CN116508922A - Preparation method of normal-temperature lactobacillus - Google Patents
Preparation method of normal-temperature lactobacillus Download PDFInfo
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- CN116508922A CN116508922A CN202310571598.9A CN202310571598A CN116508922A CN 116508922 A CN116508922 A CN 116508922A CN 202310571598 A CN202310571598 A CN 202310571598A CN 116508922 A CN116508922 A CN 116508922A
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- strain
- composite
- temperature
- tea
- lactobacillus
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Classifications
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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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/02—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation containing fruit or vegetable juices
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
- A23F3/00—Tea; Tea substitutes; Preparations thereof
- A23F3/16—Tea extraction; Tea extracts; Treating tea extract; Making instant tea
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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Abstract
The invention relates to a preparation method of normal-temperature lactobacillus, which belongs to the technical field of microorganisms and comprises the following steps: preparing composite tea powder and composite plant extract; preparing a composite strain and a strain carrier, and then packaging the composite lactobacillus strain by using the strain carrier to obtain a packaged strain; mixing sucrose and emulsion stabilizer, adding fruit juice and sweetener, pumping into a compounding tank, adding skimmed milk powder, packaging strain, compound tea powder and compound plant extract, regulating acid and quality control, and metering volume to obtain lactobacillus at room temperature. The compound tea is used for matching, the black tea is used as a main body for matching with the green tea and the oolong tea, the taste and the system clarity are balanced, the high-activity compound strain powder is used for increasing the flavor quality of the product, the taste is adjusted through sugar replacement and juice, the methanol extracts of the fruits of the shoulder pole and the seeds of the avocado are used as components of the product, the inhibition effect of the system on mixed bacteria and other microorganisms is improved, and the quality guarantee period is prolonged.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a preparation method of normal-temperature lactobacillus.
Background
The lactobacillus beverage is prepared by taking milk or dairy product as raw material, adding water into emulsion prepared by lactobacillus fermentation, and mixing with one or more of white sugar or sweetener, sour agent, fruit juice, tea, coffee, plant extract, etc. The lactobacillus beverage is characterized in that lactose in the cow milk is decomposed into galactose and lactic acid due to the fermentation of lactobacillus, and partial protein is decomposed into small peptide chains and amino acids, so that the nutrition components of the cow milk are easier to digest and absorb, the lactobacillus beverage is suitable for being drunk by lactose intolerant patients, and the utilization rate of calcium, phosphorus and the like in human bodies can be improved. Meanwhile, the lactobacillus also has the effects of improving intestinal flora, regulating gastrointestinal functions, preventing aging, maintaining beauty and keeping young, enhancing organism immunity and the like. Thus, lactic acid bacteria beverages are highly favored by consumers.
In the common lactobacillus beverage in the market, the addition amount of lactobacillus is small, the activity of strain is low, the beneficial effect on human body is not obvious after gastric acid in digestive tract is destroyed, and even the sugar, fat and additive are excessively added in the formula, so that the human body is injured. In addition, the sterilized or non-living lactobacillus product does not contain bioactive components, and the shelf life of the lactobacillus is generally about 180 days due to sterilization, heating and inactivation in the production process, while the shelf life of the product containing living bacteria is generally about 15-30 days, which is not beneficial to storage and sales.
Disclosure of Invention
The invention aims to provide a preparation method of normal-temperature lactobacillus, which uses a strain carrier to package composite lactobacillus strains to obtain packaged strains; then preparing composite tea powder and composite plant extract for later use; mixing sucrose and emulsion stabilizer, adding sweetener, pumping into a compounding tank, adding skimmed milk powder, packaging strain, compound tea powder and compound plant extract, regulating acid and quality control, and metering volume to obtain lactobacillus at room temperature. The compound tea is used for matching, the black tea is used as a main body for matching with the green tea and the oolong tea, the taste and the system clarity are balanced, the high-activity compound strain powder is used for increasing the flavor quality of the product, the taste is adjusted through sugar replacement and juice, the methanol extracts of the fruits of the shoulder pole and the seeds of the avocado are used as components of the product, the inhibition effect of the system on mixed bacteria and other microorganisms is improved, and the quality guarantee period is prolonged.
The invention aims to solve the technical problems:
the invention solves the problems that in the common lactobacillus beverage, the addition amount of lactobacillus is small, the activity of strain is low, after gastric acid in the digestive tract is destroyed, the beneficial effect on human body is not obvious, the quality guarantee period of the product containing living bacteria is short, and the storage and the sales are not facilitated.
The aim of the invention can be achieved by the following technical scheme:
a preparation method of normal temperature lactobacillus comprises the following steps:
(1) Preparing composite strain and strain carrier, then packaging the composite lactobacillus strain by using the strain carrier to obtain packaged strain, and the specific steps are as follows:
b1, inulin and pectin are added in an amount of 1:1, and placing in sterile deionized water, stirring at 30rpm at room temperature until the pectin-inulin solution is dissolved, filtering by using a 0.2 μm pore size filter, and diluting to 0.4mg/mL by using the sterile deionized water to obtain the pectin-inulin solution; adding nisin with concentration of 1-2mg/mL into pectin-inulin solution under stirring and room temperature, regulating pH to 4.0-7.0 with NaOH solution to obtain encapsulated nisin solution, and storing in sterile environment at 4deg.C;
b2, adding 1g of composite strain into 9mL of 2% (w/v) sterile sodium alginate, uniformly stirring at 30 ℃, adding into 9mL of 0.15mol/L sterile calcium chloride solution, and standing for 4-5 minutes to obtain composite strain microcapsules;
and B3, adding the composite strain microcapsule into the encapsulated nisin solution, and culturing at 25 ℃ for 4 hours by using a shaking table to obtain the encapsulated strain.
(2) Uniformly mixing sucrose and an emulsion stabilizer, adding a sweetener, then pumping into a batching tank, adding skimmed milk powder, packaging strains, compound tea powder and compound plant extracts, adjusting acid and quality control, and then fixing the volume to prepare normal-temperature lactobacillus, wherein the specific steps are as follows:
s1, uniformly mixing sucrose and an emulsion stabilizer, adding the mixture into sterile deionized water at 80-85 ℃, shearing the mixture for 15-20 minutes at 1400rpm by using a high-speed shearing emulsifier until uniform glue solution is formed, then adding fruit juice and a sweetener, uniformly stirring the mixture, reducing the temperature to below 30 ℃ by using a plate heat exchanger, and pumping the mixture into a material mixing tank for later use;
s2, adding the skim milk powder, the packaging strains, the compound tea powder and the compound plant extract into a material mixing tank, and stirring for 15-20 minutes until the materials are uniform;
s3, adding lactic acid and sterile deionized water into an acid adding tank, and starting stirring to obtain acid liquor; cooling the temperature of the batching pipe to 20 ℃, adding acid by using a spraying method, adding acid liquor into the batching tank through a atomization device in the acid adding tank after approaching preset acidity, adjusting the acid, immediately fixing the volume, and stirring for 15-20 minutes;
s4, after testing the solids, the pH value and the acidity in the milk-containing beverage, adding the yoghurt flavor, adjusting the aroma, continuously stirring for 10-15 minutes, homogenizing for 20 minutes under 18-20MPa, and filling in a sterile filling machine to obtain the normal-temperature lactobacillus.
Further, the specific steps for preparing the composite tea powder are as follows:
a1, washing composite tea leaves with low-temperature deionized water, respectively drying the composite tea leaves at 80 ℃, controlling the water content to be less than 6%, then sending the composite tea leaves into a pulverizer for pulverizing, and sieving the composite tea leaves with a 60-mesh sieve to obtain tea dust;
a2, leaching the tea dust by using deionized water at the leaching temperature of 90-100 ℃ for 15 minutes, and repeating the leaching operation twice to obtain leaching liquor;
a3, centrifuging the leaching solution at 8000rpm, discarding the precipitate, and concentrating by using a nanofiltration membrane with the molecular weight cut-off of 100-2000 to obtain a concentrated solution; and then spray drying the concentrated solution to obtain the composite tea powder.
Further, the compound tea powder is obtained by mixing black tea, green tea and oolong tea in a ratio of 10:5:2, and the mass ratio of deionized water to tea dust is 9-15:1, spray drying inlet temperature 100 ℃, outlet temperature 45 ℃ and airflow velocity 40m 3 /h。
Further, the preparation method of the composite plant extract comprises the following specific steps:
washing the shoulder pole fruits and the avocado seeds with sterile deionized water under the sterile environment, drying at 90 ℃, respectively grinding the shoulder pole fruits and the avocado seeds into powder by using a mortar, extracting sequentially by using 60% methanol solution by a supercritical CO2 method at the extraction temperature of 35-40 ℃ and the extraction pressure of 25-28MPa for 3-5 hours, collecting extract liquid, freeze-drying the extract liquid to obtain the extracts of the shoulder pole fruits and the avocado seeds, and mixing the extracts with the components of 4: mixing at a mass ratio of 1-2 to obtain the compound plant extract.
Further, the pectin starting material has a degree of polymerization between 0 and 60%.
Further, the composite strain is prepared by the following steps:
c1, centrifuging strain mixed liquor for 5 minutes at 10614 Xg centrifugal force under the aseptic environment and 4 ℃, then re-suspending in MRS culture medium, then statically incubating for 1.5 hours at 45 ℃, centrifuging the cultured fermentation liquor for 5 minutes at 4 ℃ and 10614 Xg centrifugal force, and re-suspending in sterile deionized water to obtain composite strain suspension;
c2, under the aseptic environment, mixing skimmed milk, trehalose, sucrose and yoghurt essence with 80:10:2:5, obtaining a cryoprotectant after mixing in a mass ratio, and storing at 4 ℃ under a sterile condition;
and C3, transferring the composite strain suspension into a cryoprotectant, pre-freezing for 12 hours at the temperature of minus 80 ℃, drying in a freeze dryer, wherein the temperature of a condenser is minus 60 ℃, the pressure of a cavity is less than or equal to 20Pa, and the freezing time is 36 hours, thus obtaining the composite strain.
Further, the strain mixture is 1×10 9 CFU/mL of Lactobacillus gasseri OLL2716, lactobacillus bulgaricus OLL1073R-1, streptococcus thermophilus and Lactobacillus buchneri at a ratio of 5:4:1:1 in a sterile environment.
Further, the emulsion stabilizer is prepared from soybean polysaccharide, pectin, resistant dextrin, inulin and guar gum at a ratio of 3:2:2:3:3, wherein the sweetener is erythritol.
The invention has the beneficial effects that:
(1) According to the technical scheme, the compound tea is used for matching, the black tea is used as a main body and matched with the green tea and the oolong tea, the black tea has more obvious sweet taste, polyphenols, saccharides, amino acids and the like in the black tea and saliva produce chemical action, saliva secretion can be stimulated, so that the oral cavity is moistened, the clarity of a system is improved through adding the green tea, the content of tea polyphenols, caffeine, amino acids and the like in the green tea is more, the effects of stimulating oral mucosa and promoting secretion of digestive glands are achieved, and the oolong tea is beneficial to promoting the secretion of saliva and supplementing tea fragrance. In addition, natural fruit juice and sugar substitute are used for adjusting sweetness, and the sweet taste improving agent has the characteristics of safe components and low heat.
(2) In the technical scheme of the invention, the composite strain of the combination of the lactobacillus gasseri OLL2716, the lactobacillus bulgaricus OLL1073R-1, the streptococcus thermophilus and the lactobacillus buchneri is used as zymophyte, the lactobacillus bulgaricus OLL2716 has good artificial gastric juice tolerance, is a gastric acid resistant lactobacillus, has good effect on stomach health, the lactobacillus bulgaricus OLL1073R-1 can produce more extracellular polysaccharide, can increase the content of prebiotics, and the streptococcus thermophilus and the lactobacillus buchneri can produce lactase, so that the digestion effect of lactose in the digestive tract is improved, and the lactose intolerance caused by the ingestion of dairy products by human bodies is reduced.
(3) In the technical scheme of the invention, the packaging amount of the strain can be increased by packaging the dried composite strain compared with the mode of combining the strain liquid and the packaging carrier, so that the strain brings more obvious benefits to human bodies. Pre-freezing with cryoprotectant composed of skimmed milk, trehalose, sucrose and yogurt essence before freeze drying, and freeze drying. The combination of the cryoprotectant has the functions of embedding microorganisms and locking free water around the strain, and the strain can be fully embedded into the skim milk in the pre-freezing process, so that the distribution of the strain on the surface of the dry powder is reduced, and the activity of the strain in the drying process is protected.
(4) In the technical scheme of the invention, the encapsulated nisin is combined with the composite strain, so that the encapsulated nisin can achieve the effect of further controlled release on the strain, and meanwhile, the sucrose in the cryoprotectant is isolated from contacting with the system, so that the carbon source of the strain is reduced, and the sugar burden is increased. The combination mode increases the in-situ activity of the strain in the system, increases the protection effect on the strain, and simultaneously has good killing effect on the mixed bacteria by packaging nisin and good anti-corrosion effect in the system; in the human digestive system, seaweed gel and nisin have resistance to acidic pH values, shrink in the low pH value environment of gastric juice, and strains have activity; under the environment of high pH value of intestinal juice, the glycosidic bond of seaweed gel is decomposed, nisin is inactivated in alkaline pH value, strain is released, the highest utilization rate is achieved in human body, and as many prebiotics as possible are absorbed.
(5) According to the technical scheme, the methanol extracts of the shoulder pole fruits and the avocado seeds are used as components of the product, the shoulder pole fruits have good bactericidal effect on mixed bacteria and fungi, can selectively inhibit the growth of aeromonas and paratyphoid pseudomonas, have obvious inhibition effect on pathogenic and putrefying microorganisms such as escherichia coli, increase the antibacterial spectrum range, and have synergistic effect on antibacterial effect with packaged nisin. In addition, the ethanol extract of the shoulder pole fruits contains beneficial substances such as alkaloid, triterpene, glycoside, steroid, tannin, flavonoid and the like, and has good effects in relieving fever, easing pain, resisting inflammation and resisting oxidation. The extract of the seed of the avocado has good preservative effect, is natural and nontoxic, can increase the stability of a system, prolongs the shelf life of a product, and is synergistic in preservative effect with the encapsulated nisin.
Detailed Description
The technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention, and it is apparent that the described embodiments are only some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The reagents used in the following examples and comparative examples were all of reagent grade and all procedures were performed in a sterile environment.
Example 1
The compound tea powder is prepared by the following steps:
mixing black tea, green tea and oolong tea at a ratio of 10:5:2 to obtain composite tea, and storing at 4deg.C for use;
washing the composite tea leaves with low-temperature deionized water, respectively drying the composite tea leaves at 80 ℃, controlling the water content to be less than 6%, then sending the composite tea leaves into a pulverizer for pulverizing, and sieving the composite tea leaves with a 60-mesh sieve to obtain tea dust; subsequently, using 1200g deionized water to leach 100g tea dust, wherein the leaching temperature is 95 ℃, the leaching time is 15 minutes, and the leaching operation is repeated twice to obtain leaching liquor; centrifuging the leaching solution at 8000rpm, discarding the precipitate, and concentrating with nanofiltration membrane with molecular weight cut-off of 1000 to obtain concentrated solution; and then spray drying the concentrated solution to obtain the composite tea powder.
Example 2
The encapsulated strain is prepared by the following steps:
mixing 20g of inulin and 20g of pectin with polymerization degree of 0-20%, placing in 500mL of sterile deionized water, stirring at 30rpm to dissolve at room temperature, filtering with a 0.2 μm pore size filter, and diluting to 0.4mg/mL with sterile deionized water to obtain pectin-inulin solution;
adding nisin with concentration of 1mg/mL into 100mL pectin-inulin solution under stirring and room temperature, regulating pH value of the system to 4.0-7.0 with NaOH and HCl solution to obtain encapsulated nisin solution, and storing under aseptic environment and 4deg.C;
1×10 9 CFU/mL of Lactobacillus gasseri OLL2716, lactobacillus bulgaricus OLL1073R-1, streptococcus thermophilus CICC 20378 and Lactobacillus buchneri CICC 6052 at a ratio of 5:4:1: mixing in a sterile environment according to the volume ratio of 1 to obtain a strain mixed solution, and storing for later use;
centrifuging the strain mixed solution for 5 minutes at 10614 Xg centrifugal force under the aseptic environment and the condition of 4 ℃, then re-suspending in an MRS culture medium, then statically incubating for 1.5 hours at 45 ℃, centrifuging the cultured fermentation solution for 5 minutes at the condition of 4 ℃ and the condition of 10614 Xg centrifugal force, re-suspending in sterile deionized water, and controlling the volume to be unchanged to obtain a composite strain suspension;
in a sterile environment, skimmed milk, trehalose, sucrose and yoghurt flavor are mixed in an amount of 80:10:2:5, obtaining a cryoprotectant after mixing in a mass ratio, and storing at 4 ℃ under a sterile condition;
transferring 950mL of composite strain suspension into 50mL of cryoprotectant, pre-freezing for 12 hours at-80 ℃, drying in a freeze dryer, wherein the temperature of a condenser is-60 ℃, the pressure of a cavity is less than or equal to 20Pa, and the freezing time is 36 hours, thus obtaining the composite strain;
then adding 10g of composite strain into 90mL of 2% (w/v) sterile sodium alginate, uniformly stirring at 30 ℃, adding into 90mL of 0.15mol/L sterile calcium chloride solution, and standing for 4-5 minutes to obtain composite strain microcapsules;
10g of the composite strain microcapsule is added into 50mL of encapsulated nisin solution, and shake culture is carried out at 25 ℃ for 4 hours, thus obtaining the encapsulated strain.
Example 3
This embodiment differs from embodiment 2 in that:
the polymerization degree of the pectin raw material is 20% -40%; the concentration of Streptococcus lactis was 1.5mg/mL and the amount was 25mL.
Example 4
This embodiment differs from embodiment 2 in that:
the polymerization degree of the pectin raw material is 40% -60%; the concentration of the streptococcus lactis is 2mg/mL, and the dosage is 30mL.
Example 5
The composite plant extract is prepared by the following steps:
washing the fruits and seeds of Tamarinus officinalis with sterile deionized water under aseptic condition, oven drying at 90deg.C, grinding into powder respectively with mortar, sequentially using 60% methanol solution and supercritical CO 2 Extracting at 38deg.C under 27MPa for 5 hr, collecting extractive solution, lyophilizing to obtain extract of fruits of Tamarindus indica and seeds, and mixing at a mass ratio of 8:3 to obtain compound plant extract.
Example 6
The normal temperature lactic acid bacteria are prepared by the following steps:
soy polysaccharide, pectin, resistant dextrin, inulin and guar gum were mixed at 3:2:2:3:3, mixing the materials according to the mass ratio to prepare an emulsion stabilizer, and storing the emulsion stabilizer for later use;
evaporating and concentrating 250mL of fresh apple juice and 250mL of fresh peach juice until the volume is 100mL to obtain concentrated juice, and storing for later use;
uniformly mixing 30g of sucrose with 40g of emulsion stabilizer, adding into sterile deionized water at 85 ℃, shearing for 20 minutes at 1400rpm by using a high-speed shearing emulsifier until uniform glue solution is formed, then adding 0.3g of erythritol and 100mL of concentrated juice, uniformly stirring, reducing the temperature to below 30 ℃ by a plate heat exchanger, and pumping into a batching tank for later use;
150g of skim milk powder, the encapsulated strains prepared in example 2, the compound tea powder prepared in example 1 and 11g of compound plant extract are added into a preparation tank and stirred for 20 minutes until uniform;
adding lactic acid and 1000mL of sterile deionized water into an acid adding tank, and starting stirring to obtain acid liquor; after the temperature of a batching pipe is reduced to 20 ℃, adding acid by using a spraying method, after the temperature is close to 70 ℃, adding acid liquor into the batching tank through a atomization device in the acid adding tank until the acidity is 70 ℃, immediately metering the volume after acid adjustment, and stirring for 20 minutes;
testing the solid, pH value and acidity of the milk-containing beverage, adding yoghurt flavor, adjusting aroma, continuously stirring for 15 minutes, homogenizing for 20 minutes under 20MPa, and filling in a sterile filling machine to obtain the normal-temperature lactobacillus.
Example 7
This embodiment differs from embodiment 6 in that:
the encapsulated strain used in this example was prepared as in example 3.
Example 8
This embodiment differs from embodiment 6 in that:
the encapsulated strain used in this example was prepared as in example 4.
Comparative example 1
The difference between this comparative example and example 7 is that in the encapsulated strain, the strain mixture was composed of 1X 10 9 CFU/mL of Lactobacillus gasseri OLL2716, streptococcus thermophilus, and Lactobacillus buchneri at 1:1 in a sterile environment.
Comparative example 2
The difference between this comparative example and example 7 is that the comparative example does not contain a plant extract.
Comparative example 3
The difference between this comparative example and example 7 is that the strain in this comparative example is not encapsulated.
The lactic acid bacteria products prepared in examples 6-8 and comparative examples 1-3 were now subjected to stability tests. 20mL of each group of fresh products are taken and transferred into a 50mL measuring cylinder, stored in a sealed manner at 6+/-2 ℃ for 48 hours, and then the separation index is measured. Taking each group of fresh samples, storing each sample at room temperature (about 25 ℃), and observing state change after 7 days; seven days later, 1mL of each sample was taken out, added to 9mL of PBS buffer, diluted 3 times with PBS buffer, then inoculated in a sterile petri dish, cultured in a constant temperature incubator, and after 48 hours, the colony count was measured to count the strain survival rate, and each sample was tested 5 times, and the test results are shown in table 1 below.
TABLE 1 stability test of lactic acid bacteria products prepared in examples 6-8 and comparative examples 1-3
As can be seen from the above Table 1, the separation indexes in examples 6-8 are low, the strain survival rate is higher than 98%, the system has good stability, and the results in comparative example 3 show that the strain survival rate is improved by encapsulating the strain.
The lactic acid bacteria products prepared in examples 6 to 8 and comparative examples 1 to 3 were now tested for antioxidant effect. 7mM 2, 2-azido-bis (3-ethylbenzothiazoline-6-sulfonic acid) aqueous solution was combined with 2.45mM K 2 S 2 O 8 The aqueous solution was left at room temperature and kept in the dark for 16 hours using 96% ethanol solution at 1:10, and diluting the solution to obtain a diluted solution. 200. Mu.L of the samples prepared in examples 6 to 8 and comparative examples 1 to 3 were mixed with 3.8mL of a 95% ethanol solution, 1mL of the diluted solution was added, and the mixture was stirred uniformly and kept in a dark environment for 6 minutes. The free radicals were measured using a spectrophotometer at 734 nm. Five readings were taken for each sample and averaged, and the test results are shown in table 2 below.
TABLE 2 antioxidant Effect test of lactic acid bacteria products prepared in examples 6-8 and comparative examples 1-3
From the test results of table 2 above, it is understood that the lactic acid bacteria product prepared in example 7 has the highest antioxidant activity, and from the results of comparative example 2, it is understood that the plant extract can improve the effect of the product in terms of antioxidant.
In the description of the present specification, the descriptions of the terms "one embodiment," "example," "specific example," and the like, mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the present invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiments or examples. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
The foregoing is merely illustrative and explanatory of the invention, as various modifications and additions may be made to the particular embodiments described, or in a similar manner, by those skilled in the art, without departing from the scope of the invention or exceeding the scope of the invention as defined in the claims.
Claims (10)
1. The preparation method of the normal-temperature lactobacillus is characterized by comprising the following steps of:
(1) Preparing a composite strain and a strain carrier, and then packaging the composite lactobacillus strain by using the strain carrier to obtain a packaged strain;
(2) Mixing sucrose and emulsion stabilizer, adding fruit juice and sweetener, pumping into a compounding tank, adding skimmed milk powder, packaging strain, compound tea powder and compound plant extract, regulating acid and quality control, and metering volume to obtain lactobacillus at room temperature.
2. The method for preparing normal temperature lactic acid bacteria according to claim 1, wherein the specific steps for preparing the composite tea powder are as follows:
a1, washing the composite tea leaves with low-temperature deionized water, respectively drying at 80 ℃, and then crushing and sieving to obtain tea dust;
a2, leaching the tea dust by using deionized water, wherein the leaching temperature is 90-100 ℃, the leaching time is 15 minutes, and the leaching operation is repeated twice to obtain leaching liquor;
and A3, centrifuging the leaching solution, discarding the precipitate, concentrating by using a nanofiltration membrane to obtain a concentrated solution, and spray-drying the concentrated solution to obtain the composite tea powder.
3. The preparation method of the normal-temperature lactobacillus according to claim 2, wherein the compound tea powder is obtained by mixing black tea, green tea and oolong tea in a ratio of 10:5:2, and the mass ratio of deionized water to tea dust is 9-15:1, spray drying inlet temperature 100 ℃, outlet temperature 45 ℃ and airflow velocity 40m 3 /h。
4. The method for preparing normal temperature lactic acid bacteria according to claim 1, wherein the specific operation for preparing the encapsulated strains comprises the steps of:
b1, inulin and pectin are added in an amount of 1:1, and placing the mixture into sterile deionized water, stirring the mixture at room temperature until the mixture is dissolved, filtering the mixture, and diluting the mixture to 0.4mg/mL by using the sterile deionized water to obtain pectin-inulin solution;
b2, adding nisin into the pectin-inulin solution under stirring and room temperature, regulating the pH value of the system to be between 4.0 and 7.0 by using NaOH solution to obtain an encapsulated nisin solution, and storing the encapsulated nisin solution in a sterile environment at 4 ℃;
adding the composite strain into sterile sodium alginate, uniformly stirring at 30 ℃, adding into a sterilized calcium chloride solution, and standing to obtain a composite strain microcapsule; adding the composite strain microcapsule into the encapsulated nisin solution, and culturing at 25deg.C for 4 hr by using a shaking table to obtain the encapsulated strain.
5. The method of producing a room temperature lactic acid bacterium according to claim 4, wherein the polymerization degree of the pectin raw material is between 0 and 60%.
6. The method for preparing normal temperature lactic acid bacteria according to claim 4, wherein the composite bacterial strain is prepared by the following steps:
c1, centrifuging the strain mixed solution in a sterile environment at 4 ℃, then re-suspending in an MRS culture medium, then statically incubating at 45 ℃, centrifuging the cultured fermentation liquor at 4 ℃ for 5 minutes, and re-suspending in sterile deionized water to obtain a composite strain suspension;
c2, under the aseptic environment, mixing skimmed milk, trehalose, sucrose and yoghurt essence with 80:10:2:5, obtaining a cryoprotectant after mixing in a mass ratio, and storing at 4 ℃ under a sterile condition;
and C3, transferring the composite strain suspension into a cryoprotectant, pre-freezing for 12 hours at the temperature of minus 80 ℃, drying in a freeze dryer, wherein the temperature of a condenser is minus 60 ℃, the pressure of a cavity is less than or equal to 20Pa, and the freezing time is 36 hours, thus obtaining the composite strain.
7. The method for preparing room temperature lactic acid bacteria according to claim 6, wherein the strain mixture is 1X 10 9 CFU/mL of Lactobacillus gasseri OLL2716, lactobacillus bulgaricus OLL1073R-1, streptococcus thermophilus and Lactobacillus buchneri at a ratio of 5:4:1:1 in a sterile environment.
8. The method for preparing normal temperature lactic acid bacteria according to claim 1, wherein the specific operations for preparing the composite plant extract are:
washing the fruits of Tamarinus officinalis and seeds of Butyrospermum parkii with sterile deionized water under aseptic environment, oven drying, grinding the fruits of Tamarinus officinalis and seeds of Butyrospermum parkii respectively into powder, sequentially using 60% methanol solution and supercritical CO 2 Extracting at 35-40deg.C under 25-28MPa for 3-5 hr, collecting extractive solution, and lyophilizing to obtain extract of fruits of Tamarinus officinalis and seeds of Butyrospermum parkii, with a dosage of 4: mixing at a mass ratio of 1-2 to obtain the compound plant extract.
9. The method for preparing the normal-temperature lactic acid bacteria according to claim 1, wherein the specific operation for preparing the normal-temperature lactic acid bacteria is as follows:
s1, uniformly mixing sucrose and an emulsion stabilizer, adding the mixture into sterile deionized water at 80-85 ℃, then shearing at 1400rpm until uniform glue solution is formed, then adding fruit juice and a sweetener, uniformly stirring, reducing the temperature to below 30 ℃ through a plate heat exchanger, and pumping into a batching tank for later use;
s2, adding the skim milk powder, the packaging strains, the compound tea powder and the compound plant extract into a material mixing tank, and stirring for 15-20 minutes until the materials are uniform;
s3, adding lactic acid and sterile deionized water into an acid adding tank, and starting stirring to obtain acid liquor; cooling the temperature of the batching pipe to 20 ℃, adding acid by using a spraying method, adding acid liquor into the batching tank through a atomization device in the acid adding tank after approaching preset acidity, adjusting the acid, immediately fixing the volume, and stirring for 15-20 minutes;
s4, after testing the solids, the pH value and the acidity in the milk-containing beverage, adding the yoghurt flavor, adjusting the aroma, continuously stirring for 10-15 minutes, homogenizing for 20 minutes under 18-20MPa, and filling in a sterile filling machine to obtain the normal-temperature lactobacillus.
10. The method for preparing normal temperature lactic acid bacteria according to claim 9, wherein the emulsion stabilizer is prepared from soybean polysaccharide, pectin, resistant dextrin, inulin and guar gum in a ratio of 3:2:2:3:3, wherein the sweetener is erythritol.
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