CN115960760A - Pediococcus pentosaceus with cholesterol reducing and bacteriostatic effects and high-density industrial production fermentation medium thereof - Google Patents
Pediococcus pentosaceus with cholesterol reducing and bacteriostatic effects and high-density industrial production fermentation medium thereof Download PDFInfo
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Abstract
The invention discloses Pediococcus pentosaceus with cholesterol reducing and bacteriostatic effects and a high-density industrial production fermentation medium thereof, wherein the Pediococcus pentosaceus is Pediococcus pentosaceus PG-2 with the preservation number of GDMCC No:62392. the high-density industrial fermentation medium of pediococcus pentosaceus PG-2 comprises 40g/L of sugarcane molasses liquid, 20g/L of corn steep liquor dry powder, 1.5g/L of dipotassium hydrogen phosphate, 5g/L of sodium acetate, 1mL/L of Tween80, 2g/L of triammonium citrate, 0.2g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate tetrahydrate and the balance of water. When the culture medium is used for culturing pediococcus pentosaceus, the concentration of bacteria is obviously higher than that of an MRS culture medium; effectively obtains higher density bacteria amount, improves the utilization rate of industrial byproducts, greatly reduces the production cost and improves the production efficiency.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to pediococcus pentosaceus with cholesterol-reducing and bacteriostatic effects and a high-density industrial production fermentation medium thereof.
Background
Hyperlipidemia is a metabolic disease in which the balance of lipid metabolism in the body is disrupted, resulting in higher than average levels of blood lipids. Through the investigation and research of the world health organization, the main factor of the disease onset such as atherosclerosis and coronary heart disease is hyperlipidemia. The related reports indicate that if the cholesterol level in the body is 1mmol/L higher than the normal average level, there is a 35% probability of coronary heart disease. Data show that mortality caused by hyperlipidemia and its associated diseases accounts for half of the worldwide mortality.
The regulation of cholesterol levels in the blood of the body is mainly achieved by adopting drugs to prevent the self-synthesis of endogenous cholesterol, reduce the absorption of exogenous cholesterol through diet adjustment and promote the efflux of cholesterol. In the seventies and eighty years of the last century, researches on reducing blood fat by probiotics are started abroad. Two bacterial strains deeply researched for the function of reducing blood fat in China belong to lactobacillus plantarum, namely L.plantarum P8 and ST-III. Animal experiments show that the L.plantarum P8 and ST-III strains both have obvious blood fat reducing effect. Therefore, the development of safe and effective lactobacillus for regulating blood lipid balance has wide prospects in the fields of food and medicine. Pediococcus pentosaceus (Pediococcus pentosaceus) is used as an inherent strain of animal intestinal microorganisms, inhibits the proliferation of pathogenic bacteria through self-propagation, and improves the disease resistance and the immune function of an organism. However, the viable count of the pediococcus pentosaceus produced by the existing production process is not high, and the production cost is high, so that the use effect and the application and popularization of the pediococcus pentosaceus are directly influenced. Therefore, the fermentative production of pediococcus pentosaceus is yet to be developed.
Disclosure of Invention
Aiming at the defects of low viable count and high production cost of pediococcus pentosaceus in the prior art and relieving the growing severe situation of the domestic hyperlipemia population, the invention provides the pediococcus pentosaceus with cholesterol reduction and broad-spectrum antibacterial effects and a high-density industrial production fermentation medium thereof.
The invention aims to provide a Pediococcus pentosaceus (Pediococcus pentosaceus) PG-2 with a preservation number of: GDMCC No:62392.
the Pediococcus pentosaceus PG-2 is derived from Asian otter feces. Observing the colony morphology by naked eyes, wherein the colony is milky white, smooth in surface, neat in edge and opaque; gram stain is purple, gram positive bacteria are obtained, and the shape of the bacteria is circular when observed under a microscope. The 16S rRNA sequence of the Pediococcus pentosaceus PG-2 has the length of 1356bp, the specific sequence is shown as SEQ ID NO.1, the Pediococcus pentosaceus is identified as the Pediococcus pentosaceus, and the Pediococcus pentosaceus is named as the Pediococcus pentosaceus (Pediococcus pentosaceus) PG-2.
It is a second object of the present invention to provide a preparation containing the aforementioned Pediococcus pentosaceus PG-2 or a fermentation broth thereof as an active ingredient.
The third purpose of the invention is to provide the use of the Pediococcus pentosaceus PG-2 or the preparation in at least one of the following (1) to (4):
(1) Preventing and treating pathogenic bacteria;
(2) Preparing a cholesterol-lowering medicine or health-care product;
(3) Preparing an animal feed or feed additive;
(4) Preparing food or food additive.
Preferably, the pathogenic bacteria include Staphylococcus aureus (Staphylococcus aureus), escherichia coli (Escherichia coli), and Salmonella enteritidis (Salmonella enteritidis).
The fourth purpose of the invention is to provide a fermentation medium for high-density industrial production of pediococcus pentosaceus, which contains 30-40g/L of cane molasses liquid, 20-25g/L of corn steep liquor dry powder, 1.5g/L of dipotassium hydrogen phosphate, 5g/L of sodium acetate, 80 mL/L of Tween, 2g/L of triammonium citrate, 0.2g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate tetrahydrate and water as a solvent.
Preferably, the fermentation medium contains 40g/L of sugarcane molasses, 20g/L of corn steep liquor dry powder, 1.5g/L of dipotassium phosphate, 5g/L of sodium acetate, 80 mL/L of Tween, 2g/L of triammonium citrate, 0.2g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate tetrahydrate and water as a solvent.
The fifth purpose of the invention is to provide the application of the high-density industrial production fermentation medium in high-efficiency fermentation of pediococcus pentosaceus.
The sixth object of the present invention is to provide a method for efficiently fermenting pediococcus pentosaceus by inoculating the pediococcus pentosaceus PG-2 to the high-density industrial production fermentation medium for fermentation culture.
Preferably, the method comprises the following steps: activating the preserved pediococcus pentosaceus PG-2 on an MRS agar culture medium plate, selecting a single colony, inoculating the single colony in an MRS liquid culture medium for culture, and transferring the bacterial liquid to a high-density industrial production fermentation culture medium for fermentation culture.
Preferably, the single colony is selected and inoculated in an MRS liquid culture medium for culture under the culture conditions that: culturing at 37 deg.C for 14 hr; the transferring is to transfer the bacterial liquid according to the inoculum size of 1 percent of the volume fraction; the fermentation culture conditions are as follows: fermenting at 37 deg.C for 48 hr.
Compared with the prior art, the invention has the following beneficial effects:
(1) The pediococcus pentosaceus PG-2 disclosed by the invention can generate antibacterial substances, has a good antibacterial effect on common livestock and poultry aquaculture pathogenic bacteria, comprises a good inhibition effect on staphylococcus aureus, escherichia coli and salmonella enteritidis, and simultaneously has an excellent cholesterol reduction effect, and the cholesterol clearance rate is as high as (40.66 +/-0.25)%;
(2) The high-density industrial production fermentation medium for pediococcus pentosaceus PG-2 is suitable for large-scale fermentation of pediococcus pentosaceus. The industrial by-product cane molasses is used as a carbon source, the corn steep liquor dry powder is used as a main nitrogen source to culture the pediococcus pentosaceus, the production cost is reduced, the rapid growth of the pediococcus pentosaceus is facilitated, and the viable bacteria number after fermentation is obviously improved compared with that of an MRS culture medium.
The Pediococcus pentosaceus PG-2 is preserved in Guangdong province microbial strain preservation center (GDMCC) at 18 months 4 and 2022, and the address is No. 59 building of Dazhou college No. 100 of Xieli Zhonglu in Guangzhou city, and the zip code is as follows: 510070, the preservation number is: GDMCC No:62392.
drawings
FIG. 1 is a morphological diagram of PG-2 colony of pediococcus pentosaceus of the present invention.
FIG. 2 is a microscopic image of the PG-2 of Pediococcus pentosaceus of the present invention.
FIG. 3 is a schematic diagram and bacteriostatic pictures of the supernatant after centrifugation of the fermentation broth of Pediococcus pentosaceus PG-2 of the present invention against 3 pathogenic bacteria.
FIG. 4 shows the effect of a single inorganic salt on the viable count of Pediococcus pentosaceus PG-2.
FIG. 5 shows the effect of various inorganic salts on the viable count of Pediococcus pentosaceus PG-2.
Detailed Description
The following is a further description of the present invention in conjunction with embodiments thereof, and is not intended to limit the present invention. In the following examples, all consumables, instruments and other related items and detection methods are conventional unless otherwise specified.
Example 1: separation, purification, identification and preservation of Pediococcus pentosaceus (PG-2)
(1) Separation, screening and preservation of Pediococcus pentosaceus (PG-2)
The Pediococcus pentosaceus PG-2 is separated from Asian otter excrement.
MRS agar medium: 10g of casein digest, 10g of beef extract powder, 4g of yeast extract powder, 20g of glucose, 2g of dipotassium hydrogen phosphate, 2g of triammonium citrate, 5g of sodium acetate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate tetrahydrate, 801.08g of tween-agar and 15g of agar, mixing the components, adding water to a constant volume of 1000mL, adjusting the pH value to 5.7 +/-0.2, and subpackaging into 250mL triangular bottles with 100mL of each bottle; sterilizing at 121 deg.C under high temperature and high pressure for 20min, and pouring into flat plate.
Asian otter feces are coated on a plate containing an MRS agar culture medium by a dilution coating method, and a strain PG-2 is obtained after purification. The colony morphology was visually observed, and the results are shown in FIG. 1. The bacterial colony is milky white, smooth in surface, neat in edge and non-transparent; the gram stain is purple, and is gram-positive bacteria; the shape of the cells was observed under a microscope to be circular (FIG. 2).
Extracting genome DNA of the strain PG-2, amplifying by using bacterial 16S rRNA gene amplification universal primers 27F/1492R (27F. And (3) carrying out homology comparison analysis on the sequencing result and a 16S rRNA sequence in an EzBioCloud database, and preliminarily identifying that the separated strain PG-2 is pediococcus pentosaceus by combining morphological observation. It was named: pediococcus pentosaceus (Pediococcus pentosaceus) PG-2. The strain was deposited in the Guangdong province culture Collection (GDMCC) at 18 days 4 and 4 months 2022, address: building 5 of first furnance, large yard, 100, building 59, guangdong province, guangzhou, china, zip code: 510070, with a collection number of GDMCC No:62392.
the 16S rRNA sequence (SEQ ID NO. 1) of Pediococcus pentosaceus PG-2 is specifically shown as follows:
GGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCGACTTCGTGTAGGCGAGTTGCAGCCTACAGTCCGAACTGAGAATGGTTTTAAGAGATTAGCTTAACCTCGCGGTCTCGCGACTCGTTGTACCATCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGATTTGACGTCGTCCCCACCTTCCTCCGGTTTGTCACCGGCAGTCTCACTAGAGTGCCCAACTTAATGCTGGCAACTAGTAATAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCATTCTGTCCCCGAAGGGAACCTCTAATCTCTTAGACTGTCAGAAGATGTCAAGACCTGGTAAGGTTCTTCGCGTAGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCTTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGATTACTTAATGCGTTAGCTGCAGCACTGAAGGGCGGAAACCCTCCAACACTTAGTAATCATCGTTTACGGCATGGACTACCAGGGTATCTAATCCTGTTCGCTACCCATGCTTTCGAGCCTCAGCGTCAGTTGCAGACCAGACAGCCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTTCACCGCTACACATGGAGTTCCACTGTCCTCTTCTGCACTCAAGTCTCCCAGTTTCCAATGCACTTCTTCGGTTGAGCCGAAGGCTTTCACATTAGACTTAAAAGACCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGATAACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAAATACCGTCACTGGGTAAACAGTTACTCTTACCCACGTTCTTCTTTAACAACAGAGCTTTACGAGCCGAAACCCTTCTTCACTCACGCGGCGTTGCTCCATCAGACTTGCGTCCATTGTGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCTCAGTCCCAATGTGGCCGATTACCCTCTCAGGTCGGCTACGTATCACTGCCTTGGTGAGCCTTTACCTCACCAACTAGCTAATACGCCGCGGGTCCATCCAGAAGTGATAGCAGAGCCATCTTTTAAAAGAAAACCATGCGGTTTTCTCTGTTATACGGTATTAGCATCTGTTTCCAGGTGTTATCCCCTACTTCTGGGCAGGTTACCCACGTGTTACTCACCCGTTCGCCACTCACTTCGTGTTAAAATCTCAATCAGTACAAGTACG。
example 2: pediococcus pentosaceus PG-2 antibacterial effect determination
(1) Preparing test bacteria supernatant: transferring the activated Pediococcus pentosaceus PG-2 into an MRS liquid culture medium (except that no agar is added, the formula is the same as that of the MRS agar culture medium) according to the inoculation amount of 5% of volume fraction, and placing the culture medium in a 37 ℃ three-gas culture box for culturing for 18h, wherein the oxygen content in the three-gas culture box is 5% and the carbon dioxide content is 10%; the culture fluid is centrifuged, and the supernatant is taken and placed at 4 ℃ for later use.
(2) Preparing a bacterium-containing plate: transferring the test strain into nutrient broth culture medium (formula: peptone 10.0g/L, beef extract powder 3.0g/L, sodium chloride 5.0g/L, pH 7.4 + -0.2, solvent is water), culturing in 30 deg.C constant temperature incubator (as shown in Table 1), and adjusting OD with ultraviolet spectrophotometer 600 And when the value reaches 0.1, respectively sucking 0.1mL of bacterial liquid, adding the bacterial liquid into corresponding 100mL of culture medium which is melted at about 45 ℃, fully shaking up, pouring the plate, and after the bacterial liquid is naturally solidified, uniformly punching holes (with the hole diameter of 9 mm) on the same plate by using a puncher respectively.
(3) Bacteriostasis test
And (3) sucking 100 mu L of supernatant bacterial liquid obtained by centrifuging the Pediococcus pentosaceus PG-2, respectively adding the supernatant bacterial liquid into each hole containing a test pathogenic bacterium plate, repeating the steps for three groups, culturing the cells in an incubator at 30 ℃ for about 24 hours, observing the bacteriostatic effect, and measuring the diameter of a bacteriostatic circle (the result is shown in figure 3).
TABLE 1 test strains for Pediococcus pentosaceus PG-2 bacteriostatic effect test
As can be seen from FIG. 3, the supernatant liquid of the pediococcus pentosaceus PG-2 of the invention has better inhibitory effect on staphylococcus aureus, escherichia coli and salmonella enteritidis.
Example 3: determination of BSH enzyme activity produced by Pediococcus pentosaceus PG-2
(1) Drawing of taurine standard curve
The standard curve is characterized in that the concentration (mu mol/mL) of taurine is used as an abscissa, and the light absorption value (A570 nm) is used as an ordinate, so that the standard curve of the prepared taurine is as follows: y =0.2286x-0.0041,R 2 ＝0.9923。
(2) Preparation and determination of samples
After the pediococcus pentosaceus PG-2 is activated for 3 times, the pediococcus pentosaceus PG-2 is inoculated into an MRS liquid culture medium according to the inoculation amount of 3 percent of volume fraction, cultured for 24 hours at 37 ℃, and an appropriate amount of fermentation liquor is taken for centrifugation to collect thalli. The cells were washed 2 times with 0.1mol/L (pH 7.0) phosphate buffer, the absorbance was adjusted to 1.0 at 600nm, and dithiothreitol was added to prevent oxidation of Bile Salt Hydrolase (BSH). Taking 1mL of the adjusted bacterial liquid, carrying out ultrasonic disruption, and centrifuging to obtain a cell-free extracting solution. Taking 10 mu L of cell-free extract, adding 180 mu L of 0.1mol/L phosphate buffer solution, 10 mu L of 200mmol/L sodium taurocholate and 10 mu L of paraffin oil, uniformly mixing, placing at 37 ℃ for reaction for 30min, adding trichloroacetic acid with the same volume to stop the reaction, uniformly mixing, and centrifuging to obtain supernatant. Mixing 0.1mL of supernatant with 1.9mL of ninhydrin color developing solution, heating in boiling water for 14min, cooling with tap water, and measuring light absorption value with microplate reader at 570 nm. The positive control strain adopts Lactobacillus plantarum GDMCC1.140.
Definition of BSH crude enzyme activity: the amount of the substance which binds to bile salts to produce amino acids is hydrolyzed by the crude enzyme in μmol/(h · mL) per unit time and unit volume. BSH can be hydrolyzed and combined with bile salt to generate amino acid and free bile acid, and the free bile acid can form a complex with cholesterol and is discharged out of a body, so that the content of the cholesterol in blood is reduced. Therefore, BSH can be used as an important index for screening cholesterol degrading strains.
The result shows that the crude enzyme activity of the PG-2 of the BSH-producing Pediococcus pentosaceus (Pediococcus pentosaceus) is as high as 0.75 mu mol/(h.mL), and the crude enzyme activity of the GDMCC1.140 of the positive control lactobacillus plantarum is 0.70 mu mol/(h.mL), which indicates that the PG-2 of the Pediococcus pentosaceus (Pediococcus pentosaceus) has the potential of efficiently reducing cholesterol.
Example 4: determination of PG-2 Cholesterol removing Effect of Pediococcus pentosaceus
(1) Drawing of Cholesterol Standard Curve
The standard curve is prepared by taking the concentration (mg/mL) of cholesterol as an abscissa and an absorbance (A500 nm) as an ordinate, and the standard curve of cholesterol is prepared as follows: y =24.393x-0.0125 2 ＝0.9944。
(2) Preparation and determination of samples
After activating the pediococcus pentosaceus PG-2 for 3 times, inoculating the pediococcus pentosaceus PG-2 into an MRS culture medium (MRS-CHOL culture medium) containing cholesterol according to the inoculation amount of 3 percent of volume fraction, culturing for 24 hours at 37 ℃, centrifuging 1mL of fermentation liquor to obtain supernatant, measuring the content of the cholesterol by adopting an o-phthalaldehyde method, and calculating the removal rate of the cholesterol of different strains according to the following formula. MRS-CHOL culture medium is prepared by adding 3% cholate and 1% cholesterol into MRS liquid culture medium. The positive control strain adopts Lactobacillus plantarum GDMCC1.140.
Wherein A is the concentration of cholesterol in the medium before inoculation; and B is the concentration of cholesterol in the culture medium after inoculation and fermentation.
The result shows that the cholesterol clearance rate of the PG-2 of the Pediococcus pentosaceus (Pediococcus pentosaceus) is as high as (40.66 +/-0.25)%, the cholesterol clearance rate of the positive control lactobacillus plantarum GDMCC1.140 is 45 percent, and the result shows that the PG-2 of the Pediococcus pentosaceus (Pediococcus pentosaceus) has high cholesterol reducing capability. The strain PG-2 can be applied to products with the function of degrading cholesterol, such as feed additives.
Example 5: comparison of results of inorganic salt fermentation single-factor test in culture medium
The fermentation medium comprises the following components in parts by mass:
culture medium 1: MRS liquid culture medium;
culture medium 2: 40g/L of sugarcane molasses liquid, 20g/L of corn steep liquor dry powder, 1.5g/L of dipotassium phosphate, 5g/L of sodium acetate, 801mL/L of Tween, 2g/L of triammonium citrate, 0.2g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate tetrahydrate and water as a solvent;
and (3) a culture medium: 40g/L of sugarcane molasses liquid, 20g/L of corn steep liquor dry powder and water as a solvent;
culture medium 4: 40g/L of sugarcane molasses liquid, 20g/L of corn steep liquor dry powder, 1.5g/L of dipotassium hydrogen phosphate and water as a solvent;
and (5) culture medium: 40g/L of sugarcane molasses liquid, 20g/L of corn steep liquor dry powder, 5g/L of sodium acetate and water as a solvent;
and (6) a culture medium: 40g/L of cane molasses liquid, 20g/L of corn steep liquor dry powder, 1mL/L of Tween80, and water as a solvent;
and (7) a culture medium: 40g/L of cane molasses liquid, 20g/L of corn steep liquor dry powder, 2g/L of triammonium citrate and water as a solvent;
and (3) a culture medium 8: 40g/L of cane molasses liquid, 20g/L of corn steep liquor dry powder, 0.2g/L of magnesium sulfate heptahydrate and water as a solvent;
culture medium 9: 40g/L of sugarcane molasses liquid, 20g/L of corn steep liquor dry powder, 0.05g/L of manganese sulfate tetrahydrate and water as a solvent;
the pH of the above medium was 6.0.
The preparation method of the culture medium comprises the following steps: mixing the above components, dissolving with distilled water, placing the prepared culture medium into 250mL triangular flask, stirring, and sterilizing at 115 deg.C under high temperature and high pressure for 20min.
In order to achieve the best effect of the culture medium, when the culture medium is used for fermenting pediococcus pentosaceus, the fermentation process is as follows:
(1) Activating the cryopreserved pediococcus pentosaceus PG-2 on an MRS agar culture medium plate (the components and the preparation method are the same as the embodiment 1), selecting a single colony to inoculate a test tube containing an MRS liquid culture medium, and culturing at 37 ℃ for about 14 hours until the liquid culture medium is obviously turbid to obtain a seed culture solution;
(2) Preparing 50mL of each of the 9 fermentation culture media according to a culture medium formula, filling the prepared fermentation culture media into a 250mL triangular flask, and cooling the temperature to 37 ℃ after sterilization;
(3) Inoculating the obtained seed culture solution into triangular flasks filled with different fermentation media according to the inoculation amount of 1% of the volume fraction; each set was 3 replicates.
(4) The fermentation conditions were set as follows: fermenting at 37 deg.C for 48 hr, and diluting the fermented liquid for coating.
The results of the viable cell count of the bacterial suspension obtained by the fermentation culture using the 9 types of fermentation media are shown in FIG. 4. Figure 4 results show that: pediococcus pentosaceus PG-2 was found to have the highest viable count in Medium 2, followed by Medium 1 and Medium 7. Since the viable count of medium 7 was significantly higher than that of basal medium 3, medium 7 was selected as the optimized fermentation medium.
Example 6: comparison of test results of various inorganic salt fermentation factors in culture medium
The fermentation medium comprises the following components in parts by mass:
culture medium 1: MRS liquid culture medium;
culture medium 2: 40g/L of cane molasses liquid, 20g/L of corn steep liquor dry powder, 1.5g/L of dipotassium phosphate, 5g/L of sodium acetate, 801mL/L of Tween, 2g/L of triammonium citrate, 0.2g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate tetrahydrate and water serving as a solvent;
culture medium 3: 40g/L of sugarcane molasses liquid, 20g/L of corn steep liquor dry powder and water as a solvent;
culture medium 4: 40g/L of cane molasses liquid, 20g/L of corn steep liquor dry powder, 2g/L of triammonium citrate, 1.5g/L of dipotassium phosphate and water as a solvent;
and (5) a culture medium: 40g/L of cane molasses liquid, 20g/L of corn steep liquor dry powder, 2g/L of triammonium citrate, 1mL/L of Tween80 and water as a solvent;
and 6, a culture medium: 40g/L of cane molasses liquid, 20g/L of corn steep liquor dry powder, 2g/L of triammonium citrate, 0.2g/L of magnesium sulfate heptahydrate and water as a solvent;
and (7) a culture medium: 40g/L of sugarcane molasses liquid, 20g/L of corn steep liquor dry powder, 2g/L of triammonium citrate, 0.05g/L of manganese sulfate tetrahydrate and water as a solvent;
culture medium 8: 40g/L of cane molasses liquid, 20g/L of corn steep liquor dry powder, 2g/L of triammonium citrate and water as a solvent;
the pH of the above medium was 6.0. Each set was 3 replicates.
The medium preparation method and fermentation process were the same as in example 5.
The results of the viable cell count of the bacterial suspension obtained by the fermentation culture using the above 8 types of fermentation media are shown in FIG. 5. Figure 5 results show that: pediococcus pentosaceus PG-2 has the highest viable count in Medium 2, followed by Medium 1 and Medium 4. The viable count of the culture medium 1 and the culture medium 2 is obviously higher than that of the culture medium 4, and the viable count of the culture medium 4 and the culture medium 8 is not greatly different, so that the culture medium 2 is preferably used as an industrial high-density fermentation culture medium for pediococcus pentosaceus PG-2 and is beneficial to the growth of the pediococcus pentosaceus PG-2. And the carbon source and the nitrogen source of the culture medium adopt industrial byproducts, so the fermentation cost of the culture medium is obviously lower than that of the MRS liquid culture medium.
The above are only preferred embodiments of the present invention, and it should be noted that the above preferred embodiments should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.
Claims (10)
1. Pediococcus pentosaceus (Pediococcus pentosaceus) PG-2 with a preservation number of: GDMCC No:62392.
2. a preparation comprising the Pediococcus pentosaceus (Pediococcus pentosaceus) PG-2 or a fermentation liquid thereof according to claim 1 as an active ingredient.
3. Use of Pediococcus pentosaceus (Pediococcus pentosaceus) PG-2 according to claim 1 or a formulation according to claim 2 in at least one of the following (1) to (4):
(1) Preventing and treating pathogenic bacteria;
(2) Preparing a cholesterol-lowering medicine or health-care product;
(3) Preparing an animal feed or feed additive;
(4) Preparing food or food additive.
4. The use according to claim 3, wherein the pathogenic bacteria comprise Staphylococcus aureus (Staphylococcus aureus), escherichia coli (Escherichia coli), salmonella enteritidis (Salmonella enteritidis).
5. The fermentation medium for the high-density industrial production of pediococcus pentosaceus is characterized by comprising 30-40g/L of sugarcane molasses liquid, 20-25g/L of corn steep liquor dry powder, 1.5g/L of dipotassium phosphate, 5g/L of sodium acetate, 1mL/L of Tween, 2g/L of triammonium citrate, 0.2g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate tetrahydrate and water as a solvent.
6. The high-density industrial production fermentation medium according to claim 5, wherein the medium comprises 40g/L of molasses, 20g/L of corn steep liquor dry powder, 1.5g/L of dipotassium phosphate, 5g/L of sodium acetate, 80 mL/L of Tween, 2g/L of triammonium citrate, 0.2g/L of magnesium sulfate heptahydrate, 0.05g/L of manganese sulfate tetrahydrate and water as a solvent.
7. Use of the high-density industrial production fermentation medium according to claim 5 or 6 for the high-efficiency fermentation of pediococcus pentosaceus.
8. A method for efficiently fermenting Pediococcus pentosaceus, characterized in that Pediococcus pentosaceus (PG-2) according to claim 1 is inoculated into the high-density industrial production fermentation medium according to claim 5 for fermentation culture.
9. The method according to claim 8, wherein the preserved Pediococcus pentosaceus (PG-2) is activated on a MRS agar medium plate, a single colony is selected and inoculated in an MRS liquid medium for culture, and then the bacterial liquid is transferred to a high-density industrial production fermentation medium for fermentation culture.
10. The method according to claim 9, wherein the single colony is inoculated in MRS liquid medium and cultured under the following conditions: culturing at 37 deg.C for 14 hr; the transfer is carried out by transferring the bacterial liquid according to the inoculum size of 1% of the volume fraction; the fermentation culture conditions are as follows: fermenting at 37 deg.C for 48 hr.
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| CN117586909A (en) * | 2023-10-27 | 2024-02-23 | 青岛农业大学 | Pediococcus pentosaceus LWQ1 and application thereof |
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