CN110960452A - Yeast extract containing phloretin and preparation process and application thereof - Google Patents
Yeast extract containing phloretin and preparation process and application thereof Download PDFInfo
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- CN110960452A CN110960452A CN201911364044.1A CN201911364044A CN110960452A CN 110960452 A CN110960452 A CN 110960452A CN 201911364044 A CN201911364044 A CN 201911364044A CN 110960452 A CN110960452 A CN 110960452A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9728—Fungi, e.g. yeasts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/34—Alcohols
- A61K8/345—Alcohols containing more than one hydroxy group
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
- A61K8/35—Ketones, e.g. benzophenone
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
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Abstract
The invention provides a rhizocortin-containing yeast extract and a preparation process and application thereof, wherein the content of the rhizocortin in the yeast extract is up to 20-1000mg/L, so that the application effect of the yeast extract in the field of cosmetics is greatly improved, and the yeast strain adopts the China center for type culture Collection (CCTCCNO: M2018718), so that the content of the rhizocortin can be maximally increased, and the cost of producing the rhizocortin by yeast is effectively reduced. Meanwhile, 1, 2-pentanediol or 1, 2-hexanediol is adopted as a leaching agent in the preparation process of the extract, so that more phloretin and yeast compounds which are difficult to dissolve in water after cracking can be dissolved, the extraction rate of substances after cracking of the yeast is improved, the phloretin and the yeast compounds can be used as a cosmetic moisturizing preservative, the phloretin and the yeast extracts can be kept in the yeast extract without adverse effects, and the yeast extract is promoted to be better applied to the field of cosmetics.
Description
Technical Field
The invention relates to the technical field of phloretin, in particular to a phloretin-containing yeast extract and a preparation process and application thereof.
Background
The yeast extract (yeastextract) is a powdery, pasty or liquid product which is prepared by using fresh yeast as a raw material, inducing multiple enzyme systems in the yeast body by using modern biotechnology means or changing medium environment, adding (not adding) certain exogenous enzymes to promote degradation of intracellular proteins and nucleic acid substances under proper conditions, and performing some refining procedures. The main components of the yeast extract comprise polypeptide, amino acid, nucleotide, B vitamins, trace elements, metabolic intermediates and the like, and the existence of the nutrient components causes the yeast extract to have various efficacies. The yeast extract is an excellent natural seasoning, has wide application in the food industry, is gradually applied to the skin care product industry later, and relates to the fields of whitening, anti-aging, black eye removing, acne removing products, hair washing and care and the like. However, polypeptides, amino acids, nucleotides, etc. are mainly used as nutrients in the body, and have a limited skin-care effect.
Phloretin (Phloretin) is a novel natural skin whitening agent newly researched and developed abroad, is mainly distributed on the peel and the root bark of juicy fruits such as apples, pears and the like, and is a multifunctional flavonoid compound. The activity of flavonoid compounds against lipid peroxides has been proved in the 60 s of the 20 th century, phloretin has extremely strong inoxidizability, the inoxidizability concentration of lipid is between 10 and 30PPm, free radicals in skin can be eliminated, the free radicals are substances which are generated by oxidation reaction and are harmful to human bodies, are related to human body degenerative diseases, and can prevent saccharide components from entering epidermal cells when being externally used, so that the excessive secretion of sebaceous glands is inhibited, and acne with vigorous secretion and the like can be treated.
In addition, the phloretin has the effects of inhibiting the activity of melanocytes, fading various skin color spots, inhibiting tyrosinase well, and is a very safe and effective freckle removing and whitening agent. The phloretin with the same concentration has better inhibition effect on tyrosinase than arbutin and kojic acid which are natural components of the same kind, and when the phloretin is compounded with arbutin and kojic acid, the inhibition rate of the product on tyrosinase can be greatly improved, so that the inhibition rate reaches 100%. In addition, the phloretin is a humectant which can absorb 4-5 times of the water of the phloretin per se and a transdermal penetration enhancer which can promote the absorption and utilization of other functional factors and bring the phloretin into play with better efficacy.
At present, the domestic synthesis process of phloretin is a main way of plant extraction or chemical synthesis, although the phloretin prepared by the two ways can also reach high purity, the cost is too high, the operation is too complex in terms of plant extraction, the consumption of raw materials is too much, and the sustainable development is not facilitated.
The domestic preparation of yeast extract by using saccharomyces cerevisiae at the present stage generally comprises the following process steps: 1. and (3) an autolysis stage: the pH value of yeast milk is adjusted to about 5, and the yeast milk is maintained for 24 hours at about 55 ℃; 2. enzyme inactivation stage: raising the temperature to about 70 ℃ and maintaining the temperature for half an hour; 3. a separation stage: removing cell debris; 4. and (3) an evaporation concentration stage: evaporating extract until solid content is 40-45%, and ensuring temperature not to exceed 55 deg.C; 5. drying or concentrating stage: drying the extract until the solid content is more than 94% to obtain powder, and concentrating until the solid content is 70-75% to obtain paste. The content of phloretin in the extract obtained by the method is extremely low.
Disclosure of Invention
The first object of the present invention is to provide a yeast extract containing phloretin, which has been widely used in the field of cosmetics, in view of the above-mentioned disadvantages of the prior art.
The second purpose of the invention is to provide a preparation process of the yeast extract containing the phloretin, which is used for replacing the prior art and overcoming the technical difficulty that the content of the phloretin extracted by the prior art is too low.
In order to achieve the first purpose, the invention adopts the following technical scheme:
the yeast extract containing phloretin is characterized in that the content of the phloretin in the yeast extract is 20-1000mg/L in percentage by mass.
Furthermore, the saccharomyces cerevisiae strain used for preparing the yeast extract is China center for type culture Collection, and the strain number is CCTCC NO: M2018718.
A process for preparing a rhizocortin-containing yeast extract, comprising the steps of:
s1, collecting cells, wherein the method for collecting the yeast cells containing the phloretin comprises but is not limited to removing the fermentation liquor by one or more of centrifugation and filtration;
s2, yeast autolysis, adding pure water which is 10-100% of the volume of the original fermentation liquor into the collected cells to resuspend the thalli, adjusting the pH value to 4.5-5.0, and maintaining the temperature at 55-60 ℃ for 24 hours;
s3, performing physical crushing, wherein the thalli after autolysis are thoroughly crushed by using a physical crushing method to obtain suspension, and the physical crushing method comprises but is not limited to one or more combinations of ultrasonic and high-pressure homogenization;
s4, leaching, namely adding a leaching agent accounting for 20-30% of the volume of the original fermentation liquid into the suspension, uniformly mixing, and maintaining at 70 ℃ for 30 min;
s5, removing insoluble substances, and centrifuging or filtering the liquid obtained after leaching to remove the insoluble substances to obtain clear and transparent liquid, namely the yeast extract containing the phloretin.
Further, the leaching agent is pure water.
Further, the leaching agent is one or the combination of 1, 2-pentanediol and 1, 2-hexanediol.
Further, the method for cultivating the yeast cell containing the phloretin comprises the steps of taking a saccharomyces cerevisiae strain as a raw material, adding a fermentation culture medium and a bacterial liquid into a 30L fermentation tank, supplementing after fermenting for a certain time, adding a supplemented culture medium, adding p-hydroxyphenylpropionic acid and malonic acid as substrates for synthesizing the phloretin by yeast fermentation after the supplemented culture medium is added, and adjusting the pH of the fermentation liquid by using acid and alkali before the fermentation starts and in the fermentation process to stabilize the pH of the fermentation liquid in a certain interval so as to generate the yeast cell containing the phloretin.
Another object of the invention is the use of the yeast extract containing phloretin in the cosmetic field.
Compared with the prior art, the invention has the following beneficial effects:
1. the rhizocortin-containing yeast extract provided by the invention has the rhizocortin content as high as 20-1000mg/L, so that the application effect of the yeast extract in the field of skin care is greatly improved, and the yeast strain adopts the China center for type culture Collection, has the strain number of CCTCCNO M2018718, and can maximally increase the content of the rhizocortin;
2. the preparation process of the rhizocortin-containing yeast extract is different from the prior art in that a leaching agent is added, wherein the leaching agent can be pure water, 1, 2-pentanediol or 1, 2-hexanediol, the effect of the pure water is general, the best effect is 1, 2-pentanediol or 1, 2-hexanediol, and the effect of the pure water and the effect of the 1, 2-pentanediol or 1, 2-hexanediol are far greater than that of the pure water when the pure water and the 1, 2-pentanediol or the 1, 2-hexanediol are mixed or used independently;
3. the 1, 2-pentanediol or 1, 2-hexanediol is adopted as the leaching agent in the preparation process of the phloretin-containing yeast extract, so that more phloretin and yeast insoluble compounds after cracking can be dissolved, the leaching rate of substances after cracking of yeast is improved, and the phloretin and the yeast extract are usually used as a cosmetic moisturizing preservative and are kept in the yeast extract without causing adverse effects, so that the yeast extract is better applied to the field of cosmetics.
Detailed Description
In order to make the technical means, the creation characteristics, the achievement purposes and the functions of the invention clearer and easier to understand, the invention is further described by combining the following specific embodiments:
based on the defects brought forward by the background technology of the invention, the invention provides the yeast extract containing the phloretin, and the content of the phloretin in the yeast extract is 20-1000mg/L in percentage by mass. And the saccharomyces cerevisiae strain used for preparing the yeast extract is China center for type culture Collection, and the strain number is CCTCCNO: M2018718. The content of the phloretin is as high as 20-1000mg/L, the application effect of the yeast extract in the field of cosmetics is greatly improved, and the yeast strain adopts the China center for type culture Collection, the strain number is CCTCCNO: M2018718, the content of the phloretin can be maximally increased, and the cost of producing the phloretin by yeast is effectively reduced.
The method comprises the steps of taking saccharomyces cerevisiae capable of synthesizing phloretin through fermentation as a raw material, adding a fermentation culture medium and a bacterial liquid into a 30L fermentation tank, supplementing after fermenting for a certain time, adding a supplemented culture medium, adding p-hydroxyphenylpropionic acid and malonic acid as substrates for synthesizing phloretin through yeast fermentation after the supplemented culture medium is added, adjusting the pH of fermentation liquor by using acid and alkali before the fermentation starts and in the fermentation process to stabilize the pH of the fermentation liquor in a certain interval, finally generating phloretin-containing yeast cells, collecting the cells, then collecting the cells, and then performing autolysis, physical crushing and insoluble removal to obtain the phloretin-containing yeast extract.
Wherein the reagents for forming a 30L fermentation medium system comprise: the 10L fermentation medium system contains the following reagents: 200g of glucose, 150g of ammonium sulfate, 80g of potassium dihydrogen phosphate, 61g of magnesium sulfate heptahydrate, 20g of dropfoundation, 100mL of transcetalsolution, 25g of filter-sterilized adenine, 10mL of 2g/L of calcium chloride dihydrate, 160mL of filter-sterilized vitaminolysis and 25g of histidine.
After the fermentation medium and the bacterial liquid are added into the 30L fermentation tank, the OD600 is controlled to be between 1.5 and 3, and the volume of the mixed liquid is 10L.
And a 5L supplemented medium system is formed, wherein the supplemented medium system comprises the following reagents: 3375mL of glucose sterilized alone, 57.2% of potassium dihydrogen phosphate sterilized alone, 250mL of 18%, 25.6g of magnesium sulfate heptahydrate, dropoutpowder10g, tracemetallation 50mL, 25g of filter-sterilized adenine, 5mL of calcium chloride dihydrate, 2g/L of filter-sterilized Vitamansolution 80mL, 25g of histidine, 17.5g of potassium sulfate and 1.4g of sodium sulfate. The second supplement contains the following reagents: 3475mL of glucose sterilized alone, 53.3%, 25.6g of magnesium sulfate heptahydrate, dropoutlet powder10g, tracemetallation 50mL, 25g of filter-sterilized adenine, 5mL, 2g/L of calcium chloride dihydrate, 80mL of filter-sterilized vitaminolysis, 25g of histidine, 17.5g of potassium sulfate, and 1.4g of sodium sulfate.
Feeding is carried out according to a quasi-exponential feeding equation, and is specifically calculated according to a feeding formula.
Wherein X0Is t0Biomass at time point, g/L. V0Is t0Volume at time, L. μ set is the growth rate of yeast, h-1. F (t) feed rate, L/h. Concentration of glucose in SF feed medium, g/L. YX/S Saccharomyces cerevisiae apparent coefficient, gDCW (dry cell weight)/g glucose. m is the maintenance factor, h-1. t feed time, h. The feeding can also be carried out according to a time-interval feeding process: 10 hours after the start of feeding, 50mL/h/L, 10 to 12 hours after the start of feeding, 150mL/h/L, 12 to 20 hours after the start of feeding, 200mL/h/L, 20 to 30 hours after the start of feeding, 250mL/h/L, 30 to 64 hours after the start of feeding, 150 mL/h/L.
After the completion of the feeding, the volume of the fermentation liquid in the fermentation tank is 20L. The concentration of the solution of the p-hydroxyphenylpropionic acid and the malonic acid is 0.5 to 2g/L all the time after fermentation. The acid for adjusting the pH effect is phosphoric acid, and the alkali is ammonia water.
The preparation process of the yeast extract containing phloretin comprises the following steps:
s1, collecting cells, wherein the method for collecting the yeast cells containing the phloretin comprises but is not limited to removing the fermentation liquor by one or more of centrifugation and filtration;
s2, yeast autolysis, adding the collected cells into pure water with the volume of 10-100% of the original fermentation broth for resuspending the thallus, adjusting the pH value to 4.5-5.0, and maintaining the pH value at 55-60 ℃ for 24 hours;
s3, performing physical crushing, wherein the thalli after autolysis are thoroughly crushed by using a physical crushing method to obtain suspension, and the physical crushing method comprises but is not limited to one or more combinations of ultrasonic and high-pressure homogenization;
s4, leaching, namely adding a leaching agent accounting for 20-30% of the volume of the original fermentation liquid into the suspension, uniformly mixing, and maintaining at 70 ℃ for 30 min;
s5, removing insoluble substances, and centrifuging or filtering the liquid obtained after leaching to remove the insoluble substances to obtain clear and transparent liquid, namely the yeast extract containing the phloretin.
The preparation process of the yeast extract containing phloretin is different from the prior art in that a leaching agent is added, wherein the leaching agent can be pure water, 1, 2-pentanediol or 1, 2-hexanediol, the effect of the pure water is general, the best effect is 1, 2-pentanediol or 1, 2-hexanediol, and the effect of the pure water and the effect of the 1, 2-pentanediol or the 1, 2-hexanediol are far greater than that of the pure water when the pure water and the 1, 2-pentanediol or the 1, 2-hexanediol are mixed or used independently; meanwhile, the leaching agent adopts 1, 2-pentanediol or 1, 2-hexanediol, so that more phloretin and yeast cracked compounds which are difficult to dissolve in water can be dissolved by the phloretin and the yeast cracked compounds, the leaching rate of cracked substances of the yeast is improved, the phloretin and the yeast cracked substances are usually used as a cosmetic moisturizing preservative, the phloretin and the yeast cracked substances are kept in the yeast extract and cannot generate adverse effects, and the yeast extract is promoted to be better applied to the field of cosmetics.
Example 1
In the preparation process of the yeast extract containing phloretin, pure water is adopted as a leaching agent, and the content of the pure water is 25% of the volume of the original fermentation liquor.
Example 2
In the preparation process of the yeast extract containing phloretin, 1, 2-pentanediol is adopted as a leaching agent, and the content of the 1, 2-pentanediol is 25 percent of the volume of the original fermentation liquor.
Example 3
In the preparation process of the yeast extract containing phloretin, 1, 2-hexanediol is used as a leaching agent, and the content of the hexanediol is 25% of the volume of the original fermentation liquor.
Example 4
In the preparation process of the yeast extract containing phloretin, 1, 2-pentanediol and 1, 2-hexanediol are mixed as a leaching agent, and the ratio of the two is 1: 1, the total content of the fermentation liquor is 25 percent of the volume of the original fermentation liquor.
The pure water resuspended in the above examples 1 to 4 was 28% by volume of the original fermentation broth.
And a comparison test is carried out by adjusting the content of the leaching agent and the amount of pure water resuspended bacteria, and the specific content of the specific comparative examples is shown in the following table:
it can be seen that the content of phloretin is closely related to the type and content of the leaching agent, and the higher the content, the lower the preparation cost of the phloretin as a cosmetic raw material.
Finally, the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting, although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all of them should be covered in the claims of the present invention.
Claims (8)
1. The yeast extract containing phloretin is characterized in that the content of the phloretin in the yeast extract is 20-1000mg/L in percentage by mass.
2. The rhizocortin-containing yeast extract according to claim 1, wherein the Saccharomyces cerevisiae strain used for preparing the yeast extract is China Center for Type Culture Collection (CCTCCNO) with strain number of M2018718.
3. The process for the preparation of a rhizocortin-containing yeast extract according to claim 1 or 2, comprising the following steps:
s1, collecting cells, wherein the method for collecting the yeast cells containing the phloretin comprises but is not limited to removing the fermentation liquor by one or more of centrifugation and filtration;
s2, yeast autolysis, adding the collected cells into pure water with the volume of 10-100% of the original fermentation broth for resuspending the thallus, adjusting the pH value to 4.5-5.0, and maintaining the pH value at 55-60 ℃ for 24 hours;
s3, performing physical crushing, wherein the thalli after autolysis are thoroughly crushed by using a physical crushing method to obtain suspension, and the physical crushing method comprises but is not limited to one or more combinations of ultrasonic and high-pressure homogenization;
s4, leaching, namely adding a leaching agent accounting for 20-30% of the volume of the original fermentation liquid into the suspension, uniformly mixing, and maintaining at 70 ℃ for 30 min;
s5, removing insoluble substances, and centrifuging or filtering the liquid obtained after leaching to remove the insoluble substances to obtain clear and transparent liquid, namely the yeast extract containing the phloretin.
4. The process according to claim 3, wherein the leaching agent is purified water.
5. The process according to claim 3, wherein the leaching agent is one or a combination of 1, 2-pentanediol and 1, 2-hexanediol.
6. The process according to claim 4 or 5, wherein the amount of the pure water-resuspended bacteria is 20-30% of the volume of the original fermentation broth.
7. The process for preparing a yeast extract containing phloretin according to claim 3, wherein the method for culturing the yeast cells containing phloretin comprises the steps of taking a saccharomyces cerevisiae strain as a raw material, adding a fermentation medium and a bacterial liquid into a 30L fermentation tank, supplementing after fermenting for a certain time, adding a supplemented medium, adding p-hydroxyphenylpropionic acid and malonic acid as substrates for synthesizing phloretin by yeast fermentation after the supplemented medium is added, and adjusting the pH of the fermentation liquid by using acid and alkali before the fermentation starts and during the fermentation process to stabilize the pH of the fermentation liquid in a certain interval, thereby producing the yeast cells containing phloretin.
8. Use of a rhizocortin-containing yeast extract according to claim 1 or 2 in the cosmetic field.
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| CN109913508A (en) * | 2018-06-05 | 2019-06-21 | 嘉兴欣贝莱生物科技有限公司 | A method of phloretin is synthesized using cyanobacteria |
| CN110117550A (en) * | 2019-01-09 | 2019-08-13 | 嘉兴欣贝莱生物科技有限公司 | Technique and saccharomyces cerevisiae based on fermentation by saccharomyces cerevisiae production phloretin |
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| CN103864864A (en) * | 2014-03-27 | 2014-06-18 | 江苏斯威森生物医药工程研究中心有限公司 | Method for efficiently extracting phlorizin from plants |
| CN107586795A (en) * | 2017-10-10 | 2018-01-16 | 嘉兴欣贝莱生物科技有限公司 | A kind of method of fermentation by saccharomyces cerevisiae production phloretin |
| CN109913508A (en) * | 2018-06-05 | 2019-06-21 | 嘉兴欣贝莱生物科技有限公司 | A method of phloretin is synthesized using cyanobacteria |
| CN109180457A (en) * | 2018-08-08 | 2019-01-11 | 嘉兴欣贝莱生物科技有限公司 | A kind of process for separating and purifying of biosynthesis phloretin |
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