CN116676226B - Lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application - Google Patents
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- CN116676226B CN116676226B CN202310645941.XA CN202310645941A CN116676226B CN 116676226 B CN116676226 B CN 116676226B CN 202310645941 A CN202310645941 A CN 202310645941A CN 116676226 B CN116676226 B CN 116676226B
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- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000007269 microbial metabolism Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000000926 neurological effect Effects 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 235000019260 propionic acid Nutrition 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000009323 psychological health Effects 0.000 description 1
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 description 1
- 229910052704 radon Inorganic materials 0.000 description 1
- SYUHGPGVQRZVTB-UHFFFAOYSA-N radon atom Chemical compound [Rn] SYUHGPGVQRZVTB-UHFFFAOYSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000035806 respiratory chain Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 235000021391 short chain fatty acids Nutrition 0.000 description 1
- 150000004666 short chain fatty acids Chemical class 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 230000004622 sleep time Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
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Abstract
The invention belongs to the technical field of microorganisms, and relates to a Lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application, wherein the LP-28 strain is preserved in China general microbiological culture collection center (CGMCC) with the preservation number of 7.463 in the year 6 and 25 of 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3. The lactobacillus plantarum LP-28 strain can be directly metabolized in the intestinal environment of a human body to produce gamma-aminobutyric acid, can be applied to products for improving sleep quality, improves the balance of intestinal flora, and enhances the immunity of organisms. Meanwhile, the yogurt prepared by fermenting the probiotic EM28 formed by combining the lactobacillus plantarum LP-28 strain and lactobacillus rhamnosus LR-28 strain is proper in sweetness and sourness and good in flavor.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus plantarum LP-28 for relieving anxiety, yoghourt and application thereof.
Background
Investigation by the world health organization data shows that 27% of people worldwide have sleep disorders, with china accounting for 38.2%. From 2013 to 2018, the average sleep time of Chinese is shortened from 8.8 hours to 6.5 hours. China becomes a country with high sleep problems, and insomnia becomes a health problem of particular concern to modern people.
Poor sleep quality is a risk factor of various diseases, has close relation with hypertension, coronary heart disease, diabetes and the like, and can influence the psychological health condition of people, so that people are tired, anxious and depressed, negative emotion is increased, and the attention degree and satisfaction degree to the external matters are reduced. Anxiety and depression symptoms increase the prevalence and incidence of cardiovascular diseases.
Gamma-aminobutyric acid (Gamma-aminobutyric acid, GABA) is an important central nervous system inhibitory neurotransmitter, and GABA as a small molecular weight non-protein amino acid has edible safety, and can be used for the production of foods such as beverages. Research shows that the intake of a certain amount of GABA has the physiological effects of improving anxiety emotion of organisms and improving sleep quality.
Currently, GABA is synthesized by utilizing microbial metabolism to convert sodium glutamate or glutamic acid into GABA, and sodium glutamate or glutamic acid substrate is additionally provided in the process. In the prior art, strains for directly converting and producing GABA are cultured by microbial fermentation, but the fermentation process for producing GABA is carried out in an in-vitro fermentation tank, the culture medium used under the fermentation condition and the culture environment have great difference from the complex human intestinal environment, and the strains cannot be directly metabolized in the human intestinal environment to produce GABA. Therefore, the advantage of GABA production by the above strain needs to be exerted by means of in vitro fermentation, but is not necessarily applicable as a product of direct metabolism in human intestinal environment for GABA production.
On the other hand, yogurt is a common dairy product obtained by fermentation with lactic acid bacteria, and the sour taste of the yogurt is derived from acid substances generated in the fermentation process of lactic acid bacteria. The acidogenesis of lactobacillus plantarum and lactobacillus rhamnosus in the metabolic process is too much, so that the acidity of the yoghurt after reaching the fermentation end point is too high (the acidity is 170-250 DEG T), and the mouthfeel is seriously influenced. Therefore, the yogurt sold in the market at present does not adopt the two strains for fermentation, and the streptococcus thermophilus and the lactobacillus bulgaricus are adopted for synergistic fermentation to control the acidity, but generally only have fermentation effect, and are not probiotics.
Disclosure of Invention
Aiming at the technical problems, the invention provides lactobacillus plantarum LP-28, yoghourt and application thereof, and the lactobacillus plantarum LP-28 can regulate the balance of intestinal microbial flora, enhance the immunity of organisms and directly metabolize and produce GABA in the intestinal environment of human bodies. Meanwhile, the yoghurt prepared by the combined fermentation of the lactobacillus plantarum LP-28 strain and the lactobacillus rhamnosus LR-28 strain is improved by a novel process, and has proper sour taste and good flavor.
In order to achieve the aim of the invention, the invention adopts the following technical scheme:
A lactobacillus plantarum LP-28 strain, which is classified as lactobacillus plantarum (Lactobacillus plantarum) and preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.7.463 in the year 6 and 25 of 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
Compared with the prior art, the lactobacillus plantarum LP-28 strain with the preservation number of CGMCC No.7.463 provided by the invention has the following advantages:
(1) The lactobacillus plantarum LP-28 strain provided by the invention belongs to lactobacillus and can effectively resist gastric acid and bile salts, and is subjected to field planting in intestinal tracts so as to improve intestinal microbial flora, regulate the balance of the intestinal microbial flora and strengthen the immunity of organisms.
(2) The living bacterium of the lactobacillus plantarum LP-28 provided by the invention can produce high-activity glutamate decarboxylase (GAD) in an intestinal environment, and Glu is decarboxylated to form GABA by utilizing the GAD activity. GABA absorbs and enters the human body, enters nerve cells along with blood, and combines with GABA receptors among neurons, thereby playing a role in inhibiting nerve excitation, thereby playing roles in relieving dysphoria and improving sleep quality.
The 16S rRNA sequence is shown below:
AGGACGTGCCGCGTGCCTATGATGCAAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGCTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAACCAGCCGCCTAAGGGGGCCCATTGC
in a second aspect, the invention also provides the use of the lactobacillus plantarum LP-28 strain described above for the production of gamma-aminobutyric acid.
In a third aspect, the invention also provides application of the lactobacillus plantarum LP-28 strain in preparing health care products for improving sleep quality.
The lactobacillus plantarum LP-28 strain provided by the invention can produce high-activity glutamate decarboxylase, and can directly utilize intestinal contents to produce GABA through a metabolic process in a complex intestinal environment of a human body, so that the lactobacillus plantarum LP-28 strain can be prepared into a sleep-aiding product, and the GABA produced by in vivo metabolism of the lactobacillus plantarum LP-28 strain is utilized to achieve the aim of improving sleep quality.
In a fourth aspect, the invention also provides a fermentation product of the above strain of Lactobacillus plantarum LP-28, said fermentation product being obtained by fermentation of a strain of Lactobacillus plantarum LP-28. The fermented product can be further processed to improve sleep quality.
The lactobacillus plantarum LP-28 strain can also be cultured by a culture medium, and Glu is decarboxylated to form GABA by utilizing the high activity of GAD. Therefore, the product for improving sleep quality can also be prepared by adopting an in-vitro fermentation culture mode and utilizing a fermentation product or a fermentation extract thereof.
In a fifth aspect, the present invention also provides a method for producing gamma-aminobutyric acid by using the lactobacillus plantarum LP-28 strain, which specifically comprises the following steps: seed culture is carried out on lactobacillus plantarum LP-28 strain in an improved MRS liquid culture medium, and anaerobic fermentation culture is carried out on the cultured fermentation broth on an intestinal simulated culture medium.
Further, the anaerobic fermentation culture conditions are as follows: the fermentation temperature is 35-37 ℃, the pH is 5-7, and the culture is carried out for more than 48 hours.
Alternatively, the method may specifically employ the following procedure:
Seed culture stage: inoculating lactobacillus plantarum LP-28 seed solution into an improved MRS liquid culture medium according to an inoculum size of 3%, shaking and mixing uniformly, culturing for 24 hours at 37 ℃, sequentially and continuously activating for three generations, taking 100ml of the cultured fermentation liquid, and obtaining a fermentation liquid stock solution sample, and refrigerating at 4 ℃ for later use.
CMGM composition (g/L): starch, 5.0; pectin, 2.0; guar gum, 1.0; mucin (porcine stomach type III), 4.0; xylan, 2.0; arabinogalactan, 2.0; inulin, 1.0; casein, 3.0; peptone water, 5.0; tryptone, 5.0; bile salts, 0.4; yeast extract, 4.5; feSO 4·7H2 O,0.005; sodium chloride, 4.5; potassium chloride ,4.5;KH2PO4,0.5;MgSO4·7H2O,1.25;CaCl2·6H2O,0.15;NaHCO3,1.5; cysteine, 0.8; heme, 0.05; tween 80,1.0.
The intestinal tract simulation culture medium is placed in an intestinal tract in-vitro fermentation simulation system fermentation tank and is used for carrying out fermentation culture on the fermentation liquid: the human colon three-stage continuous culture intestinal model system consists of three fermentors, a simulated proximal fermentor (V1, 280 mL), a transverse fermentor (V2, 300 mL) and a distal colon fermentor (V3, 320, mL), respectively. Three fermenters in series were maintained at 37 ℃ and pH values were maintained at 5.5 (V1), 6.2 (V2) and 6.8 (V3), respectively, and anaerobic conditions were created by continuous N 2 introduction. V1, V2 and V3 are all introduced CMGM by peristaltic pumps. Fresh healthy human fecal samples were collected, stored in an anaerobic cabinet (10% H 2、10% CO2、80% N2), and 1:5 (w/w) fecal dilutions in anaerobic PBS (0.1 mol/L PBS, pH 7.4) were prepared within 15 minutes after collection. Each fermenter of the intestinal model was inoculated with 100 mL faeces dilutions. The total system transit time was set to 48 hours based on the average retention time of healthy individuals. After inoculation with fecal dilutions, the intestinal model was run for 24 hours to stabilize the bacterial population prior to injection into the culture medium. After 24 hours, the system was run for 8 full capacity cycles to achieve steady state, and then the above fermentation broth was added to V1 in an amount of 1% (w/V) of the volume of the contents in the tank, followed by eight volume cycles to reach steady state, and further culture for 24 hours.
In a sixth aspect, the invention provides a probiotic EM28 comprising the above strain of Lactobacillus plantarum LP-28 and Lactobacillus rhamnosus LR-28.
In a seventh aspect, the present invention provides a yoghurt product prepared from the probiotic group EM28 as described above after fermentation of milk.
Lactobacillus rhamnosus LR-28 is preserved in China general microbiological culture Collection center (CGMCC) with a preservation number of 7.464 in the 6 th month of 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
Microorganisms increase the expression of the proton respiratory chain complex while producing GABA, promote ATP synthesis, and up-regulate the activity of F1F0-ATP hydrolase, promote the ATP-dependent H + excretion process under acidic conditions, thereby changing the pH of the extracellular environment. More importantly, GABA is zwitterionic in the physiological environment and therefore plays a role in acid-base regulation.
The probiotic EM28 formed by combining lactobacillus rhamnosus LR-28 and lactobacillus plantarum LP-28 can produce synergistic effect in the process of fermenting milk, and the GABA yield is far higher than that of the pure superposition of the two bacteria. Therefore, the acidity of the yoghurt provided by the invention is finally between 90 and 100 ℃ and the pH value is between 4.25 and 4.35, and compared with the unflavoured yoghurt obtained by fermentation of other lactic acid bacteria, the yoghurt provided by the invention has proper acidity and does not need to be additionally added with additives to improve the flavor.
Further, the ratio of the viable count of the lactobacillus plantarum LP-28 strain to the lactobacillus rhamnosus LR-28 strain is: 1-3:4-7.
In different periods of yoghurt production, due to different pH values in the yoghurt, the metabolic states of the yoghurt and the yoghurt are different, and only if the two strains keep a good symbiotic relationship and a synergistic effect, the yoghurt can obtain good flavor. The lactobacillus plantarum LP-28 strain and lactobacillus rhamnosus LR-28 strain can maintain a good symbiotic relationship within the above proportion range, so that a synergistic effect is generated, and the yoghurt has good flavor.
Further, the total inoculation amount of the lactobacillus plantarum LP-28 strain and the lactobacillus rhamnosus LR-28 strain is 0.002-0.006%; the fermentation conditions are as follows: culturing at 38-43deg.C for 10-20 hr.
In the production of the yoghourt, the primary fermentation is mainly acid production, and the strain can utilize lactose metabolism to produce lactic acid, acetic acid, propionic acid and other acids to endow the yoghourt with certain acidity and cool mouthfeel. Therefore, the inoculation amount and fermentation conditions need to be strictly controlled to achieve the optimal acidity.
Further, one or more of food materials, stabilizers or sweeteners may be added to the yogurt material. The food material comprises fruits, grains, etc.
In an eighth aspect, the invention provides a yoghurt powder, which is prepared by spray drying the yoghurt product.
Drawings
FIG. 1 shows the bacterial morphology of Lactobacillus plantarum LP-28 strain in example 1 of the present invention;
FIG. 2 (a) shows the bacteriostatic ability of Lactobacillus plantarum LP-28 strain against E.coli;
FIG. 2 (b) shows the bacteriostatic ability of Lactobacillus plantarum LP-28 strain against Staphylococcus aureus;
FIG. 2 (c) shows the antibacterial ability of Lactobacillus plantarum LP-28 strain against Salmonella;
FIG. 3 is a sleep survey of group A people in example 7 of the present invention;
FIG. 4 is a sleep survey of group B people in example 7 of the present invention;
FIG. 5 is a sleep survey of group C people in example 7 of the present invention;
FIG. 6 shows the improvement of Hamiltonian anxiety among groups of people in example 7 of the present invention.
Detailed Description
The present invention will be described in further detail with reference to specific embodiments in order to make the objects, technical solutions and advantages of the present invention more apparent. It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.
The sources of the strains used in the following examples of the invention are as follows:
Lactobacillus plantarum LP-28, which is classified under the name Lactobacillus plantarum (Lactobacillus plantarum), is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.7.463 at the date of 25 and 6 in 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
Lactobacillus rhamnosus LR-28, which is classified as lactobacillus rhamnosus (Lactobacillus rhamnosus) and is preserved in the China general microbiological culture Collection center (CGMCC) with the preservation number of 7.464 in the 6 th month of 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
Modified MRS liquid Medium (g/L): peptone 8.0-10.0; 9.0 to 10.0 portions of beef powder; 3.0 to 5.0 percent of yeast powder; glucose 15.0-20.0; magnesium sulfate, 0.08-0.1; sodium acetate 1.8-2.0; ammonium citrate, 4.0-5.0; 1.6 to 2.0 portions of radon dipotassium phosphate; manganese sulfate, 0.03-0.05; tween 80,0.8-1.0; l-cysteine hydrochloride, 0.05-0.07 (pH 6.2.+ -. 0.2).
CMGM composition (g/L): starch, 5.0; pectin, 2.0; guar gum, 1.0; mucin (porcine stomach type III), 4.0; xylan, 2.0; arabinogalactan, 2.0; inulin, 1.0; casein, 3.0; peptone water, 5.0; tryptone, 5.0; bile salts, 0.4; yeast extract, 4.5; feSO 4·7H2 O,0.005; sodium chloride, 4.5; potassium chloride ,4.5;KH2PO4,0.5;MgSO4·7H2O,1.25;CaCl2·6H2O,0.15;NaHCO3,1.5; cysteine, 0.8; heme, 0.05; tween 80,1.0.
Intestinal tract in vitro fermentation simulation system: the human colon three-stage continuous culture intestinal model system consists of three fermentors, a simulated proximal fermentor (V1, 280 mL), a transverse fermentor (V2, 300 mL) and a distal colon fermentor (V3, 320, mL), respectively. Three fermenters in series were maintained at 37 ℃, pH was maintained at 5.5 (V1), 6.2 (V2) and 6.8 (V3), and anaerobic conditions were created by continuous N 2 introduction. V1, V2 and V3 are all introduced CMGM by peristaltic pumps. Fresh healthy human fecal samples were collected, stored in an anaerobic cabinet (10% H 2、10% CO2、80% N2), and 1:5 (w/w) fecal dilutions in anaerobic PBS (0.1 mol/L PBS pH 7.4) were prepared within 15 minutes after collection. Each fermenter of the intestinal model was inoculated with 100 mL faeces dilutions. The total system transit time was set to 48 hours based on the average retention time of healthy individuals. After inoculation with fecal dilutions, the intestinal model was run for 24 hours to stabilize the bacterial population prior to injection into the culture medium. After 24 hours, the system was run for 8 full capacity cycles to achieve steady state, and then the above fermentation broth was added to V1 in an amount of 1% (w/V) of the volume of the contents in the tank, followed by eight volume cycles to reach steady state, and further culture for 24 hours.
Other experimental materials and instruments were commercially available unless otherwise specified.
Example 1
The present example provides a strain of Lactobacillus plantarum LP-28 with reduced anxiety, which strain was selected by the following procedure:
1. isolation and screening of Lactobacillus plantarum LP-28
1.1 Bacterial source:
9 months 2019, tashikuer, tajike county, traditional yogurt in herlands.
1.2 Separation
Taking 10g of traditional yoghurt in the herdsman's home, adding the traditional yoghurt into 100ml of physiological saline, fully and uniformly mixing, and diluting the mixture, wherein the dilution gradient is 10 -1,10-2,10-3,10-4,10-5; each gradient was applied in 100. Mu.L to modified MRS solid medium supplemented with calcium carbonate, and each gradient was repeated. Placing in a constant temperature incubator at 37 ℃ for culturing for 48 hours.
Selecting a proper gradient (10 -5) from the above culture concentration gradients, selecting 15-25 single colonies with clear boundaries and transparent circles, subculturing the selected colonies on a newly prepared MRS solid culture medium by a streaking method for 24-48h, and repeating the steps three times again to obtain the monoclonal strain. After the strain is cultured for 24 hours by an MRS liquid culture medium, bacterial liquid and 50% glycerol are uniformly mixed according to a ratio of 1:1 (the final concentration of the glycerol is 25%), and the strain is preserved at-75 ℃, and simultaneously inoculated to an MRS solid culture medium test tube inclined plane for temporary preservation.
1.3 Preliminary screening of Strain
The glycerol-preserved bacterial liquid was inoculated into 1.5mL of MRS liquid medium at an inoculum size of 2% and placed in a constant temperature incubator at 37℃for 10 hours as seed bacterial liquid for the next experiment.
Inoculating the seed bacterial liquid into 2mL EP tube filled with MRS liquid culture medium according to the inoculation amount of 2%, and culturing for 48h at 37 ℃; picking single colony on MRS liquid culture medium, and culturing at 37deg.C for 16 hr; transferring to GYP liquid culture medium, standing at 37deg.C for 48 hr; taking 1 mu L of supernatant fluid of the fermentation liquor, spotting the supernatant fluid on filter paper (each 10 samples of +1 standard/piece of filter paper), centrifuging the supernatant fluid for 10min at 2000rpm, drying the filter paper after 4h of chromatography in a developing agent, spraying 8g/L ninhydrin on the filter paper, developing the color in a baking oven at 70 ℃ for 5min, and primarily screening 22 strains with gamma-aminobutyric acid (GABA) production.
1.4 Strain double Screen
Inoculating the seed bacterial liquid of 22 strains obtained by primary screening into 2mL EP tube filled with MRS liquid culture medium according to the inoculum size of 2%, and culturing for 48h at 37 ℃; picking single colony on MRS liquid culture medium, and culturing at 37deg.C for 16 hr; then transferring to GYP liquid culture medium, standing and culturing for 48h at 37 ℃. Centrifuging to obtain supernatant, and quantitatively determining GABA production capacity of 18 strains by adopting high performance liquid chromatography. The 22 strains obtained by separation have GABA producing capacity, wherein the strain numbered LP-28 has the strongest GABA producing capacity, and GABA yield is up to 3.653g/L.
1.5 Identification of LP-28 Strain
1.5.1 Colony morphology and fungus morphology characteristics of LP-28 Strain
Gram staining: positive; colony morphology observation: the surface is smooth and glossy; cell morphology:
The polymorphic bacilli are shown in FIG. 1.
1.5.2 Sequencing analysis of 16S rRNA of LP-28 Strain
TABLE 1 LP-28 similarity of 16S rRNA sequences of strains with reference strains
In summary, the identification of LP-28 as a Lactobacillus plantarum (Lactobacillus plantarum) strain was performed in combination with the results of 16S rRNA sequence analysis and physiological and biochemical identification.
The lactobacillus plantarum LP-28 strain is preserved in China general microbiological culture Collection center (CGMCC) at 25-6-2022, and the preservation number is CGMCC No.7.463; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
2. Gastrointestinal digestion resistance of Lactobacillus plantarum LP-28 Strain
Gastric juice is simulated manually: taking 16.4mL of dilute hydrochloric acid, adding 800mL of water and 10g of protease, shaking uniformly, adding water to dilute into 1000mL, mixing uniformly, and filtering by a sterile filter membrane with the thickness of 0.22 mu m.
Manually simulating intestinal juice: taking 6.8g of monopotassium phosphate, adding 500mL of water to dissolve the monopotassium phosphate, and adjusting the pH to 6.8 by using 0.1mol/L sodium hydroxide solution; and (3) taking 10g of pancreatin, adding a proper amount of water to dissolve, mixing the two solutions, adding water to dilute to 1000mL, uniformly mixing, and filtering with a 0.22 mu m sterile filter membrane to obtain the pancreatic enzyme.
The lactobacillus plantarum LP-28 screened in the example 1 is cultured and activated with common commercial lactobacillus plantarum to obtain an inoculation liquid with the bacterial liquid concentration of 1 multiplied by 10 9 CFU/mL.
Gastric acid tolerance test: 1mL of Lactobacillus plantarum LP-28 strain inoculum and common commercial Lactobacillus plantarum inoculum (strain No. ATCC 8014) were inoculated and respectively preheated to a test tube filled with 9mL of simulated gastric fluid, cultured in an incubator at 37℃and 80r/min, and viable counts of 0, 2 and 4 hours were determined (plate count method).
Bile salt resistance test: 1mL of Lactobacillus plantarum LP-28 strain inoculum and common commercial Lactobacillus plantarum inoculum (strain No. ATCC 8014) were inoculated and respectively preheated to a test tube filled with 9mL of simulated intestinal fluid, and cultured in an incubator at 37℃and 80r/min, and viable counts of 0, 2 and 4 hours were measured (plate count method).
TABLE 2 survival rate of strains by simulation of gastrointestinal digests (100 mL bacteria count)
Compared with common commercial strains, the lactobacillus plantarum LP-28 has very high acid resistance and bile salt resistance, can resist artificial gastric juice and artificial intestinal juice, and ensures that enough viable bacteria are planted in intestinal tracts after the gastric and intestinal digestions.
3. High adhesion capability of lactobacillus plantarum LP-28 to Caco-2 cells
Inoculating Caco-2 cells with the concentration of 1X 10/mL at one time to a 12-well plate, culturing to form single-layer cells under the condition of 37 ℃ and 5% CO, discarding culture solution, lightly washing three times by using sterile PBS buffer solution, adding 1mL of DMEM stock solution, respectively adding lactobacillus plantarum LP-28 strain inoculating solution and 100 mu L of common commercial lactobacillus plantarum (strain number ATCC 8014) bacterial suspension (suspended by DMEM, the concentration is about 10 0 cfu/mL), making three holes for each strain, after culturing for 2 hours, discarding supernatant, washing three times by using sterile PBS, adding 300 mu L of 0.25% pancreatin into each hole to digest for 5-8min, adding 300 mu L of incomplete DMEM culture medium (containing serum without double antibodies), and then performing plate dilution coating to calculate the number of adhered strains.
Adhesion (%) = number of adhesion strains/initial inoculation number x 100%.
TABLE 3 cell adhesion ability of strains
Compared with common commercial lactobacillus plantarum, the lactobacillus plantarum LP-28 can be more effectively adhered to intestinal mucosa to form a biological barrier, protect intestinal tracts and provide important preconditions for colonisation in human intestinal tracts.
4. Antibacterial ability of Lactobacillus plantarum LP-28 Strain
The activated Lactobacillus plantarum LP-28 was inoculated at an inoculum size of 2% into 2mL EP tubes containing MRS liquid medium and incubated in a 37℃incubator for 18h. Taking 500g of lactobacillus plantarum LP-28 culture solution, and centrifuging for 10min to obtain a supernatant for later use.
Coli (ATCC 25922), staphylococcus aureus (ATCC 43300) and salmonella (ATCC 50760) were used as indicator bacteria. Taking 100 mu L of activated indicator bacteria, adjusting the concentration of the indicator bacteria to 1.0X10 5 CFU/mL, coating a plate on a corresponding culture medium, placing the coated culture dish on a semi-open working table for 10min, adding 200 mu L of lactobacillus plantarum LP-28 supernatant into oxford cups placed on a solid culture medium, culturing at 37 ℃ for 16h steadily, observing and measuring the diameter of a bacteriostasis ring of each bacterial liquid, and performing three groups of experiments in parallel.
The results show that the lactobacillus plantarum LP-28 can inhibit the growth of escherichia coli, staphylococcus aureus and salmonella, and has antibacterial effect. The lactobacillus plantarum LP-28 can be effectively adhered to human intestinal tracts, short-chain fatty acids (such as acetic acid and lactic acid), hydrogen peroxide, bacteriocin and the like generated by the lactobacillus plantarum LP-28 can effectively inhibit the growth of putrefying bacteria and the generation of harmful metabolites such as amine, phenol, indole and the like, and reduce the pH value in the intestinal tracts.
Example 2
The embodiment provides a fermentation product of lactobacillus plantarum LP-28 strain, which is prepared by the following steps: inoculating seed solution of lactobacillus plantarum LP-28 strain into an improved MRS liquid culture medium according to an inoculum size of 3%, shaking and mixing uniformly, culturing for 24 hours at 37 ℃, and sequentially and continuously activating for three generations to obtain fermentation liquor.
Taking the cultured fermentation liquor, adding the fermentation liquor into a fermentation tank (V1) of an intestinal in-vitro fermentation simulation system which is connected with CMGM and fecal diluent and completes 8 full-capacity turnover by adding the fermentation liquor into the tank in an adding amount of 1% (w/V) of the volume of the content, performing fermentation culture, performing volume turnover for eight times, reaching a steady state, centrifuging, and taking supernatant.
Considering the operating volume (900 mL) and retention time (48 h) of the colon model system, fermentation broth was added to V1 at 1% (w/V) per day, followed by eight additional volume turnovers to steady state, centrifugation, and supernatant removal.
The GABA content in the fermentation broth obtained in example 2 was quantitatively determined by high performance liquid chromatography, and the GABA content was 10.+ -. 4.2mg/100ml. Lactobacillus plantarum LP-28 was shown to be capable of producing GABA under in vitro simulated fermentation conditions in the intestinal tract.
Example 3
The present example provides a probiotic EM28 comprising Lactobacillus plantarum LP-28 strain obtained in example 1 in combination with Lactobacillus rhamnosus LR-28 strain. The mass ratio of the lactobacillus plantarum LP-28 strain to the lactobacillus rhamnosus LR-28 strain is 1:2.5.
Example 4
This example provides a yoghurt prepared from milk fermented with the probiotic EM28 of example 3.
Is prepared from the following raw materials: raw milk: 865.5g; white granulated sugar: 110g; cream: 5g; starch: 16g; sodium glutamate: 1g; sodium bicarbonate: 1.3g; probiotic EM28:0.035g (Lactobacillus plantarum LP-28 strain: 0.01g; lactobacillus rhamnosus LR-28 strain: 0.025 g).
Preparing yoghourt: mixing raw milk, white granulated sugar, cream and starch, sterilizing at 95deg.C for 5min, cooling to 40deg.C, adding sodium glutamate and sodium bicarbonate, inoculating lactobacillus plantarum LP-28 strain and lactobacillus rhamnosus LR-28 strain, and sealing for fermentation. The fermentation is carried out for 7h for the first time, the fermentation termination time is predicted according to the acid rising condition, the total fermentation time is about 10-11h, and the fermentation is terminated when the pH value is 4.30+/-0.05 (the pH value is used as the standard for judging the fermentation end point). Demulsification stirring after fermentation, cooling and filling. The final viable bacteria content of the obtained yoghourt is as follows: 1.0X10 8 CFU/ml; the acidity is: 98 deg.t.
Example 5
This example provides a yoghurt prepared from milk fermented with the probiotic EM28 of example 3.
The preparation method comprises the following steps: whole milk powder: 103g; water: 762.5g; white granulated sugar: 110g; cream: 5g; starch: 16g; sodium glutamate: 1g; sodium bicarbonate: 1.3g; probiotic EM28:0.035g (Lactobacillus plantarum LP-28 strain: 0.01g; lactobacillus rhamnosus LR-28 strain: 0.025 g).
Preparing yoghourt: mixing whole milk powder, water, white sugar, cream and starch, sterilizing at 95deg.C for 5min, cooling to 40deg.C, adding sodium glutamate and sodium bicarbonate, inoculating lactobacillus plantarum strain LP-28 and lactobacillus rhamnosus strain LR-28, and sealing for fermentation. The fermentation is carried out for 7h for the first time, the fermentation termination time is predicted according to the acid rising condition, the total fermentation time is about 10-11h, and the fermentation is terminated when the pH value is 4.30+/-0.05 (the pH value is used as the standard for judging the fermentation end point). Demulsification stirring after fermentation, cooling and filling. The final viable bacteria content of the obtained yoghourt is as follows: 1.0X10 8 CFU/ml; the acidity is: 97T.
Example 6
This example provides a yoghurt prepared from milk fermented with the probiotic EM28 of example 3.
Is prepared from the following raw materials: skimmed milk powder: 98g; water: 767.5g; white granulated sugar: 110g; cream: 5g; starch: 16g; sodium glutamate: 1g; sodium bicarbonate: 1.3g; probiotic EM28:0.035g (Lactobacillus plantarum LP-28 strain: 0.01g; lactobacillus rhamnosus LR-28 strain: 0.025 g).
Preparing yoghourt: mixing skimmed milk powder, water, white sugar, cream and starch, sterilizing at 95deg.C for 5min, cooling to 40deg.C, adding sodium glutamate and sodium bicarbonate, inoculating lactobacillus plantarum strain LP-28 and lactobacillus rhamnosus strain LR-28, and sealing for fermentation. The fermentation is carried out for 7h for the first time, the fermentation termination time is predicted according to the acid rising condition, the total fermentation time is about 10-11h, and the fermentation is terminated when the pH value is 4.30+/-0.05 (the pH value is used as the standard for judging the fermentation end point). Demulsification stirring after fermentation, cooling and filling. The final viable bacteria content of the obtained yoghourt is as follows: 1.0X10 8 CFU/ml; the acidity is: 95T.
Example 7
The present example provides the use of lactobacillus plantarum LP-28 strain as a raw material for the preparation of a product for improving sleep quality.
Hamilton anxiety scale (Hamilton Anxiey Scale, HAMA), which was one of the scales commonly used in psychiatric clinics, was compiled by Hamilton in 1959 and was used primarily to assess the severity of anxiety in neurological and other patients.
1. Test procedure
Insomnia patients enrolled 195 for anxiety symptoms (screening criteria: HAMA (Hamiltonian anxiety) > 7 population screening into groups), group A (20-35 years) were grouped by age: 60 people; group B (35-59 years): 70 people; group C (over 60 years): 65 persons. The yogurt obtained in example 5 was administered to patients suffering from anxiety and insomnia, 250ml per day, with a period of 2 weeks. The anxiety symptoms of the three groups are investigated, and the hamiltonian anxiety scale is investigated.
2. Test results and analysis
2.1 Sleep findings for 2 weeks:
group A has 60 anxiety symptoms, 38 people have obviously improved sleep quality, 22 people have no obvious effect, and the effective rate is 63.30 percent (shown in figure 1);
group B has 70 anxiety symptoms, 32 people have obviously improved sleep quality, 38 people have no obvious effect, and the effective rate is 45.70 percent (shown in figure 2);
Group C has 65 anxiety symptoms, 36 people have obviously improved sleep quality, 29 people have no obvious effect, and the effective rate is 55.30 percent (shown in figure 3);
2.2A, B, C results of three group of symptomatic improvement studies
Table 4A group results of symptomatic improvement study
Table 5B group results of symptomatic improvement study
Table 6C group results of symptomatic improvement study
2.3 Improvement amount was determined using hamiltonian anxiety scale (HAMA) investigation
The results are shown in Table 7 and FIG. 4.
Tables 7A and B, C three groups were examined using hamiltonian anxiety scale (HAMA) to determine improvement
From the above tables 5 to 7, it can be seen that the yogurt provided by the invention has good improving effects in relieving dysphoria and improving sleep quality.
In conclusion, the lactobacillus plantarum LP-28 strain provided by the invention has the capability of high-yielding GABA in the human intestinal environment, can improve and regulate the balance of intestinal microbial flora, and can enhance the immunity of organisms.
The foregoing description of the preferred embodiments of the invention is not intended to be limiting, but rather is intended to cover all modifications, equivalents, or alternatives falling within the spirit and principles of the invention.
Claims (7)
1. The application of the lactobacillus plantarum LP-28 strain in preparing health care products for improving sleep quality is characterized in that the lactobacillus plantarum LP-28 strain is classified as lactobacillus plantarum (Lactobacillus plantarum) which is preserved in China general microbiological culture Collection center (CGMCC) No.7.463 in the 6 th month 25 of 2022; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
2. Use of the lactobacillus plantarum LP-28 strain as claimed in claim 1, for the production of gamma-aminobutyric acid.
3. A method for producing gamma-aminobutyric acid using lactobacillus plantarum LP-28 strain as claimed in claim 1, characterized in that the method comprises in particular the following operations: seed culture is carried out on lactobacillus plantarum LP-28 strain in an improved MRS liquid culture medium, and anaerobic fermentation culture is carried out on the cultured fermentation broth on an intestinal simulated culture medium.
4. A method for producing gamma-aminobutyric acid by lactobacillus plantarum LP-28 strain as claimed in claim 3, wherein the anaerobic fermentation culture conditions are: the fermentation temperature is 35-40 ℃, the pH is 4.0-4.7, and the culture is carried out for more than 18 hours.
5. A yoghurt product, which is prepared by fermenting a milk product with the lactobacillus plantarum LP-28 strain and the lactobacillus rhamnosus (Lactobacillus rhamnosus) LR-28 strain of claim 1, wherein the lactobacillus rhamnosus LR-28 strain is preserved in the general microbiological center of the China general microbiological culture collection center (cmv) at the month 26 of 2022, and the preservation number is CGMCC No.7.464; the preservation address is the Beijing Chaoyang district North Star West Leu 1, 3.
6. Yoghurt preparation in accordance with claim 5, characterized in that the ratio of viable count of lactobacillus plantarum LP-28 strain to lactobacillus rhamnosus LR-28 strain is: 1-3:4-7; and/or
The total inoculation amount of the lactobacillus plantarum LP-28 strain and lactobacillus rhamnosus LR-28 strain is 0.002-0.006%; the fermentation conditions are as follows: culturing at 38-43deg.C for 10-20 hr; and/or
The raw materials for preparing the yoghurt product also comprise one or more than two of food raw materials, stabilizers or sweeteners.
7. Yoghurt powder, characterized in that it is obtained from a yoghurt product as claimed in claim 5 by spray drying.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109136130A (en) * | 2018-08-27 | 2019-01-04 | 南昌大学 | One plant of Lactobacillus rhamnosus NCU2217 |
| CN111728111A (en) * | 2020-06-28 | 2020-10-02 | 武汉康复得生物科技股份有限公司 | Probiotic composition for relieving anxiety or depression and application thereof |
| CN112501063A (en) * | 2020-12-03 | 2021-03-16 | 兰州大学 | Lactobacillus plantarum LZU-J-TSL6 capable of highly producing GABA and application thereof |
| CN115161250A (en) * | 2022-09-06 | 2022-10-11 | 广东益可维生物技术有限公司 | Lactobacillus plantarum with functions of improving sleep, strengthening brain and improving intelligence and application thereof |
| CN115521889A (en) * | 2022-10-30 | 2022-12-27 | 江南大学 | Lactobacillus plantarum WL02 capable of producing gamma-aminobutyric acid and application thereof |
| CN116676225A (en) * | 2023-05-23 | 2023-09-01 | 河北源民生物科技有限公司 | Lactobacillus rhamnosus LR-28 strain with nerve soothing and sleep aiding effects, fermentation product, hypnotic fungus group mixture and application |
-
2023
- 2023-05-23 CN CN202310645941.XA patent/CN116676226B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109136130A (en) * | 2018-08-27 | 2019-01-04 | 南昌大学 | One plant of Lactobacillus rhamnosus NCU2217 |
| CN111728111A (en) * | 2020-06-28 | 2020-10-02 | 武汉康复得生物科技股份有限公司 | Probiotic composition for relieving anxiety or depression and application thereof |
| CN112501063A (en) * | 2020-12-03 | 2021-03-16 | 兰州大学 | Lactobacillus plantarum LZU-J-TSL6 capable of highly producing GABA and application thereof |
| CN115161250A (en) * | 2022-09-06 | 2022-10-11 | 广东益可维生物技术有限公司 | Lactobacillus plantarum with functions of improving sleep, strengthening brain and improving intelligence and application thereof |
| CN115521889A (en) * | 2022-10-30 | 2022-12-27 | 江南大学 | Lactobacillus plantarum WL02 capable of producing gamma-aminobutyric acid and application thereof |
| CN116676225A (en) * | 2023-05-23 | 2023-09-01 | 河北源民生物科技有限公司 | Lactobacillus rhamnosus LR-28 strain with nerve soothing and sleep aiding effects, fermentation product, hypnotic fungus group mixture and application |
Non-Patent Citations (1)
| Title |
|---|
| 石慧等.《食品分子微生物学》.中国农业大学出版社,2019,(2019年5月第1版),第79页. * |
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