CN116555075A - Lactobacillus plantarum JF1 and application thereof in preparation of anti-aging food and drug - Google Patents
Lactobacillus plantarum JF1 and application thereof in preparation of anti-aging food and drug Download PDFInfo
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- CN116555075A CN116555075A CN202310234091.4A CN202310234091A CN116555075A CN 116555075 A CN116555075 A CN 116555075A CN 202310234091 A CN202310234091 A CN 202310234091A CN 116555075 A CN116555075 A CN 116555075A
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- lactobacillus plantarum
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- A—HUMAN NECESSITIES
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- 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
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Abstract
The invention belongs to the technical field of probiotics and application thereof, and particularly relates to lactobacillus plantarum JF1 and application thereof in preparation of anti-aging food and drugs. In order to further develop and utilize the probiotic functions of lactobacillus plantarum, the invention separates a lactobacillus plantarum (Lactplatinum sp lan) JF1 strain from the excrement of a healthy adult, and the strain has various probiotic effects of producing protease, 3-hydroxybutyric acid, gamma-aminobutyric acid, hyaluronic acid, inhibiting alpha-glucosidase activity, inhibiting acetylcholinesterase activity and the like. Therefore, the JF1 strain has various functions of improving protein allergy, resisting inflammation, improving intestinal flora, resisting depression, dispelling alcohol, resisting aging, resisting wrinkle, reducing blood glucose, promoting intestinal absorption and peristalsis and the like by promoting digestion and absorption of protein food, can be used in the fields of resisting bacteria, inflammation, aging and the like, and has important application value and economic value.
Description
Technical Field
The invention belongs to the technical field of probiotics and application thereof, and particularly relates to lactobacillus plantarum JF1 and application thereof in preparation of anti-aging food and drugs.
Background
Lactobacillus plantarum (Lactobacilli), also known as Lactobacillus plantarum, belongs to the genus Lactobacillus of the family Lactobacillus, and is anaerobic or facultative anaerobic, with an optimal culture pH of 6.5, and the cells are in the form of straight or curved rods, singly, sometimes in pairs or in chains. Lactobacillus plantarum is a probiotic in human gastrointestinal tract, can produce various natural antibacterial substances such as organic acid, bacteriocin, hydrogen peroxide, diethyl phthalide and the like, and has various functions of maintaining balance of intestinal flora, reducing cholesterol level, improving organism immunity, promoting nutrient absorption and the like.
Lactobacillus plantarum is an edible lactobacillus, is closely related to human life, is commonly used in dairy products, meat and fermented products of a plurality of vegetables, can play a beneficial role by being planted in intestinal tracts through stomach, has important influence on intestinal microorganisms, and has wide application in the fields of food fermentation, industrial lactic acid fermentation, medical care and the like. It is well known that lactobacillus is a part of the normal flora of the human body, and is an important component of the normal microbial system of the intestinal tract and is accompanied by a host for life, and has important significance for maintaining the microecological balance of the intestinal tract. The lactobacillus plantarum is also an important intestinal probiotics serving as edible lactobacillus, can play a role in assisting intestinal peristalsis, can also enhance the digestion capacity of a human body, improve immunity and the like. In recent years, lactobacillus plantarum has become a hotspot for research as a probiotic lactobacillus with great potential, and is being continuously made into probiotic preparations suitable for human and animals.
At present, different strains of lactobacillus plantarum have been reported to have different probiotic functions: (1) anti-pathogenic bacteria: it has been found that Lactobacillus plantarum has a strong inhibitory effect on Salmonella and on helicobacter pylori. (2) modulating immunity: studies show that the lactobacillus plantarum HK-LP has strong induction effect on IL-12 of mice, and the administration of the HK-LP can inhibit the generation of immunoglobulin, resist food allergy of the mice caused by natural diet, and inhibit the growth of tumors and the transplantation of tumor cells in the bodies of the mice. (3) cadmium removal effect: the research shows that the lactobacillus plantarum CCFM8610 has acid resistance, has good tolerance function and adsorption capacity to cadmium ions in vitro, can reduce the cadmium content in the liver and kidney of the cadmium-exposed mice, and has the effect of promoting the cadmium discharge of the cadmium-exposed mice through feces. (4) inhibition of Escherichia coli: the research shows that the lactobacillus plantarum N13 has acid resistance and cholate resistance, has very high adhesion to the intestinal epithelial cell strain HT-29 cells, can inhibit the growth of intestinal invasive escherichia coli, and can prevent the invasion of the intestinal epithelial cells to HT-29. (5) has proteolytic activity: studies show that the lactobacillus plantarum CW006 has proteolytic activity, and can ferment raw milk to generate angiotensin converting enzyme inhibitory peptide, thereby achieving the antihypertensive effect.
The diversity of the sources of lactobacillus plantarum leads to its diversity of genes and functions. However, the current studies on the isolation and identification, the probiotics characteristics and the metabolic mechanism of the lactobacillus plantarum are still relatively few, which also affects the development and utilization of the lactobacillus plantarum to a certain extent. Therefore, it is necessary to make lactobacillus plantarum function better according to different sources, such as determining the efficacy according to the functions of the strain or the probiotics metabolite, and defining the application prospect. In conclusion, the research and application of the probiotic lactobacillus plantarum have a relatively wide development space.
Disclosure of Invention
In order to overcome the defects in the prior art, the invention separates a lactobacillus plantarum (Lactobacillus plantarum) JF1 strain from the excrement of a healthy adult, and the lactobacillus plantarum JF1 strain has various probiotics and mainly comprises the functions of producing 3-hydroxybutyric acid, producing hyaluronic acid, inhibiting acetylcholinesterase activity and the like, thereby having great potential application value.
In order to achieve the above purpose, the present invention is realized by the following technical scheme:
the first aspect of the present invention provides a lactobacillus plantarum (lactobacillus plantarum) JF1 strain, wherein the lactobacillus plantarum JF1 strain is deposited with the China center for type culture collection at the year 09 and 27 of 2022, and the deposit number is: ccccccm 20221503; the full sequence of the 16SrDNA of the lactobacillus plantarum JF1 strain is shown in SEQ ID No: 1.
In a second aspect, the invention provides the use of a lactobacillus plantarum (Lactobacilli splanostarum) JF1 strain according to the first aspect for the production of 3-hydroxybutyric acid.
The research shows that the probiotic lactobacillus plantarum JF1 strain can produce 3-hydroxybutyric acid (3-HB), which suggests that the lactobacillus plantarum JF1 strain is expected to be used for producing 3-HB, and can be used for providing energy for various activities of the body, resisting osteoporosis, preventing and treating chronic syndrome, improving brain cognitive functions, improving lipid metabolism and other fields through the characteristic of producing 3-HB.
In a third aspect, the invention provides the use of a lactobacillus plantarum (lactobacillus plantarum) JF1 strain according to the first aspect for hyaluronic acid production.
The research shows that the probiotics lactobacillus plantarum JF1 strain can produce Hyaluronic Acid (HA), which suggests that the lactobacillus plantarum JF1 strain is hopeful to be used for producing HA, and is used for the fields of anti-inflammatory, anti-angiogenesis, anti-aging, wound inflammation promotion, healing and the like through the characteristic of producing HA.
In a fourth aspect, the invention provides the use of a lactobacillus plantarum (lactobacillus plantarum) JF1 strain according to the first aspect for the preparation of an acetylcholinesterase inhibitor.
The research shows that the probiotics lactobacillus plantarum JF1 strain can effectively inhibit the activity of acetylcholinesterase, and suggests that the lactobacillus plantarum JF1 strain is expected to be used in the fields of enhancing cognitive ability and memory, dilating blood vessels, exciting skeletal muscles and smooth muscles, promoting gastrointestinal absorption and peristalsis and the like.
In a fifth aspect, the invention provides the use of a lactobacillus plantarum (lactobacillus plantarum) JF1 strain according to the first aspect for the production of proteases.
The research shows that the lactobacillus plantarum JF1 strain can produce protease, which suggests that the lactobacillus plantarum JF1 strain is expected to be used for producing the protease, and the characteristic of the protease is used for promoting the digestion and absorption of human body on protein in food, improving the absorption of small peptide and amino acid, resisting allergy and other fields.
In a sixth aspect, the invention provides the use of a lactobacillus plantarum (lactobacillus plantarum) JF1 strain according to the first aspect for the production of gamma-aminobutyric acid.
The research shows that the lactobacillus plantarum JF1 strain can produce gamma-aminobutyric acid (GABA), which suggests that the lactobacillus plantarum JF1 strain is expected to be used for producing GABA, and the lactobacillus plantarum JF1 strain can be used for improving the sleeping quality of organisms, resisting depression, resisting anxiety, reducing blood pressure, improving lipid metabolism, enhancing memory capacity, improving brain activity and other fields through the characteristic of producing GABA.
In a seventh aspect, the invention provides the use of a lactobacillus plantarum (lactobacillus plantarum) JF1 strain according to the first aspect for the preparation of an alpha-glucosidase inhibitor.
Through researches, the probiotic lactobacillus plantarum JF1 strain can effectively inhibit the activity of alpha-glucosidase, the alpha-glucosidase is related to type 2 diabetes, and the inhibition of the alpha-glucosidase is one of methods for controlling postprandial hyperglycemia, so that the lactobacillus plantarum JF1 strain is suggested to be expected to be applied to the fields of reducing blood sugar, inhibiting obesity and the like.
According to an eighth aspect of the present invention there is provided a microbial agent for use in a probiotic apparatus, said microbial agent comprising a lactobacillus plantarum (lactobacillus plantarum) JF1 strain according to the first aspect.
Preferably, the lactobacillus plantarum (lactobacillus plantarum) JF1 strain is a fermented thallus mixture. The culture medium used for fermentation is MRS culture medium.
Preferably, in the field of pharmaceutical application, the microbial agent further comprises a pharmaceutically acceptable carrier and/or excipient.
More preferably, the above excipients refer to diluents, binders, lubricants, disintegrants, co-solvents, stabilizers, etc. and some pharmaceutically acceptable bases which are useful in the pharmaceutical field. The carrier is a functional pharmaceutical adjuvant available in the pharmaceutical field and comprises a surfactant, a suspending agent, an emulsifying agent and a plurality of novel pharmaceutical polymer materials, such as cyclodextrin, chitosan, polylactic acid (PLA), polyglycolic acid-polylactic acid copolymer (PLGA), hyaluronic acid and the like.
Preferably, in the field of medical application, the dosage forms of the microbial inoculum comprise injection, tablet, granule, capsule, dripping pill, sustained release agent and oral liquid preparation.
More preferably, the above-mentioned dosage forms refer to clinically usual dosage forms. Pharmaceutical formulations may be administered orally or parenterally (e.g., intravenously, subcutaneously, intraperitoneally, or topically), and if some drugs are unstable under gastric conditions, they may be formulated as enteric coated tablets.
Compared with the prior art, the invention has the beneficial effects that:
the invention separates a lactobacillus plantarum (Lactiplantibacillus plantarum) JF1 strain from the feces of a healthy adult, and the lactobacillus plantarum JF1 strain has various probiotics effects, including excellent protease activity, capability of producing and secreting 3-hydroxybutyric acid, capability of producing and secreting gamma-aminobutyric acid, capability of producing and secreting hyaluronic acid, capability of inhibiting alpha-glucosidase activity and capability of inhibiting acetylcholinesterase activity. Therefore, the lactobacillus plantarum JF1 strain has the effects of promoting digestion and absorption of protein foods and improving protein allergy; anti-inflammatory and improving intestinal flora; anti-depression and anti-alcohol; anti-aging and anti-wrinkle; reducing blood glucose; promoting intestinal absorption and peristalsis. Therefore, the lactobacillus plantarum JF1 strain newly separated by the invention has various probiotics effects, can be used in the fields of antibiosis, anti-inflammation, anti-aging and the like, for example, can be manufactured into anti-aging food and medicine, and has important application value and economic value.
Drawings
FIG. 1 is a phylogenetic tree of Lactobacillus plantarum JF1 and other homologous strains;
FIG. 2 shows the degradation experiment (left, blank control; right, experimental group) of Lactobacillus plantarum JF1 on milk plates;
FIG. 3 shows that Lactobacillus plantarum JF1 can produce and secrete 3-hydroxybutyric acid;
FIG. 4 shows that Lactobacillus plantarum JF1 can produce and secrete gamma-aminobutyric acid;
FIG. 5 shows that Lactobacillus plantarum JF1 can produce and secrete hyaluronic acid;
FIG. 6 shows that the secretion of the fermentation broth of Lactobacillus plantarum JF1 significantly inhibits the activity of alpha-glucosidase;
fig. 7 shows that the secretion of lactobacillus plantarum JF1 fermentation broth significantly inhibited acetylcholinesterase activity.
Detailed Description
The following describes the invention in more detail. The description of these embodiments is provided to assist understanding of the present invention, but is not intended to limit the present invention. In addition, the technical features of the embodiments of the present invention described below may be combined with each other as long as they do not collide with each other.
The experimental methods in the following examples, unless otherwise specified, are conventional, and the experimental materials used in the following examples, unless otherwise specified, are commercially available.
The following examples relate to the following experimental materials:
(1) Strains: lactobacillus plantarum (Lactipiantibacillus splantarum) JF1 strain is isolated from feces of a healthy adult in Guangdong of China by a laboratory of a national institute of university of middle mountain student's academy of life intestinal microbiome research and moisturizing laboratory and stored in a glycerol tube at-80 ℃ in a low-temperature freezing manner. Typically, it is inoculated onto the surface of an MRS solid medium plate and cultured upside down in a thermostatic anaerobic incubator at 37℃for 24 hours to obtain colonies, or cultured in an MRS liquid medium with shaking in a thermostatic anaerobic incubator at 37℃for 24-48 hours to obtain a fermentation broth.
(2) The kit comprises: 3-hydroxybutyric acid (3-HB) detection kit (Cloud-CloneCorp., cat: CEB022 Ge), gamma-aminobutyric acid (GABA) detection kit (Cloud-CloneCorp., cat: CEA900 Ge), hyaluronic acid (also known as hyaluronic acid, HA) detection kit (Cloud-CloneCorp., cat: CEA182 Ge), alpha-glucosidase inhibitor screening kit (abcam, cat: ab 284520), acetylcholinesterase inhibitor screening kit (abnova, cat: KA 6219).
(3) MRS plate: 10g of beef extract, 10g of peptone, 5g of yeast extract, 2g of triammonium citrate, 5g of sodium acetate, 20g of glucose, 2g of dipotassium hydrogen phosphate, 1mL of Tween 80, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 15g of agar and ddH 2 Filling O to 1L, adjusting pH to 6.2-6.6, and autoclaving at 121deg.C for 20min to obtain MRS plate.
(4) MRS liquid medium: 10g of beef extract, 10g of peptone, 5g of yeast extract, 2g of triammonium citrate, 5g of sodium acetate, 20g of glucose, 2g of dipotassium hydrogen phosphate, 1mL of Tween 80, 0.58g of magnesium sulfate, 0.25g of manganese sulfate and ddH 2 Adding O to 1L, adjusting pH to 6.2-6.6, and autoclaving at 121deg.C for 20min to obtain MRS liquid culture medium.
(5) MP plate: 10g of skimmed milk powder, 1g of sodium chloride, 10g of beef extract, 10g of peptone, 5g of yeast extract, 20g of glucose, 2g of tri-ammonium citrate, 5g of sodium acetate, 2g of dipotassium hydrogen phosphate, 0.5mL of Tween 80, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 15g of agar and ddH 2 Filling O to 1L, adjusting pH to 6.2-6.6, autoclaving at 121deg.C for 20min, and making into MP plate.
Example 1 isolation and characterization of Lactobacillus plantarum (Lactobacilli splanostarum) JF1
Lactobacillus plantarum lactplatinum lasticillum flutarum jf1 strain was isolated from faeces of a healthy adult (male, 32 years, bmi=22.5) in the sea bead region of guangzhou province, guangdong, china, as follows:
the fecal sample was repeatedly washed 3 times with sterile water, placed in a mortar, 500uL of sterile water was added per 100mg of fecal sample, thoroughly ground to a homogenate, and then an appropriate amount of the grinding fluid was pipetted, spread on an MRS plate, and incubated at room temperature for 3 days. The colonies to be streaked and purified in the separation experiment plates were numbered with a marker and strain numbers were marked on the plates accordingly. Colonies were picked and inoculated onto MRS plates and the strains were purified by plate streaking. If the strain cannot be separated by the method, colonies need to be picked from the enrichment plate, and the colonies are coated on the MRS plate after being subjected to gradient dilution by the MRS liquid culture medium. Reference is made to the "Berger's Manual of bacteria identification" (eighth edition) and the "manual of fungus classification identification", which identify strains belonging to bacteria first. The primary separation is carried out to obtain a purified strain, the strain number is JF1, and after 24 hours of culture, the colony is observed to be white round and convex in the center, and the surface of the colony is fine and smooth.
Next, after molecular identification by 16SrDNA universal primer (27F: AGAGTTTGATCTGGCTCAG, 1492R: TACGGCTACCTTGTTACGACCTT), the isolated L.plantarumJF1 strain was subjected to whole genome sequencing by Beijing Baimeike Biotechnology Co. The resulting sequence 16SrDNAsequence (SEQ ID No: 1) was subjected to BLAST alignment at the Genome database of NCBI. The results showed that the JF1 strain had >99% homology with the known lactobacillus plantarum 16SrDNA sequence and was analyzed by evolution with the homologous strain to confirm that the JF1 strain was a different strain of lactobacillus plantarum (fig. 1).
Finally, strain JF1 is preserved, and the preservation information is as follows: preservation time: 2022, 09, 27; preservation unit name: china Center for Type Culture Collection (CCTCC); deposit number: ccccccm 20221503; deposit unit address: chinese university of Wuhan; classification naming: lactplatinum lanostarum.
Lactobacillus plantarum is a probiotic bacterial strain which can be used for food, has wide probiotic effects, such as anti-inflammatory, antihypertensive and antipathogenic bacteria, and the like, but different strains from different sources have different effects, which indicates that the novel lactobacillus plantarum JF1 separated from human excrement can be used as probiotic bacteria, and possibly has novel effects and functions.
16SrDNAsequence (1439bp,SEQ ID No:1) of planarumjf 1:
ATAATGCAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGTTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTAGGAACCAGCCGCCT。
example 2 function of Lactobacillus plantarum (Lactobacilli splantarum) JF1 and application thereof
(1) Protease capable of producing degradable milk protein by lactobacillus plantarum JF1 strain
The identification and measurement of the ability of lactobacillus plantarum JF1 to secrete proteolytic proteins were performed according to an agar well diffusion test using a skim milk plate medium (MP plate). In the test, 3uL of lactobacillus plantarum JF1 bacteria solution with the concentration of 10Abs is dripped into the test group, and 3uL of blank MRS culture medium is dripped into the control group. The cells were cultured in an anaerobic incubator at 37℃for 3 days in an inverted manner. The results showed that JF1 significantly degraded protein and formed a distinct degradation circle (fig. 2) compared to the control with the blank medium, indicating that lactobacillus plantarum JF1 produced protease.
Therefore, the lactobacillus plantarum JF1 can promote the digestion and absorption of human body to protein in food and improve the absorption of small peptide and amino acid when used as a probiotic bacterial strain. And can be used for resisting allergy (improving food allergy caused by protein dyspepsia or non-absorption). In addition, the method can also be used for extracting protease and is applied to the production in the protease fields of food industry, washing industry and the like; can also be used in microbial feed to help animals digest and absorb nutrition, and improve the utilization rate of the feed.
(2) Lactobacillus plantarum JF1 strain can produce and secrete 3-hydroxybutyric acid (3-HB)
Lactobacillus plantarum JF1 strain cultured with MRS liquid medium to stationary phase was expanded into new MRS liquid medium at dilution ratio of 1:30, bacterial suspension was collected at 24h of stationary phase, and fermentation broth supernatant was collected after centrifugation at 10,000Xg and 4℃for 10min, and 3-HB concentration of the fermentation broth supernatant was measured by 3-HB specific ELISA kit (CEB 022 Ge). The results showed that the concentration of 3-HB in the supernatant of JF1 was 1.752. Mu.g/mL compared to the absence of 3-HB in the blank medium MRS, indicating that Lactobacillus plantarum JF1 can produce and secrete 3-hydroxybutyric acid during the stationary phase (FIG. 3).
3-HB can provide energy for various physical activities and is a potential energy/functional food that has been added to athlete drinks, so the probiotic Lactobacillus plantarum JF1 strain can be used as an additive to energy foods. Meanwhile, 3-HB can also effectively resist osteoporosis, prevent and treat chronic syndromes (hypertension, alcoholic fatty liver, enteritis, intestinal cancer and the like), improve brain cognitive functions (improving learning and memory capacity, protecting glial cells, improving Alzheimer's disease and the like), and improve lipid metabolism. Thus, the probiotic Lactobacillus plantarum JF1 strain can exert the above multiple purposes through the effect of producing 3-hydroxybutyric acid.
In addition, 3-hydroxybutyric acid is an endogenous small molecule substance naturally produced by the body, has an important role in maintaining the integrity of colorectal tissues, and has the functions of maintaining intestinal health, preventing colonic diseases and diminishing inflammation and productivity. The 3-HB treatment can promote the proliferation of beneficial intestinal bacteria, relieve the symptoms of multiple sclerosis, and has great potential in the aspects of regulating flora and improving health. Therefore, the probiotic lactobacillus plantarum JF1 strain also helps to improve intestinal flora and relieve intestinal inflammation.
(3) Lactobacillus plantarum JF1 strain can produce and secrete gamma-aminobutyric acid (GABA)
Lactobacillus plantarum JF1 cultured with MRS liquid culture medium to a stationary phase is expanded and cultured into a new MRS liquid culture medium at a dilution ratio of 1:30, bacterial suspension is collected when the culture is carried out to the stationary phase for 24 hours, fermentation broth supernatant is collected after centrifugation at 10,000Xg and 4 ℃ for 10 minutes, and GABA concentration of the fermentation broth supernatant is measured by a GABA specific ELISA kit (CEA 900 Ge). The results showed that the concentration of GABA in the fermentation supernatant of JF1 was significantly increased compared to the low concentration of GABA in the blank medium MRS, and the accumulated amount was 206.4pg/mL, indicating that lactobacillus plantarum JF1 can produce and secrete gamma-aminobutyric acid in the stationary phase (fig. 4).
Gamma-aminobutyric acid is an important central nervous system inhibitory neurotransmitter, and is widely present in animals, plants and microorganisms. It has been demonstrated that GABA, a small molecular weight non-protein amino acid, is food safe and can be used as a food additive. Research shows that intake of a certain amount of GABA has the physiological effects of improving sleeping quality of organisms, resisting depression, resisting anxiety, reducing blood pressure, improving lipid metabolism, enhancing memory and brain activity, accelerating brain metabolism, strengthening liver and kidney, promoting ethanol metabolism (dispelling alcohol effect), improving climacteric syndrome and the like.
Thus, the probiotic Lactobacillus plantarum JF1 strain can exert the above multiple purposes through the effect of producing gamma-aminobutyric acid.
(4) Lactobacillus plantarum JF1 strain can produce and secrete Hyaluronic Acid (HA)
Lactobacillus plantarum JF1 cultured with MRS liquid culture medium to a stationary phase is expanded and cultured into a new MRS liquid culture medium at a dilution ratio of 1:30, bacterial suspension is collected when the culture is carried out to the stationary phase for 24 hours, fermentation broth supernatant is collected after centrifugation at 10,000×g and 4 ℃ for 10 minutes, and HA concentration of the fermentation broth supernatant is measured by a hyaluronic acid (also called hyaluronic acid, HA) specific ELISA kit (CEA 182 Ge). The results showed that the concentration of HA in the fermentation supernatant of JF1 was significantly increased compared to the low concentration of HA in the blank medium MRS, with an accumulated amount of 95.2ng/mL, indicating that lactobacillus plantarum JF1 can produce and secrete hyaluronic acid in the stationary phase (fig. 5).
Hyaluronic acid, also known as hyaluronic acid, is a biodegradable, biocompatible, non-toxic, non-allergenic polymer with a variety of biological functions. Has anti-inflammatory and anti-angiogenesis effects, and has strong anti-aging, moisturizing and wrinkle smoothing abilities. As the anti-wrinkle agent is favorable for skin anti-wrinkle, promotes wound anti-inflammation and healing, can be used as an anti-wrinkle agent, and has the potential of developing skin cosmetics. In addition, HA HAs high lubricating, water absorbing and retaining capacity, and may affect various cell functions, such as migration, adhesion, proliferation, etc. and is used widely in ophthalmic surgery, arthritis treatment, wound healing rack, tissue engineering, implantation material, etc.
Thus, the probiotic lactobacillus plantarum JF1 strain may serve several purposes as above by producing hyaluronic acid.
(5) The lactobacillus plantarum JF1 strain fermentation liquor can effectively inhibit the activity of alpha-glucosidase
Lactobacillus plantarum JF1 cultured by using an MRS liquid culture medium to a stationary phase is expanded and cultured into a new MRS liquid culture medium at a dilution ratio of 1:30, bacterial suspension is collected when the culture is carried out to the stationary phase for 24 hours, fermentation broth supernatant is collected after centrifugation at 10,000Xg and 4 ℃ for 10 minutes, and the influence of the fermentation broth supernatant on the enzyme activity ability of alpha-glucosidase to hydrolyze glucose is measured by an alpha-glucosidase inhibitor screening kit (ab 284520). The results showed that the fermentation supernatant of JF1 significantly inhibited the ability of α -glucosidase to hydrolyze glucose compared to the non-inhibitory effect of the blank medium MRS, with an inhibition rate of about 88.24%, indicating that the fermentation broth of lactobacillus plantarum JF1 can effectively inhibit the activity of α -glucosidase (fig. 6).
a-glucosidase, an enzyme that plays a role in carbohydrate breakdown, is associated with type 2 diabetes, and inhibition of α -glucosidase is one of the methods of controlling postprandial hyperglycemia, thereby contributing to the treatment of diabetes. Thus, α -glucosidase inhibitors help to maintain blood glucose levels and can improve diabetic complications. Furthermore, inhibition of α -glucosidase activity may control obesity.
Thus, the probiotic lactobacillus plantarum JF1 strain has an inhibitory activity on a-glucosidase, which makes it a potential hypoglycemic and obesity inhibiting probiotic.
(6) The lactobacillus plantarum JF1 strain fermentation liquor can effectively inhibit the activity of acetylcholinesterase
Lactobacillus plantarum JF1 cultured by using an MRS liquid culture medium to a stationary phase is expanded and cultured into a new MRS liquid culture medium at a dilution ratio of 1:30, bacterial suspension is collected when the culture medium is cultured to the stationary phase for 24 hours, fermentation broth supernatant is collected after centrifugation at 10,000×g and 4 ℃ for 10 minutes, and the inhibitory capacity of the fermentation broth supernatant on acetylcholinesterase (AchE) activity is measured by an acetylcholinesterase inhibitor screening kit (KA 6219). The results showed that the fermentation supernatant of JF1 significantly inhibited the activity of acetylcholinesterase compared to the non-inhibitory effect of the blank medium MRS, with an inhibition rate of about 13.28%, indicating that the fermentation broth of lactobacillus plantarum JF1 can effectively inhibit the activity of acetylcholinesterase (fig. 7).
The acetylcholinesterase inhibitor can enable cholinergic to accumulate at synapses under the action of acetylcholine released from nerve fiber tips through reversible inhibition of acetylcholinesterase, thereby exciting cholinergic receptors, and prolonging and increasing the action of acetylcholine. While acetylcholinesterase inhibitors can enhance cognition and memory (improve Alzheimer's disease, improve cognitive dysfunction, etc.), dilate blood vessels (improve tachycardia, etc.), excite skeletal and smooth muscles (improve constipation, postoperative abdominal distension, postoperative urinary retention, etc.).
Thus, the probiotic lactobacillus plantarum JF1 strain excites skeletal and intestinal smooth muscle by inhibiting acetylcholinesterase activity, which makes it a potential probiotic for promoting gastrointestinal absorption and peristalsis.
Taken together, the novel isolated lactobacillus plantarum JF1 strain of the present invention has a variety of probiotic effects: (1) has excellent protease activity; (2) 3-hydroxybutyric acid may be produced and secreted; (3) gamma-aminobutyric acid can be produced and secreted; (4) can produce and secrete hyaluronic acid; (5) can inhibit alpha-glucosidase activity; (6) can inhibit acetylcholinesterase activity. Therefore, the lactobacillus plantarum JF1 strain has important application value and economic value.
The embodiments of the present invention have been described in detail above, but the present invention is not limited to the described embodiments. It will be apparent to those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, and yet fall within the scope of the invention.
Claims (9)
1. A lactobacillus plantarum (JF 1 strain) characterized in that the lactobacillus plantarum JF1 strain was deposited with the chinese collection of typical cultures at month 27 of 2022, accession number: ccccccm 20221503; the full sequence of the 16SrDNA of the lactobacillus plantarum JF1 strain is shown in SEQ ID No: 1.
2. Use of a lactobacillus plantarum (lactplatinum splantarum) JF1 strain according to claim 1 for the production of 3-hydroxybutyric acid.
3. Use of lactobacillus plantarum (lactplantibilum) JF1 strain according to claim 1 for the production of hyaluronic acid.
4. Use of a lactobacillus plantarum (lactplatinum) JF1 strain according to claim 1 for the preparation of an acetylcholinesterase inhibitor.
5. Use of a lactobacillus plantarum (lactplatinum) JF1 strain according to claim 1 for the production of proteases.
6. Use of lactobacillus plantarum (lactplatinum splantarum) JF1 strain according to claim 1 for the production of gamma-aminobutyric acid.
7. Use of a lactobacillus plantarum (lactplatinum) JF1 strain according to claim 1 for the preparation of an alpha-glucosidase inhibitor.
8. A microbial agent for use in a probiotic function, comprising the lactobacillus plantarum (JF 1) strain of claim 1.
9. A bacterial agent for use in a probiotic apparatus according to claim 8, wherein the lactobacillus plantarum (lactobacillus plantarum) JF1 strain is a fermented bacterial mixture.
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