CN117551568A - Lactobacillus plantarum TS with phenyllactic acid production capability - Google Patents
Lactobacillus plantarum TS with phenyllactic acid production capability Download PDFInfo
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- CN117551568A CN117551568A CN202311334712.2A CN202311334712A CN117551568A CN 117551568 A CN117551568 A CN 117551568A CN 202311334712 A CN202311334712 A CN 202311334712A CN 117551568 A CN117551568 A CN 117551568A
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- lactobacillus plantarum
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- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 42
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 35
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 35
- NWCHELUCVWSRRS-SECBINFHSA-N (2r)-2-hydroxy-2-phenylpropanoic acid Chemical compound OC(=O)[C@@](O)(C)C1=CC=CC=C1 NWCHELUCVWSRRS-SECBINFHSA-N 0.000 title claims abstract description 22
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 5
- 239000003242 anti bacterial agent Substances 0.000 claims abstract description 9
- 238000004321 preservation Methods 0.000 claims abstract description 9
- 239000001963 growth medium Substances 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 11
- 238000012258 culturing Methods 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 7
- 239000003814 drug Substances 0.000 claims description 5
- 238000003786 synthesis reaction Methods 0.000 claims description 5
- 230000003115 biocidal effect Effects 0.000 claims description 4
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- 230000000968 intestinal effect Effects 0.000 claims description 4
- 230000008855 peristalsis Effects 0.000 claims description 4
- 230000001737 promoting effect Effects 0.000 claims description 3
- 230000005764 inhibitory process Effects 0.000 abstract description 9
- 229940088710 antibiotic agent Drugs 0.000 abstract description 7
- 241000193830 Bacillus <bacterium> Species 0.000 abstract description 5
- 241000588724 Escherichia coli Species 0.000 abstract description 5
- 241000589517 Pseudomonas aeruginosa Species 0.000 abstract description 5
- 241000191967 Staphylococcus aureus Species 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 230000007774 longterm Effects 0.000 abstract description 4
- 239000002002 slurry Substances 0.000 abstract description 4
- 244000005700 microbiome Species 0.000 abstract description 3
- 238000000746 purification Methods 0.000 abstract description 3
- 206010059866 Drug resistance Diseases 0.000 abstract description 2
- 238000000926 separation method Methods 0.000 abstract description 2
- 230000001580 bacterial effect Effects 0.000 description 11
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 10
- 108020004465 16S ribosomal RNA Proteins 0.000 description 7
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 7
- 239000008103 glucose Substances 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 229920001817 Agar Polymers 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- 230000000844 anti-bacterial effect Effects 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000001888 Peptone Substances 0.000 description 5
- 108010080698 Peptones Proteins 0.000 description 5
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 5
- 235000015278 beef Nutrition 0.000 description 5
- 229940041514 candida albicans extract Drugs 0.000 description 5
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 5
- 239000012153 distilled water Substances 0.000 description 5
- 239000000284 extract Substances 0.000 description 5
- 235000013305 food Nutrition 0.000 description 5
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 5
- 235000019341 magnesium sulphate Nutrition 0.000 description 5
- 229940099596 manganese sulfate Drugs 0.000 description 5
- 239000011702 manganese sulphate Substances 0.000 description 5
- 235000007079 manganese sulphate Nutrition 0.000 description 5
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 5
- 235000019319 peptone Nutrition 0.000 description 5
- 239000001632 sodium acetate Substances 0.000 description 5
- 235000017281 sodium acetate Nutrition 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 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 5
- 239000001393 triammonium citrate Substances 0.000 description 5
- 235000011046 triammonium citrate Nutrition 0.000 description 5
- 239000012138 yeast extract Substances 0.000 description 5
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 4
- 150000007524 organic acids Chemical class 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 238000001308 synthesis method Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 238000003794 Gram staining Methods 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- PYMYPHUHKUWMLA-WDCZJNDASA-N arabinose Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)C=O PYMYPHUHKUWMLA-WDCZJNDASA-N 0.000 description 2
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000004310 lactic acid Substances 0.000 description 2
- 235000014655 lactic acid Nutrition 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 2
- 229920000053 polysorbate 80 Polymers 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 230000002335 preservative effect Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- VOXXWSYKYCBWHO-QMMMGPOBSA-N (S)-3-phenyllactic acid Chemical compound OC(=O)[C@@H](O)CC1=CC=CC=C1 VOXXWSYKYCBWHO-QMMMGPOBSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010256 biochemical assay Methods 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 238000007036 catalytic synthesis reaction Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 238000010227 cup method (microbiological evaluation) Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 231100000086 high toxicity Toxicity 0.000 description 1
- 238000013095 identification testing Methods 0.000 description 1
- 230000007413 intestinal health Effects 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 239000010871 livestock manure Substances 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229960005190 phenylalanine Drugs 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- C12P7/00—Preparation of oxygen-containing organic compounds
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- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
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Abstract
The invention provides a lactobacillus plantarum TS with phenyllactic acid production capability, which belongs to the technical field of microorganisms, wherein the lactobacillus plantarum is lactobacillus plantarum Lactobacillusplantarum TS which is preserved in China Center for Type Culture Collection (CCTCC) at the preservation number of NO: m2023796. The lactobacillus plantarum TS obtained through separation and purification in the slurry can produce the phenyllactic acid, has a good inhibition effect on staphylococcus aureus, escherichia coli, bacillus and pseudomonas aeruginosa, can be used for preparing fermented feed, and further solves the problems of safety, drug resistance and the like caused by long-term use of antibiotics in the prior art.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to lactobacillus plantarum TS with phenyllactic acid production capability.
Background
The long-term use of antibiotics leads to the generation of drug-resistant pathogenic bacteria, and the addition and use of antibiotics in foods and animal feeds are greatly limited, so that the problem of 'tikang syndrome' in the industries of foods, feeds and the like is a primary task, and organic acid can be used as a substitute of antibiotics within a few years.
And the phenyllactic acid is a novel organic acid antibacterial substance. Studies have shown that phenyllactic acid is a novel natural preservative which is expected to be applied to the fields of food, medicine, feed and the like. The phenyllactic acid added into the food and the feed can prolong the shelf life and inhibit pathogenic microorganisms, and the like, and provides a brand-new 'alternate thinking' for people in a forbidden large environment. However, the phenyllactic acid is expensive as a natural preservative, and the main synthesis methods are chemical synthesis methods and biological synthesis methods, wherein the chemical synthesis methods generate pollution, are time-consuming and labor-consuming, and a solvent with high toxicity is added in the extraction process of the phenyllactic acid, so that the phenyllactic acid cannot be used in industries such as food and feed; the biosynthesis method is divided into a whole cell catalytic synthesis method and a microorganism growth metabolism synthesis method, and is a novel method for synthesizing phenyllactic acid.
Thus, obtaining a strain that produces phenyllactic acid by the microbial growth metabolic synthesis method is a necessary trend for solving the problems of safety and drug resistance caused by the long-term use of antibiotics.
Disclosure of Invention
In view of the above, the invention aims to provide a lactobacillus plantarum TS with the capacity of producing phenyllactic acid, and the lactobacillus plantarum TS is obtained by separating and purifying from slurry, not only can produce phenyllactic acid, but also has a good antibacterial effect, can be used as an antibiotic substitute, is applied to the preparation of fermented feed, and further solves the problems of safety and the like caused by long-term use of antibiotics in the prior art.
In order to achieve the above purpose, the present invention provides the following technical solutions:
a lactobacillus plantarum which is lactobacillus plantarum Lactobacillus plantarum TS and is preserved in the China center for type culture collection (cctccc) NO: m2023796.
The invention also provides a method for culturing the lactobacillus plantarum, which comprises the step of inoculating the lactobacillus plantarum Lactobacillus plantarum TS to a culture medium for culturing.
Preferably, the temperature of the culture is 30-40 ℃, and the time of the culture is 20-28 h.
The invention also provides application of the lactobacillus plantarum in synthesis of phenyllactic acid.
The invention also provides application of the lactobacillus plantarum in preparing fermented feed.
The invention also provides application of the lactobacillus plantarum in preparation of a medicine for promoting intestinal peristalsis.
The invention also provides application of the lactobacillus plantarum in preparation of antibiotic substitutes.
Compared with the prior art, the invention has the following beneficial effects:
bacterial colony of strain TS obtained by separating and purifying in slurry is milky white, the center is convex, the surface is moist, and the diameter is 1-2 mm; microscopic examination results show that the strains are in a short rod shape and are arranged singly or in pairs; arabinose, mannose, glucose and the like can be utilized; has good inhibition effect on staphylococcus aureus, escherichia coli, bacillus and pseudomonas aeruginosa, can produce phenyllactic acid, and can be used for preparing fermented feed instead of antibiotics. When the strain is applied to fermented feed, the utilization rate of the feed can be improved, the intestinal peristalsis of animals is promoted, and the edible safety is higher.
Drawings
FIG. 1 shows amplification products of the rDNA gene of strain 16S, wherein the PCR results of strain TS, strain TS1, strain TS2, strain DBb4, strain YB1, strain QJS 1.1.1, strain QJS 1.2.2, strain QJS 2.1.1 and marker are shown in sequence from left to right;
FIG. 2 is a phylogenetic tree of the strain based on 16S rDNA;
FIG. 3 shows colony morphology of strains on a plate;
FIG. 4 shows the results of gram staining of strains;
FIG. 5 shows the results of a bacterial strain bacteriostasis test against Staphylococcus aureus;
FIG. 6 shows the results of a bacterial strain bacteriostasis test on E.coli;
FIG. 7 shows the results of a bacterial strain bacteriostasis test on Bacillus;
FIG. 8 shows the results of a bacterial inhibition test of the strain Pseudomonas aeruginosa.
Description of biological preservation
Lactobacillus plantarum Lactobacillus plantarum TS is preserved in China Center for Type Culture Collection (CCTCC), the preservation address is China, wuhan, university of Wuhan, the preservation date is 2023, 5 and 22 days, and the preservation number is CCTCC NO: m2023796.
Detailed Description
The invention provides a lactobacillus plantarum which is lactobacillus plantarum Lactobacillus plantarum TS and is preserved in China Center for Type Culture Collection (CCTCC) in 2023, 5 and 22 days, wherein the preservation number is CCTCC NO: m2023796.
The invention also provides a method for culturing the lactobacillus plantarum, which comprises the step of inoculating the lactobacillus plantarum Lactobacillus plantarum TS to a culture medium for culturing.
In the present invention, the medium is preferably an MRS solid medium; the formula of MRS solid culture medium is preferably glucose 18-22 g, peptone 8-12 g, beef extract 8-12 g, yeast extract 3-7 g, crystalline sodium acetate 3-7 g, triammonium citrate 1-3 g, dipotassium hydrogen phosphate 1-3 g, tween-80.5-1.5 mL, magnesium sulfate 0.50-0.60 g, manganese sulfate 0.2-0.3 g, agar 15-20 g, distilled water 800-1200 mL and pH value 6.2-6.4; further preferably, the feed comprises 19 to 21g of glucose, 9 to 11g of peptone, 9 to 11g of beef extract, 4 to 6g of yeast extract, 4 to 6g of crystalline sodium acetate, 2g of triammonium citrate, 2g of dipotassium hydrogen phosphate, 800.7 to 1.3mL of tween-800.7, 0.54 to 0.56g of magnesium sulfate, 0.23 to 0.27g of manganese sulfate, 16 to 19g of agar, 900 to 1100mL of distilled water and pH value of 6.25 to 6.35; still more preferably, 20g of glucose, 10g of peptone, 10g of beef extract, 5g of yeast extract powder, 5g of crystalline sodium acetate, 3g of tri-ammonium citrate, 2g of dipotassium hydrogen phosphate, 1.0mL of tween-80, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 17-18 g of agar, 1000mL of distilled water and pH value of 6.3; the temperature of the culture is preferably 30 to 40 ℃, more preferably 32 to 38 ℃, still more preferably 34 to 37 ℃, still more preferably 35 to 36 ℃; the time for the culture is preferably 20 to 28 hours, more preferably 22 to 26 hours, still more preferably 24 to 25 hours.
The invention also provides application of the lactobacillus plantarum in synthesis of phenyllactic acid.
The invention also provides application of the lactobacillus plantarum in preparing fermented feed.
The invention also provides application of the lactobacillus plantarum in preparation of a medicine for promoting intestinal peristalsis.
The invention also provides application of the lactobacillus plantarum in preparation of antibiotic substitutes.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
1.1 isolation and purification of Strain
Taking 1mL of water from Tianshui City of Gansu province Qin An county, xingguo province and Lin Cunjiang, inoculating the water into an enrichment medium, and culturing at 35 ℃ for 24 hours to enrich strains; 0.1mL of diluted solution was takenThe concentration of the bacterial cells was 10 8 The bacterial liquid of CFU/mL is inoculated into a primary screening lower layer culture medium, cultured for 24 hours at 35 ℃, injected into a primary screening upper layer culture medium, continuously cultured for 24 hours, colonies with the size of 4-5 mm and a calcium dissolving ring are selected to be inoculated into an MRS solid culture medium, streaked and purified for 5 times, microscopic examination is carried out, the strain morphology is observed, and the strain is placed into a refrigerator at 4 ℃ for inclined plane preservation. The results are shown in FIG. 3.
Wherein, the formula of MRS liquid culture medium is: 20g of glucose, 10g of peptone, 10g of beef extract, 5g of yeast extract powder, 5g of crystalline sodium acetate, 2g of tri-ammonium citrate, 2g of dipotassium hydrogen phosphate, 801mL of Tween-801 g of magnesium sulfate, 0.58g of manganese sulfate, 0.25g of pH value of 6.4 and 1000mL of distilled water, and sterilizing at 121 ℃ for 20 min;
the formula of MRS solid culture medium is as follows: 20g of glucose, 10g of peptone, 10g of beef extract, 5g of yeast extract powder, 5g of crystalline sodium acetate, 2g of tri-ammonium citrate, 2g of dipotassium hydrogen phosphate, 1mL of tween-80, 0.58g of magnesium sulfate, 0.25g of manganese sulfate, 18g of agar, 6.4 of pH value and 1000mL of distilled water, and sterilizing at 121 ℃ for 20 min;
the formula of the enrichment medium is that 2.0g/L phenyllactic acid is added into MRS liquid medium;
the formula of the primary screening lower culture medium is as follows: 2.0g/L phenylalanine is added into MRS agar culture medium;
the formula of the primary screening upper layer culture medium is as follows: 30g/L of calcium carbonate was added to MRS agar medium.
The results showed that the bacterial strains obtained by separation and purification had colony sizes of about 1-2mm, wet surfaces, raised centers and microscopic examination showed that the strains were in the form of short rods, singly or in pairs. And the strain was named TS.
1.2 physiological Biochemical assay
The isolated and purified strain was subjected to gram staining and physiological and biochemical identification tests using a bacterial biochemical identification tube. The experimental results are shown in fig. 4 and table 1.
TABLE 1 physiological and Biochemical identification results
As can be seen from fig. 4 and table 1, the result of the gram staining is bluish violet, and the strain belongs to gram positive bacteria and is a facultative anaerobe; the strain can utilize arabinose, mannose, glucose and the like, reduce nitrate to be negative, and can grow on 6.5% high-salt broth.
1.3 16S rDNA molecular identification
The isolated and purified strain was subjected to molecular characterization using the 16S rDNA method. 16S rDNA amplification and sequencing were performed by Shanghai Biotechnology Co.
The 16S rDNA nucleotide sequence of the strain TS is as follows:
AGGCTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGCTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAACCAGCCGCCTAAGGTGGGACAGATGATTAGGGTGAAGTCGTAA(SEQ ID No.1)。
after sequencing, the 16S rDNA sequences of strain TS were aligned in NCBI (National Center for Biotechnology Information ) database, and it was found that the gene sequence of strain TS was in the same cluster as Lactobacillus plantarum (Lactobacillus plantarum) GVAP in all similar sequences. The identified strain TS is Lactobacillus plantarum in combination with the physiological and biochemical characteristics described above.
The nucleotide sequence of the 16S rDNA of Lactobacillus plantarum GVIP is:
CTGGCTCAGGACGAACGCTGGCGGCGTGCCTAATACATGCAAGTCGAACGAACTCTGGTATTGATTGGTGCTTGCATCATGATTTACATTTGAGTGAGTGGCGAACTGGTGAGTAACACGTGGGAAACCTGCCCAGAAGCGGGGGATAACACCTGGAAACAGATGCTAATACCGCATAACAACTTGGACCGCATGGTCCGAGCTTGAAAGATGGCTTCGGCTATCACTTTTGGATGGTCCCGCGGCGTATTAGCTAGATGGTGGGGTAACGGCTCACCATGGCAATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACATATCTGAGAGTAACTGTTCAGGTATTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTCAACCGAAGAAGTGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGTATGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCATACCGTAAACGATGAATGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGGCCGCAAGGCTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATACTATGCAAATCTAAGAGATTAGACGTTCCCTTCGGGGACATGGATACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATTATCAGTTGCCAGCATTAAGTTGGGCACTCTGGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGATGGTACAACGAGTTGCGAACTCGCGAGAGTAAGCTAATCTCTTAAAGCCATTCTCAGTTCGGATTGTAGGCTGCAACTCGCCTACATGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAACCAGCCGCCTAAGGTGGGACAGATGATTAGGGTGAAGTCGTAACAAGGGTAACCGTAAGGG(SEQ ID No.2)。
the lactobacillus plantarum (Lactobacillus plantarum) strain TS separated from the slurry is preserved in China center for type culture collection (China center for type culture collection) at the month 22 of 2023, and the strain preservation number is CCTCC NO: m2023796.
Example 2 bacteriostasis test
The bacteriostasis test is carried out by adopting an oxford cup method, and specifically comprises the following steps: placing 5 oxford cups on 5 MRS solid culture media (same as example 1) respectively containing indicator bacteria of staphylococcus aureus, bacillus, pseudomonas aeruginosa and escherichia coli, and injecting 0.1mL of thallus with concentration of 10 8 The CFU/mL bacterial solution was cultured in a constant temperature incubator at 35℃for 24 hours, and the culture was repeated 3 times, and the average value was obtained from the observation result. The results are shown in Table 2.
The results were evaluated by reference to the criteria described in "Zhang Suhui, cao Guowen, xu Dengfeng, etc.. The study of the isolation and identification of lactic acid bacteria in piglet manure [ J ]. Feed industry, 2007 (14): 31-33 ], specifically: the diameter of the inhibition zone is smaller than 4mm and is insensitive, 5-10 mm is low-sensitivity, 11-15 mm is medium-sensitivity, and more than 15mm is high-sensitivity.
TABLE 2 antibacterial test results
The results show that the inhibition effect of the strain on staphylococcus aureus reaches high sensitivity (the diameter of a inhibition zone is more than 15.00 mm), and the inhibition effect on escherichia coli, bacillus and pseudomonas aeruginosa is moderate sensitivity (11.00 mm < the diameter of the inhibition zone is less than 15.00 mm). Therefore, the organic acid such as lactic acid and the like generated in the culture process can enable the bacterial liquid to be acidic, acid inhibition is generated on the growth of the indicator bacteria, meanwhile, the strain can generate a substance phenyllactic acid with an antibacterial effect, and the phenyllactic acid can play a synergistic effect with the organic acid to generate stronger antibacterial effect.
The strain provided by the invention is a probiotic capable of producing phenyllactic acid, has good antibacterial capability, and can play a role in prolonging the shelf life of feed while improving the intestinal health of animals. The lactobacillus plantarum provided by the invention has the characteristics that the lactobacillus plantarum can be applied to the aspect of tibody.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (7)
1. The lactobacillus plantarum is Lactobacillusplantarum TS, and is preserved in China Center for Type Culture Collection (CCTCC) in 2023, 5 and 22 days, and the preservation number is CCTCC NO: m2023796.
2. The method for culturing lactobacillus plantarum according to claim 1, wherein lactobacillus plantarum Lactobacillusplantarum TS is inoculated in a culture medium for culturing.
3. The method according to claim 2, wherein the temperature of the culture is 30 to 40℃and the time of the culture is 20 to 28 hours.
4. Use of lactobacillus plantarum according to claim 1 for the synthesis of phenyllactic acid.
5. Use of lactobacillus plantarum according to claim 1 for the preparation of a fermented feed.
6. Use of lactobacillus plantarum according to claim 1 for the manufacture of a medicament for promoting intestinal peristalsis.
7. Use of lactobacillus plantarum according to claim 1 for the preparation of an antibiotic substitute.
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| CN117903998A (en) * | 2024-03-01 | 2024-04-19 | 朗恒科技集团有限公司 | Lactobacillus plantarum ST10-12 for high-yield phenyllactic acid and application thereof |
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