CN114891663B - Lactobacillus plantarum LP1406 and isolated culture method - Google Patents
Lactobacillus plantarum LP1406 and isolated culture method Download PDFInfo
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- CN114891663B CN114891663B CN202210388926.7A CN202210388926A CN114891663B CN 114891663 B CN114891663 B CN 114891663B CN 202210388926 A CN202210388926 A CN 202210388926A CN 114891663 B CN114891663 B CN 114891663B
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- A—HUMAN NECESSITIES
- 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
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/60—Edible seaweed
-
- A—HUMAN NECESSITIES
- 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
- A23L17/00—Food-from-the-sea products; Fish products; Fish meal; Fish-egg substitutes; Preparation or treatment thereof
- A23L17/65—Addition of, or treatment with, microorganisms or enzymes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N1/02—Separating microorganisms from their culture media
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/90—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in food processing or handling, e.g. food conservation
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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Abstract
The invention belongs to the field of microorganism culture, and particularly relates to a method for culturing lactobacillus plantarum capable of fermenting and degrading kelp. The lactobacillus plantarum is preserved in China center for type culture Collection, with the preservation number of 2021, 11 and 29: cctccc No. M20211500. The invention provides a method for culturing plant milk stalks, which comprises the following steps: s1: separating and culturing lactobacillus plantarum; s2: screening lactobacillus plantarum fermented kelp; s3: fermenting kelp; s4, evaluating fermentation effect. The lactobacillus plantarum provided by the invention can utilize natural kelp to grow rapidly, has excellent performance of the fermented kelp and increased yield of metabolites, and provides a novel microorganism treatment method for improving the bioactivity of the fermented kelp; the strain disclosed by the invention has an obvious effect of fermenting kelp, can improve the antioxidant and hypoglycemic activities in a fermented sample, has the inhibition capacity of ABTS of 82%, and has the inhibition capacity of alpha-glucosidase of 75.40%.
Description
Technical Field
The invention belongs to the field of microorganism culture, and particularly relates to lactobacillus plantarum and a method for culturing the lactobacillus plantarum.
Background
Lactobacillus plantarum is a genus of Lactobacillus in the family of Lactobacillus, and can survive in various environments, and is closely related to human life. Lactobacillus plantarum has strong acid resistance, and most of the Lactobacillus plantarum can tolerate NaCl with concentration higher than 5%. Has the characteristics of rapid propagation, short growth period, rich nutrition of thalli and the like. The biological functions of the strain mainly comprise immunoregulation, antagonism of pathogenic bacteria, cholesterol reduction, blood sugar reduction, cardiovascular and cerebrovascular prevention and the like, and the strain is a probiotic with wide application. In addition, the lactobacillus plantarum has rich enzyme system and vigorous metabolism, and has the potential of fermenting by utilizing kelp components. The mechanism of reducing blood sugar of lactobacillus mainly reduces blood sugar level by regulating intestinal tract balance of human body and immunity function of organism, improving antioxidant capacity, regulating sugar metabolism and lipid metabolism.
Regarding the use of lactobacillus plantarum, the following patent documents disclose:
CN114231473A is a probiotic lactobacillus plantarum and application thereof in preparing low-salt fermented meat food; CN114196600a lactobacillus plantarum with effect of alleviating irritable bowel syndrome and application thereof; CN114134089a is a high-activity composite microbial inoculum and application thereof in food production. There is little disclosure about the application of lactobacillus plantarum in kelp degradation.
The kelp contains rich chemical components and bioactive substances, also contains rich fucoidin in the kelp, has high value in the processing application of functional foods and medicines, and also contains various vitamins, minerals and the like. Researches prove that the kelp has various biological functions of reducing blood fat, reducing blood sugar, regulating immunity, resisting blood coagulation, resisting tumor, expelling lead, removing toxin, resisting oxidation and the like. However, the kelp industry in China is still in a primary state with low added value and low benefit mainly comprising traditional rough processing and primary processed products, and the development space of the kelp industry is still far from being explored.
Therefore, the kelp processing and fermentation industry is innovatively developed, kelp products with nutrition and health care values are developed, the requirements of health China strategy are met, and the continuous development of kelp industry chains is promoted. If a proper lactobacillus plantarum is searched and cultured, the lactobacillus plantarum can ferment and degrade kelp and improve the functions of the fermentation on reducing blood sugar and resisting oxidization of the kelp, so that a foundation for large-scale production and application is necessarily provided for the deep processing of the kelp.
Disclosure of Invention
In order to solve the technical problems, the invention provides the lactobacillus plantarum LP1406 which can utilize natural kelp to grow rapidly and has the alginic acid degrading capability, and the lactobacillus plantarum LP1406 can improve the glucose reducing and antioxidant capability of kelp fermentation samples, thereby providing a good foundation for the fine and deep processing of kelp.
The invention also provides a specific method for applying the strain in the fermentation of kelp, and the hypoglycemic and antioxidant capacities of the products obtained after application are evaluated.
Lactobacillus plantarum LP1406 described in the present invention was deposited at the chinese collection for typical cultures at 2021, 11 and 29, accession number: chinese armed, preservation number cctccc NO: m20211500, class designation: lactobacillus plantarum LP1406Lactiplantibacillus plantarumLP1406。
The nucleotide sequence of the lactobacillus plantarum LP1406 (hereinafter referred to as lactobacillus plantarum) is shown as SEQ ID NO.1, and after being cultured for 24 hours at 37 ℃ in an MRS culture medium, the lactobacillus plantarum LP1406 has the surface colony diameter of about 3mm, is convex, is round, has smooth surface, is fine and compact, has white color, and occasionally has light yellow or dark yellow color; the colony has uniform texture and uniform color on the front and back surfaces, edges and the central part; the somatic cells are rod-shaped, single or chain-formed; gram staining positive, H 2 O 2 Positive experiments, positive methyl red experiments, positive gelatin liquefaction experiments, negative V.P. experiments, negative hydrogen sulfide experiments, negative oxidase experiments, and positive glucose, maltose, galactose and mannose experiments; has obvious antibacterial effect on staphylococcus aureus, escherichia coli and salmonella typhimurium; the pH tolerance range is 2.0-6.0.
In the isolation and culture of lactobacillus plantarum, a solid MRS medium is used, containing: 10g/L of peptone, 10g/L of beef extract, 5g/L of yeast powder, 20g/L of glucose, 2g/L of ammonium citrate, 0.58g/L of magnesium sulfate heptahydrate, 3.12g/L of sodium acetate, 1.63g/L of disodium hydrogen phosphate, 2.75g/L of potassium acetate, 1mL of Tween 80 and 0.25g/L of manganese sulfate tetrahydrate.
The application of lactobacillus plantarum LP1406 in fermented seaweed products, in particular in fermented kelp, is the subject of the important protection of the present invention.
The application of lactobacillus plantarum LP1406 in fermenting kelp comprises the following specific steps: firstly, screening lactobacillus plantarum for fermenting kelp, and then, fermenting the kelp;
when lactobacillus plantarum fermented kelp is screened, single colony on a solid flat plate is inoculated into a seed culture medium for culture, and then inoculated into a kelp enzymolysis liquid culture medium for culture; the seed culture medium is a liquid MRS culture medium and contains: 10g/L of peptone, 10g/L of beef extract, 5g/L of yeast powder, 20g/L of glucose, 2g/L of ammonium citrate, 0.58g/L of magnesium sulfate heptahydrate, 3.12g/L of sodium acetate, 1.63g/L of disodium hydrogen phosphate, 2.75g/L of potassium acetate, 1mL of Tween 80 and 0.25g/L of manganese sulfate tetrahydrate;
the preparation method of the kelp enzymolysis liquid culture medium comprises the following steps: washing kelp with water, drying, grinding, sieving the powder with a 40-mesh sieve, collecting, dissolving in distilled water at a ratio of 1:50 (w/v), adjusting the pH to 4.8, adding cellulase and pectase for enzymolysis, wherein the addition amounts of the cellulase and the pectase are respectively 2.5wt% and 0.26 and wt%, stirring the reaction solution at 50 ℃ for 2 hours, adjusting the pH to 8.0, adding 0.3wt% alkaline pectase, incubating at 60 ℃ for 1.5 hours, centrifuging the reaction product, and collecting the enzymolysis supernatant as kelp enzymolysis solution;
when the kelp is fermented, single colony on an MRS flat plate is inoculated into a seed culture medium, and the kelp is cultured for 24 hours at 37 ℃; then inoculating the strain into kelp enzymolysis liquid culture medium according to the inoculum size of 5% -10%, and culturing at 37 ℃ for 48 hours.
When the kelp is fermented, single colony on an MRS flat plate is inoculated into a seed culture medium, and the kelp is cultured for 24 hours at 37 ℃; and then inoculating the kelp into a kelp fermentation culture medium according to the inoculum size of 5% -10%, supplementing glucose in the kelp enzymolysis liquid to 3%, sterilizing at 121 ℃ for 20min, inoculating lactobacillus plantarum with 3% content, and standing at 37 ℃ for 48h.
A method for fermenting kelp using lactobacillus plantarum LP1406 comprising the steps of:
s1: isolation and culture of lactobacillus plantarum
The method comprises the following specific steps of:
(1) Taking 1mL of milk product, putting into a test tube filled with 9mL of physiological saline, and fully and vortex oscillating by a vortex oscillator to ensure that samples are fully and uniformly mixed;
(2) 1mL of the mixed sample is fully sucked by a pipette, added into a test tube containing 9mL of physiological saline, and the like to prepare 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 Is a suspension of (a);
(3) From a concentration of 10 with a pipette -2 ，10 -4 ，10 -6 Fully sucking 0.1mL of mixed solution in a test tube, respectively diluting and coating the mixed solution in a sterilized solid MRS culture medium, and culturing for 24 hours at 37 ℃;
(4) After the culture is finished, observing the colony morphology, and picking a milky white smooth moist single colony for microscopic examination;
(5) Performing separation and purification culture on the rod-shaped bacterial colony by microscopic examination, after the bacterial colony grows well, checking whether the bacterial colony has mixed bacteria, and if so, performing separation and purification again until a pure bacterial strain culture is obtained;
s2: screening of Lactobacillus plantarum fermented kelp
Screening lactobacillus plantarum of fermented kelp, which comprises the following specific steps: inoculating single colony on the solid plate into a seed culture medium, culturing at 37 ℃ for 24 hours, then inoculating the single colony into a screening culture medium according to the inoculum size of 5-10%, and culturing at 37 ℃ for 40-60 hours;
s3: fermented kelp
The fermented kelp comprises the following specific operation steps: inoculating single colony on MRS plate into seed culture medium, and culturing at 37deg.C for 24 hr; and then inoculating the kelp into a kelp fermentation culture medium according to the inoculum size of 5% -10%, supplementing glucose in the kelp enzymolysis liquid to 3%, sterilizing at 121 ℃ for 20min, inoculating lactobacillus plantarum with 3% content, and standing for culturing at 37 ℃ for 48h.
The beneficial effects of the invention are as follows:
(1) The lactobacillus plantarum LP1406 provided by the invention can be used for rapidly fermenting natural kelp, and has strong alginic acid degradation capability in the fermentation process;
(2) The obtained fermentation product has excellent hypoglycemic and antioxidant capacities, and the results of in-vitro hypoglycemic evaluation and in-vitro antioxidant experiments show that the inhibition capacity of the fermentation product to alpha-glucosidase reaches 75.40 percent and the inhibition rate to ABTS is 82 percent, which is an effect which cannot be achieved by common lactobacillus plantarum;
(3) Compared with the traditional method, the lactobacillus plantarum provided by the invention has the advantages of high efficiency, energy conservation, time saving, simple operation and high added value production.
Drawings
FIG. 1 is a photograph showing colony morphology of Lactobacillus plantarum;
FIG. 2 is a photograph of Lactobacillus plantarum by microscopic examination.
Detailed Description
The present invention will now be further described in connection with specific embodiments in order to enable those skilled in the art to better understand the invention.
The lactobacillus plantarum provided by the invention is preserved in China Center for Type Culture Collection (CCTCC) in 2021, 11 and 29 days, and the preservation number is CCTCC NO: m20211500.
Example 1
A method for culturing lactobacillus plantarum, comprising the steps of:
s1: isolation and culture of lactobacillus plantarum
The method comprises the following specific steps of:
(1) Taking 1mL of milk product, putting into a test tube filled with 9mL of physiological saline, and fully and vortex oscillating by a vortex oscillator to ensure that samples are fully and uniformly mixed;
(2) 1mL of the mixed sample is fully sucked by a pipette, added into a test tube containing 9mL of physiological saline, and the like to prepare 10 -2 、10 -3 、10 -4 、10 -5 、10 -6 Is a suspension of (a);
(3) From a concentration of 10 with a pipette -2 ，10 -4 ，10 -6 Fully sucking 0.1mL of mixed solution in a test tube, respectively diluting and coating the mixed solution in a sterilized solid MRS culture medium, and culturing for 24 hours at 37 ℃;
in a solid MRS culture medium, 10g/L of peptone, 10g/L of beef extract, 5g/L of yeast powder, 20g/L of glucose, 2g/L of ammonium citrate, 0.58g/L of magnesium sulfate heptahydrate, 3.12g/L of sodium acetate, 1.63g/L of disodium hydrogen phosphate, 2.75g/L of potassium acetate, 1mL of tween 80 and 0.25g/L of manganese sulfate tetrahydrate;
after the inoculating loop is burnt and cooled by the alcohol lamp, a loop of bacterial colony is picked, the flat plate is placed beside the alcohol lamp, and 3-4 continuous straight lines are drawn by the inoculating loop according to Z. Then the ring of burned strain is cooled, and the rotating plate is scribed in the same way from the tail end of the first scribing area to the other area, and the scribing is carried out for 3-4 areas. After scribing, culturing in an inverted manner in a constant temperature incubator at 37 ℃ for 24 hours;
(4) After the culture is finished, observing the colony morphology, and picking a milky white smooth moist single colony for microscopic examination;
(5) And (3) performing separation and purification culture on the rod-shaped bacterial colony by microscopic examination, checking whether the bacterial colony grows well, and if so, performing separation and purification again until a pure bacterial strain culture is obtained.
(6) The colonies meeting the conditions were subjected to 16S rDNA sequencing analysis, and sequencing results were aligned with BLAST to obtain Lactobacillus plantarum.
Example 2
The preservation method of the lactobacillus plantarum comprises the following steps:
lactobacillus plantarum is inoculated in a liquid culture medium to be cultivated to a logarithmic phase, and 80% of glycerol is sterilized by high-pressure steam for later use. Taking 1mL of sterilized glycerol, putting the sterilized glycerol into the sterilized glycerol and fully and uniformly mixing the sterilized glycerol and the bacterial liquid, so that the concentration of the glycerol is about 10% -30%, and freezing the sterilized glycerol at the temperature of minus 80 ℃.
The liquid culture medium of the preservation method comprises the following components: 10g/L of peptone, 10g/L of beef extract, 5g/L of yeast powder, 20g/L of glucose, 2g/L of ammonium citrate, 0.58g/L of magnesium sulfate heptahydrate, 3.12g/L of sodium acetate, 1.63g/L of disodium hydrogen phosphate, 2.75g/L of potassium acetate, 1mL of Tween 80 and 0.25g/L of manganese sulfate tetrahydrate.
Example 3
The purification method of the lactobacillus plantarum comprises the following steps:
and (3) picking single colonies obtained by separation on an ultra-clean bench through an inoculating loop burnt by alcohol, inoculating the single colonies to a lactobacillus plantarum solid culture medium plate, and culturing at the constant temperature of 37 ℃ for 24 hours to stop culturing. Checking whether there is other bacteria by observing colony morphology and microscopic examination, if there is other bacteria, continuing to pick single colony for purification until obtaining pure culture of lactobacillus plantarum.
The lactobacillus plantarum obtained by the method has the following characteristics:
gram staining is positive, H2O2 experiment is positive, methyl red experiment is positive, gelatin liquefaction experiment is positive, V.P. experiment is negative, hydrogen sulfide experiment is negative, oxidase experiment is negative, and glucose, maltose, galactose and mannose experiments are positive. Has obvious antibacterial effect on staphylococcus aureus, escherichia coli and salmonella typhimurium; the pH tolerance range is 2.0-6.0.
Example 4.1
The lactobacillus plantarum is added into the kelp enzymolysis liquid to verify the effects of the lactobacillus plantarum on oxidization resistance and blood sugar reduction after kelp fermentation.
The specific steps are as follows:
s2: screening of fermented kelp Lactobacillus plantarum
Screening lactobacillus plantarum of fermented kelp, which comprises the following specific steps: inoculating single colony on solid plate into seed culture medium, culturing at 37deg.C for 24 hr, inoculating into screening culture medium (kelp enzymolysis solution) according to 5-10% inoculum size, and culturing at 37deg.C for 40-60 hr;
the seed culture medium is a liquid MRS culture medium, which comprises: 10g/L of peptone, 10g/L of beef extract, 5g/L of yeast powder, 20g/L of glucose, 2g/L of ammonium citrate, 0.58g/L of magnesium sulfate heptahydrate, 3.12g/L of sodium acetate, 1.63g/L of disodium hydrogen phosphate, 2.75g/L of potassium acetate, 1mL of Tween 80 and 0.25g/L of manganese sulfate tetrahydrate;
s3: fermented kelp
The fermented kelp comprises the following specific operation steps: inoculating single colony on MRS plate into seed culture medium, and culturing at 37deg.C for 24 hr; then inoculating the kelp into a kelp fermentation culture medium according to the inoculum size of 5% -10%, supplementing glucose in kelp enzymolysis liquid to 3%, sterilizing at 121 ℃ for 20min, inoculating lactobacillus plantarum with 3% content, placing at 37 ℃, and standing for culturing for 48h;
in the above steps S2 and S3, the specific preparation process of the kelp enzymatic hydrolysate is as follows: washing kelp with water, drying, grinding, sieving the powder with a 40-mesh sieve, collecting, dissolving in distilled water at a ratio of 1:50 (w/v), adjusting pH to 4.8, adding cellulase and pectase for enzymolysis, wherein the addition amounts of the cellulase and the pectase are respectively 2.5wt% and 0.26 and wt%, stirring the reaction solution at 50 ℃ for 2 hours, adjusting pH to 8.0, adding 0.3wt% alkaline pectase, incubating at 60 ℃ for 1.5 hours, centrifuging the reaction product, and collecting the enzymolysis supernatant as kelp enzymolysis solution.
The fermentation product obtained by the above process was evaluated for fermentation effect.
The fermentation effect evaluation is mainly carried out from two aspects of in-vitro blood sugar reduction evaluation and in-vitro antioxidation:
regarding the in vitro hypoglycemic evaluation and in vitro antioxidant evaluation, the specific steps are as follows:
the specific operation method for in-vitro blood sugar reduction evaluation comprises the following steps: respectively dissolving a fermented kelp sample in PBS (phosphate buffer solution) with pH of 6.8, diluting the sample into different concentrations in test tubes, respectively adding 0.2U/mL of alpha-glucosidase solution, preheating the test tubes in a water bath at 37 ℃ for 40min, adding PNPG substrate into the test tubes after the preheating is finished to react, wherein the reaction temperature is 37 ℃ and the reaction time is 20min, and finally adding 1mol/L sodium carbonate solution with the volume of 3 times of liquid into the test tubes to terminate the reaction. Determining the inhibition rate of alpha-glucosidase;
the specific operation method of the in vitro antioxidation experiment comprises the following steps: by mixing ABTS stock solution (7 mM) and potassium persulfate solution (4.9 mM) at 1:1, preparing a working solution by mixing in proportion; incubating the mixture in the dark at room temperature overnight; then, the mixture was diluted with distilled water, and the absorbance at 734nm was 0.7.+ -. 0.02; the samples were mixed with ABTS working solution, left in the dark at room temperature for 8 minutes, then absorbance was recorded with a spectrophotometer at 734nm, and blank and ascorbic acid positive controls were placed.
The specific steps are as follows:
the removal efficiency of the fermentation sample on the ABTS can reach 87.63 percent respectively, and the inhibition capacity on the alpha-glucosidase can reach 80.40 percent respectively.
The results show that the lactobacillus plantarum is used for fermenting the kelp, and the antioxidant and blood sugar reducing capabilities of the fermented sample are obviously improved.
Example 4.2
Glucose is added into the kelp enzymolysis liquid to 3%, after sterilization for 20min at 121 ℃, lactobacillus plantarum with 3% content is inoculated, and the kelp enzymolysis liquid is placed into a shaking table for culture at 37 ℃ and 180 r/min, and the fermentation time is 36h. The removal efficiency of the fermentation sample on the ABTS can reach 80.63 percent respectively, and the inhibition capacity on the alpha-glucosidase can reach 76.40 percent respectively.
Example 4.3
Glucose is added into the kelp enzymolysis liquid to 3%, after sterilization for 20min at 121 ℃, lactobacillus plantarum with 3% content is inoculated, and the kelp enzymolysis liquid is placed at 37 ℃ for static culture, and the fermentation time is 72h. The cleaning efficiency of the fermentation sample on the ABTS can reach 77.21 percent respectively, and the inhibition capacity on the alpha-glucosidase can reach 73.25 percent respectively.
Comparative example
From the data in the above table, it can be seen that the product of the present invention is far superior to other species in both the efficiency of ABTS elimination and the ability to inhibit α -glucosidase.
Sequence listing
<110> Shandong Xiao's Biotechnology Co., ltd
Qilu University of Technology
Shandong zhuoran Biotechnology Co.,Ltd.
Sheng Rong biotechnology (Shandong) Limited
SHANDONG CHENZHANG BIOTECHNOLOGY Co.,Ltd.
<120> Lactobacillus plantarum LP1406 and isolated culture method
<141> 2022-04-13
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1455
<212> DNA
<213> Lactobacillus plantarum LP1406 (Lactiplantibacillus plantarum LP 1406)
<400> 1
cctaatacat gcagtcgaac gaactctggt attgattggt gcttgcatca tgatttacat 60
ttgagtgagt ggcgaactgg tgagtaacac gtgggaaacc tgcccagaag cgggggataa 120
cacctggaaa cagatgctaa taccgcataa caacttggac cgcatggtcc gagcttgaaa 180
gatggcttcg gctatcactt ttggatggtc ccgcggcgta ttagctagat ggtggggtaa 240
cggctcacca tggcaatgat acgtagccga cctgagaggg taatcggcca cattgggact 300
gagacacggc ccaaactcct acgggaggca gcagtaggga atcttccaca atggacgaaa 360
gtctgatgga gcaacgccgc gtgagtgaag aagggtttcg gctcgtaaaa ctctgttgtt 420
aaagaagaac atatctgaga gtaactgttc aggtattgac ggtatttaac cagaaagcca 480
cggctaacta cgtgccagca gccgcggtaa tacgtaggtg gcaagcgttg tccggattta 540
ttgggcgtaa agcgagcgca ggcggttttt taagtctgat gtgaaagcct tcggctcaac 600
cgaagaagtg catcggaaac tgggaaactt gagtgcagaa gaggacagtg gaactccatg 660
tgtagcggtg aaatgcgtag atatatggaa gaacaccagt ggcgaaggcg gctgtctggt 720
ctgtaactga cgctgaggct cgaaagtatg ggtagcaaac aggattagat accctggtag 780
tccataccgt aaacgatgaa tgctaagtgt tggagggttt ccgcccttca gtgctgcagc 840
taacgcatta agcattccgc ctggggagta cggccgcaag gctgaaactc aaaggaattg 900
acgggggccc gcacaagcgg tggagcatgt ggtttaattc gaagctacgc gaagaacctt 960
accaggtctt gacatactat gcaaatctaa gagattagac gttcccttcg gggacatgga 1020
tacaggtggt gcatggttgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa 1080
cgagcgcaac ccttattatc agttgccagc attaagttgg gcactctggt gagactgccg 1140
gtgacaaacc ggaggaaggt ggggatgacg tcaaatcatc atgcccctta tgacctgggc 1200
tacacacgtg ctacaatgga tggtacaacg agttgcgaac tcgcgagagt aagctaatct 1260
cttaaagcca ttctcagttc ggattgtagg ctgcaactcg cctacatgaa gtcggaatcg 1320
ctagtaatcg cggatcagca tgccgcggtg aatacgttcc cgggccttgt acacaccgcc 1380
cgtcacacca tgagagtttg taacacccaa agtcggtggg gtaacctttt aggaaccagc 1440
ctgctaaggt gacaa 1455
Claims (1)
1. Lactobacillus plantarum (L.) KummerLactiplantibacillus plantarum) The application of the LP1406 in fermenting kelp is characterized in that the lactobacillus plantarum LP1406 is utilized to ferment the kelp to improve the effects of antioxidation and blood sugar reduction of a fermentation product;
the lactobacillus plantarum LP1406 is preserved in China Center for Type Culture Collection (CCTCC) at the preservation number of NO: m20211500, class designation: lactobacillus plantarum (L.) KummerLactiplantibacillus plantarum）LP1406；
The specific steps of the fermented kelp are as follows:
lactobacillus plantarum on MRS plateLactiplantibacillus plantarum) Single colony of LP1406 is inoculated in seed culture medium and cultured at 37 deg.c for 24 hr; then inoculating the kelp fermentation medium according to the inoculum size of 5% -10%, supplementing glucose to 3% in the kelp enzymolysis liquid, sterilizing at 121 ℃ for 20min, inoculating lactobacillus plantarum LP1406, and standing at 37 ℃ for culturing for 48h;
the specific preparation process of the kelp enzymolysis liquid comprises the following steps: washing kelp with water, drying, grinding, sieving the powder with a 40-mesh sieve, collecting, dissolving in distilled water at a ratio of 1:50 (w/v), adjusting pH to 4.8, adding cellulase and pectase for enzymolysis, wherein the addition amounts of the cellulase and the pectase are respectively 2.5wt% and 0.26 and wt%, stirring the reaction solution at 50 ℃ for 2 hours, adjusting pH to 8.0, adding 0.3wt% alkaline pectase, incubating at 60 ℃ for 1.5 hours, centrifuging the reaction product, and collecting the enzymolysis supernatant as kelp enzymolysis solution.
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