CN111454863A - Lactobacillus fermentum and application thereof in preparation of lactobacillus fermentation liquor with anti-aging function - Google Patents
Lactobacillus fermentum and application thereof in preparation of lactobacillus fermentation liquor with anti-aging function Download PDFInfo
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Abstract
The invention discloses a lactobacillus fermentum and application thereof in preparing lactobacillus fermentation liquor, the applicant screens and obtains the lactobacillus fermentum L F26, the lactobacillus fermentation liquor prepared by the strain has the anti-aging effect, can be used for skin care, and has good maintenance effect on the skin surface.
Description
Technical Field
The invention relates to lactobacillus fermentum and application thereof in preparing lactobacillus fermentation liquor with an anti-aging function. Belongs to the technical field of biological medicine processing.
Background
Human skin has a transient, temporary indigenous flora of staphylococcus epidermidis which is beneficial to health, as intestinal bacteria are essential to the health of the host. The skin microecological flora uses skin lipid as a nutrient, maintains skin acidity, and prevents other microorganisms from growing on the skin. Most of these microorganisms are gram-positive, residing on the skin surface and in the hair follicles. Hosts have a range of structures, molecules and mechanisms to confine these temporary and transient occupants, and also to control their population and to predominate certain microorganisms. The indigenous flora occupying a given niche is beneficial to the host. And may be harmful or infectious when it moves about.
Today research on skin micro-ecological flora is still in its infancy. Since the microecological flora of human skin is unique and complex, consisting of a mixture of different classes of microorganisms: facultative anaerobes (such as propionibacterium acnes), aerobes (such as staphylococcus epidermidis) and fungi (such as malassezia furfur), which are rarely present in the flora of other organs of the human body, indicate that the skin establishes a unique host-parasitic relationship defensive system.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide lactobacillus fermentum and application thereof in preparing lactobacillus fermentation liquor with an anti-aging function.
In order to achieve the purpose, the invention adopts the following technical scheme:
the lactobacillus fermentum is lactobacillus fermentum (L actinobacillus fermentum) L F26, which is preserved in China general microbiological culture Collection center (CGMCC) at 24/2/2020 with the preservation number of CGMCC No. 19431.
Preferably, the 16S rDNA sequence of the lactobacillus fermentum is shown as SEQ ID No. 1.
GCTCAGGATGAACGCCGGCGGTGTGCCTAATACATGCAAGTCGAACGCGTTGGCCCAATTGATTGATGGTGCTTGCACCTGATTGATTTTGGTCGCCAACGAGTGGCGGACGGGTGAGTAACACGTAGGTAACCTGCCCAGAAGCGGGGGACAACATTTGGAAACAGATGCTAATACCGCATAACAACGTTGTTCGCATGAACAACGCTTAAAAGATGGCTTCTCGCTATCACTTCTGGATGGACCTGCGGTGCATTAGCTTGTTGGTGGGGTAANGGCCTACCAAGGCGATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACAATGGGACTGAGACACGGCCCATACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGGCGCAAGCCTGATGGAGCAACACCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAGCTCTGTTGTTAAAGAAGAACACGTATGAGAGTAACTGTTCATACGTTGACGGTATTTAACCAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGAGAGTGCAGGCGGTTTTCTAAGTCTGATGTGAAAGCCTTCGGCTTAACCGGAGAAGTGCATCGGAAACTGGATAACTTGAGTGCAGAAGAGGGTAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTACCTGGTCTGCAACTGACGCTGAGACTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGGAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATCTTGCGCCAACCCTAGAGATAGGGCGTTTCCTTCGGGAACGCAATGACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTACTAGTTGCCAGCATTAAGTTGGGCACTCTAGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGACGACGTCAGATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACGGTACAACGAGTCGCGAACTCGCGAGGGCAAGCAAATCTCTTAAAACCGTTCTCAGTTCGGACTGCAGGCTGCAACTCGCCTGCACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAGCCAGCCGCCTAAGGTGGGACAGATGATTAGGGTGAAGTCGTAACAAGGTAGCCGTAGGAGAACCTGCG, as shown in SEQ ID NO. 1.
The lactobacillus fermentum is applied to preparing lactobacillus fermentation liquor with an anti-aging function.
The lactobacillus fermentum is applied to preparing skin care daily chemical products, preferably anti-aging skin care daily chemical products.
Preferably, the daily chemical product for skin care is a mask or a facial cleanser.
A lactobacillus fermentation liquid with anti-aging function is prepared by fermenting the lactobacillus fermentum.
The preparation method of the lactobacillus fermentation liquor with the anti-aging function is to inoculate the lactobacillus fermentum in a fermentation substrate and ferment for 4-48 hours at 34-45 ℃.
Preferably, the fermentation substrate comprises: 2-8 g of whey protein powder, 5-15 g of white granulated sugar, 0-6 g of skimmed milk powder, 0.5-2 g of yeast extract and 100g of water.
Preferably, the fermentation substrate is sterilized before inoculation, and the specific process conditions are as follows: sterilizing for 5-30 minutes at 70-90 ℃.
Preferably, the Lactobacillus fermentum is inoculated so that the number of original bacteria is 1.0 × 104cfu/ml～1.0×105cfu/ml。
Preferably, after the fermentation is finished, the sterilization treatment is carried out, and the specific process conditions are as follows: sterilizing for 3-5 minutes at 80-90 ℃.
Preferably, the specific steps are as follows:
(1) blending fermentation substrates: 2-8 g of whey protein powder, 5-15 g of white granulated sugar, 0-6 g of skimmed milk powder, 0.5-2 g of yeast extract and 100g of water;
(2) and (3) sterilization: sterilizing for 5-30 minutes at 70-90 ℃;
(3) inoculating the Lactobacillus fermentum to the culture medium to obtain fermented lactobacillus with original bacteria number of 1.0 × 104cfu/ml～1.0×105cfu/ml；
(4) Fermentation: fermenting for 4-48 hours at 34-45 ℃;
(5) and (3) sterilization: sterilizing for 3-5 minutes at 80-90 ℃;
(6) and (6) filling.
The application of the lactobacillus fermentation liquor in the daily chemical products for skin care is preferably the application in the anti-aging daily chemical products for skin care.
Preferably, the daily chemical product for skin care is a mask or a facial cleanser.
Preferably, the weight content of the lactobacillus fermentation liquor in the daily chemical products for skin care is 1-10%.
The lactobacillus fermentation liquor prepared by the strain has an anti-aging effect, can be used for skin care, and has a good maintenance effect on the skin surface.
Drawings
FIG. 1 is a graph of the results of an anti-aging experiment on cells of fermentation broth of Lactobacillus fermentum L F26.
Preservation information
And (3) classification and naming: lactobacillus fermentum
Name of Latin literature L actinobacillus fermentum
The name of the depository: china general microbiological culture Collection center (CGMCC)
The address of the depository: xilu No.1 Hospital No. 3 of Beijing market facing Yang district
The preservation date is as follows: 24 days 2 month in 2020
Detailed Description
The present invention will be further illustrated by the following examples, which are intended to be merely illustrative and not limitative.
Reference example:
the lactobacillus fermentum L F26 is separated from dairy products and is obtained by the following method:
(1) preparation of MRS Medium
The MRS culture medium comprises the following specific components:
the preparation method comprises adding the above materials into 1000m L distilled water, heating to dissolve, adjusting pH, packaging, and autoclaving at 121 deg.C for 18 min.
(2) Lactobacillus fermentum L F26 isolated from yogurt
The separation method comprises diluting yogurt sample in sterilized normal saline to a certain gradient, placing in MRS agar plate at 0.1m L, coating on surface with L bar, performing anaerobic culture at 37 deg.C for 48 hr, picking out characteristic bacterial colony on the plate with inoculating ring, and streaking on new MRS agar plate.
The characteristics of the lactobacillus fermentum L F26 obtained by the method are as follows:
gram-positive, non-sporulating, immotile obligate anaerobic bacteria that ferment glucose produce primarily acetate and L (+) lactate, but also produce small amounts of formate and succinate, and produce no CO2Propionic acid and butyric acid. Catalase negative, benzidine reaction and indole reaction negative, nitrate is not reduced. The molar content of G + C is between 57.2 and 64.5 percent.
The obtained 16S rDNA sequence of the lactobacillus fermentum L F26 has a sequence structure shown in SEQ ID No.1, wherein the specific structure of the SEQ ID No.1 is as follows:
GCTCAGGATGAACGCCGGCGGTGTGCCTAATACATGCAAGTCGAACGCGTTGGCCCAATTGATTGATGGTGCTTGCACCTGATTGATTTTGGTCGCCAACGAGTGGCGGACGGGTGAGTAACACGTAGGTAACCTGCCCAGAAGCGGGGGACAACATTTGGAAACAGATGCTAATACCGCATAACAACGTTGTTCGCATGAACAACGCTTAAAAGATGGCTTCTCGCTATCACTTCTGGATGGACCTGCGGTGCATTAGCTTGTTGGTGGGGTAANGGCCTACCAAGGCGATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACAATGGGACTGAGACACGGCCCATACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGGCGCAAGCCTGATGGAGCAACACCGCGTGAGTGAAGAAGGGTTTCGGCTCGTAAAGCTCTGTTGTTAAAGAAGAACACGTATGAGAGTAACTGTTCATACGTTGACGGTATTTAACCAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGAGAGTGCAGGCGGTTTTCTAAGTCTGATGTGAAAGCCTTCGGCTTAACCGGAGAAGTGCATCGGAAACTGGATAACTTGAGTGCAGAAGAGGGTAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTACCTGGTCTGCAACTGACGCTGAGACTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAGTGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGGAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCTACGCGAAGAACCTTACCAGGTCTTGACATCTTGCGCCAACCCTAGAGATAGGGCGTTTCCTTCGGGAACGCAATGACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTTACTAGTTGCCAGCATTAAGTTGGGCACTCTAGTGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGACGACGTCAGATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACGGTACAACGAGTCGCGAACTCGCGAGGGCAAGCAAATCTCTTAAAACCGTTCTCAGTTCGGACTGCAGGCTGCAACTCGCCTGCACGAAGTCGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGTTTGTAACACCCAAAGTCGGTGGGGTAACCTTTTAGGAGCCAGCCGCCTAAGGTGGGACAGATGATTAGGGTGAAGTCGTAACAAGGTAGCCGTAGGAGAACCTGCG
example 1:
(1) blending: performed according to the following recipe:
8g of whey protein powder; 15g of white granulated sugar; 6g of skimmed milk powder; 2g of yeast extract; make up to 100g with water.
(2) And (3) sterilization: at 70 deg.C for 30 min.
(3) Inoculation, Lactobacillus fermentum L F26 obtained in reference example was inoculated so that the number of original bacteria was 1.0 × 105cfu/ml。
(4) Fermentation: the fermentation temperature is 45 ℃ and the fermentation time is 48 h.
(5) And (3) sterilization: 90 ℃ for 3min
(6) And (6) filling.
(7) The lactobacillus fermentum L F26 prepared in the step (6) is applied to the preparation of daily chemical products such as films, facial cleanser and the like, and the usage amount of the lactobacillus fermentum is 10% of the total weight.
Example 2:
(1) blending: performed according to the following recipe:
2g of whey protein powder; 5g of white granulated sugar; 0.5g of yeast extract; make up to 100g with water.
(2) And (3) sterilization: 90 ℃ for 5 min.
(3) Inoculation, Lactobacillus fermentum L F26 obtained in reference example was inoculated so that the number of original bacteria was 1.0 × 104cfu/ml
(4) Fermentation: the fermentation temperature is 34 ℃, and the fermentation time is 4 h.
(5) And (3) sterilization: 80 ℃ for 3min
(6) And (6) filling.
(7) The lactobacillus fermentum L F26 prepared in the step (6) is applied to the preparation of daily chemical products such as films, facial cleanser and the like, and the usage amount of the lactobacillus fermentum is 1 percent of the total weight.
Example 3:
(1) blending: performed according to the following recipe:
3g of whey protein powder; 15g of white granulated sugar; 6g of skimmed milk powder; 0.5g of yeast extract; make up to 100g with water.
(2) And (3) sterilization: 90 ℃ for 30 min.
(3) Inoculation, Lactobacillus fermentum L F26 obtained in reference example was inoculated so that the number of original bacteria was 5.0 × 104cfu/ml。
(4) Fermentation: the fermentation temperature is 40 ℃, and the fermentation time is 32 h.
(5) And (3) sterilization: 85 ℃ for 4min
(6) And (6) filling.
(7) The lactobacillus fermentum L F26 prepared in the step (6) is applied to the preparation of daily chemical products such as films, facial cleanser and the like, and the usage amount of the lactobacillus fermentum is 5% of the total weight.
Example 4:
(1) blending: performed according to the following recipe:
6g of whey protein powder; 10g of white granulated sugar; 3g of skimmed milk powder; 2g of yeast extract; make up to 100g with water.
(2) And (3) sterilization: 70 ℃ for 5 min.
(3) Inoculation, Lactobacillus fermentum L F26 obtained in reference example was inoculated so that the number of original bacteria was 1.0 × 105cfu/ml。
(4) Fermentation: the fermentation temperature is 45 ℃ and the fermentation time is 4 h.
(5) And (3) sterilization: 90 ℃ for 3min
(6) And (6) filling.
(7) The lactobacillus fermentum L F26 prepared in the step (6) is applied to the preparation of daily chemical products such as films, facial cleanser and the like, and the usage amount of the lactobacillus fermentum is 5% of the total weight.
Test example:
anti-aging experiment of fermentation broth cells of lactobacillus fermentum L F26
Studies have shown that the positive rate of β -galactosidase staining in fibroblasts cultured in vitro increases with increasing numbers of cell divisions, β -galactosidase is expressed in senescent fibroblasts as well as keratinocytes, but not in quiescent cells as well as terminally differentiated cells, so β -galactosidase expression can be considered as one of the important markers for determining cellular senescence, since it is only possible to occur in senescent cells.
By further studying the cell protection effect of HE L F of aging on aging substances, the experiment design is that SA- β -gal activity level is examined by β -galactosidase staining kit (the same method is: J. Jamal, MR. mustafa, P. -F. Wong. Paeonol protectants against aging expression sensing in end cells [ J ] Journal of ethiofemoralgraphic science, 2014,154: 428. 436.). control of fermented milk, the positive ratio of β -galactosidase in the model group is obviously increased, VC is adopted in the positive control group, the positive ratio is obviously reduced, and simultaneously fermented lactobacillus L F26 fermented liquid, lactobacillus helveticus L HR43 fermented liquid and lactobacillus L F14 fermented liquid are studied, and the lowest positive ratio of L F26 is found (FIG. 1).
The preparation method and the preparation method of the lactobacillus helveticus L HR43 fermentation broth and the lactobacillus fermentum L F14 fermentation broth are the same as those in example 1, and the lactobacillus helveticus L HR43 and the lactobacillus fermentum L F14 are all purchased from Hemike Biotech GmbH (No. 66 North Ling of Lophatherum gracile, Gaoyang, China).
Although the present invention has been described with reference to the specific embodiments, it is not intended to limit the scope of the present invention, and various modifications and variations can be made by those skilled in the art without inventive changes based on the technical solution of the present invention.
Sequence listing
<110> Biotechnology (Yangzhou) Ltd
<120> lactobacillus fermentum and application thereof in preparation of lactobacillus fermentation liquor with anti-aging function
<141>2020-04-07
<160>1
<170>SIPOSequenceListing 1.0
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gctcaggatg aacgccggcg gtgtgcctaa tacatgcaag tcgaacgcgt tggcccaatt 60
gattgatggt gcttgcacct gattgatttt ggtcgccaac gagtggcgga cgggtgagta 120
acacgtaggt aacctgccca gaagcggggg acaacatttg gaaacagatg ctaataccgc 180
ataacaacgt tgttcgcatg aacaacgctt aaaagatggc ttctcgctat cacttctgga 240
tggacctgcg gtgcattagc ttgttggtgg ggtaanggcc taccaaggcg atgatgcata 300
gccgagttga gagactgatc ggccacaatg ggactgagac acggcccata ctcctacggg 360
aggcagcagt agggaatctt ccacaatggg cgcaagcctg atggagcaac accgcgtgag 420
tgaagaaggg tttcggctcg taaagctctg ttgttaaaga agaacacgta tgagagtaac 480
tgttcatacg ttgacggtat ttaaccagaa agtcacggct aactacgtgc cagcagccgc 540
ggtaatacgt aggtggcaag cgttatccgg atttattggg cgtaaagaga gtgcaggcgg 600
ttttctaagt ctgatgtgaa agccttcggc ttaaccggag aagtgcatcg gaaactggat 660
aacttgagtg cagaagaggg tagtggaact ccatgtgtag cggtggaatg cgtagatata 720
tggaagaaca ccagtggcga aggcggctac ctggtctgca actgacgctg agactcgaaa 780
gcatgggtag cgaacaggat tagataccct ggtagtccat gccgtaaacg atgagtgcta 840
ggtgttggag ggtttccgcc cttcagtgcc ggagctaacg cattaagcac tccgcctggg 900
gagtacgacc gcaaggttga aactcaaagg aattgacggg ggcccgcaca agcggtggag 960
catgtggttt aattcgaagc tacgcgaaga accttaccag gtcttgacat cttgcgccaa 1020
ccctagagat agggcgtttc cttcgggaac gcaatgacag gtggtgcatg gtcgtcgtca 1080
gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc gcaacccttg ttactagttg 1140
ccagcattaa gttgggcact ctagtgagac tgccggtgac aaaccggagg aaggtgggga 1200
cgacgtcaga tcatcatgcc ccttatgacc tgggctacac acgtgctaca atggacggta 1260
caacgagtcg cgaactcgcg agggcaagca aatctcttaa aaccgttctc agttcggact 1320
gcaggctgca actcgcctgc acgaagtcgg aatcgctagt aatcgcggat cagcatgccg 1380
cggtgaatac gttcccgggc cttgtacaca ccgcccgtca caccatgaga gtttgtaaca 1440
cccaaagtcg gtggggtaac cttttaggag ccagccgcct aaggtgggac agatgattag 1500
ggtgaagtcg taacaaggta gccgtaggag aacctgcg 1538
Claims (10)
1. The lactobacillus fermentum is lactobacillus fermentum (L actinobacillus fermentum) L F26, which is preserved in China general microbiological culture Collection center (CGMCC) at 24/2/2020 with the preservation number of CGMCC No. 19431.
2. A Lactobacillus fermentum according to claim 1, wherein the Lactobacillus fermentum has a 16S rDNA sequence as set forth in SEQ ID No. 1.
3. Use of a lactobacillus fermentum according to claim 1 or 2 for the preparation of a lactic acid bacteria fermentation broth.
4. Use of a lactobacillus fermentum according to claim 1 or 2 for the preparation of a daily chemical product for skin care.
5. A lactic acid bacteria fermentation broth, which is prepared by fermentation of the Lactobacillus fermentum of claim 1 or 2.
6. The method for producing a lactic acid bacterium fermentation broth according to claim 5, wherein the fermentation substrate is inoculated with the above-mentioned Lactobacillus fermentum and fermented at 34 to 45 ℃ for 4 to 48 hours.
7. The method of claim 6, wherein the fermentation substrate comprises: 2-8 g of whey protein powder, 5-15 g of white granulated sugar, 0-6 g of skimmed milk powder, 0.5-2 g of yeast extract and 100g of water.
8. The method according to claim 6, wherein the fermentation substrate is sterilized before inoculation, and the specific process conditions are as follows: sterilizing for 5-30 minutes at 70-90 ℃.
9. The method according to claim 6, wherein the Lactobacillus fermentum is inoculated so that the number of original bacteria is 1.0 × 104cfu/ml～1.0×105cfu/ml。
10. Use of a lactic acid bacteria fermentation broth according to claim 5 in a daily chemical product for skin care.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112940968A (en) * | 2021-02-07 | 2021-06-11 | 山东凤凰生物有限公司 | Lactobacillus fermentum, lactobacillus fermentum culture and preparation method thereof |
| CN113832050A (en) * | 2021-08-13 | 2021-12-24 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Lactobacillus fermentum for efficiently synthesizing nicotinamide and resisting photoaging and application thereof |
| CN117736942A (en) * | 2024-02-20 | 2024-03-22 | 山东中科嘉亿生物工程有限公司 | Fermented lactobacillus mucilaginosus JYLF-315 for improving skin aging and metagen preparation and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113832050A (en) * | 2021-08-13 | 2021-12-24 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Lactobacillus fermentum for efficiently synthesizing nicotinamide and resisting photoaging and application thereof |
| CN117736942A (en) * | 2024-02-20 | 2024-03-22 | 山东中科嘉亿生物工程有限公司 | Fermented lactobacillus mucilaginosus JYLF-315 for improving skin aging and metagen preparation and application thereof |
| CN117736942B (en) * | 2024-02-20 | 2024-05-31 | 山东中科嘉亿生物工程有限公司 | Fermented lactobacillus mucilaginosus JYLF-315 for improving skin aging, and metagen preparation and application thereof |
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