CN114525319A - Heat shock protein-ectoin composition and preparation method and application thereof - Google Patents
Heat shock protein-ectoin composition and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a heat shock protein-ectoine composition, a preparation method and application thereof, wherein the preparation method comprises the following steps: s101: inoculating thermophilic thermus strain into the seed culture medium A, and performing enrichment culture to obtain a seed culture solution; s102: inoculating the seed culture solution into a fermentation culture medium A containing an ectoin extracting solution, and performing fermentation culture to obtain a thermus thermophilus fermentation liquid; s103: the thermophilic thermus strain fermentation liquor is crushed, and the obtained thermophilic thermus strain lysate is the heat shock protein-ectoin composition. The heat shock protein-ectoin composition obtained by the application has excellent anti-photoaging and photodamage repairing effects.
Description
Technical Field
The invention relates to the field of microbial fermentation products, in particular to a heat shock protein-ectoine composition and a preparation method and application thereof.
Background
Heat Shock Proteins (HSPs) are a group of special proteins with extremely conserved structures, which are produced by an organism rapidly making a stress response to external stimuli such as high temperature and the like, closing normal gene expression and opening the expression of a group of stress genes. It is used as important buffering factor in cell mutation process to regulate the spatial structure and mutation of important protein in physiological and biochemical change of cell. So that the organism can survive in the adverse environments and states of heat, cold, high salt, organic poison, heavy metal ions, oxygen free radical, ultraviolet ray, ischemia, anoxia, infection, gene injury, tissue injury, etc.
Due to the special living environment and the unique high-temperature fermentation process, the heat shock protein generated in the thermophilic thermus strain has excellent photoaging, photodamage and inflammation repairing effects. However, the yield of heat shock proteins in Thermus thermophilus is not high, which greatly limits the application of heat shock proteins.
Disclosure of Invention
The application provides a heat shock protein-ectoin composition, a preparation method and an application thereof, and the preparation method can effectively promote the content of the heat shock protein in a thermophilic thermus strain, thereby promoting the improvement of the light injury repair effect of a strain fermentation product.
In a first aspect, the present application provides a method for preparing a heat shock protein-ectoin composition, comprising the steps of:
s101: inoculating thermophilic thermus strain into the seed culture medium A, and performing enrichment culture to obtain a seed culture solution;
s102: inoculating the seed culture solution into a fermentation culture medium A containing an ectoin extracting solution, and performing fermentation culture to obtain a thermus thermophilus fermentation liquid;
s103: the thermophilic thermus strain fermentation liquor is crushed, and the obtained thermophilic thermus strain lysate is the heat shock protein-ectoin composition.
In the technical scheme, the seed culture solution of the thermus thermophilus is placed in the fermentation culture medium containing the ectoin extracting solution for fermentation culture, so that the content of the heat shock protein in the thermus thermophilus is improved by about 25.3 percent compared with the conventional fermentation culture medium.
The ectoine (tetrahydromethylpyrimidine carboxylic acid) is synthesized by halophilic or halotolerant bacteria, and is a main compatible solute capable of resisting external high-salt stress. It is accumulated in cells in large quantity, can resist the impact of high osmotic pressure, simultaneously stabilizes the hydration layer structure of protein (especially under the conditions of freezing, drying and high temperature), belongs to organic micromolecules with characteristics of hydrophilicity and zwitterion, is stable to heat, is not easy to decompose, is easy to dissolve in water, is used in cosmetics, and can reduce sunburn and the like of skin caused by ultraviolet irradiation. The protein is added into a fermentation culture medium, can regulate and control a signal path expressed by the heat shock protein, and induces and promotes the increase of the protein production amount.
No special requirement is imposed on the thermophilic thermus strain, and strains with the strain numbers of CICC 2489 or BAA-163 are preferably adopted. In addition, the time for enrichment culture is preferably 12-30 hours; fermentation culture operation, wherein the inoculation amount is preferably 1-10%, and the culture time is preferably 12-30 hours; the disruption treatment is preferably carried out by sonication to achieve sufficient release of the intracellular lysate.
Preferably, the ectoin extract comprises one or more of ectoin, amino acid, polypeptide, polysaccharide and polyalcohol.
From the above, it can be seen that the use of ectoin as a component of the medium can promote the expression of heat shock proteins of Thermus thermophilus and increase the content of heat shock proteins in the cells. However, the applicant found that the heat shock protein content in the cells obtained by fermentation was further increased by about 10.4% by using a cell lysate containing ectoin, amino acids, polypeptides, polysaccharides and polyols as it is, as compared with the case where the fermentation culture of thermophilic bacteria is carried out by adding ectoin alone. The reason for this may be that amino acids, polypeptides, polysaccharides, polyols and ectoin in halophilic bacteria can synergistically promote the expression and generation of heat shock proteins in the fermentation process of thermophilic thermus bacteria, improve the content of heat shock proteins, enhance the repair efficacy and slow down photoaging.
Meanwhile, the thermophilic thermus strain is crushed to obtain lysate containing the components, complex separation and purification processes are not needed, the production cost can be effectively reduced,
preferably, in the step S101 and the step S102, the temperature of the enrichment culture and the fermentation culture is 45-85 ℃, and the culture time is 12-30 hours.
By adopting the technical scheme, the thermophilic thermus inhabiting bacteria can be fully fermented under the high-temperature environment, so that sufficient heat shock proteins can be obtained; the suitable culture time is favorable for guaranteeing the survival rate of the thalli.
Preferably, the seed culture medium A comprises the following components in percentage by weight:
yeast powder: 5-15 g/L;
polypeptone: 5-25 g/L;
NaCl：1-5g/L。
by adopting the technical scheme, the logarithmic growth of halophilic bacteria is effectively guaranteed, and the synthesis of ectoin is promoted. In addition, the pH of the seed culture medium A is preferably 7.0-7.2.
Preferably, the fermentation medium A comprises the following components in percentage by weight:
yeast powder: 5-15 g/L;
polypeptone: 5-25 g/L;
FeCl3：0.05～0.2g/L；
CaSO4·2H2O：0.1～0.5g/L；
MgSO4·2H2O：0.1～0.5g/L；
KNO3：0.1～0.5g/L；
NaNO3：1～3g/L；
Na2HPO4：0.1～0.5g/L；
MnSO4：1～3g/L；
ZnSO4·7H2O：0.3～1g/L。
by adopting the technical scheme, the halophilic bacteria and the thermophilic thermus bacteria are subjected to double-bacteria high-temperature combined fermentation, and compared with a single fermentation product, the fermentation product has higher anti-inflammation and photodamage repair functions. The ectoin extracting solution can promote the expression of a thermus thermophilus heat shock protein pathway, and a composition of ectoin and heat shock protein is obtained, so that a more excellent photodamage repairing effect is achieved. In addition, the pH of the fermentation medium A in the application is preferably 7.0-7.2.
Preferably, the ectoin extract is prepared by the following method:
s201: inoculating halophilic bacteria into the seed culture medium B, and performing enrichment culture to obtain a seed culture solution;
s202: inoculating the seed culture solution into a fermentation culture medium B, and performing fermentation culture to obtain halophilic bacteria fermentation liquor;
s203: and centrifuging the halophilic bacteria fermentation liquor, removing thalli to obtain fermentation supernatant, and desalting the fermentation liquor to obtain the ectoin extracting solution.
The halophilic bacteria are Halomonas elongata, and preferably, the strain with the strain number CICC 2489 is adopted. Wherein, the time of enrichment culture is preferably 16-30 hours; fermentation culture operation, wherein the inoculation amount is preferably 1-10%, and the culture time is preferably 16-30 hours; the desalting treatment adopts a dialysis bag with the specification of 100-500nm for dialysis purification, and the salt can be fully removed after circulation for 2-4 times, so as to obtain a crude extract mainly containing the ectoin.
Preferably, in step S201 and step S202, the temperature of the enrichment culture and the fermentation culture are both 26 to 32 ℃, and the culture time is both 16 to 30 hours.
By adopting the technical scheme, the logarithmic growth of halophilic bacteria and the synthesis of ectoin can be effectively guaranteed at proper temperature and fermentation time.
Preferably, the seed culture medium B comprises the following components in percentage by weight:
NaCl：40～60g/L；
MgCl2·6H2O：5～15g/L；
MgSO4·7H2O：2～5g/L；
CaCl2：0.1～0.5g/L；
KCl：1～3g/L；
NaHCO3：0.01～0.1g/L；
NaBr：0.01～0.1g/L；
peptone: 5-10 g/L;
yeast powder: 5-10 g/L;
glucose: 1-3 g/L.
By adopting the technical scheme, the yeast powder and the halophilic bacteria are mixed and fermented, so that the content of substances such as amino acid, polysaccharide and the like in the ectoin extracting solution can be improved, and the ectoin can be synergistically acted on the synthesis of the thermus thermophilus HSPs. In addition, the pH of the seed culture medium B is preferably 7.0-7.2.
Preferably, the fermentation medium B comprises the following components in percentage by weight:
NaCl：40～70g/L；
MgCl2·6H2O：5～20g/L；
MgSO4·7H2O：2～10g/L；
CaCl2：0.1～1g/L；
KCl：1～5g/L；
NaHCO3：0.01～0.1g/L；
peptone: 5-20 g/L;
yeast powder: 5-20 g/L;
glucose: 1-5 g/L.
By adopting the technical scheme, the ectoin extracting solution prepared by halophilic bacteria fermentation can reach 0.1-1 g/L after primary desalting treatment, and in addition to the ectoin, the extracting solution also contains amino acid, polypeptide, polysaccharide, polyol and other components through HPLC analysis, and can act synergistically to promote the expression of heat shock protein in a thermophilic thermus thermophilus fermentation product. In addition, the pH of the fermentation medium B is preferably 7.0 to 7.2.
In a second aspect, the present application provides a heat shock protein-ectoin composition made by any of the methods described above.
In a third aspect, the present application provides a heat shock protein-ectoin composition for use in cosmetics.
By adopting the technical scheme, the heat shock protein-ectoin composition contains active ingredients such as ectoin, heat shock protein, amino acid, polypeptide and the like, and can play a synergistic effect in cosmetic application.
When the addition amount is 1-5%, the effect on the photodamage repairing layer surface is more prominent. Secondly, at the level of cell efficacy evaluation, the relative proliferation rate of keratinocytes (Hacat) after UV irradiation decreased to 50%, and with the addition of the heat shock protein-ectoin composition, the injury repair effect gradually increased, and the Hacat cell proliferation rate also increased. In addition, the skin care product can also obviously promote the proliferation of fibroblasts (HSF), further promote the synthesis of collagen, enhance the skin elasticity, fade wrinkles and slow down photoaging.
Still further, Matrix Metalloproteinases (MMPs) are important in the photoaging evaluation of 3D skin models, and when ROS is overproduced, the expression of MMPs is induced, which in turn breaks down collagen, producing aging symptoms such as wrinkles, laxity, and fine lines. The addition of the heat shock protein-ectoin composition can effectively reduce the expression of MMP-1 and MMP-9 and further damage caused by oxidative stress.
In the evaluation of human body efficacy, the heat shock protein-ectoin composition can effectively improve the percutaneous water loss, repair the damaged barrier and improve the skin hydration ability; meanwhile, the skin aging phenomenon caused by the degradation of skin elastin and the loss of collagen can be well improved, the skin elasticity is enhanced, the skin smoothness is improved, and the aging roughness is reduced; in addition, the compound can accelerate melanin catabolism, repair ultraviolet sunburn, reduce the phenomena of color spot deposition, dark yellow and the like, and has very positive effects on the aspects of light protection and light aging resistance.
When the heat shock protein-ectoin composition is applied to a cosmetic formula, p-hydroxyacetophenone and butanediol can be compounded to be used as preservatives so as to keep a raw material system stable. Other raw materials can be added according to the requirement to prepare the finished product formula.
In summary, the present application has the following beneficial effects:
1. the method adopts a fermentation substrate containing an ectoin extracting solution to carry out fermentation culture on the thermus thermophilus, and the ectoin, amino acid, polysaccharide, polypeptide, polyalcohol and the like contained in the ectoin extracting solution can promote the synthesis of the heat shock protein of the thermus thermophilus and promote thalli obtained by fermentation to have higher heat shock protein content. The heat shock protein-ectoin composition formed by mixing the thallus lysate and ectoin can play a more excellent role in photodamage repair on one hand; on the other hand, the extraction process of the ectoin extract can be effectively simplified.
2. The halophilic bacteria and the thermophilic thermus are adopted for carrying out double-bacteria high-temperature mixed fermentation, which is beneficial to improving the light injury repair effect of the fermentation product.
Detailed Description
Preparation example
Preparation example 1, an ectoin extract was prepared as follows:
s201: inoculating halophilic bacteria (the strain number is CICC 2489) into the seed culture medium B, placing the seed culture medium B into a shaking table for enrichment culture, and culturing for 25 hours under the conditions that the rotating speed is 100 and 150rpm and the culture temperature is 26-32 ℃ to obtain a seed culture solution;
wherein, the seed culture medium B comprises the following components in percentage by weight: NaCl: 50 g/L; MgCl2·6H2O：10g/L；MgSO4·7H2O：3g/L；CaCl2：0.2g/L；KCl：2g/L；NaHCO3: 0.03 g/L; NaBr: 0.05 g/L; peptone: 8 g/L; yeast powder: 6 g/L; glucose: 2 g/L; the pH of the seed culture medium B is 7.0-7.2.
S202: inoculating the obtained seed culture solution into a fermentation medium B (5L fermentation tank) according to the inoculation amount of 10%, and performing enrichment culture at the rotation speed of 200-250rpm and the culture temperature of 26-32 ℃ for 20 hours to obtain halophilic bacteria fermentation broth;
wherein the fermentation medium B comprises the following components in percentage by weight: NaCl: 45 g/L; MgCl2·6H2O：15g/L；MgSO4·7H2O：6g/L；CaCl2：0.5g/L；KCl：4g/L；NaHCO3: 0.07 g/L; peptone: 15 g/L; yeast powder: 12 g/L; glucose: 3 g/L; the pH of the seed culture medium B is 7.0-7.2.
S203: centrifuging the halophilic bacteria fermentation liquid obtained in the last step at 6000-; then, desalting treatment is carried out for 6h by using a dialysis bag (100nm), and circulation is carried out for 2-4 times to obtain an ectoin extracting solution.
The ectoin extract mainly contains ectoin, amino acids and their derivatives, polypeptide, polysaccharide, and polyalcohol and its derivatives.
Preparation example 2, an ectoin extract was prepared as follows:
s201: inoculating halophilic bacteria (the strain number is CICC 2489) into the seed culture medium B, placing the seed culture medium B into a shaking table for enrichment culture, and culturing for 16 hours under the conditions that the rotating speed is 100 and 150rpm and the culture temperature is 26-32 ℃ to obtain a seed culture solution;
wherein, the seed culture medium B comprises the following components in percentage by weight: NaCl: 40 g/L; MgCl2·6H2O：5g/L；MgSO4·7H2O：5g/L；CaCl2：0.4g/L；KCl：1g/L；NaHCO3: 0.05 g/L; NaBr: 0.08 g/L; peptone: 5 g/L; yeast powder: 10 g/L; glucose: 3 g/L; the pH of the seed culture medium B is 7.0-7.2.
S202: inoculating the obtained seed culture solution into a fermentation medium B (5L fermentation tank) according to the inoculation amount of 5%, and performing enrichment culture at the rotation speed of 200-250rpm and the culture temperature of 26-32 ℃ for 30 hours to obtain halophilic bacteria fermentation broth;
wherein the fermentation medium B comprises the following components in percentage by weight: NaCl: 45 g/L; MgCl2·6H2O：15g/L；MgSO4·7H2O：6g/L；CaCl2：0.5g/L；KCl：4g/L；NaHCO3: 0.07 g/L; peptone: 15 g/L; yeast powder: 12 g/L; glucose: 3 g/L; the pH of the seed culture medium B is 7.0-7.2.
S203: centrifuging the halophilic bacteria fermentation liquid obtained in the last step at 6000-; then, desalting with dialysis bag (200nm) for 8 hr, and circulating for 2-4 times to obtain ectoin extractive solution.
The ectoin extract mainly contains ectoin, amino acids and their derivatives, polypeptide, polysaccharide, and polyalcohol and their derivatives.
Preparation example 3, an ectoine extract, was different from preparation example 1 in that no yeast powder was added to both the seed medium B and the fermentation medium B.
Examples
Example 1
The application discloses a preparation method of a heat shock protein-ectoin composition, which comprises the following steps:
s101: inoculating thermophilic Thermus (strain number is CICC 2489) into seed culture medium A, and shake culturing at rotation speed of 100-;
wherein, the seed culture medium A comprises the following components in percentage by weight: 5g/L of yeast powder, 10g/L of polypeptone, 2g/L of NaCl and 7.0-7.2 of pH.
S102: the obtained seed culture solution was inoculated into a fermentation medium A (5L fermenter) containing the crude extract of ectoin (prepared in preparation example 1) at a rate of 200-250rpm for 20 hours at a cultivation temperature of 80 ℃;
wherein the fermentation medium A comprises the following components in percentage by weight: ectoin extract: 300 g/L; yeast powder: 15 g/L; polypeptone: 20 g/L; FeCl3：0.05g/L；CaSO4·2H2O：0.2g/L；MgSO4·2H2O：0.1g/L；KNO3：0.5g/L；NaNO3：2g/L；Na2HPO4：0.4g/L；MnSO4：1g/L；ZnSO4·7H2O: 0.5 g/L; the pH value of the fermentation medium A is 7.0-7.2.
S103: carrying out ultrasonic crushing treatment on the thermophilic thermus strain fermentation liquor to promote the lysate of the thermophilic thermus strain to be fully released so as to obtain the heat shock protein-ectoin composition.
Example 2
The application discloses a preparation method of a heat shock protein-ectoin composition, which comprises the following steps:
s101: inoculating thermophilic Thermus (strain number is CICC 2489) into seed culture medium A, and shake culturing at rotation speed of 100-;
wherein, the seed culture medium A comprises the following components in percentage by weight: 10g/L of yeast powder, 15g/L of polypeptone, 5g/L of NaCl and 7.0-7.2 of pH.
S102: the obtained seed culture solution is inoculated into a fermentation culture medium A (5L fermentation tank) containing the ectoine crude extract (prepared in the preparation example 2) according to the inoculation amount of 5 percent, the rotation speed is 200-250rpm, the culture temperature is 75 ℃, and the culture time is 15 hours, so as to obtain the thermus thermophilus fermentation liquor;
wherein the fermentation medium A comprises the following components in percentage by weight: ectoin extract: 500 g/L; yeast powder: 6 g/L; polypeptone: 20 g/L; FeCl3：0.1g/L；CaSO4·2H2O：0.1g/L；MgSO4·2H2O：0.3g/L；KNO3：0.2g/L；NaNO3：1g/L；Na2HPO4：0.5g/L；MnSO4：2g/L；ZnSO4·7H2O: 0.8 g/L; the pH value of the fermentation medium A is 7.0-7.2.
S103: carrying out ultrasonic crushing treatment on the thermophilic thermus strain fermentation liquor to promote the lysate of the thermophilic thermus strain to be fully released so as to obtain the heat shock protein-ectoin composition.
Example 3
The present application discloses a method for preparing a heat shock protein-ectoin composition, which is different from example 1 in that the amount of ectoin extract is 100g/L in step S102.
Example 4
The application discloses a preparation method of a heat shock protein-ectoin composition, which is different from the embodiment 1 in that the dosage of the ectoin extracting solution in the step S102 is 600 g/L.
Example 5
The present application discloses a method for preparing a heat shock protein-ectoin composition, which is different from example 1 in that in step S102, an ectoin extract is prepared in preparation example 3.
Example 6
The present application discloses a method for preparing a heat shock protein-ectoin composition, which is different from example 1 in that in step S102, the ectoin extract is a 1g/L aqueous solution of ectoin.
Example 7
The application discloses a preparation method of a heat shock protein-ectoin composition, which is different from the preparation method of example 1 in that no yeast powder is added into a seed culture medium A and a fermentation culture medium A.
Comparative example
Comparative example 1
The present application discloses a method for preparing a heat shock protein-ectoin composition, which is different from example 1 in that no ectoin extract is added to the fermentation medium a in step S102.
Performance test
Test 1: heat shock protein content assay
The test method comprises the following steps: ELISA detection principle: the basis of ELISA is immobilization of antigen or antibody and enzyme labeling. The antigen or antibody is bound on the surface of the solid phase carrier to maintain the immunological activity, and the enzyme labeled antigen or antibody has both the immunological activity and the enzyme activity. In the measurement, the specimen to be tested (the antibody or antigen to be measured therein) is reacted with the antigen or antibody on the surface of the solid carrier. And then separating the antigen-antibody complex on the solid phase carrier from other substances in the liquid by a washing method. Then adding enzyme-labeled antigen or antibody, and binding on the solid phase carrier through reaction. The amount of enzyme on the solid phase will then be proportional to the amount of the substance to be detected in the sample. After the substrate of enzyme reaction is added, the substrate is catalyzed by enzyme into colored product, the amount of the product has direct relation with the amount of the detected substance in the sample, so that qualitative or quantitative analysis can be carried out according to the color generation degree.
The operation flow of the kit is as follows: adopts a full-automatic enzyme immunoassay analyzer and applies the experimental principle of a double-antibody sandwich method. At the detection wavelength of 450nm, an HSP monoclonal antibody is pre-coated on a microporous plate, a sample to be detected is added into a coating plate, and then an anti-HSP monoclonal antibody marked by horseradish peroxidase (HRP) is added to incubate in micropores of the coating plate together to form an antibody-antigen-enzyme-labeled antibody compound. Subsequently, washing the plate to remove the unbound additives; finally, 3',5,5' -Tetramethylbenzidine (TMB) is added for color reaction to generate a blue product, and the product becomes yellow after the reaction is ended. The absorbance (OD value) of each well was measured with shaking for 5 seconds at a wavelength of 450nm, and the measurement was repeated three times within 10min after the addition of the stop solution.
And (4) making a standard curve according to the concentration of the HSPs in the standard substance and the corresponding absorbance value (OD value), and calculating the heat shock protein concentration in the sample according to the standard curve.
TABLE 1 test results (%)
Test 2: heat shock protein-ectoin composition efficacy test method: screening out the poor skin barrier (the transepidermal water loss rate TEWL is more than 25 g/h.m)2) The age range of 20 Chinese healthy women as test volunteers in the age range of 30-55 years in the face wrinkle population (30-45 years), and the subjects respectively perform related tests before, after 2 weeks, 4 weeks and 8 weeks of using the product on the face, wherein the related tests comprise: the indexes of transdermal water loss, skin tightness, skin elasticity, skin smoothness, and melanin value, and can be used for indicating light repairing, photoaging, etcThe product has good therapeutic effect.
According to the technical safety of cosmetics (2021 edition), sunburn caused by photodamage is characterized by the skin melanin index (MI value), and is measured by a skin melanin detector (a parameter for characterizing the melanin content in the skin by measuring the absorption of a specific wavelength spectrum on the skin surface).
The melanin reduction rate (4 weeks) is (melanin content before cosmetics application-melanin content after cosmetics 28 are continuously applied)/melanin content before cosmetics application x 100%. The subject did not use other whitening products during the test, and the higher the measured value, the higher the melanin content in the skin.
TABLE 2 test results
And (3) analyzing test results:
(1) as can be seen by combining examples 1-4 and comparative example 1 and combining tables 1 and 2, the content of heat shock protein in the obtained fermentation product is increased by about 25.3% compared with that in comparative example 1 when the fermentation substrate containing the ectoine extracting solution is adopted for culture in examples 1-4; the fermentation product of the heat shock protein-ectoin composition can also obtain more excellent photodamage repairing effect when being used in cosmetics.
The reason for the above phenomenon may be that ectoin, amino acids, polypeptides, polysaccharides and polyols contained in the ectoin extract synergistically act on thermus thermophilus to promote synthesis of heat shock proteins and increase the content of heat shock proteins in the fermentation product. And because the heat shock protein can effectively prevent photodamage and the generation of photoaging appearance (spots, wrinkles, dry skin and the like), resist UV, protect the DNA structure of cells and enhance the integrity of the skin. Thus, the increase in heat shock protein content helps to enhance its photodamage repair efficacy in cosmetics.
(2) As can be seen by combining example 1 and example 4 and tables 1 and 2, the ectoin extract of example 1 contains ectoin, amino acids, polypeptides, polysaccharides and polyols, while the ectoin extract of example 4 contains only ectoin. Finally, the content of the heat shock protein in the fermentation product obtained in example 1 is increased by about 10.4% compared with that in example 4; the fermentation product of the heat shock protein-ectoin composition can also obtain more excellent photodamage repairing effect when being used in cosmetics.
The above reasons may be that amino acids, polypeptides, polysaccharides and polyols have a synergistic effect on the expression of heat shock proteins in thermus thermophilus, which is beneficial to increase the content of heat shock proteins in fermentation products.
The present embodiment is only for explaining the present application, and it is not limited to the present application, and those skilled in the art can make modifications of the present embodiment without inventive contribution as needed after reading the present specification, but all of them are protected by patent law within the scope of the claims of the present application.
Claims (10)
1. The preparation method of the heat shock protein-ectoin composition is characterized by comprising the following steps:
s101: inoculating thermophilic thermus strain into the seed culture medium A, and performing enrichment culture to obtain a seed culture solution;
s102: inoculating the seed culture solution into a fermentation culture medium A containing an ectoin extracting solution, and performing fermentation culture to obtain a thermus thermophilus fermentation liquid;
s103: the thermophilic thermus strain fermentation liquor is crushed, and the obtained thermophilic thermus strain lysate is the heat shock protein-ectoin composition.
2. The method of preparing a heat shock protein-ectoin composition according to claim 1, wherein the ectoin extract comprises one or more of ectoin, amino acids, polypeptides, polysaccharides, and polyols.
3. The method of claim 1, wherein the temperature of the enrichment culture and the fermentation culture are 45-85 ℃ and the time of the enrichment culture and the fermentation culture are 12-30 hours in steps S101 and S102.
4. The method of producing a heat shock protein-ectoine composition according to claim 1, wherein the seed culture medium A comprises the following components:
yeast powder: 5-15 g/L;
polypeptone: 5-25 g/L;
NaCl：1-5g/L。
5. the method of producing a heat shock protein-ectoin composition according to claim 1, wherein the fermentation medium a comprises the following components:
ectoin extract: 300-500 g/L;
yeast powder: 5-15 g/L;
polypeptone: 5-25 g/L;
FeCl3：0.05～0.2g/L；
CaSO4·2H2O：0.1～0.5g/L；
MgSO4·2H2O：0.1～0.5g/L；
KNO3：0.1～0.5g/L；
NaNO3：1～3g/L；
Na2HPO4：0.1～0.5g/L；
MnSO4：1～3g/L；
ZnSO4·7H2O：0.3～1g/L。
6. the method of producing a heat shock protein-ectoin composition according to claim 1, wherein the ectoin extract is produced by the following method:
s201: inoculating halophilic bacteria into the seed culture medium B, and performing enrichment culture to obtain a seed culture solution;
s202: inoculating the seed culture solution into a fermentation culture medium B, and performing fermentation culture to obtain halophilic bacteria fermentation liquor;
s203: and centrifuging the halophilic bacteria fermentation liquor, removing thalli to obtain fermentation supernatant, and desalting the fermentation liquor to obtain the ectoin extracting solution.
7. The method of claim 6, wherein the temperature of the enrichment culture and the fermentation culture are both 26-32 ℃ and the time of the enrichment culture and the fermentation culture are both 16-30 hours in steps S101 and S102.
8. The method for preparing a heat shock protein-ectoin composition according to claim 6, wherein the fermentation medium B comprises the following components in amounts:
NaCl：40～70g/L；
MgCl2·6H2O：5～20g/L；
MgSO4·7H2O：2～10g/L；
CaCl2：0.1～1g/L；
KCl：1～5g/L；
NaHCO3：0.01～0.1g/L；
peptone: 5-20 g/L;
yeast powder: 5-20 g/L;
glucose: 1-5 g/L.
9. A heat shock protein-ectoin composition prepared according to any one of claims 1 to 8.
10. Use of the heat shock protein-ectoin composition of any one of claims 1 to 9 for application to a cosmetic raw material.
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