CN113995128A - Preparation method of citrus lactic acid bacteria capsule with high flavone utilization degree - Google Patents

Preparation method of citrus lactic acid bacteria capsule with high flavone utilization degree Download PDF

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CN113995128A
CN113995128A CN202111357942.1A CN202111357942A CN113995128A CN 113995128 A CN113995128 A CN 113995128A CN 202111357942 A CN202111357942 A CN 202111357942A CN 113995128 A CN113995128 A CN 113995128A
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citrus
lactic acid
acid bacteria
fermentation
orange peel
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曹雪丹
方修贵
赵凯
王天玉
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Zhejiang Citrus Research Institute
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Priority to PCT/CN2022/113880 priority patent/WO2023087822A1/en
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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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • 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
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/065—Microorganisms
    • 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

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Abstract

The invention belongs to the technical field of citrus processing, and relates to a preparation method of citrus lactic acid bacteria capsules with high flavone utilization degree, which comprises the following steps: (1) processing raw materials, grinding dried orange peel raw materials, adding soaked rice, stirring, steaming in water, and cooling; (2) adding yeast for saccharification, namely adding sweet distiller yeast into the orange peel raw material cooled to the saccharification temperature while the orange peel raw material is hot, uniformly stirring, and saccharifying at constant temperature; (3) heating to inactivate enzyme, heating the citrus peel saccharide to above 65 deg.C after saccharification, inactivating enzyme, and cooling; (4) adding lactobacillus, adding the cooled mandarin orange peel saccharide into the activated lactobacillus in the RMS liquid culture medium, stirring, and semi-solid fermenting at constant temperature under sealed state; (5) squeezing and separating; (6) and (5) post-treatment. The preparation method of the citrus lactic acid bacteria capsule with high flavone utilization degree provided by the invention improves the bioavailability of citrus flavone products and also gives consideration to the health care effect of intestinal tracts.

Description

Preparation method of citrus lactic acid bacteria capsule with high flavone utilization degree
Technical Field
The invention belongs to the technical field of citrus processing, and relates to a preparation method of citrus lactobacillus capsules with high flavone utilization degree.
Background
Citrus is the first fruit in the world and produces about 1.46 million t annually. It is not only sour, sweet and tasty and rich in nutrition, but also is one of the important sources of human dietary flavone. The citrus peel, a byproduct of the citrus processing industry, accounts for about 1/4 of the total weight of citrus fruit, and has a much higher flavone content than fruit juice. However, the citrus peels are bitter and astringent in taste and are difficult to eat directly, and except for a small amount of citrus peels used as seasonings (such as thirteen spices) and pastry fillings (such as dried orange peel moon cakes), most citrus peels are abandoned, so that the resource waste is caused.
Flavone is one of the most important active ingredients in citrus, and has various physiological activities of dilating coronary artery, increasing coronary blood flow, reducing blood viscosity, improving or inhibiting obesity, resisting inflammation, relieving cough, relieving pain, etc. However, most of the natural flavone exists in a glucoside form, is difficult to be absorbed by small intestinal cells and is not easy to be utilized by organisms. Researches show that only less than 30 percent of the main flavonoid compound in citrus, namely hesperidin, can be absorbed by small intestines after the hesperidin is taken into a human body.
The lactobacillus is a microorganism beneficial to the life health of a host, and has the functions of protecting the host mucous membrane, enhancing the immunologic function, promoting the nutrient absorption, keeping the intestinal tract healthy and the like. It has been found that lactic acid bacteria can improve common metabolic diseases such as obesity, high cholesterol, hypertension and hyperglycemia by regulating the balance of the bacterial flora in the intestinal tract.
The current technical status is as follows:
the citrus flavonoid compound is mainly extracted and purified by using citrus peel residues, young fruits and the like in industry, is used as a medicine or health product additive in a raw material form, has a complex extraction process and relates to various organic solvents, and has large equipment investment and low environmental friendliness.
The citrus lactic acid bacteria products in the market are mostly prepared by taking fruit and vegetable juice such as citrus as a raw material and adding probiotics for fermentation (or non-fermentation) blending, the number of viable bacteria in the sterilized products is almost zero, and the cold chain transportation cost of the non-sterilized products is high.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a preparation method of citrus lactic acid bacteria capsules with high flavone utilization degree, which improves the bioavailability of citrus flavone products and gives consideration to the health care effect of intestinal tracts.
In order to solve the above technical problems, the object of the present invention is achieved by the following technical solutions:
a preparation method of citrus lactic acid bacteria capsules with high flavone utilization degree comprises the following steps:
(1) processing raw materials, grinding dried orange peel raw materials, adding soaked rice, stirring, steaming in water, and cooling; preferably, the water content of the citrus peel raw material is less than 10%; soaking the rice in water, wherein the mass ratio of the water to the rice is 0.5-2: 1; the steaming time is 10-30 min. Preferably, the citrus peel raw material is ground and sieved by using a sieve with 60-100 meshes, and the citrus peel raw material can be citrus peel, can also be citrus young fruit slices, and can also be a mixture of the citrus peel raw material and the citrus young fruit slices.
(2) Adding yeast for saccharification, namely adding sweet distiller yeast into the orange peel raw material cooled to the saccharification temperature while the orange peel raw material is hot, uniformly stirring, and saccharifying at constant temperature; preferably, the saccharification temperature is 35 +/-5 ℃; the addition amount of the sweet distiller's yeast is 2-5% of the total mass of the raw materials.
(3) Heating to inactivate enzyme, heating the citrus peel saccharide to above 65 deg.C after saccharification, inactivating enzyme, and cooling; preferably, the enzyme deactivation time is 5-15 min; the content of soluble solid in the saccharified liquid is more than 8 percent.
(4) Adding lactobacillus, adding the cooled mandarin orange peel saccharide into the activated lactobacillus in the RMS liquid culture medium, stirring, and semi-solid fermenting at constant temperature under sealed state; preferably, the number of lactic acid bacteria in the RMS liquid medium is > 5.0log CFU/g; the constant-temperature fermentation condition is 30 +/-5 ℃, and the fermentation time is 3-7 d; the room temperature referred to in the present invention is 20. + -. 5 ℃.
(5) Squeezing and separating, squeezing the fermented citrus peel, and respectively collecting fermentation liquor and fermentation residues for later use.
(6) Post-treatment, namely respectively treating the fermentation liquor and the fermentation residues, mixing and preparing into capsules.
The step (6) further comprises:
(6a) freeze-drying and crushing, freeze-drying the fermentation residues, crushing the dried fermentation residues by using a roller crusher, and collecting fermentation residue powder.
(6b) Spray drying, spray drying the fermentation liquor, and collecting fermentation liquor powder.
(6c) Mixing and filling, namely fully mixing the fermentation residue powder and the fermentation liquid powder, sieving and filling into capsules; sieving with a 60-100 mesh sieve.
Compared with the prior art, the invention has the following beneficial effects:
1. the invention avoids the use of organic solvent in the process of extracting the flavonoid compound in the whole process. The rice starch saccharification and the slight alcohol fermentation are carried out under the action of microorganisms such as rhizopus, mucor and saccharomycetes and amylase and pectinase generated by the microorganisms in the distiller's yeast, the decomposition of macromolecules such as pectin and cellulose in orange peel is accelerated, the system viscosity is reduced, the release of flavone is promoted, and the biotransformation efficiency of the subsequent probiotic fermentation process on the flavone is improved.
2. The invention greatly improves the bioavailability of the flavonoid compounds in organisms. In the proliferation process of probiotics, flavonoid glycoside bonds are exclusively hydrolyzed by beta-glucosidase, alpha-rhamnosidase and the like generated by metabolism, complex biotransformations such as deglycosylation, demethylation and the like are carried out on the flavonoid, and the generated micromolecule flavonoid aglycone, glucoside and phenolic acid substances are easier to be absorbed and utilized by organisms, thereby promoting the effective exertion of the bioactivity of the flavonoid.
3. The invention reasonably enriches the probiotic content and fermentation flavor of the citrus flavonoid capsule. The active lactobacillus is continuously proliferated under the semi-solid fermentation condition, so that the quantity of probiotics is improved to a great extent, and a large amount of flavor substances are accumulated in the fermentation process. By adopting the technical scheme of separating fermentation residues from liquid and freeze-drying at low temperature, the viable bacteria quantity in the citrus probiotic capsule is ensured, the unique fermentation flavor of the product is endowed, and the homogenization tendency of common citrus flavonoid products in the market is avoided.
4. The invention makes full use of other components in the raw materials such as orange peel, rice and the like. Under the action of enzyme, the citrus peel pectin is hydrolyzed to generate a large amount of low molecular pectin, galacturonic acid and other monosaccharides, the low molecular pectin has various pharmacological effects of resisting tumors and aging, improving the immune function and the like, and finally the low molecular pectin and the galacturonic acid are dried and crushed to be retained in a citrus probiotic capsule product. The latter, like the rice saccharified product, can provide a carbon source for the proliferation and fermentation of microorganisms, and ensure the normal metabolism and proliferation of microorganisms such as yeast, lactic acid bacteria and the like. Meanwhile, cellulose in the orange peel is partially depolymerized and then changed from long-chain molecules to short-chain molecules, so that a good supporting material is provided for spray drying of fermentation liquor.
Drawings
FIG. 1 is a process flow diagram of the present invention;
Detailed Description
The present invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. Further, modifications to the invention, as would be obvious to those of ordinary skill in the art, are intended to be included within the scope of the present invention as defined by the following claims and their equivalents.
Example 1
Weighing 1000g of fresh orange peel, placing the orange peel in a constant-temperature air-blowing drying oven, drying the orange peel by hot air at 70 ℃ for 4h to measure the water content to be 8.4%, rolling the orange peel by using a roller type crusher, and sieving the orange peel by using a 60-mesh sieve to obtain 300g of orange peel powder for later use.
Weighing 100g of rice, cleaning, adding 100g of water, soaking overnight, mixing with the orange peel powder, steaming in a pot for 15min with water, cooling to below 40 ℃, adding 10g of sweet distiller's yeast while hot, stirring uniformly, and placing in a water bath at 35 ℃ for heat preservation and saccharification.
Saccharifying for 8-12 h, heating to 70 ℃, inactivating enzyme, sterilizing for 5min, cooling to room temperature (20 +/-5 ℃), adding 50g of lactobacillus acidophilus L.acidophilus LA85 culture solution which is activated in MRS liquid culture medium, and measuring the viable count of the lactobacillus acidophilus L.acidophilus LA85 to reach 6.2log CFU/g.
Covering and standing for fermentation for 5d at constant temperature of 30 ℃, wrapping with 100-mesh gauze, squeezing and separating, and respectively collecting 430g of fermentation residues and 220g of fermentation liquor for later use.
The fermentation residue is laid in a freezing tray, the thickness is about 0.5-0.8 cm, the fermentation residue is placed in a cold trap of a freeze dryer to be rapidly frozen to-40 ℃ for about 8 hours, then the fermentation residue is subjected to vacuum freeze drying until the water content is less than 5%, and the fermentation residue is crushed and sieved by a 100-mesh sieve to obtain 131g of fermentation residue freeze-dried powder for later use.
And transferring the fermentation liquor into a spray dryer, and adjusting the temperature to 150-220 ℃ to obtain 47g of fermentation liquor spray-dried powder for later use.
Mixing the two dry powders, encapsulating, packaging, and inspecting to obtain the final product.
Example 2
Weighing 1000g of fresh orange peel, placing the orange peel in a constant-temperature air-blast drying oven, drying the orange peel by hot air at 65 ℃ for 6h to measure the water content to be 7.6%, rolling the orange peel by using a roller type crusher, and sieving the orange peel by using a 60-mesh sieve to obtain 315g of orange peel powder for later use.
Weighing 95g of rice, cleaning, adding 95g of water, soaking overnight, mixing with the orange peel powder, steaming in a pot for 15min with water, cooling to below 40 ℃, adding 10.1g of sweet distiller's yeast while hot, stirring uniformly, and saccharifying in a water bath at 35 ℃.
Saccharifying for 8-12 h, heating to 70 ℃, inactivating enzyme, sterilizing for 5min, cooling to room temperature (20 +/-5 ℃), adding 50g of lactobacillus plantarum L.plantarum N13 culture solution activated in MRS liquid culture medium, and measuring the viable count to 5.1log CFU/g.
Covering and standing for fermentation for 7d at the constant temperature of 35 ℃, wrapping with 100-mesh gauze, squeezing and separating, and respectively collecting 390g of fermentation residues and 230g of fermentation liquor for later use.
The fermentation residue is laid in a freezing tray, the thickness is about 0.5-0.8 cm, the fermentation residue is placed in a cold trap of a freeze dryer to be rapidly frozen to-40 ℃ for about 8 hours, then the fermentation residue is subjected to vacuum freeze drying until the water content is less than 5%, and the fermentation residue is crushed and sieved by a 100-mesh sieve to obtain 108g of fermentation residue freeze-dried powder for later use.
And transferring the fermentation liquor into a spray dryer, and adjusting the temperature to 150-220 ℃ to obtain 41g of fermentation liquor spray dry powder for later use.
Mixing the two dry powders, encapsulating, packaging, and inspecting to obtain the final product.
Example 3
10kg of dried orange peel and 10kg of dried young orange slices are respectively ground by a roller type grinder and sieved by a 60-mesh sieve to obtain 19kg of orange peel powder, and a little sample is taken and stored in a sealed and frozen state at (-18 ℃) for testing.
Weighing 3kg of rice, cleaning, adding 3kg of water, soaking overnight, mixing with the orange peel powder, steaming in a pot with water for 30min, cooling to below 40 ℃, adding 0.5kg of sweet distiller's yeast while hot, stirring, and saccharifying at 35 ℃ in a sandwich tank.
Saccharifying for 18-24 h, heating to 70 ℃, inactivating enzyme, sterilizing for 10min, cooling to 35 ℃, adding 0.5kg of MRS liquid culture medium activating solution of Lactobacillus brevis LB01, and measuring the viable count of the strain to 5.3log CFU/g.
And (3) covering, standing and fermenting for 3-7 days at the constant temperature of 32-35 ℃, separating by using a screw squeezer, and respectively collecting 20.5kg of fermentation residues and 10.2kg of fermentation liquor for later use. Taking a little sample, sealing, freezing (-18 ℃) and storing to be tested.
Drying the reserved sample to constant weight, precisely weighing 0.1G, putting the weighed sample into a 50mL measuring flask, adding about 25mL of methanol, carrying out ultrasonic extraction for 3min, adding methanol to scale, shaking up, filtering by using a 0.22-micrometer microporous filter membrane, and carrying out qualitative analysis by adopting a G6500 type Agilent liquid-mass combination-flight time mass spectrometer: chromatography column Waters BEH C18(2.1 mm. times.10 mm, 1.7 μm); the column temperature is 25 ℃; the flow rate is 0.3 mL/min; the sample size was 5. mu.L. Mobile phase a 0.1% acetic acid solution; mobile phase B acetonitrile; gradient elution (A: 0-5 min, 95-80%, 5-10 min, 80-65%, 10-15 min, 65-40%, 15-18 min, 40% -5%, 18-18.5 min, 5-95%, 18.5-20 min, 95%); ion source ESI-(ii) a Capillary voltage 2500V; the gas flow rate is 10L/min; the ion source temperature is 350 ℃; collision energy 30 eV; collision diffusion energy 20 eV; the ion range was scanned at 100-1500 m/z. Quantitative analysis is carried out by adopting a 1200 model Agilent high performance liquid chromatograph: detector DAD, wavelength 200-. Mobile phase A0.1% formic acid solution, mobile phase B acetonitrile, gradient elution (A: 0-12 min, 95-80%, 12-22 min, 80-65%, 22-36 min, 65-40%, 36-41 min, 40%, 41-42 min, 0-95%, 42-46 min, 95%). The content of partial flavonoids and the content of conversion products were varied as shown in Table 1.
The fermentation residue is laid in a freezing tray, the thickness is about 1.0-1.5 cm, the fermentation residue is placed in a cold trap of a freeze dryer to be rapidly frozen to-60 ℃ for about 4 hours, then the fermentation residue is subjected to vacuum freeze drying until the water content is less than 5%, and the fermentation residue is crushed and sieved by a 100-mesh sieve to obtain 7.3kg of fermentation residue freeze-dried powder for later use.
And (3) screening the fermentation liquor by a vibrating screen of 80 meshes, transferring the fermentation liquor into a spray dryer, adjusting the temperature to 150-220 ℃ to obtain 2.7kg of fermentation liquor spray-dried powder for later use. (about 0.9kg of filter residue is collected at the position of the vibrating screen, MRS liquid culture medium is added back, and the filter residue can be used for preparing probiotic activated liquid)
Mixing the two dry powders, encapsulating, packaging, and inspecting to obtain the final product.
TABLE 1 changes in the content of flavonoids before and after fermentation of lactic acid bacteria
Figure BDA0003358014630000081

Claims (7)

1. A preparation method of citrus lactic acid bacteria capsules with high flavone utilization degree is characterized by comprising the following steps:
(1) processing raw materials, grinding dried orange peel raw materials, adding soaked rice, stirring, steaming in water, and cooling;
(2) adding yeast for saccharification, namely adding sweet distiller yeast into the orange peel raw material cooled to the saccharification temperature while the orange peel raw material is hot, uniformly stirring, and saccharifying at constant temperature;
(3) heating to inactivate enzyme, heating the citrus peel saccharide to above 65 deg.C after saccharification, inactivating enzyme, and cooling;
(4) adding lactobacillus, adding the cooled mandarin orange peel saccharide into the activated lactobacillus in the RMS liquid culture medium, stirring, and semi-solid fermenting at constant temperature under sealed state;
(5) squeezing and separating, namely squeezing the fermented citrus peel, and respectively collecting fermentation liquor and fermentation residues for later use;
(6) post-treatment, namely respectively treating the fermentation liquor and the fermentation residues, mixing and preparing into capsules.
2. The method for preparing citrus lactic acid bacteria capsule with high flavone availability according to claim 1, wherein the post-treatment of the step (6) comprises:
(6a) freeze-drying and crushing, namely freeze-drying the fermentation residues, crushing the dried fermentation residues by using a roller crusher, and collecting fermentation residue powder;
(6b) spray drying, namely spray drying the fermentation liquor, and collecting fermentation liquor powder;
(6c) mixing, filling, mixing the fermented residue powder and fermented liquid powder, sieving, and making into capsule.
3. The method for preparing citrus lactic acid bacteria capsules with high flavone availability according to claim 1, wherein the water content of the citrus peel raw material in step (1) is less than 10%; soaking the rice in water, wherein the mass ratio of the water to the rice is 0.5-2: 1; steaming for 10-30 min.
4. The method for preparing citrus lactic acid bacteria capsules with high flavone availability according to claim 1, wherein the saccharification temperature in step (2) is 35 ± 5 ℃; the addition amount of the sweet distiller's yeast is 2-5% of the total mass of the raw materials.
5. The method for preparing citrus lactic acid bacteria capsules with high flavone availability according to claim 1, wherein the enzyme deactivation time in the step (3) is 5-15 min; the content of soluble solid in the saccharified liquid is more than 8 percent.
6. The method for preparing citrus lactic acid bacteria capsule with high flavone availability according to claim 1, wherein the number of lactic acid bacteria in the RMS liquid medium in step (4) is more than 5.0log CFU/g; the constant temperature fermentation condition is 30 + -5 deg.C, and the fermentation time is 3-7 d.
7. The method for preparing citrus lactic acid bacteria capsules with high flavone availability according to claim 1, wherein the citrus peel raw material is ground and sieved in step (1), and a 60-100 mesh sieve is used in step (1) and step (6).
CN202111357942.1A 2021-11-16 2021-11-16 Preparation method of citrus lactic acid bacteria capsule with high flavone utilization degree Pending CN113995128A (en)

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