of sauce pickled radish comprises the following steps:
(1) slitting: cleaning fresh radish, cutting into strips, and air drying at normal temperature for 20 h;
(2) pickling: mixing the dried radish and salt according to the mass ratio of 15:1, and sealing and pickling for 3 months;
(3) desalting and dehydrating: ultrasonically cleaning pickled radish in clear water for 20min, then cleaning for 20min in a flowing manner, squeezing to remove salt water, taking out and airing for 24 h;
(4) preparing materials: adding seasonings into the aired radish, wherein the seasonings comprise the following components in parts by weight: 5 parts of soy sauce, 5 parts of vinegar, 3 parts of chilli powder, 3 parts of pepper powder, 3 parts of white granulated sugar, 3 parts of monosodium glutamate, 2 parts of sesame oil and 2 parts of peanut oil; the mass ratio of the added seasoning to the radish is 1: 80; mixing and stirring the materials, and then carrying out pasteurization, wherein the sterilization temperature is 80 ℃, and the sterilization time is 10 min; then adding probiotic powder with the mass ratio of 1:200 to the radish and uniformly mixing;
(5) subpackaging: in a volume ratio of 1:0.1 of N2And CO2Subpackaging under the protection of mixed gas to obtain the sauce pickled radish, and refrigerating and storing at the temperature below 0 ℃.
The preparation method of the probiotic powder comprises the following steps:
A) adding chitosan into a mixed solvent with the volume ratio of water to ethanol being 1:1, then adding epoxy chloropropane, stirring and reacting for 4 hours at 70 ℃, filtering, washing and drying a product to obtain epoxy chloropropane modified chitosan; wherein the adding proportion of the chitosan, the mixed solvent and the epichlorohydrin is 1 g: 40mL of: 12 mL;
B) adding L-aspartic acid and glycine into water for dissolving, adding sodium carbonate to adjust the pH of the solution to 10.5, then adding epichlorohydrin modified chitosan, and reacting for 18h at 75 ℃; then adding sodium carbonate to adjust the pH value of the solution to 12, continuously reacting for 8 hours at 75 ℃, filtering, washing and drying the product to obtain amino acid modified chitosan, wherein the mass ratio of the L-aspartic acid to the glycine to the epichlorohydrin modified chitosan is 4:1: 1;
C) dissolving amino acid modified chitosan in an acetic acid-sodium acetate buffer solution with the pH value of 5.5 to obtain a modified chitosan solution;
D) activating bacillus coagulans, inoculating the activated bacillus coagulans into an MRS culture medium, and performing anaerobic culture at 37 ℃ to obtain a seed solution; after the multi-generation seed liquid amplification culture, inoculating the seed liquid in a fermentation tank, and carrying out anaerobic culture in an MRS culture medium at 37 ℃ to the late logarithmic phase to obtain probiotic fermentation liquid; centrifuging the fermentation liquor at 4000r/min and 4 ℃ for 10min, collecting bacterial sludge, and washing out the bacterial sludge by using a phosphate buffer solution with the pH value of 6.5 to obtain a probiotic bacterial suspension;
E) mixing the modified chitosan solution with a gelatin solution, a probiotic suspension, a xylo-oligosaccharide solution and an emulsifier Tween 80 at 35 ℃, and uniformly stirring to obtain an emulsion, wherein the mass ratio of the modified chitosan to the gelatin to the probiotic suspension to the xylo-oligosaccharide to the Tween 80 in the emulsion is 6:8:1:0.1:0.1, and the mass concentration of the modified chitosan is 6%; adjusting the pH of the emulsion to 4.0 with acetic acid solution, stirring at constant temperature and reacting for 20 min;
F) and cooling the reacted mixed solution to 4 ℃, adding glutaraldehyde with the volume ratio of 1:8 to the emulsion, continuing to stir for reaction for 20min, adjusting the pH of the system to 8 by using a sodium hydroxide solution, continuing to stir for 15min, and filtering, washing and drying the product to obtain the probiotic powder.
Example 3:
a preparation method of sauce pickled radish comprises the following steps:
(1) slitting: cleaning fresh radish, cutting into strips, and airing at normal temperature for 30 h;
(2) pickling: mixing the dried radish and salt according to the mass ratio of 20:1, and sealing and pickling for 12 months;
(3) desalting and dehydrating: ultrasonically cleaning pickled radish in clear water for 30min, then cleaning for 10min in a flowing manner, squeezing to remove salt water, taking out and airing for 18 h;
(4) preparing materials: adding seasonings into the aired radish, wherein the seasonings comprise the following components in parts by weight: 10 parts of soy sauce, 10 parts of vinegar, 5 parts of chilli powder, 5 parts of pepper powder, 5 parts of white granulated sugar, 5 parts of monosodium glutamate, 3 parts of sesame oil and 5 parts of peanut oil; the mass ratio of the added seasoning to the radish is 1: 100; mixing and stirring materials, and performing pasteurization at 85 deg.C for 8 min; then adding probiotic powder with the mass ratio of 1:300 to the radish and uniformly mixing;
(5) subpackaging: n in a volume ratio of 1:0.32And CO2Subpackaging under the protection of mixed gas to obtain the sauce pickled radish, and refrigerating and storing at the temperature below 0 ℃.
The preparation method of the probiotic powder comprises the following steps:
A) adding chitosan into a mixed solvent with the volume ratio of water to ethanol being 1:2, then adding epoxy chloropropane, stirring and reacting for 3 hours at 75 ℃, filtering, washing and drying a product to obtain epoxy chloropropane modified chitosan; wherein the adding proportion of the chitosan, the mixed solvent and the epichlorohydrin is 1 g: 60mL of: 13 mL;
B) adding L-aspartic acid and glycine into water for dissolving, adding sodium carbonate to adjust the pH of the solution to 11.5, then adding epichlorohydrin modified chitosan, and reacting for 12 hours at 85 ℃; then adding sodium carbonate to adjust the pH value of the solution to 13, continuously reacting for 6 hours at 85 ℃, filtering, washing and drying the product to obtain amino acid modified chitosan, wherein the mass ratio of the L-aspartic acid to the glycine to the epichlorohydrin modified chitosan is 5:2: 1;
C) dissolving amino acid modified chitosan in acetic acid-sodium acetate buffer solution with pH of 6.5 to obtain modified chitosan solution;
D) activating bacillus coagulans, inoculating the activated bacillus coagulans into an MRS culture medium, and performing anaerobic culture at 37 ℃ to obtain a seed solution; after the multi-generation seed liquid amplification culture, inoculating the seed liquid in a fermentation tank, and carrying out anaerobic culture in an MRS culture medium at 37 ℃ to the late logarithmic phase to obtain probiotic fermentation liquid; centrifuging the fermentation liquor at 4000r/min and 4 ℃ for 10min, collecting bacterial sludge, and washing out the bacterial sludge by using a phosphate buffer solution with the pH value of 7.0 to obtain a probiotic bacterial suspension;
E) mixing the modified chitosan solution with a gelatin solution, a probiotic bacterial suspension, xylo-oligosaccharide, fructo-oligosaccharide solution and an emulsifier Tween 80 at 45 ℃, and uniformly stirring to obtain an emulsion, wherein the mass ratio of the modified chitosan to the gelatin to the probiotic bacterial suspension to the xylo-oligosaccharide to the fructo-oligosaccharide to the Tween 80 is 8:10:1:0.2:0.3:0.3, and the mass concentration of the modified chitosan is 8%; adjusting the pH of the emulsion to 4.2 by using an acetic acid solution, and stirring and reacting at constant temperature for 30 min;
F) and cooling the reacted mixed solution to 8 ℃, adding glutaraldehyde with the volume ratio of 1:10 to the emulsion, continuing to stir for reaction for 30min, adjusting the pH of the system to 9 by using a sodium hydroxide solution, continuing to stir for 20min, and filtering, washing and drying the product to obtain the probiotic powder.
Comparative example 1 (no modification of chitosan):
the preparation method of the probiotic powder used in comparative example 1 was:
A) dissolving chitosan in 1 wt% acetic acid solution to obtain chitosan solution;
B) activating bacillus coagulans, inoculating the activated bacillus coagulans into an MRS culture medium, and performing anaerobic culture at 37 ℃ to obtain a seed solution; after the multi-generation seed liquid amplification culture, inoculating the seed liquid in a fermentation tank, and carrying out anaerobic culture in an MRS culture medium at 37 ℃ to the late logarithmic phase to obtain probiotic fermentation liquid; centrifuging the fermentation liquor at 4000r/min and 4 ℃ for 10min, collecting bacterial sludge, and washing out the bacterial sludge by using a phosphate buffer solution with the pH value of 6.8 to obtain a probiotic bacterial suspension;
C) mixing a chitosan solution, a gelatin solution, a probiotic bacterial suspension, xylo-oligosaccharide, fructo-oligosaccharide solution and an emulsifier Tween 80 at 40 ℃, and uniformly stirring to obtain an emulsion, wherein the mass ratio of the chitosan to the gelatin to the probiotic bacterial suspension to the xylo-oligosaccharide to the fructo-oligosaccharide to the Tween 80 is 7:9:1:0.1:0.2:0.2, and the mass concentration of the chitosan is 7%; adjusting the pH of the emulsion to 6.5 by using a sodium hydroxide solution, and stirring and reacting at constant temperature for 25 min;
D) and cooling the reacted mixed solution to 5 ℃, adding glutaraldehyde with the volume ratio of 1:9 to the emulsion, continuing to stir for reaction for 25min, adjusting the pH of the system to 4.0 by using an acetic acid solution, continuing to stir for 18min, and filtering, washing and drying the product to obtain the probiotic powder.
The rest is the same as in example 1.
Comparative example 2 (modification of chitosan with L-aspartic acid only):
the preparation method of the probiotic powder used in comparative example 2 was:
A) adding chitosan into a mixed solvent of water and ethanol in a volume ratio of 1:1.5, then adding epoxy chloropropane, stirring and reacting for 3.5h at 72 ℃, filtering, washing and drying a product to obtain epoxy chloropropane modified chitosan; wherein the adding proportion of the chitosan, the mixed solvent and the epichlorohydrin is 1 g: 50mL of: 12.5 mL;
B) adding L-aspartic acid into water for dissolving, adding sodium carbonate to adjust the pH value of the solution to 12.5, reacting for 24 hours at 80 ℃, filtering, washing and drying a product to obtain amino acid modified chitosan, wherein the mass ratio of the L-aspartic acid to the epichlorohydrin modified chitosan is 6: 1;
C) dissolving amino acid modified chitosan in acetic acid-sodium acetate buffer solution with pH of 6.0 to obtain modified chitosan solution;
D) activating bacillus coagulans, inoculating the activated bacillus coagulans into an MRS culture medium, and performing anaerobic culture at 37 ℃ to obtain a seed solution; after the multi-generation seed liquid amplification culture, inoculating the seed liquid in a fermentation tank, and carrying out anaerobic culture in an MRS culture medium at 37 ℃ to the late logarithmic phase to obtain probiotic fermentation liquid; centrifuging the fermentation liquor at 4000r/min and 4 ℃ for 10min, collecting bacterial sludge, and washing out the bacterial sludge by using a phosphate buffer solution with the pH value of 6.8 to obtain a probiotic bacterial suspension;
E) mixing the modified chitosan solution with a gelatin solution, a probiotic bacterial suspension, xylo-oligosaccharide, fructo-oligosaccharide solution and an emulsifier Tween 80 at 40 ℃, and uniformly stirring to obtain an emulsion, wherein the mass ratio of the modified chitosan to the gelatin to the probiotic bacterial suspension to the xylo-oligosaccharide to the fructo-oligosaccharide to the Tween 80 is 7:9:1:0.1:0.2:0.2, and the mass concentration of the modified chitosan is 7%; adjusting the pH of the emulsion to 4.1 by using an acetic acid solution, and stirring at constant temperature for reaction for 25 min;
F) and cooling the reacted mixed solution to 5 ℃, adding glutaraldehyde with the volume ratio of 1:9 to the emulsion, continuing to stir for reaction for 25min, adjusting the pH of the system to 8.5 by using a sodium hydroxide solution, continuing to stir for 18min, and filtering, washing and drying the product to obtain the probiotic powder.
The rest is the same as in example 1.
Comparative example 3 (chitosan modified with glycine only):
the preparation method of the probiotic powder used in comparative example 3 was:
A) adding chitosan into a mixed solvent of water and ethanol in a volume ratio of 1:1.5, then adding epoxy chloropropane, stirring and reacting for 3.5h at 72 ℃, filtering, washing and drying a product to obtain epoxy chloropropane modified chitosan; wherein the adding proportion of the chitosan, the mixed solvent and the epichlorohydrin is 1 g: 50mL of: 12.5 mL;
B) adding glycine into water to dissolve, adding sodium carbonate to adjust the pH value of the solution to 11.1, then adding epoxy chloropropane modified chitosan, reacting for 23 hours at 80 ℃, filtering, washing and drying a product to obtain amino acid modified chitosan, wherein the mass ratio of the glycine to the epoxy chloropropane modified chitosan is 6: 1;
C) dissolving amino acid modified chitosan in acetic acid-sodium acetate buffer solution with pH of 6.0 to obtain modified chitosan solution;
D) activating bacillus coagulans, inoculating the activated bacillus coagulans into an MRS culture medium, and performing anaerobic culture at 37 ℃ to obtain a seed solution; after the multi-generation seed liquid amplification culture, inoculating the seed liquid in a fermentation tank, and carrying out anaerobic culture in an MRS culture medium at 37 ℃ to the late logarithmic phase to obtain probiotic fermentation liquid; centrifuging the fermentation liquor at 4000r/min and 4 ℃ for 10min, collecting bacterial sludge, and washing out the bacterial sludge by using a phosphate buffer solution with the pH value of 6.8 to obtain a probiotic bacterial suspension;
E) mixing the modified chitosan solution with a gelatin solution, a probiotic bacterial suspension, xylo-oligosaccharide, fructo-oligosaccharide solution and an emulsifier Tween 80 at 40 ℃, and uniformly stirring to obtain an emulsion, wherein the mass ratio of the modified chitosan to the gelatin to the probiotic bacterial suspension to the xylo-oligosaccharide to the fructo-oligosaccharide to the Tween 80 is 7:9:1:0.1:0.2:0.2, and the mass concentration of the modified chitosan is 7%; adjusting the pH of the emulsion to 4.1 by using an acetic acid solution, and stirring at constant temperature for reaction for 25 min;
F) and cooling the reacted mixed solution to 5 ℃, adding glutaraldehyde with the volume ratio of 1:9 to the emulsion, continuing to stir for reaction for 25min, adjusting the pH of the system to 8.5 by using a sodium hydroxide solution, continuing to stir for 18min, and filtering, washing and drying the product to obtain the probiotic powder.
The rest is the same as in example 1.
Comparative example 4 (changing the reaction conditions of amino acid and chitosan):
the preparation method of the probiotic powder used in comparative example 4 was:
A) adding chitosan into a mixed solvent of water and ethanol in a volume ratio of 1:1.5, then adding epoxy chloropropane, stirring and reacting for 3.5h at 72 ℃, filtering, washing and drying a product to obtain epoxy chloropropane modified chitosan; wherein the adding proportion of the chitosan, the mixed solvent and the epichlorohydrin is 1 g: 50mL of: 12.5 mL;
B) adding L-aspartic acid and glycine into water to dissolve, adding sodium carbonate to adjust the pH value of the solution to 11.1, then adding epoxy chloropropane modified chitosan, reacting at 80 ℃ for 23h, filtering, washing and drying the product to obtain amino acid modified chitosan, wherein the mass ratio of the L-aspartic acid to the glycine to the epoxy chloropropane modified chitosan is 4.5:1.5: 1;
C) dissolving amino acid modified chitosan in acetic acid-sodium acetate buffer solution with pH of 6.0 to obtain modified chitosan solution;
D) activating bacillus coagulans, inoculating the activated bacillus coagulans into an MRS culture medium, and performing anaerobic culture at 37 ℃ to obtain a seed solution; after the multi-generation seed liquid amplification culture, inoculating the seed liquid in a fermentation tank, and carrying out anaerobic culture in an MRS culture medium at 37 ℃ to the late logarithmic phase to obtain probiotic fermentation liquid; centrifuging the fermentation liquor at 4000r/min and 4 ℃ for 10min, collecting bacterial sludge, and washing out the bacterial sludge by using a phosphate buffer solution with the pH value of 6.8 to obtain a probiotic bacterial suspension;
E) mixing the modified chitosan solution with a gelatin solution, a probiotic bacterial suspension, xylo-oligosaccharide, fructo-oligosaccharide solution and an emulsifier Tween 80 at 40 ℃, and uniformly stirring to obtain an emulsion, wherein the mass ratio of the modified chitosan to the gelatin to the probiotic bacterial suspension to the xylo-oligosaccharide to the fructo-oligosaccharide to the Tween 80 is 7:9:1:0.1:0.2:0.2, and the mass concentration of the modified chitosan is 7%; adjusting the pH of the emulsion to 4.1 by using an acetic acid solution, and stirring at constant temperature for reaction for 25 min;
F) and cooling the reacted mixed solution to 5 ℃, adding glutaraldehyde with the volume ratio of 1:9 to the emulsion, continuing to stir for reaction for 25min, adjusting the pH of the system to 8.5 by using a sodium hydroxide solution, continuing to stir for 18min, and filtering, washing and drying the product to obtain the probiotic powder.
Comparative example 5 (no oligosaccharide added):
the preparation method of the probiotic powder used in comparative example 5 was:
A) adding chitosan into a mixed solvent of water and ethanol in a volume ratio of 1:1.5, then adding epoxy chloropropane, stirring and reacting for 3.5h at 72 ℃, filtering, washing and drying a product to obtain epoxy chloropropane modified chitosan; wherein the adding proportion of the chitosan, the mixed solvent and the epichlorohydrin is 1 g: 50mL of: 12.5 mL;
B) adding L-aspartic acid and glycine into water for dissolving, adding sodium carbonate to adjust the pH of the solution to 11.1, then adding epichlorohydrin modified chitosan, and reacting for 16h at 80 ℃; then adding sodium carbonate to adjust the pH value of the solution to 12.5, continuously reacting for 7 hours at the temperature of 80 ℃, filtering, washing and drying the product to obtain amino acid modified chitosan, wherein the mass ratio of the L-aspartic acid to the glycine to the epoxy chloropropane modified chitosan is 4.5:1.5: 1;
C) dissolving amino acid modified chitosan in acetic acid-sodium acetate buffer solution with pH of 6.0 to obtain modified chitosan solution;
D) activating bacillus coagulans, inoculating the activated bacillus coagulans into an MRS culture medium, and performing anaerobic culture at 37 ℃ to obtain a seed solution; after the multi-generation seed liquid amplification culture, inoculating the seed liquid in a fermentation tank, and carrying out anaerobic culture in an MRS culture medium at 37 ℃ to the late logarithmic phase to obtain probiotic fermentation liquid; centrifuging the fermentation liquor at 4000r/min and 4 ℃ for 10min, collecting bacterial sludge, and washing out the bacterial sludge by using a phosphate buffer solution with the pH value of 6.8 to obtain a probiotic bacterial suspension;
E) mixing the modified chitosan solution with a gelatin solution, a probiotic suspension and an emulsifier Tween 80 at 40 ℃, and uniformly stirring to obtain an emulsion, wherein the mass ratio of the modified chitosan to the gelatin to the probiotic suspension to the Tween 80 is 7:9:1:0.2, and the mass concentration of the modified chitosan is 7%; adjusting the pH of the emulsion to 4.1 by using an acetic acid solution, and stirring at constant temperature for reaction for 25 min;
F) and cooling the reacted mixed solution to 5 ℃, adding glutaraldehyde with the volume ratio of 1:9 to the emulsion, continuing to stir for reaction for 25min, adjusting the pH of the system to 8.5 by using a sodium hydroxide solution, continuing to stir for 18min, and filtering, washing and drying the product to obtain the probiotic powder.
The rest is the same as in example 1.
The embedding rate, stability, acid resistance and enteric solubility of the probiotic powder prepared in the above examples and comparative example 1 were tested, and the results are shown in tables 1 to 3.
The method for testing the embedding rate comprises the following steps: adding 0.1g of probiotic powder into 50mL of artificial intestinal juice (potassium dihydrogen phosphate 6.8g is dissolved in 500mL of water, the pH is adjusted to 6.8 by using 0.4% sodium hydroxide solution, 10g of pancreatin is dissolved in a proper amount of water, the two solutions are mixed and added with water to a constant volume of 1000mL), disintegrating for 1h in a shaking table incubator at 37 ℃ and 180rpm, and then absorbing 1mL of probiotic powder to perform constant temperature culture at 37 ℃ for 48h to determine the viable count of the bacillus coagulans. The above experiment was performed in triplicate. The encapsulation rate is the number of living bacteria in the microcapsule/the number of added living bacteria x 100%.
The stability test method comprises the following steps: weighing 10g of probiotic powder, sealing with platinum paper, placing in a constant-temperature incubator with relative humidity of 60% -65% and temperature of 37 ℃ for 3 months, taking 1 part (1g) per month, and counting viable bacteria by adopting a 10-fold dilution method.
The acid resistance test method comprises the following steps: taking 0.1g of probiotic powder, respectively placing the probiotic powder into triangular flasks containing 50mL of artificial gastric juice (hydrochloric acid 16.4mL, pepsin 10g, water is added and the volume is adjusted to 1000mL) with the pH value of 1.2, culturing for 3h in a shaking incubator at 37 ℃ and the rotating speed of 180rpm, taking the artificial gastric juice as a blank sample, sampling every hour, measuring the light transmittance at 600nm, and analyzing the acid resistance of the microcapsule according to the change of the light transmittance.
The enteric solubility determination method comprises the following steps: adding 0.1g of probiotic powder into 50mL of artificial intestinal juice with the pH value of 6.8, disintegrating for 1h in a shaking incubator at 37 ℃ and the rotation speed of 180rpm, taking the artificial intestinal juice as a blank sample, sampling every 15min, measuring the light transmittance at 600nm, and analyzing the disintegration condition of the microcapsules in the artificial intestinal juice according to the change of the light transmittance.
Table 1: and testing the embedding rate and stability of the probiotic powder.
Figure BDA0002980721710000121
Table 2: and (5) testing the acid resistance of the probiotic powder.
Figure BDA0002980721710000122
Table 3: results of the probiotic powder enteric test.
Figure BDA0002980721710000123
As can be seen from tables 1 to 3, the probiotic powder prepared by the method of the invention in examples 1 to 3 has high embedding rate and good storage stability; the light transmittance is basically kept unchanged in the process of treating in the artificial gastric juice for 3 hours, which shows that the probiotic powder is basically not disintegrated and released in the gastric juice; when the probiotic powder is treated in the artificial intestinal juice, the light transmittance is obviously reduced in the first 30min, the light transmittance is basically kept unchanged after 30min, and the observation shows that the solution does not contain solid particles after 30min, which indicates that the probiotic powder can be completely disintegrated in the artificial intestinal juice for 30min, releases probiotics and is beneficial to the effect of the probiotics.
In the comparative example 1, unmodified chitosan and gelatin are used for complex coacervation to serve as a wall material, and the light transmittance of the probiotic powder is reduced after the probiotic powder is treated in the artificial gastric juice for 1 hour, which indicates that the probiotic powder is not resistant to gastric acid when the unmodified chitosan is used as the wall material. In the comparative example 2, only L-aspartic acid is used for modifying chitosan, so that the embedding rate of the probiotic powder is low, and thalli cannot be effectively protected; this is probably because the ratio of carboxyl groups introduced by modification with only L-aspartic acid is too large, and the modified chitosan has low solubility at pH around 4 and is difficult to efficiently undergo complex coacervation and crosslinking with gelatin. In the comparative example 3, when only glycine is used for modifying chitosan, the embedding rate of the probiotic powder is also low, the gastric acid resistance is poor, and the disintegration rate in intestinal juice is reduced; it is possible that it is difficult to balance the positive charge formed by protonation of the amino group in chitosan due to insufficient carboxyl groups introduced when modified with glycine alone. In comparative example 4, conditions of the reaction of L-aspartic acid and glycine with chitosan are changed, the reaction is carried out at a pH of about 11.0, and the optimum pH of the reaction of L-aspartic acid and epichlorohydrin cannot be reached, so that the grafting rate of L-aspartic acid is low, the introduced carboxyl is insufficient, and the embedding rate and the gastric acid resistance of the probiotic powder are reduced. In contrast, the core material of the probiotic powder of comparative example 5 was not added with oligosaccharide, and the storage stability of the probiotic was significantly reduced compared to that of example 1, indicating that oligosaccharide is beneficial to improving the storage stability of the probiotic.
The sensory properties of the pickled radishes obtained in the above example were scored by randomly selecting 50 volunteers, and the final average score was taken, and the results are shown in table 4.
Table 4: and evaluating the sensory performance of the pickled radishes.
Figure BDA0002980721710000131
The pickled vegetables prepared by the method are fresh, fragrant, crisp and tender, have delicious flavor and are good in eating feeling; meanwhile, the probiotic bacteria also have the nutrition and health care effects of the probiotics, and are beneficial to the health of human bodies.

Claims (10)

1. The preparation method of the pickles is characterized by comprising the following steps:
(1) slitting: cleaning fresh vegetables, cutting and airing;
(2) pickling: mixing the dried vegetables with salt, and sealing and pickling;
(3) desalting and dehydrating: placing the pickled vegetables in clear water, ultrasonically cleaning for 20-30 min, then carrying out flowing cleaning for 10-20 min, squeezing out salt water, taking out and airing;
(4) preparing materials: adding seasonings into the aired vegetables, mixing and stirring the seasonings, sterilizing, and then adding probiotic powder and uniformly mixing;
(5) subpackaging: in N2And CO2And (5) subpackaging under the protection of mixed gas to obtain the pickled vegetables.
2. The method for preparing pickles according to claim 1, wherein the vegetables in the step (1) are selected from one or more of radish, cucumber, mustard, lettuce and turnip cabbage; and the airing time is 20-30 h.
3. The method for preparing pickles according to claim 2, wherein the vegetable in step (1) is radish or cucumber.
4. The method for preparing pickles according to claim 1, wherein the seasoning in the step (4) comprises the following components in parts by weight: 5-10 parts of soy sauce, 5-10 parts of vinegar, 3-5 parts of chili powder, 3-5 parts of pepper powder, 3-5 parts of white granulated sugar, 3-5 parts of monosodium glutamate, 2-3 parts of sesame oil and 2-5 parts of vegetable oil; the mass ratio of the added seasonings to the vegetables is 1: 80-100.
5. The method for preparing pickles according to claim 1 or 4, wherein the pasteurization in the step (4) is carried out at 80-85 ℃ for 8-10 min.
6. The method for preparing pickles according to claim 1, wherein the probiotic powder in the step (4) is prepared by the following steps:
A) adding chitosan into a mixed solvent of water and ethanol in a volume ratio of 1: 1-2, then adding epoxy chloropropane, stirring and reacting for 3-4 h at 70-75 ℃, filtering, washing and drying a product to obtain epoxy chloropropane modified chitosan; wherein the adding proportion of the chitosan, the mixed solvent and the epichlorohydrin is 1 g: 40-60 mL: 12-13 mL;
B) adding L-aspartic acid and glycine into water for dissolving, adding sodium carbonate to adjust the pH value of the solution to 10.5-11.5, then adding epoxy chloropropane modified chitosan, and reacting for 12-18 h at 75-85 ℃; then adding sodium carbonate to adjust the pH value of the solution to 12-13, continuously reacting for 6-8 h at 75-85 ℃, filtering, washing and drying the product to obtain amino acid modified chitosan, wherein the mass ratio of L-aspartic acid to glycine to epichlorohydrin modified chitosan is 4-5: 1-2: 1;
C) dissolving amino acid modified chitosan in an acetic acid-sodium acetate buffer solution with the pH value of 5.5-6.5 to obtain a modified chitosan solution;
D) mixing the modified chitosan solution with a gelatin solution, a probiotic suspension, an oligosaccharide solution and an emulsifier at 35-45 ℃, and uniformly stirring to obtain an emulsion, wherein the mass ratio of the modified chitosan to the gelatin to the probiotic suspension to the oligosaccharide to the emulsifier is 6-8: 8-10: 1: 0.1-0.5: 0.1-0.3, and the mass concentration of the modified chitosan is 6-8%; adjusting the pH of the emulsion to 4.0-4.2 by using an acetic acid solution, and stirring and reacting at a constant temperature for 20-30 min;
E) and cooling the reacted mixed solution to 4-8 ℃, adding glutaraldehyde with the volume ratio of 1: 8-10 to the emulsion, continuing to stir for reaction for 20-30 min, adjusting the pH of the system to 8-9 by using a sodium hydroxide solution, continuing to stir for 15-20 min, and filtering, washing and drying the product to obtain the probiotic powder.
7. The method for preparing the pickled vegetables according to claim 1 or 6, wherein the mass ratio of the probiotic powder to the vegetables added in the step (4) is 1: 200-300; the probiotic is selected from one or more of Bifidobacterium lactis, Bifidobacterium adolescentis, Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium longum, Lactobacillus acidophilus, Lactobacillus casei and Bacillus coagulans.
8. The method for preparing pickles according to claim 7, wherein the probiotic is Bacillus coagulans.
9. The method for preparing pickles according to claim 6, wherein the oligosaccharide in step D) is one or more selected from fructo-oligosaccharide, xylo-oligosaccharide, galacto-oligosaccharide, malto-oligosaccharide, stachyose, raffinose, soybean oligosaccharide and isomalto-oligosaccharide.
10. The method for preparing pickles according to claim 1, wherein N in the step (5)2And CO2The volume ratio of (A) to (B) is 1: 0.1-0.3.
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