CN116536186B - Lactobacillus brevis capable of fermenting soybean oligosaccharide and application thereof - Google Patents
Lactobacillus brevis capable of fermenting soybean oligosaccharide and application thereof Download PDFInfo
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- CN116536186B CN116536186B CN202310260013.1A CN202310260013A CN116536186B CN 116536186 B CN116536186 B CN 116536186B CN 202310260013 A CN202310260013 A CN 202310260013A CN 116536186 B CN116536186 B CN 116536186B
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- 239000000284 extract Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 235000001497 healthy food Nutrition 0.000 description 1
- 239000003228 hemolysin Substances 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 210000002429 large intestine Anatomy 0.000 description 1
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 1
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 1
- CDUFCUKTJFSWPL-UHFFFAOYSA-L manganese(II) sulfate tetrahydrate Chemical compound O.O.O.O.[Mn+2].[O-]S([O-])(=O)=O CDUFCUKTJFSWPL-UHFFFAOYSA-L 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 239000002777 nucleoside Substances 0.000 description 1
- 150000003833 nucleoside derivatives Chemical class 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 1
- 229920000053 polysorbate 80 Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002213 purine nucleotide Substances 0.000 description 1
- 150000004671 saturated fatty acids Chemical class 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 235000003687 soy isoflavones Nutrition 0.000 description 1
- 229940001941 soy protein Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229950000244 sulfanilic acid Drugs 0.000 description 1
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- 230000001502 supplementing effect Effects 0.000 description 1
- 238000003239 susceptibility assay Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
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Abstract
The strain is lactobacillus brevis grx-SOS04, is obtained by screening old Beijing acid bean juice of a traditional fermented bean product, can ferment sucrose, raffinose and stachyose in the soybean oligosaccharide, especially indigestible raffinose and stachyose, has good growth condition in MRS culture medium taking raffinose or stachyose as a sole carbon source, can be used for preparing fermented soybean milk, has 86.9 DEG T of acidity, has 72.8% of water holding capacity after being stored for 14 days, has 8.84lg (CFU/ml) of viable bacteria number, can also be compounded with other lactobacillus for preparing fermented milk beverage and the like, and can be used for preparing probiotic powder after drying a culture of the lactobacillus.
Description
Technical Field
The application belongs to the technical field of microorganisms, and particularly relates to lactic acid bacteria capable of fermenting soybean oligosaccharides and application thereof.
Background
With the increasing change of dietary nutrition concept and the increasing demand for healthy foods, plant-based fermented milk is receiving increasing consumer attention as a healthy substitute and a nutrition supplementing source for cow milk products. At present, no influencing bean plant-based fermented milk product is marketed in the domestic market.
The fermented milk based on bean plants contains rich functional substances such as soy protein, soy isoflavone and the like, has high nutritive value, does not contain animal-derived components such as cow milk allergen, lactose, cholesterol, saturated fatty acid and the like, and can meet the requirements of special crowds such as lactose intolerance, fat reduction groups and the like. However, sucrose, raffinose and stachyose in soybean milk are used as main carbon sources, and because of lacking of digestive enzyme alpha-D-galactosidase for hydrolyzing the raffinose and stachyose in human bodies, the raffinose and stachyose in the soybean plant-based fermented milk are not digested and absorbed in small intestines, enter large intestines and are fermented by intestinal microorganisms in colon to generate gas, so that adverse reactions such as dyspepsia, abdominal distention and borygmus of human bodies are easily caused, and further development and utilization of the soybean plant-based fermented milk are limited.
Disclosure of Invention
This section is intended to outline some aspects of embodiments of the application and to briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section as well as in the description of the application and in the title of the application, which may not be used to limit the scope of the application.
The present application has been made in view of the above and/or problems occurring in the prior art.
Therefore, the application aims to overcome the defects in the prior art and provide lactobacillus brevis capable of fermenting soybean oligosaccharides.
In order to solve the technical problems, the application provides the following technical scheme: a lactobacillus brevis capable of fermenting soybean oligosaccharides, characterized in that the strain is lactobacillus brevis grx-SOS04.
As a preferable embodiment of the Lactobacillus brevis of the present application, there is provided: the lactobacillus brevis is separated from the old Beijing acid bean juice of the traditional fermented bean product.
As a preferable embodiment of the Lactobacillus brevis of the present application, there is provided: the strain can ferment soybean oligosaccharides such as sucrose, raffinose, stachyose and the like, in particular non-digestible raffinose and stachyose.
As a preferable embodiment of the Lactobacillus brevis of the present application, there is provided: the lactobacillus brevis is obtained by adopting a turbidity measurement method and screening in an MRS culture medium with raffinose or stachyose as a unique carbon source.
As a preferable embodiment of the Lactobacillus brevis of the present application, there is provided: the Lactobacillus brevis has OD in MRS culture medium with raffinose as unique carbon source 600 Reaching 0.603, OD in MRS culture medium with stachyose as unique carbon source 600 Up to 0.643.
As a preferable embodiment of the Lactobacillus brevis of the present application, there is provided: the Lactobacillus brevis is treated with adenylate (500 g/L) DeltaOD in MRS Medium as sole carbon Source 600 Reaching 0.201, delta OD in MRS culture medium with inosinic acid (5.00 g/L) as unique carbon source 600 Reaching 0.262 delta OD in MRS culture medium with guanylic acid (5.00 g/L) as unique carbon source 600 Up to 1.1762; and can completely degrade guanylic acid, adenosine, inosine and guanosine.
As a preferable embodiment of the Lactobacillus brevis of the present application, there is provided: the strain is sensitive to 5 antibiotics of cefazolin, ampicillin-sulbactam, rifampin, chloramphenicol and penicillin, and resistant to 5 antibiotics of compound neonomine, vancomycin, streptomycin, clindamycin and tetracycline. The amino acid decarboxylase activity, nitroreductase activity and hemolytic activity of the strain are all determined to be negative, which indicates that the strain is a safe strain.
It is a further object of the present application to solve the deficiencies of the prior art and to provide an application of Lactobacillus brevis capable of fermenting soybean oligosaccharides.
In order to solve the technical problems, the application provides the following technical scheme: use of lactic acid bacteria capable of fermenting soy oligosaccharides, comprising: application of lactobacillus brevis in fermented soybean milk, lactobacillus beverage and related functional foods; the pH of the fermented soybean milk at 8h is 4.14, the acidity value is 86.9 ℃, the viable count is 9.01log (CFU/ml), the water holding capacity is 75.3%, the viscosity is 2523mPa.s, the hardness is 0.363N, and after 14d storage, the viable count is 8.84log (CFU/ml), so that the fermented soybean milk has good acid production property, growth property and texture property.
The application has the beneficial effects that:
(1) Providing a lactobacillus brevis capable of fermenting soybean oligosaccharide, wherein the lactobacillus strain is lactobacillus brevis grx-SOS04, and is preserved in China general microbiological culture collection center (CGMCC) No.25444, address: beijing, chaoyang area, north Chen Xili No. 1, 3, china academy of sciences, microbiological institute.
(2) The primary screening selects the strain with short curdling time and excellent acid production and texture characteristics of the fermented soybean milk, and the secondary screening selects the strain which can convert raffinose and stachyose highly.
(3) The strain of the application can be used for preparing fermented milk, in particular fermented soybean milk and bean plant-based fermented mixed milk; can also be used for preparing probiotic powder and applied to food.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings that are needed in the description of the embodiments will be briefly described below, it being obvious that the drawings in the following description are only some embodiments of the present application, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art. Wherein:
FIG. 1 is a colony morphology of the strain on a normal glucose MRS plate in example 1 of the present application.
FIG. 2 is a diagram showing the morphology of the strain of example 1 according to the present application observed by a conventional optical microscope.
FIG. 3 is a graph showing the amino acid decarboxylase assay of example 1 of the present application, wherein A is the tyrosine detection result and B is the histidine detection result.
FIG. 4 is a graph showing the detection of nitroreductase in example 1 of the present application.
FIG. 5 is a graph showing the measurement of hemolytic activity in example 1 of the present application.
Detailed Description
In order that the above-recited objects, features and advantages of the present application will become more apparent, a more particular description of the application will be rendered by reference to specific embodiments thereof which are illustrated in the appended drawings.
In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application, but the present application may be practiced in other ways other than those described herein, and persons skilled in the art will readily appreciate that the present application is not limited to the specific embodiments disclosed below.
Further, reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic can be included in at least one implementation of the application. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.
Example 1
Preparation method of lactobacillus brevis
The application adopts a coating plate separation method to separate lactobacillus from the old Beijing acid soybean juice sample of the traditional fermented soybean product, and screens out lactobacillus fermented soybean strains with high acid production and excellent quality by measuring indexes such as curdling time, pH, viable count, water holding capacity, viscosity, hardness and the like of the lactobacillus fermented soybean milk; by measuring the absorbance value (OD) of lactobacillus at 600nm in MRS medium containing raffinose and stachyose as the sole carbon source 600 ) Screening out strain of fermentable soybean oligosaccharide; the lactic acid bacteria strain of high conversion soybean oligosaccharide was determined by measuring the content of soybean oligosaccharide in the soybean milk before and after fermentation.
1. Isolation of lactic acid bacteria
The strain is lactobacillus brevis capable of fermenting soybean oligosaccharide, which is obtained by screening the traditional fermented soybean product, namely the old Beijing acid soybean juice, and is identified to belong to lactobacillus brevis (Lactobacillus), named grx-SOS04, and is preserved in China general microbiological culture Collection center with the preservation number of CGMCC No.25444.
2. Acid production characteristics, growth characteristics and texture characteristics of lactic acid bacteria in soybean milk culture medium
Soy milk medium: selecting normal soybean, cleaning, soaking in distilled water overnight, grinding with 80deg.C distilled water, removing residues, adjusting the ratio of bean to water to 1:10, making into soybean milk, sieving with 100 mesh sieve, boiling, packaging in fermentation bottle, sterilizing at 105deg.C for 15min, cooling to room temperature, and standing in a refrigerator at 4deg.C.
The screened strain is inoculated into a 100mL soybean milk culture medium fermentation bottle according to the addition amount of 3 percent after being activated, the soybean milk is fermented in a 37 ℃ incubator, the soybean milk curds are observed every 0.5h, and the curds time is recorded. Sampling during fermentation for 8h, and performing pH and titration acidity measurement on the fermentation sample (weighing about 5.0g of fermentation sample, recording mass m, diluting with 40ml of distilled water, adding two drops of phenolphthalein indicator, titrating to micro-powder red with 0.1mol/L NaOH, keeping color unchanged within 30s, recording volume V), and measuring viable count, water holding capacity, viscosity and hardness in parallel for three times, and taking an average value.
The results show that the strain has the advantages of 5.5h of milk coagulation time in a soybean milk culture medium, 4.14 pH at 8h, 86.9 DEG T of acidity value, 9.01log (CFU/ml) of viable count, 75.3% of water holding capacity, 2523mPa.s of viscosity, 0.363N of hardness, and good acid production property, growth property and texture property.
3. Growth of lactic acid bacteria in raffinose and stachyose media
MRS liquid medium: 20.00g of glucose, 5.00g of sodium acetate, 2.00g of dipotassium hydrogen phosphate, 10.00g of peptone, 1.00mL of tween-80, 10.00g of beef extract, 0.05g of manganese sulfate tetrahydrate, 5.00g of yeast extract, 2.00g of diammonium citrate, 0.20g of magnesium sulfate heptahydrate, adding distilled water to 1000mL of volume, and sterilizing at 121 ℃ for 15min.
MRS-Glc+Raf medium: glucose in MRS culture medium is changed into raffinose, and the rest is sterilized at 121deg.C for 15min.
MRS-Glc+Sta Medium: the glucose in MRS culture medium is changed into stachyose, and the rest is sterilized at 121deg.C for 15min for use.
MRS-Glc Medium: glucose in MRS culture medium is removed, no carbon source exists, and the rest is sterilized at 121deg.C for 15min for use.
Inoculating the strain into MRS-Glc+Raf and MRS-Glc+Sta liquid culture medium at 3%, culturing at 37deg.C, sampling at 0 hr and 20 hr, and measuring absorbance (OD) of fermentation broth at 600nm with enzyme-labeled instrument 600 ) The growth of lactic acid bacteria in MRS-Glc+Raf and MRS-Glc+Sta liquid medium was recorded, with a blank medium without sugar MRS-Glc medium as a negative control.
The results show that the strain has better growth condition and OD than other strains in MRS-Glc+Raf culture medium and MRS-Glc+Sta culture medium 600 raffinose Reaching 0.603, OD 600 stachyose Up to 0.643.
4. Evaluation of lactic acid bacterium fermentation soybean oligosaccharide ability
After lactobacillus is activated in MRS liquid culture medium for 2 generations, respectively inoculating MRS-Glc+Raf culture medium and MRS-Glc+Sta culture medium which take raffinose and stachyose as unique carbon sources, fermenting in a 37 ℃ incubator, sampling for 0h, 12h and 24h, and measuring the raffinose and stachyose content. Taking 1ml of lactobacillus fermentation liquor, centrifuging at 8000r/min for 2min, filtering the supernatant with a 0.22 μm water phase filter membrane, and then detecting by HPLC, and analyzing the content of raffinose and stachyose in the supernatant.
Meanwhile, lactobacillus is inoculated into a soybean milk culture medium, and the soybean milk is fermented in an incubator at 37 ℃ for 8 hours. Sample treatment: taking 2mL of fermented soybean milk sample, adding 1.5mL of sterile deionized water for dissolution, and carrying out water bath at 60 ℃ for 10min; 0.25mL of Carrez I solution (0.5 mol/L potassium ferrocyanide aqueous solution aqueous potassium ferrocyanide), 0.25mL of Carrez II solution (0.5 mol/L zinc acetate aqueous solution aqueous zinc acetate) and 1mL of acetonitrile were added, mixed well, left to stand at room temperature for 1h, centrifuged at 10000r/min for 8min, and the supernatant was filtered through a 0.22 μm aqueous filter membrane for HPLC detection.
The results show that: the strain is fermented for 24 hours in an MRS-Glc+Raf culture medium and an MRS-Glc+Sta culture medium which take raffinose and stachyose as unique carbon sources, and the content of the raffinose and stachyose is respectively reduced to 3.1g/L and 4.6g/L from 20 g/L; in the soybean milk fermented by the strain of the application, the stachyose content is reduced from 15.6g/L to 3.2g/L, and the raffinose content is also reduced to undetectable levels after 8 hours.
5. Evaluation of ability of lactic acid bacteria to degrade purine nucleotides
After lactobacillus is activated in MRS liquid culture medium for 2 generations, respectively inoculating MRS-Glc+AMP culture medium, MRS-Glc+IMP culture medium and MRS-Glc+GMP culture medium which take adenylate, inosinic acid or guanylic acid as unique carbon sources, fermenting in a 37 ℃ incubator, sampling after 20 hours, and measuring the content of the adenylate, the inosinic acid and the guanylic acid. Taking 1ml of lactobacillus fermentation liquor, centrifuging at 8000r/min for 2min, filtering the supernatant with a 0.22 μm water phase filter membrane, and then detecting by HPLC, and analyzing the content of adenylate, inosinic acid and guanylic acid in the supernatant.
Meanwhile, extracting lactobacillus thallus, inoculating the lactobacillus thallus into a phosphate buffer system mixed by 0.2g/L adenosine, inosine and guanosine, incubating for 4 hours at 37 ℃, and sampling and measuring the content of each nucleoside in the system. Taking 1ml of lactobacillus fermentation liquor, centrifuging at 8000r/min for 2min, taking supernatant, filtering with a 0.22 μm water phase filter membrane, and then detecting by HPLC, and analyzing the content of adenosine, inosine and guanosine in the supernatant.
The results show that: in MRS-Glc+AMP medium, MRS-Glc+IMP medium, MRS-Glc+GMP medium, and MRS-Glc+GMP medium with adenylate, inosinic acid, and guanylate as the only carbon sources, after fermentation for 20h, the adenylate and inosinic acid contents were reduced from 5.00g/L to 4.60g/L and 3.54g/L, respectively, and guanylate was reduced to undetectable levels. After 4 hours of culture in a mixed phosphate buffer system of adenosine, inosine and guanosine (0.20 g/L) at 37 ℃, the adenosine, the inosine and the guanosine can be completely degraded.
6. Strain safety detection
a) Antibiotic resistance sensitivity assay
The sensitivity of the lactic acid bacteria to antibiotics was examined by using a drug sensitive paper agar diffusion method. The 10 antibiotics include ampicillin, vancomycin, cefazolin, compound neonomine, streptomycin, rifampin, clindamycin, tetracycline, chloramphenicol and penicillin. After the strain of the application is activated for two generations, 1ml of bacterial liquid is taken and is centrifugated in a 1.5ml centrifuge tube at 8000r/min for 5min, and the centrifugated bacterial liquid is diluted to OD by sterile physiological saline 600 200 μl of the dilutions were spread in MRS solid medium, and the drug sensitive paper sheets were removed under aseptic conditions and applied to plates of 3 drug sensitive paper sheets each. Placing in a room for 30min, then placing in a incubator at 37 ℃ for culturing for 24h, and measuring and recording the diameter of the inhibition zone. Results referring to table 1, lactic acid bacteria were evaluated for resistance to various antibiotics according to antimicrobial susceptibility test performance criteria established by CLSI in the united states.
TABLE 1 criterion for determining resistance of lactic acid bacteria to antibiotics
TABLE 2 results of resistance of inventive strains to antibiotics
The results show that: the strain is sensitive to 5 antibiotics of cefazolin, ampicillin-sulbactam, rifampin, chloramphenicol and penicillin, and is resistant to 5 antibiotics of compound neonomine, vancomycin, streptomycin, clindamycin and tetracycline.
b) Determination of toxic substance production
(1) And (3) generating biogenic amine, diluting activated lactobacillus of 2 generations with normal saline, coating the diluted lactobacillus in an amino acid decarboxylase detection medium added with tyrosine and histidine, culturing for 3 days at 37 ℃, observing the color change condition of the medium, wherein the color change is positive, the color change is negative, and meanwhile, using staphylococcus aureus as a control experiment.
(2) And (3) generating nitrite, inoculating activated 2-generation lactobacillus into a nitrate culture medium according to the addition amount of 3%, culturing for 4d at 37 ℃, sequentially dripping 3-5 drops of alpha-naphthylamine solution and sulfanilic acid solution into the culture solution, simultaneously using staphylococcus aureus as a control experiment, and observing an experimental result.
(3) And (3) generating hemolysin, namely marking the lactobacillus to be detected and control positive bacteria staphylococcus aureus after activating for 2 generations in a Columbia blood agar plate, culturing in a 37 ℃ incubator for 48 hours, and observing whether a hemolytic ring appears around a colony.
The results are shown in figures 3, 4 and 5, and the amino acid decarboxylase assay, nitroreductase assay and hemolytic activity assay of the strain are all negative, which indicates that the strain is a safe strain.
6. Identification of lactic acid bacteria
6.1 morphological observations and gram staining
The colony characteristics on the MRS solid plate were observed visually, and single colonies were picked on a glass slide, and after gram staining, the strain morphology was observed by a biological microscope.
As shown in figures 1 and 2, the bacterial colony of the strain is milky white, nearly round, convex and smooth, and has regular edges. After gram staining, the gram staining is positive, long rod-shaped and free of spores.
6.2 identification of Strain 16S rDNA
(1) Extracting genome: after the strain is activated, 1ml of bacterial liquid is taken for centrifugation, and a bacterial genome DNA extraction kit is selected for extracting DNA;
(2) And (3) PCR amplification: amplifying by using the genome DNA of the strain as a template, wherein the primers are (27F: AGAGTTTGATCCTGGGCTCAG;
14992R GGTTACCTTGTTACGACTT), PCR amplification procedure: 95 ℃ for 5min;95 ℃ for 30s;55 ℃ for 15s; the temperature is 72 ℃ and 2min, and the extension is continued for 5min at 72 ℃ after 30 cycles.
The PCR products were electrophoretically detected and sent to Shanghai Biotechnology Co.Ltd for sequencing. Sequencing results: the nucleotide length is about 1500bp, and the sequence obtained by sequencing is subjected to homologous comparison analysis with a GenBank database of NCBI, so that the result shows that the strain is lactobacillus brevis, and the 16S rDNA sequence consistency is 98%. The 16S rDNA sequence is shown below,
AGGGCCTGGGGGGTTGCCTATAGATGCAAGTCGAACGAGCTTCCGTTGAATGACGTGCTTGCACTGATTTCAACAATGAAGCGAGTGGCGAACTGGTGAGTAACACGTGGGGAATCTGCCCAGAAGCAGGGGATAACACTTGGAAACAGGTGCTAATACCGTATAACAACAAAATCCGCATGGATTTTGTTTGAAAGGTGGCTTCGGCTATCACTTCTGGATGATCCCGCGGCGTATTAGTTAGTTGGTGAGGTAAAGGCCCACCAAGACGATGATACGTAGCCGACCTGAGAGGGTAATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGAAAGTCTGATGGAGCAATGCCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAACTCTGTTGTTAAAGAAGAACACCTTTGAGAGTAACTGTTCAAGGGTTGACGGTATTTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCTTCGGCTTAACCGGAGAAGTGCATCGGAAACTGGGAGACTT GAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTAGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGAGGGTTTCCGCCCTTCAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGACCCTACCAAGTCTTGACATCTTCTGCCATTCTAGAGATAGACGTTCCCCTTCGGGGACCGAATGACAGGTGGTGCATGTTGTCGTCAGCTTCGTGTCTGAGATGTGGTTAAGTCCCGCACTAGCCCACCCTTATTATCCAGCTGCTCAGCATATCGATGGCGCACTCTAGTGA
TABLE 3 homology alignment of lactic acid bacteria 16S rDNA Gene sequences
Example 2
2. Inventive strain application examples
1. The strain of the application can be used for preparing fermented soybean milk.
Selecting normal soybean, cleaning, soaking in distilled water overnight, grinding with 80deg.C distilled water, removing residues, adjusting the ratio of soybean water to 1:10, making into soybean milk, sieving with 100 mesh sieve, boiling, optionally adding sucrose,
packaging in fermentation bottle, sterilizing at 105deg.C for 15min. The strain of the application is taken, activated and inoculated to a soybean milk culture medium for fermentation, the inoculum size is 3 percent, and the strain can be refrigerated for eating after fermentation for 8 hours at 37 ℃.
2. The strain of the application can be used for preparing Cheng Yi raw fungus powder and is applied to food.
Freezing the lactobacillus to-40deg.C to-60deg.C, lyophilizing to obtain powder, and adding the powder into various beverages or foods as supplement.
The application provides a lactobacillus capable of fermenting soybean oligosaccharide, which is further developed and utilized. The lactobacillus strain is lactobacillus brevis grx-SOS04, and is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.25444 and the address: beijing, chaoyang area, north Chen Xili No. 1, 3, china academy of sciences, microbiological institute.
The strain is separated from traditional fermented bean product acid pulp water, and experiments prove that the strain grows OD in MRS culture medium with raffinose as unique carbon source 600 Reaching 0.603, and growing OD in MRS culture medium with stachyose as unique carbon source 600 The pH reached 0.643 at 8h of fermented soy milk was 4.14, an acidity value of 86.9℃T, a viable count of 9.01log CFU/ml, a water holding capacity of 75.3%, a viscosity of 2523mPa.s, a hardness of 0.363N, and a viable count of 8.84log CFU/ml after 14d storage.
The strain of the application can be used for preparing fermented milk, in particular fermented soybean milk and bean plant-based fermented mixed milk; can also be used for preparing probiotic powder and applied to food.
It should be noted that the above embodiments are only for illustrating the technical solution of the present application and not for limiting the same, and although the present application has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solution of the present application may be modified or substituted without departing from the spirit and scope of the technical solution of the present application, which is intended to be covered in the scope of the claims of the present application.
Claims (3)
1. A lactobacillus capable of fermenting soybean oligosaccharide is characterized in that the strain is Lactobacillus brevisLactobacillus brevis) The grx-SOS04 has a preservation number of CGMCC No.25444 and is preserved in the China general microbiological culture Collection center (China Committee for culture Collection of microorganisms).
2. Use of lactic acid bacteria capable of fermenting soy oligosaccharides as claimed in claim 1, characterized in that: the lactobacillus is applied to fermenting soybean milk and producing lactobacillus beverage and functional food.
3. The use according to claim 2, wherein: the pH of the fermented soybean milk at 8 hours is 4.14, the acidity value is 86.9 ℃, the viable count is 9.01log CFU/ml, the water holding capacity is 75.3%, the viscosity is 2523mPa.s, the hardness is 0.363N, and the viable count is 8.84log CFU/ml after 14d storage.
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CN111944711A (en) * | 2020-07-07 | 2020-11-17 | 浙大宁波理工学院 | Lactobacillus brevis capable of degrading purine nucleosides and application thereof |
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CN111154697A (en) * | 2020-02-12 | 2020-05-15 | 江南大学 | Leuconostoc mesenteroides intestinal membrane subspecies for producing galactosidase and application thereof |
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