CN111321096A - Method for improving bioavailability of apple juice polyphenol - Google Patents
Method for improving bioavailability of apple juice polyphenol Download PDFInfo
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- CN111321096A CN111321096A CN202010243654.2A CN202010243654A CN111321096A CN 111321096 A CN111321096 A CN 111321096A CN 202010243654 A CN202010243654 A CN 202010243654A CN 111321096 A CN111321096 A CN 111321096A
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- lactobacillus rhamnosus
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Images
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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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
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
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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
- 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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- 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
- A23V2400/175—Rhamnosus
Abstract
The invention discloses a method for improving bioavailability of polyphenol in apple juice. Belongs to the technical field of food fermentation. The technical scheme of the invention comprises the following steps: activation of lactobacillus rhamnosus zrx01 → strain acclimation → fermentation of apple juice. Compared with the prior art, the invention has the following beneficial effects: (1) the lactobacillus rhamnosus belongs to anaerobic bacteria, is not easy to grow, and can be rancid when the apple juice is not fermented well, and the lactobacillus rhamnosus zrx01 is facultative anaerobic bacteria and is easier to propagate and grow in the apple juice; (2) the adaptability of the strain is increased by continuously domesticating the strain; (3) the lactobacillus rhamnosus fermented apple juice can convert apple polyphenol into substances which are easy to absorb by small molecules, so that the bioavailability of the polyphenol in vivo is improved.
Description
Technical Field
The invention relates to the technical field of food fermentation, in particular to a method for improving bioavailability of polyphenol in apple juice.
Background
The apple is rich in nutrition, and contains various active substances such as vitamins, minerals, dietary fibers, polysaccharides and the like. Polyphenol is a general term for polyphenol substances contained in apples, and consists of one or more aromatic ring compounds and one or more hydroxyl groups, and mainly comprises phenolic acid, flavonoid, p-diphenylethylene, coumarin, tannin and the like. The polyphenol has antioxidant, anticancer, blood pressure lowering, blood sugar lowering, antibacterial, health promoting and pharmacological effects.
The average intake of polyphenol in human body is 1g/d, but after the human body takes 50mg aglycone, the total metabolite content in the blood plasma is 0-4 μmol/L, and the urine contains 0.3% -43% of the intake dose, thus the bioavailability of natural polyphenol in human body is very low. The polyphenol exists in food in the form of glycosylation, esters or polymers, is hydrolyzed by intestinal enzymes (such as lactase-phlorizin hydrolase) or microbial flora before absorption, is widely metabolized under the action of metabolic enzymes such as cytochrome P450 family, catechol-O-methyltransferase, sulfate transferase, glucuronyl transferase and the like, the original structure of the polyphenol is changed, then a part of polyphenol enters blood circulation, and a part of polyphenol is discharged back to the intestinal cavity by intestinal efflux transporters such as P-glycoprotein, multidrug resistance related protein-2, breast cancer drug resistance protein and the like, so that the bioavailability is reduced.
A small amount of intestinal flora (bifidobacterium and lactobacillus) can generate a plurality of enzymes related to polyphenol metabolism, such as glucosylcharase, α -rhamnosidase, β -glucosidase, β -galactosidase, esterase, tannase and the like, and can degrade polyphenol into micromolecular phenolic acid substances and improve the utilization rate of polyphenol.
In summary, it is an urgent need to solve the problem of providing a method for fermenting apple juice to improve the bioavailability of apple polyphenol.
Disclosure of Invention
In view of the above, the present invention provides a method for improving bioavailability of polyphenols in apple juice.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for improving bioavailability of polyphenol in apple juice comprises the following steps:
(1) activating strains: inoculating lactobacillus rhamnosus zrx01 to MRS liquid culture medium for strain activation to obtain lactobacillus rhamnosus activation solution;
the Lactobacillus rhamnosus zrx01 is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, the preservation number is CGMCC No.15076, the Lactobacillus rhamnosus is classified and named as Lactobacillus rhamnosus, and the preservation address is as follows: west road No.1, north american academy of sciences, north kyo, yang ward, No. 3, date of deposit: 12 months and 18 days 2017;
the lactobacillus rhamnosus belongs to anaerobic bacteria, is not easy to grow, and can be rancid when the apple juice is not fermented well, and the lactobacillus rhamnosus zrx01 in the invention is facultative anaerobic bacteria, so that the apple juice can be easily propagated and grown in the apple juice.
(2) Domesticating strains: sequentially inoculating the lactobacillus rhamnosus activating solution into milk apple juice culture mediums with different proportions for acclimatization culture, and then inoculating into pure apple juice culture mediums for acclimatization culture to obtain lactobacillus rhamnosus acclimatization culture solution;
the apple is rich in nutrition, contains various active substances such as vitamins, mineral substances, dietary fibers, polysaccharides and the like, and the nutrient substances contained in the apple juice can be used as a fermentation substrate, however, due to the high acidity and low pH value of the apple juice, the strains cannot be well proliferated and fermented in the fermented apple juice. The technical scheme of the invention increases the adaptability of the strain by continuously domesticating the strain.
(3) Fermentation: inoculating lactobacillus rhamnosus domestication culture solution into apple juice for fermentation.
The enzymes produced by the metabolism of lactobacillus rhamnosus enter the circulatory system of human body by releasing the combination of polyphenol and decomposing polyphenol into simpler and easily absorbed phenols.
Preferably, the specific operation of step (1) is as follows:
(11) inoculating lactobacillus rhamnosus zrx01 into an MRS liquid culture medium, wherein the inoculation amount is 1-2%, and culturing for 24-28 h at the temperature of 37 +/-1 ℃ at 180-200 r/min in a shaking table to obtain a lactobacillus rhamnosus primary activation solution;
(12) inoculating the primary activating solution of lactobacillus rhamnosus into an MRS liquid culture medium, wherein the inoculation amount is 1-2%, and culturing for 18-22 h at the temperature of 37 +/-1 ℃ at 180-200 r/min in a shaking table to obtain a secondary activating solution of lactobacillus rhamnosus;
(13) inoculating the lactobacillus rhamnosus secondary activation solution into an MRS liquid culture medium, wherein the inoculation amount is 1-2%, and culturing for 18-22 h at the temperature of 37 +/-1 ℃ at 180-200 r/min in a shaking table to obtain the lactobacillus rhamnosus activation solution.
Preferably, the volume ratio gradient of the milk to the apple juice in the milk apple juice culture medium in the step (2) is set to be 9:1, 8:2, 6:4, 4:6, 2:8 and 1:9 in sequence.
Preferably, the domestication culture conditions in step (2) are: the inoculation amount is 1-2%, and the culture is carried out for 24-28 h at the temperature of 37 +/-1 ℃ at 180-200 r/min in a shaking bed.
Preferably, the fermentation conditions in the step (3) are as follows: the inoculation amount is 1%, and the mixture is cultured for 36-38 h at a speed of 180-200 r/min in a shaking table.
According to the technical scheme, compared with the prior art, the invention has the following beneficial effects: the lactobacillus rhamnosus belongs to anaerobic bacteria, is not easy to grow, and can be rancid when the apple juice is not fermented, and the lactobacillus rhamnosus zrx01 in the invention is facultative anaerobic bacteria, so that the lactobacillus rhamnosus can be easily propagated and grown in the apple juice; (2) the adaptability of the strain is increased by continuously domesticating the strain; (3) the lactobacillus rhamnosus fermented apple juice can convert apple polyphenol into substances which are easy to absorb by small molecules, so that the biological activity of the polyphenol in vivo is improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.
FIG. 1 is a HPLC chart of unfermented apple juice polyphenols in experiment 4 of the present invention; wherein the chromatographic peaks represent: 1. protocatechuic acid; 2. procyanidin B2(ii) a 3. Chlorogenic acid; 4. epicatechin; 5. caffeic acid; 6. ferulic acid; phlorizin; 8. ellagic acid;
FIG. 2 is a HPLC chart of apple juice polyphenol fermented by Lactobacillus rhamnosus in experiment 4 of the present invention; wherein the chromatographic peaks represent: 2. procyanidin B2(ii) a 3. Chlorogenic acid; 5. caffeic acid; 6. ferulic acid; 7. phlorizin;
FIG. 3 is a polyphenol HPLC chart of Lactobacillus rhamnosus fermented apple juice digested by gastrointestinal fluids in experiment 4 of the present invention; wherein the chromatographic peaks represent: 2. procyanidin B2(ii) a 3. Chlorogenic acid; 5. caffeic acid; 6. ferulic acid; 7. phlorizin; and a, b and c are newly appeared peaks.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The required medicament is a conventional experimental medicament purchased from a market channel; the unrecited experimental method is a conventional experimental method, and the detailed description is omitted.
Example 1
(1) Activating strains:
(11) inoculating lactobacillus rhamnosus zrx01 into MRS liquid culture medium with the inoculation amount of 1%, and culturing in a shaker at 37 deg.C for 24h at 180r/min to obtain lactobacillus rhamnosus primary activation solution;
(12) inoculating the primary activating solution of lactobacillus rhamnosus into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing for 18h at 37 ℃ at 180r/min in a shaking table to obtain a secondary activating solution of lactobacillus rhamnosus;
(13) inoculating the secondary activating solution of lactobacillus rhamnosus into an MRS liquid culture medium, wherein the inoculation amount is 1%, and culturing for 18h at 37 ℃ at 180r/min in a shaking table to obtain the lactobacillus rhamnosus activating solution.
(2) Domesticating strains:
sequentially inoculating the lactobacillus rhamnosus activation solution into milk apple juice culture media with different apple juice ratios for acclimatization culture (the volume ratio of milk to apple juice is 9:1, 8:2, 6:4, 4:6, 2:8 and 1:9 in sequence), and then performing acclimatization culture in pure apple juice culture media under the culture conditions: inoculating 1% of lactobacillus rhamnosus, and culturing in a shaker at 37 deg.C for 24 hr at 180r/min to obtain lactobacillus rhamnosus acclimation culture solution;
(3) fermentation:
inoculating the lactobacillus rhamnosus domestication culture solution into apple juice for fermentation under the condition that the inoculation amount is 1%, and culturing for 36h at 180r/min in a shaking table to obtain the lactobacillus rhamnosus fermented apple juice.
Example 2
(1) Activating strains:
(11) inoculating lactobacillus rhamnosus zrx01 into MRS liquid culture medium, with the inoculum size of 2%, culturing in a shaker at 37 deg.C for 28h at 200r/min to obtain lactobacillus rhamnosus primary activation solution;
(12) inoculating the primary activating solution of lactobacillus rhamnosus into an MRS liquid culture medium, wherein the inoculation amount is 2%, and culturing for 22h at the temperature of 37 ℃ at the speed of 200r/min in a shaking table to obtain a secondary activating solution of lactobacillus rhamnosus;
(13) inoculating the lactobacillus rhamnosus secondary activation solution into an MRS liquid culture medium, wherein the inoculation amount is 2%, and culturing for 22h at 37 ℃ in a shaking table at 200r/min to obtain the lactobacillus rhamnosus activation solution.
(2) Domesticating strains:
sequentially inoculating the lactobacillus rhamnosus activation solution into milk apple juice culture media with different apple juice ratios for acclimatization culture (the volume ratio of milk to apple juice is 9:1, 8:2, 6:4, 4:6, 2:8 and 1:9 in sequence), and then performing acclimatization culture in pure apple juice culture media under the culture conditions: inoculating 2% of lactobacillus rhamnosus, and culturing in a shaker at 37 deg.C for 28h at 200r/min to obtain lactobacillus rhamnosus acclimation culture solution;
(3) fermentation:
inoculating the lactobacillus rhamnosus domestication culture solution into apple juice for fermentation under the condition that the inoculation amount is 1%, and culturing for 38h at 200r/min in a shaking table to obtain the lactobacillus rhamnosus fermented apple juice.
Example 3
(1) Activating strains:
(11) inoculating lactobacillus rhamnosus zrx01 into MRS liquid culture medium with the inoculation amount of 1.5%, and culturing in a shaker at 37 deg.C for 26h at 190r/min to obtain lactobacillus rhamnosus primary activation solution;
(12) inoculating the primary activating solution of lactobacillus rhamnosus into an MRS liquid culture medium, wherein the inoculation amount is 1.5%, and culturing for 20h at the temperature of 37 ℃ at the speed of 190r/min in a shaking table to obtain a secondary activating solution of lactobacillus rhamnosus;
(13) inoculating the lactobacillus rhamnosus secondary activation solution into an MRS liquid culture medium, wherein the inoculation amount is 1.5%, and culturing for 20h at the temperature of 37 ℃ at the speed of 190r/min in a shaking table to obtain the lactobacillus rhamnosus activation solution.
(2) Domesticating strains:
sequentially inoculating the lactobacillus rhamnosus activation solution into milk apple juice culture media with different apple juice ratios for acclimatization culture (the volume ratio of milk to apple juice is 9:1, 8:2, 6:4, 4:6, 2:8 and 1:9 in sequence), and then performing acclimatization culture in pure apple juice culture media under the culture conditions: inoculating with an inoculum size of 1.5%, and culturing in a shaker at 37 deg.C for 26h at 190r/min to obtain Lactobacillus rhamnosus acclimation culture solution;
(3) fermentation:
inoculating the lactobacillus rhamnosus domestication culture solution into apple juice for fermentation under the condition that the inoculation amount is 1%, and culturing for 37h at 190r/min in a shaking table to obtain the lactobacillus rhamnosus fermented apple juice.
Comparative example 1
Lactobacillus plantarum zrx03(GenBank NO. MN784485) is used for fermenting apple juice, the inoculation amount is 1%, and the apple juice is cultured for 18h at 37 ℃ to obtain the Lactobacillus plantarum fermented apple juice.
Comparative example 2
Lactobacillus acidophilus zrx02(GenBank NO. KP325412.1) is used for fermenting apple juice, the inoculation amount is 3%, and the apple juice is cultured at 37 ℃ for 18h to obtain the Lactobacillus acidophilus fermented apple juice.
(1) Experimental samples: untreated apple juice, lactobacillus rhamnosus fermented apple juice prepared in example 1, lactobacillus plantarum fermented apple juice prepared in comparative example 1, lactobacillus acidophilus fermented apple juice prepared in comparative example 2.
(2) The experimental method comprises the following steps:
(21) the DRS-IIPlus in vitro bionic mouse stomach system is opened, and the temperature is adjusted to 37 ℃. Device operating parameters: simulating the injection speed of gastric juice to 25L/min; the simulated intestinal fluid injection speed is 30L/min, the chyme evacuation speed is 100L/min, the gastric extrusion frequency is 3cpm, the gastric rolling frequency is 12cpm, and the intestinal rolling frequency is 36 cpm.
(22) The digestion solution was prepared according to the instructions of the apparatus, and 6mL of the sample was aspirated by a syringe and used. 15mL of simulated digestive juice is sucked by intestinal juice and gastric juice injectors, the simulated digestive juice is installed on corresponding injection pumps, 0.6mL of simulated gastric juice is injected into a new bionic rat stomach model to serve as fasting gastric juice, then 6mL of sample is injected, the rat stomach model is accurately placed under a stomach roller pressing plate, a simulated pyloric tube is connected with a simulated duodenal tube, and intestinal juice is filled into the simulated intestinal juice tube in front of an intestinal roller.
(23) After the external digestion starts, the anterior stomach of the bionic rat stomach model is compressed up and down by the pressing plate, the glandular stomach part is extruded from the antrum to the pylorus by the two rollers, the sample is continuously ground in the rat stomach model, namely, the sample is physically digested, and in the process, simulated gastric juice flows into the rat stomach model from the inner surface of the upper wall of the glandular stomach part at a certain speed and is mixed with the sample to generate chemical digestion reaction.
(24) The digestion time is 30min, 60min, 90min, 120min and 180min, and a certain amount of digested samples are taken from the tee joint at the tail end of the duodenum model and placed into a 1.5mL centrifuge tube to stand. And after digestion is finished for 3 hours, stopping the DRS-IIPlus device, and collecting chyme (stomach chyme) remained in the rat stomach model and chyme (duodenum chyme) remained in the duodenum model respectively and storing the collected chyme in a 5mL centrifuge tube to obtain the apple juice after gastrointestinal fluid digestion.
(1) And (3) determining a sample: untreated apple juice before and after gastrointestinal fluid digestion, lactobacillus rhamnosus fermented apple juice prepared in example 1, lactobacillus plantarum fermented apple juice prepared in comparative example 1, lactobacillus acidophilus fermented apple juice prepared in comparative example 2.
(2) The determination method comprises the following steps: and (3) a folin phenol colorimetric method.
(21) The use of gallic acid as a standard solution was chosen to give the following regression equation:
Y＝0.9758X+0.0033
R＝0.9992，Y:OD765and X is the mass concentration (mg/mL) of gallic acid.
(22) And (3) measuring the absorbance value of the sample to be measured at 765nm by using a spectrophotometer, and calculating the content of polyphenol in the sample through a standard curve.
(3) And (3) measuring results: the measurement results are shown in table 1.
TABLE 1 concentration of apple polyphenols before and after gastrointestinal fluid digestion for different samples
Note that a, b, c, d showed significant differences at α -0.05 significance levels
As can be seen from the results in Table 1, the apple polyphenols were significantly reduced after fermentation and digestion, indicating that the polyphenols had been converted and absorbed. And after being fermented by lactobacillus rhamnosus, the lactobacillus rhamnosus is easier to digest and absorb.
(1) Measurement indexes are as follows: oral bioavailability F (%) is generally estimated by the concentration of the active ingredient in the blood circulation.
(2) And (3) determining a sample: untreated apple juice, lactobacillus rhamnosus fermented apple juice prepared in example 1, lactobacillus plantarum fermented apple juice prepared in comparative example 1, lactobacillus acidophilus fermented apple juice prepared in comparative example 2.
(3) The calculation method comprises the following steps:
F＝(AUCis administered orally× oral dose)/(AUCIntravenous injection× intravenous dose) × 100%
(4) As a result: as shown in table 2.
TABLE 2 oral bioavailability of different samples
Note that a, b, c, d showed significant differences at α -0.05 significance levels
The results in table 2 show that the oral bioavailability of the fermented apple juice is significantly improved, and the bioavailability of the fermented apple juice is higher than that of the fermented apple juice obtained by the other two strains.
(1) Measurement target: unfermented apple juice, lactobacillus rhamnosus fermented apple juice prepared in example 1, and lactobacillus rhamnosus fermented apple juice prepared in example 1 were subjected to gastrointestinal fluid digestion.
(2) The determination method comprises subjecting a chromatographic column to Diamonsil C18(250mm × 4.6.6 mm, phi 5 μm) combined with Diamonsil C18(20mm × 4.0.0 mm, phi 5 μm) protective column;
mobile phase A: 1% volume fraction acetic acid aqueous solution, mobile phase B: methanol;
gradient elution was used, gradient elution time program: 0-10 min, 5-30% of B, 10-25 min, 30-50% of B, 25-35 min, 50-70% of B, 35-40 min and 70-5% of B;
the column temperature is 30 ℃;
the flow rate is 1.0 mL/min;
sample introduction amount: 20 mu L of the solution;
a detector: diode array detector, detection wavelength: 280nm and 320 nm.
(3) And (3) measuring results: the measurement results are shown in FIGS. 1 to 3 and Table 3.
TABLE 3 Polyphenol content in different samples
Note that in Table 1, unfermented apple juice, 2 Lactobacillus rhamnosus fermented apple juice prepared in example 1, 3, a sample of Lactobacillus rhamnosus fermented apple juice prepared in example 1 after gastrointestinal fluid digestion, nd was not detected, and a, b, c indicated that there was a significant difference at a level of α -0.05
The results show that the unfermented apple juice contains more polyphenol types and contents, but the contents of various polyphenols are reduced after lactobacillus rhamnosus fermentation, the polyphenol content is further reduced after gastrointestinal fluid digestion, and a new peak appears before 8min, which shows that the polyphenols are decomposed and converted into micromolecular substances and are easier to digest and absorb, and the results correspond to the results in table 1.
The lactobacillus rhamnosus fermented apple juice can convert apple polyphenol into substances which are easy to absorb by small molecules, so that the biological activity of the polyphenol in vivo is improved.
The embodiments in the present description are described in a progressive manner, each embodiment focuses on differences from other embodiments, and the same and similar parts among the embodiments are referred to each other.
The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (5)
1. A method for improving bioavailability of polyphenol in apple juice is characterized by comprising the following steps:
(1) activating strains: inoculating lactobacillus rhamnosus zrx01 to MRS liquid culture medium for strain activation to obtain lactobacillus rhamnosus activation solution;
the lactobacillus rhamnosus zrx01 is preserved in the China general microbiological culture Collection center, the preservation number is CGMCC No.15076, and the preservation address is as follows: west road No.1, north west of the morning area, beijing, institute of microbiology, china academy of sciences, date of deposit: 12 months and 18 days 2017;
(2) domesticating strains: sequentially inoculating the lactobacillus rhamnosus activation solution into milk apple juice culture mediums with different proportions for acclimatization culture, and then inoculating into pure apple juice culture mediums for acclimatization culture to obtain lactobacillus rhamnosus acclimatization culture solution;
(3) fermentation: inoculating lactobacillus rhamnosus domestication culture solution into apple juice for fermentation.
2. The method for improving bioavailability of polyphenol in apple juice according to claim 1, wherein the specific operation of the step (1) is as follows:
(11) inoculating lactobacillus rhamnosus zrx01 into an MRS liquid culture medium, wherein the inoculation amount is 1-2%, and culturing for 24-28 h at the temperature of 37 +/-1 ℃ at 180-200 r/min in a shaking table to obtain a lactobacillus rhamnosus primary activation solution;
(12) inoculating the primary activating solution of lactobacillus rhamnosus into an MRS liquid culture medium, wherein the inoculation amount is 1-2%, and culturing for 18-22 h at the temperature of 37 +/-1 ℃ at 180-200 r/min in a shaking table to obtain a secondary activating solution of lactobacillus rhamnosus;
(13) inoculating the lactobacillus rhamnosus secondary activation solution into an MRS liquid culture medium, wherein the inoculation amount is 1-2%, and culturing for 18-22 h at the temperature of 37 +/-1 ℃ at 180-200 r/min in a shaking table to obtain the lactobacillus rhamnosus activation solution.
3. The method for improving the bioavailability of polyphenol in apple juice according to claim 1, wherein in the step (2), the volume ratio gradient of milk to apple juice in the milk apple juice culture medium is set to be 9:1, 8:2, 6:4, 4:6, 2:8 and 1:9 in sequence.
4. The method for improving the bioavailability of polyphenol in apple juice according to claim 1, wherein the domesticated culture conditions in the step (2) are as follows: the inoculation amount is 1-2%, and the culture is carried out for 24-28 h at the temperature of 37 +/-1 ℃ at 180-200 r/min in a shaking table.
5. The method for improving the bioavailability of polyphenol in apple juice according to claim 1, wherein the fermentation conditions in the step (3) are as follows: the inoculation amount is 1%, and the culture is carried out for 36-38 h at a speed of 180-200 r/min in a shaking table.
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