CN117987482A - Enterococcus faecalis XY7 and application thereof in preparation of anti-aging and sleep-aiding foods and medicines - Google Patents
Enterococcus faecalis XY7 and application thereof in preparation of anti-aging and sleep-aiding foods and medicines Download PDFInfo
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Abstract
The invention belongs to the technical field of probiotics and application thereof, and particularly relates to enterococcus faecalis XY7 and application thereof in preparation of anti-aging and sleep-aiding food and drugs. In order to further develop and utilize the probiotics function of enterococcus faecalis, the invention separates and purifies a fecal sample of a healthy adult in Guangdong area to obtain a enterococcus faecalis (Enterococcus faecalis) XY7 strain, the strain can produce and secrete gamma-aminobutyric acid, can produce and secrete hyaluronic acid, can inhibit acetylcholinesterase activity, has superoxide dismutase activity, is hopeful to be applied to the fields of anti-depression, anti-alcohol and sleep-aiding, whitening and anti-aging, promoting intestinal absorption and peristalsis, enhancing immunity, protecting liver and the like, and has important application value and economic value in the fields of anti-aging, sleep-aiding and the like.
Description
Technical Field
The invention belongs to the technical field of probiotics and application thereof, and particularly relates to enterococcus faecalis XY7 and application thereof in preparation of anti-aging and sleep-aiding food and drugs.
Background
Enterococcus faecalis (Enterococcus faecalis) is a gram-positive bacterium and a hydrogen peroxide-negative bacterium, and is one of the main flora in the human and animal intestinal tracts. Enterococcus faecalis (E.faecalis) is a facultative anaerobic gram-positive lactic acid bacterium, the form of the bacterium is spherical or chain, most of the bacterium is arranged in double or short chain, the bacterium is usually not moving, the diameter is 0.5-1.0 microns, the bacterium is free of capsules and spores, the bacterium has strong adaptability and resistance to the environment, and the bacterium can resist various antibiotics such as tetracycline, kanamycin and gentamicin, and the requirement on growth conditions is not strict. In addition, enterococcus faecalis can use arginine as an energy source to ferment sorbitol, but cannot ferment arabinose.
Enterococcus faecalis is a normal colonising bacteria of the human intestinal tract and is present in approximately 90-95% of the human body. This bacterium is often one of the first microorganisms to colonize the human gastrointestinal tract early in life and plays an important role in the development of the intestinal immune system. The feed additive catalog (2013) divides enterococcus faecalis into species that can be added to feed, which can form a biofilm in the animal's gut to adhere to the mucosa of the animal's gut and develop, grow and reproduce thereon. In addition, enterococcus faecalis can soften fibers in the feed and improve the conversion rate of the feed.
Enterococcus faecalis is one of main flora in intestinal tracts of human beings and animals, can generate natural antibiotics, and is beneficial to body health; bacteriostasis substances such as bacteriocin and the like can be produced, the growth of pathogenic bacteria such as escherichia coli, salmonella and the like can be inhibited, and the intestinal microenvironment can be improved; can inhibit the proliferation of urease-producing bacteria and putrefying bacteria in intestinal tract, reduce the content of urease and endotoxin in intestinal tract, and reduce the content of ammonia and endotoxin in blood. Meanwhile, enterococcus faecalis is a type of microorganism normally existing in the digestive tract, has strong tolerance and colonization capacity in intestinal mucosa, is facultative anaerobic lactobacillus, is suitable for production and application, is also a good feeding microorganism, and has wide application in aquatic products and livestock and poultry. In addition, enterococcus faecalis is not only a normal flora in human and animal intestinal tracts, but also an important lactobacillus, has better intestinal adhesion capability, can generate lactic acid and some antibacterial substances, inhibit the reproduction of intestinal putrefying bacteria and pathogenic bacteria and intestinal infection, reduce diarrhea rate, promote the digestion and absorption of nutrient substances, enhance immunity, improve feed intake and feed conversion rate, promote animal health, and is one of the probiotic bacteria used first choice in livestock production and food industry. According to researches of scholars, enterococcus faecalis and cellulase are added into the corn straw silage, so that the color, smell, texture and the like of the silage can be obviously improved, the disappearance rate of dry matters is improved by 8%, the mass ratio of ammoniacal nitrogen to total nitrogen is reduced by 33%, the mass fraction of butyric acid is reduced by 82%, and meanwhile, the contents of neutral washing fibers and acidic washing fibers are respectively reduced by 10% and 7%, and the quality of the silage is obviously improved. Meanwhile, researches prove that enterococcus faecalis can generate a plurality of antibacterial substances, and the antibacterial substances have good inhibition effects on pathogenic bacteria such as salmonella, escherichia coli, staphylococcus aureus and the like. Research has also shown that enterococcus faecalis is capable of producing vodkamycin and can effectively inhibit the growth and reproduction of listeria, staphylococcus aureus and spoilage microorganisms. Enterococcus faecalis also enhances the activity of macrophages, promotes the immune response in animals, and increases antibody levels. Enterococcus faecalis can form a biological film in the intestinal tract of an animal to be adhered to the intestinal mucosa of the animal, and can form a lactobacillus barrier to resist the negative effects of external germ viruses, mycotoxins and the like during development, growth and reproduction.
A great deal of research shows that different strains of enterococcus faecalis have different probiotic functions. Such as: (1) Enterococcus faecalis strain T-110 was used as A probiotic (BIO-THREE) in combination with Bacillus subtilis TO-A and Clostridium butyricum TO-A. Together, these three strains can reduce post-operative infections in surgical patients and shorten hospitalization times in infants suffering from acute diarrhea. (2) A crossover study found that a single administration of enterococcus faecalis YM0831 strain to healthy human subjects significantly improved the response of blood glucose to sucrose tolerance tests.
Since enterococcus faecalis sources are diverse, genes thereof are diverse and functionally diverse. However, the separation and identification, the probiotics and the metabolism mechanism of enterococcus faecalis are still less studied at present, and the development and the utilization of the enterococcus faecalis are affected to a certain extent. Therefore, more enterococcus faecalis with a probiotic function needs to be excavated, and different functions of the enterococcus faecalis need to be further excavated according to different sources of the enterococcus faecalis, so that the enterococcus faecalis can better play roles, for example, the efficacy of the enterococcus faecalis is determined according to the functions of the strain or the probiotic metabolic products, and the application prospect of the enterococcus faecalis is clear. In conclusion, the research and application of enterococcus faecalis have a very wide development space.
Disclosure of Invention
In order to overcome the defects in the prior art, the enterococcus faecalis (Enterococcus faecalis) XY7 strain is obtained by separating and purifying faeces samples of a healthy adult in Guangdong region, can produce and secrete gamma-aminobutyric acid, can produce and secrete hyaluronic acid, can inhibit acetylcholinesterase activity, has superoxide dismutase activity, and has important potential application value in the fields of anti-aging, sleep aiding and the like.
In order to achieve the above purpose, the technical scheme adopted by the invention is as follows:
The first aspect of the invention provides an enterococcus faecalis (Enterococcus faecalis) XY7 strain, wherein the enterococcus faecalis XY7 strain is preserved in China center for type culture Collection (China, accession number: CCTCC NO: M20231166; the 16S rDNA complete sequence of the enterococcus faecalis XY7 strain is shown in SEQ ID No: 1.
In a second aspect, the invention provides the use of the enterococcus faecalis (Enterococcus faecalis) XY7 strain of the first aspect in the production of gamma-aminobutyric acid.
The research shows that the probiotics enterococcus faecalis XY7 strain can produce gamma-aminobutyric acid (GABA), which suggests that the enterococcus faecalis XY7 strain can be used for producing GABA and is used for improving the fields of sleeping quality of organisms, resisting depression, resisting anxiety, reducing blood pressure, improving lipid metabolism and the like through the characteristic of producing GABA.
In a third aspect, the invention provides the use of the enterococcus faecalis (Enterococcus faecalis) XY7 strain of the first aspect in the production of hyaluronic acid.
The research shows that the probiotics enterococcus faecalis XY7 strain can produce Hyaluronic Acid (HA), which suggests that the enterococcus faecalis XY7 strain is hopeful to be used for producing HA, and the characteristics of producing HA are applied to the fields of anti-inflammatory and anti-angiogenesis effects, anti-aging, moisturizing, smoothing wrinkles, promoting wound inflammation and healing and the like.
In a fourth aspect, the invention provides the use of an enterococcus faecalis (Enterococcus faecalis) XY7 strain according to the first aspect in the preparation of an acetylcholinesterase inhibitor.
The study shows that the probiotics enterococcus faecalis XY7 strain can effectively inhibit the activity of acetylcholinesterase, and the probiotics enterococcus faecalis XY7 strain can be used for enhancing the cognitive ability and the memory, dilating blood vessels, exciting skeletal muscles, smooth muscles and other fields by inhibiting the activity of acetylcholinesterase.
In a fifth aspect, the invention provides the use of the enterococcus faecalis (Enterococcus faecalis) XY7 strain of the first aspect for producing superoxide dismutase.
The research shows that the probiotics enterococcus faecalis XY7 strain can produce superoxide dismutase (SOD), and the fermentation liquor has the activity of superoxide dismutase (SOD), which suggests that the enterococcus faecalis XY7 strain is hopeful to be used for preparing the SOD, and is applied to the fields of antioxidation, anti-inflammation, immunity enhancement, aging resistance, blood fat reduction, liver function enhancement, vision improvement, blood sugar reduction and the like through the function with the activity of the SOD.
In a sixth aspect, the present invention provides a method for preparing hyaluronic acid, specifically: inoculating the enterococcus faecalis (Enterococcus faecalis) XY7 strain in the first aspect into MRS liquid culture medium, culturing until the bacteria suspension is recovered after the stationary phase, and finally separating and purifying hyaluronic acid in the bacteria suspension.
Of course, the same method can be used for preparing gamma-aminobutyric acid, superoxide dismutase and the like after different separation and purification steps.
Preferably, the conditions of the culture are: anaerobic at a constant temperature of 37 ℃.
Preferably, enterococcus faecalis (Enterococcus faecalis) XY7 strain is cultured with MRS liquid medium to stationary phase, and then expanded into new MRS liquid medium at dilution ratio of 1:20-1000.
Compared with the prior art, the invention has the beneficial effects that:
The invention separates and purifies faeces samples of a healthy adult in Guangdong area to obtain a strain of enterococcus faecalis (Enterococcus faecalis) XY7, which has various probiotics effects including producing and secreting gamma-aminobutyric acid, producing and secreting hyaluronic acid, inhibiting acetylcholinesterase activity and superoxide dismutase activity. Thus, enterococcus faecalis XY7 strain has antidepressant, anti-alcohol and sleep-aiding effects; whitening and anti-aging; promoting intestinal absorption and peristalsis; enhancing immunity and protecting liver. Therefore, the enterococcus faecalis XY7 strain newly separated by the invention has various probiotics, can be used in the fields of anti-aging, sleep-aiding and the like, for example, can be prepared into anti-aging and sleep-aiding medicines, and has important application value and economic value.
Drawings
FIG. 1 is a phylogenetic tree of enterococcus faecalis XY7 strains (the built strain is derived from the Genome database of NCBI);
FIG. 2 shows that enterococcus faecalis XY7 strain may produce and secrete gamma-aminobutyric acid;
FIG. 3 shows that enterococcus faecalis XY7 strain can produce and secrete hyaluronic acid;
FIG. 4 shows that the secretable material of the fermentation broth of enterococcus faecalis XY7 strain significantly inhibited the activity of acetylcholinesterase;
FIG. 5 shows SOD activity of fermentation broth of enterococcus faecalis XY7 strain.
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) Enterococcus faecalis (Enterococcus faecalis) XY7 strain was isolated from an intestinal fecal sample of a healthy adult (BMI=19.5) in Guangzhou area of China by an intestinal microbiome study moisturizing laboratory and placed in a glycerol tube for cryogenic storage at-80 ℃. In general, the strain is inoculated on the surface of an MRS solid culture medium flat plate and is cultured for 24 hours in an inverted way in a constant temperature anaerobic incubator at 37 ℃ to obtain a bacterial colony, or is cultured for 24-48 hours in an MRS liquid culture medium in an oscillation way in the constant temperature anaerobic incubator at 37 ℃ to obtain a fermentation liquor.
(2) The kit comprises: gamma-aminobutyric acid (GABA) detection kit (Cloud-Clone corp., cat: CEA900 Ge), hyaluronic acid (also known as hyaluronic acid, HA) detection kit (Cloud-Clone corp., cat: CEA182 Ge), acetylcholinesterase inhibitor screening kit (abnova, cat: KA 6219), superoxide dismutase (SOD) assay kit (Nanjing build, cat: a 001-3).
(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, 1L of ddH 2 O, 6.2-6.6 of pH value, and autoclaving at 121 ℃ for 20min to prepare an 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, supplementing ddH 2 O to 1L, adjusting the pH to 6.2-6.6, and carrying out high-pressure sterilization at 121 ℃ for 20min to prepare the MRS liquid culture medium.
EXAMPLE 1 isolation and identification of enterococcus faecalis (Enterococcus faecalis) XY7 Strain
Enterococcus faecalis (Enterococcus faecalis) XY7 strain is isolated from faeces of a healthy adult in Guangdong province of China, and is specifically 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 homogenate, and an appropriate amount of the grinding fluid was pipetted, spread on an MRS plate, and incubated at room temperature for 3 days. Colonies to be streaked and purified in the separation assay plates were then numbered with a marker and strain numbers were marked on the plates accordingly. After labelling, 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 a culture medium after being subjected to gradient dilution by MRS liquid culture medium. Finally, reference is made to the "Berger's Manual of bacteria identification" (eighth edition) and the "manual of fungus classification identification", first to distinguish strains belonging to bacteria, and then to observe the growth conditions of colonies. The primary separation is carried out to obtain a purified strain, the strain number is XY7, and after 24 hours of culture, bacterial colonies of the strain are observed to be round or oval, moist, convex, smooth in edge and flat in surface.
Next, the isolated XY7 strain was subjected to molecular characterization by a 16S rDNA universal primer (27F: AGAGTTTGATCCTGGCTCAG,1492R: TACGGCTACCTTGTTACGACTT), and then the XY7 strain was subjected to whole genome sequencing by Beijing Baimeike Biotechnology Co. The resulting sequence 16S rDNA sequence (SEQ ID No: 1) was subjected to BLAST alignment at NCBI' S Genome database. The results showed >99% homology with the known 16S rDNA sequence of enterococcus faecalis (Enterococcus faecalis) and evolutionary analysis with homologous strains (FIG. 1) confirmed that XY7 was a different strain of enterococcus faecalis.
Finally, strain XY7 was deposited with the following information: preservation time: 2023, 7, 3; preservation unit name: china Center for Type Culture Collection (CCTCC); deposit number: CCTCC NO: M20231166; deposit unit address: chinese university of Wuhan; classification naming: enterococcus faecalis.
Enterococcus faecalis has wide probiotic effects such as anti-inflammatory, anti-aging and cholesterol reducing effects, but strains from different sources have different effects, which indicates that a novel enterococcus faecalis XY7 separated from human faeces can be used as a probiotic and possibly has novel effects and functions.
Enterococcus faecalis XY7 16S rDNA sequence(1425bp，SEQ ID No：1)：
GCTATACTGCAGTCGAACGCTTCTTTCCTCCCGAGTGCTTGCNCTCNATTGGAAAGAGGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTACCCATCAGAGGGGGATAACACTTGGAAACAGGTGCTAATACCGCATAACAGTTTATGCCGCATGGCATAAGAGTGAAAGGCGCTTTCGGGTGTCGCTGATGGATGGACCCGCGGTGCATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCCACGATGCATAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCGGCAATGGACGAAAGTCTGACCGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAACTCTGTTGTTAGAGAAGAACAAGGACGTTAGTAACTGAACGTCCCCTGACGGTATCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGAGACTTGAGTGCAGAAGAGGAGAGTGGAATTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAGGAACACCAGTGGCGAAGGCGGCTCTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCAAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTTTGACCACTCTAGAGATAGAGCTTTCCCTTCGGGGACAAAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTGTTAGTTGCCATCATTTAGTTGGGCACTCTAGCGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGGAAGTACAACGAGTCGCTAGACCGCGAGGTCATGCAAATCTCTTAAAGCTTCTCTCAGTTCGGATTGCAGGCTGCAACTCGCCTGCATGAAGCCGGAATCGCTAGTAATCGCGGATCAGCACGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTTGG.
Example 2 Functions of enterococcus faecalis (Enterococcus faecalis) XY7 Strain and uses thereof
(1) Enterococcus faecalis XY7 strain can produce and secrete gamma-aminobutyric acid (GABA)
Enterococcus faecalis XY7 cultured in 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, fermentation broth supernatant was collected after centrifugation at 10,000Xg and 4℃for 10min, and GABA concentration of fermentation broth supernatant was measured by GABA specific ELISA kit (CEA 900 Ge). The results showed that the concentration of GABA in the fermentation supernatant of strain XY7 was significantly increased compared to the low concentration of GABA in the blank medium MRS, and the accumulated amount was 75.59pg/mL, indicating that enterococcus faecalis XY7 can produce and secrete gamma-aminobutyric acid in the stationary phase (fig. 2).
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 enterococcus faecalis XY7 strain may serve several of the above purposes by producing gamma-aminobutyric acid.
(2) Enterococcus faecalis XY7 strain can produce and secrete Hyaluronic Acid (HA)
Enterococcus faecalis XY7 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 culture to stationary phase, fermentation broth supernatant was collected after centrifugation at 10,000Xg at 4℃for 10min, and then HA concentration of fermentation broth supernatant was determined by hyaluronic acid (also known as hyaluronic acid, HA) specific ELISA kit (CEA 182 Ge). The results showed that the concentration of HA in the fermentation supernatant of strain XY7 was significantly increased compared to the low concentration of HA in the blank medium MRS, with an accumulated amount of 106.03ng/mL, indicating that enterococcus faecalis XY7 can produce and secrete hyaluronic acid during stationary phase (fig. 3).
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. The anti-wrinkle agent is beneficial to 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 ability, and can affect various cell functions such as migration, adhesion and proliferation, so that HA is also widely used in biomedical fields such as ophthalmic surgery, arthritis treatment, wound healing scaffolds, tissue engineering, implant materials, and the like.
Thus, the probiotic enterococcus faecalis XY7 strain may serve several of the above purposes by having the effect of producing hyaluronic acid.
(3) Enterococcus faecalis XY7 strain fermentation liquor can effectively inhibit activity of acetylcholinesterase
Enterococcus faecalis XY7 cultured in MRS liquid culture medium to stationary phase is spread into new MRS liquid culture medium at dilution ratio of 1:30, bacterial suspension is collected when culturing to stationary phase for 24h, fermentation broth supernatant is collected after centrifugation at 10,000Xg and 4 ℃ for 10min, and then the inhibitory capacity of fermentation broth supernatant on acetylcholinesterase (AchE) activity is measured by acetylcholinesterase inhibitor screening kit (KA 6219). The results showed that the fermentation supernatant of strain XY7 significantly inhibited the activity of acetylcholinesterase compared to the non-inhibitory effect of the blank medium MRS, with an inhibition rate of about 20.38%, indicating that the fermentation broth of enterococcus faecalis XY7 was effective in inhibiting the activity of acetylcholinesterase (FIG. 4).
The acetylcholinesterase inhibitor can continuously accumulate at synapses under the action of acetylcholine released by nerve fiber tips through reversible inhibition of acetylcholinesterase, excite cholinergic receptors, and prolong and increase the action of acetylcholine. Acetylcholinesterase inhibitors can enhance cognitive ability and memory (improve Alzheimer's disease, improve cognitive dysfunction), dilate blood vessels (improve heart rate overspeed), excite skeletal and smooth muscles (improve constipation, postoperative abdominal distension, postoperative urinary retention).
Thus, the probiotic enterococcus faecalis XY7 strain can inhibit acetylcholinesterase activity, and can excite skeletal muscle and smooth muscle of intestinal tract, which makes it a potential probiotic for promoting gastrointestinal absorption and peristalsis.
(4) Enterococcus faecalis XY7 strain with superoxide dismutase (SOD) activity
Enterococcus faecalis XY7 strain cultured by MRS liquid culture medium to stationary phase is spread into new MRS liquid culture medium at dilution ratio of 1:30, bacterial suspension is collected when culturing to stationary phase for 24h, fermentation broth supernatant is collected after centrifugation at 10,000Xg and 4 ℃ for 10min, and SOD activity of fermentation broth supernatant is measured by superoxide dismutase (SOD) measuring kit (Nanjing built, cat: A001-3). Wherein, when the SOD inhibition rate in each milliliter of reaction solution reaches 50 percent, the corresponding SOD amount is one SOD activity unit (U). The results showed that the SOD activity in the fermentation supernatant of strain XY7 was 13.36U/mg compared to the placebo MRS, indicating that enterococcus faecalis XY7 can produce SOD, and that the fermentation broth has better SOD activity and shows good antioxidant capacity (FIG. 5).
Superoxide dismutase (SOD) is an antioxidant enzyme, and has antioxidant, antiinflammatory, and immunity enhancing effects. The main expression is as follows: (1) antioxidant: SOD can remove free radical in cells, relieve damage of oxidative stress to cells, and protect cells from free radical attack. (2) anti-inflammatory: SOD can inhibit inflammatory response, reduce inflammatory cell release, and reduce the degree and duration of inflammation. (3) enhancing immunity: SOD can promote the production of immune cells, improve the immunity of the body, prevent the occurrence of diseases and restore the health of the body. (4) anti-aging: can prevent lipid peroxidation, effectively remove free radicals, keep cells active, and prevent aging. (5) reducing blood fat: can remove intravascular resistance, soften blood vessel, reduce blood viscosity, and gradually restore blood lipid level. (6) enhancing liver function: can reduce toxin in blood and reduce damage of oxygen free radical to liver. (7) improving vision: is helpful for scavenging excessive free radicals in eyes, improving vision, and preventing cataract. (8) reducing blood sugar: can restore activity of islet cells, accelerate division speed, and further achieve the purpose of reducing blood sugar.
Thus, the probiotic enterococcus faecalis XY7 strain can exert the above effects by having the function of superoxide dismutase (SOD) activity.
Taken together, the newly isolated enterococcus faecalis XY7 strain of the present invention has a variety of probiotic effects: (1) gamma-aminobutyric acid can be produced and secreted; (2) can produce and secrete hyaluronic acid; (3) can inhibit acetylcholinesterase activity; (4) has superoxide dismutase activity. Therefore, the enterococcus faecalis XY7 strain obtained by the novel separation has important application value and economic value in the fields of anti-aging, sleep aiding and the like.
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 (7)
1. The application of enterococcus faecalis (Enterococcus faecalis) XY7 strain in producing gamma-aminobutyric acid is characterized in that the enterococcus faecalis XY7 strain is preserved in China center for type culture Collection (China, with the preservation number: CCTCC NO: M20231166; the 16S rDNA complete sequence of the enterococcus faecalis XY7 strain is shown in SEQ ID No: 1.
2. The application of enterococcus faecalis (Enterococcus faecalis) XY7 strain in hyaluronic acid production, which is characterized in that the enterococcus faecalis XY7 strain is preserved in China center for type culture Collection (China, with the preservation number: CCTCC NO: M20231166; the 16S rDNA complete sequence of the enterococcus faecalis XY7 strain is shown in SEQ ID No: 1.
3. The application of enterococcus faecalis (Enterococcus faecalis) XY7 strain in preparing acetylcholinesterase inhibitor is characterized in that the enterococcus faecalis XY7 strain is preserved in China center for type culture collection (China) with the preservation number of 2023, 7 and 3: CCTCC NO: M20231166; the 16S rDNA complete sequence of the enterococcus faecalis XY7 strain is shown in SEQ ID No: 1.
4. The application of enterococcus faecalis (Enterococcus faecalis) XY7 strain in producing superoxide dismutase is characterized in that the enterococcus faecalis XY7 strain is preserved in China center for type culture collection (China, with the preservation number: CCTCC NO: M20231166; the 16S rDNA complete sequence of the enterococcus faecalis XY7 strain is shown in SEQ ID No: 1.
5. A method for preparing hyaluronic acid is characterized in that enterococcus faecalis (Enterococcus faecalis) XY7 strain is inoculated in MRS liquid culture medium, bacterial suspension is collected after the culture is carried out until the stationary phase, and finally hyaluronic acid in the bacterial suspension is separated and purified to obtain the hyaluronic acid; the enterococcus faecalis XY7 strain is preserved in China center for type culture Collection (China, accession number: CCTCC NO: M20231166; the 16S rDNA complete sequence of the enterococcus faecalis XY7 strain is shown in SEQ ID No: 1.
6. The method for producing hyaluronic acid according to claim 5, wherein the conditions of the culture are: anaerobic at a constant temperature of 37 ℃.
7. The method for preparing hyaluronic acid according to claim 5, wherein enterococcus faecalis (Enterococcus faecalis) XY7 strain is cultured in MRS liquid medium to stationary phase, and then expanded into new MRS liquid medium at dilution ratio of 1:20-1000.
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