CN111956707A - Method for improving growth speed of saccharomyces cerevisiae and content of effective components in plants by using red light - Google Patents

Method for improving growth speed of saccharomyces cerevisiae and content of effective components in plants by using red light Download PDF

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CN111956707A
CN111956707A CN202010021634.0A CN202010021634A CN111956707A CN 111956707 A CN111956707 A CN 111956707A CN 202010021634 A CN202010021634 A CN 202010021634A CN 111956707 A CN111956707 A CN 111956707A
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plant
saccharomyces cerevisiae
skin
fermentation
concentration
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林咏翔
何政育
李韶郁
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Bayote Biotechnology Shanghai Co ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/77—Sapindaceae (Soapberry family), e.g. lychee or soapberry
    • 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • 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
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • 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
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/10—Preparation or pretreatment of starting material
    • A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
    • 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

Abstract

The invention relates to the field of plant extracts, in particular to a method for improving the growth speed of saccharomyces cerevisiae and the content of effective components in plants by using red light. The present invention provides a method for fermenting a plant, comprising the steps of: extracting a plant with water to obtain a plant extract; and sequentially fermenting the plant extract with saccharomyces cerevisiae, germ lactobacillus and acetic acid bacteria to obtain a plant fermented product, wherein the fermentation process comprises irradiating the plant extract, the saccharomyces cerevisiae, the germ lactobacillus and the acetic acid bacteria with a red light source with the wavelength of 620-750 nm. The method can be used for increasing the growth speed of the saccharomyces cerevisiae and the content of effective components in the plants. The invention also discloses a plant fermentation product prepared by the method, and the fermentation product can be used for preparing a composition for reducing skin fat.

Description

Method for improving growth speed of saccharomyces cerevisiae and content of effective components in plants by using red light
Technical Field
The invention relates to the field of plant extracts, in particular to a method for improving the growth speed of saccharomyces cerevisiae and the content of effective components in plants by using red light.
Background
Fat is an essential component in the human body, but excessive fat components cause damage to the human body. In countries with increasing national economy, the obesity problem is only increased but not reduced, so that in view of Asian regions, China can be a country with the increased amplitude of the next wave of obesity problem. Obesity should become a problem that needs to be solved at present.
In addition, glycation (glycation) is associated with skin and in vivo aging, and is more associated with diabetes which is a health hazard to people. The nature of the saccharification reaction is known as the Maillard reaction or carbonyl-amine browning, which is a non-enzymatic saccharification of the aldehyde (ketone) group of a sugar with an amino group-containing substance (e.g., a protein, peptide, amino acid, phospholipid, nucleic acid, or derivative thereof). Prolonged exposure to hyperglycemic conditions can lead to glucose autooxidation and protein glycation, leading to the formation of advanced glycation end products (AGEs). AGEs can cause various aging diseases such as skin wrinkles, cataract, atherosclerosis, kidney failure, etc.
The skin tissue is composed of epidermis, dermis and hypodermis, wherein the dermis contains a large amount of collagen and hyaluronic acid, and is closely related to the water retention and elasticity of the skin. The human skin can have the phenomena of oxidation, aging, rough skin or wrinkle generation and the like along with age, physiological factors or environmental factors, for example, the skin of normal young people has certain elasticity and tension, and when the expression muscle is relaxed, the skin can be quickly recovered to eliminate the wrinkle; but in the middle age, the skin begins to age obviously, becomes thin, hard, dry and has reduced tension; dermal collagen is reduced, elastic fibers are denatured and broken, so that the tension and elasticity of the skin are reduced, and therefore, the skin cannot recover quickly after the expression muscles are relaxed, and wrinkles are formed after the skin relaxes for a long time; and with the increase of age, the skin and subcutaneous tissues are more relaxed, and in addition, the atrophy or the loss of the facial supporting tissues and the softness of muscles, the skin slides and falls under the action of gravity to form deeper wrinkles. The rough skin is a skin trouble caused by an external important factor such as dryness, ultraviolet rays, an irritant substance such as a detergent or a chemical substance, or an internal important factor such as disturbance of hormone balance, and is accompanied by phenomena such as reduction of barrier function of stratum corneum, reduction of moisture content of stratum corneum, acceleration of epidermal turnover, and roughening of cutin caused by generation of scales. Therefore, if the cells on the skin lose their elasticity and moisture-retaining function, they may cause wrinkles, dryness and loss of luster.
The production of fermented food always relies on microorganisms which are inoculated into raw materials to produce required enzymes in the growth process, and the process of decomposing organic matters into beneficial substances in human life is called fermentation; so that the enzyme produced by the microorganism actually catalyzes the metabolic activity. The range of products obtained by fermentation is very wide, and the products range from pharmaceutical-grade products with very high unit price, such as hypolipidemic drugs or antibiotics, to industrial enzymes, such as amylolytic, cellulolytic, proteolytic enzymes and the like; also useful microorganisms for food, such as lactic acid bacteria, monascus, bacillus natto, edible and medicinal mushroom, etc., seasoning of monosodium glutamate, biological pesticide for agriculture, such as bacillus thuringiensis, or microorganisms of antagonistic bacteria such as bacillus subtilis, actinomycetes, trichoderma, etc.; further, the enzyme may be any of microorganisms added to animal feeds or directly added to feeds, microorganisms used for waste recovery, various pollution treatments and cleaning, and other special enzymes. In particular, in recent years, researchers have begun to perform fermentation treatment on plants (such as vegetables and fruits), and the obtained plant fermentation products can be used for improving human health. Accordingly, there has been an increasing demand for fermented foods.
However, conventional fermentation processes for producing fermentation products often require a significant amount of time for the microorganisms to grow and require industrial-scale fermentation equipment, such as fermentation tanks, which is very costly. Therefore, if a novel method for fermenting plants can be developed and can be used to increase the growth rate of saccharomyces cerevisiae and the content of effective components in plants, it would bring a great breakthrough to the art and benefit the wide population in need.
Disclosure of Invention
In view of the above, it is an object of the present invention to provide a method for fermenting plants, comprising the steps of: (a) extracting a plant with water to obtain a plant extract; and (b) sequentially performing a fermentation process on the plant extract, a Saccharomyces cerevisiae (Saccharomyces cerevisiae), a Lactobacillus plantarum (Lactobacillus plantarum) and an acetic acid bacterium (Acetobacter aceti) to obtain a plant fermentation product, wherein the fermentation process comprises irradiating the plant extract, the Saccharomyces cerevisiae, the Lactobacillus plantarum and the acetic acid bacterium with a red light source with a wavelength of 620-750 nm.
In one embodiment of the invention, the plant is rambutan (Nephelium lappacium).
In one embodiment of the present invention, prior to the extraction, a glucose having a concentration of 10% (w/w) based on the total weight is added.
In one embodiment of the present invention, the fermentation time of the saccharomyces cerevisiae is 1 to 2.5 days, the fermentation time of the lactobacillus plantarum is 1 to 3 days, and the fermentation time of the acetic acid bacterium is 3 to 10 days.
In an embodiment of the present invention, the concentration of the brewer's yeast is between 0.01-0.5% (v/v), the concentration of the germ lactic acid bacteria is between 0.01-0.25% (v/v), and the concentration of the acetic acid bacteria is between 3-10% (v/v).
In an embodiment of the present invention, the weight ratio of the plant to water is 1: 5-10.
In one embodiment of the present invention, in step (a), the temperature of the extraction is between 50 and 100 ℃, and the time of the extraction is between 0.5 and 1.5 hours.
Another object of the present invention is to provide a method for increasing the growth rate of Saccharomyces cerevisiae and the content of effective components in plants.
In one embodiment of the invention, the plant is rambutan (Nephelium lappacium) and the active ingredient is a polyphenol (polyphenol).
Another object of the present invention is to provide a plant fermentation product, which is prepared by the method of the above technical scheme.
Another object of the present invention is to provide a use of the plant fermentation product as described above for preparing a skin-care and fat-reducing composition.
In one embodiment of the present invention, the skin care includes enhancing the antioxidant capacity of the skin, enhancing the anti-glycation activity of the skin, and promoting elastin proliferation.
In one embodiment of the invention, the fat reduction includes promoting lipolysis and increasing leptin secretion.
In one embodiment of the present invention, the composition is in the form of a pharmaceutical, a nutraceutical, or a food product.
It is another object of the present invention to provide a method for increasing the growth rate of microorganisms, comprising the steps of: (a) extracting a plant with water to obtain a plant extract; and (b) performing a fermentation process on the plant extract and a microorganism to obtain a plant fermentation product, wherein the microorganism is Saccharomyces cerevisiae (Saccharomyces cerevisiae) or Acetobacter aceti (Acetobacter aceti), and the fermentation process comprises irradiating the plant extract and the microorganism with a red light source with a wavelength of 620-750 nm.
In one embodiment of the invention, the plant is rambutan (Nephelium lappacium).
In one embodiment of the present invention, prior to the extraction, a glucose having a concentration of 10% (w/w) based on the total weight is added.
In one embodiment of the present invention, the fermentation time of the microorganism is 1 to 3 days.
In one embodiment of the present invention, the concentration of the brewer's yeast is between 0.01-0.5% (v/v), and the concentration of the acetic acid bacteria is between 5% (v/v).
In one embodiment of the present invention, the growth rate of the brewers' yeast is increased by at least 47% and the growth rate of the acetic acid bacteria is increased by at least 22%.
In summary, the method for fermenting plants and the plant fermentation product prepared by the method of the present invention have the following effects: can increase the growth rate of microorganisms (such as Saccharomyces cerevisiae and Acetobacter) and the content of effective components in plants, and can improve the antioxidant capacity of skin, improve the anti-saccharification activity of skin, and promote the proliferation of elastin to achieve the effects of caring skin and beautifying skin. On the other hand, the plant fermentation product can achieve the effect of reducing fat by promoting lipolysis and increasing leptin secretion.
The following examples are presented to illustrate the present invention and are not to be construed as limiting the scope of the invention, which is intended to be limited only by the appended claims.
Drawings
FIG. 1 is a graph of data from a microorganism growth rate test;
figure 2 is a data plot of the analysis of the content of active ingredients in plants, wherein p represents <0.001 compared to the control group;
FIG. 3 is a data plot of the effectiveness of the plant fermentation product prepared by the method of the present invention in enhancing the antioxidant capacity of skin, wherein p is <0.05 as compared to a control;
FIG. 4 is a data plot of the efficacy of the plant fermentations prepared according to the present invention in enhancing anti-glycation activity of the skin, wherein p is <0.05 compared to a control;
FIG. 5 is a graph of data showing the efficacy of the plant fermentation product of the present invention in promoting elastin proliferation;
FIGS. 6A and 6B are cell staining graphs showing the effect of the plant fermentation product prepared by the method of the present invention on promoting elastin proliferation, wherein the dots represent the nucleus and the rest represent elastin;
FIG. 7 is a graph of data showing the efficacy of the plant fermentations made by the method of the present invention in promoting lipolysis, wherein p is <0.05 compared to a control;
FIG. 8 is a data plot of the utility of plant fermentations made by the present invention in increasing leptin secretion;
FIG. 9 is a data plot showing the efficacy of the method for increasing the growth rate of microorganisms of the present invention in increasing the growth rate of Saccharomyces cerevisiae.
Detailed Description
Definition of
As used herein, the numerical values are approximations and all numerical data are reported to be within the 20 percent range, preferably within the 10 percent range, and most preferably within the 5 percent range.
Statistical analysis was performed using Excel software. Data are presented as mean ± Standard Deviation (SD), and differences between individual data are analyzed by student's t-test (student's t-test).
According to the present invention, rambutan (Nephelus lappacium) is a large tropical fruit tree of the genus Nephelus (Nephelus) of the family Sapindaceae (Sapindaceae), called "rambutan" in Malaysia, which means "hairy antler". The ripe rambutan fruits are not all red, but have yellow fruits. Some rambutan has a nucleus similar in size to sesame. Rambutan has a taste similar to that of litchi, white pulp (aril), transparent and juicy pulp, sweet and sour taste, and contains 1 seed and is flat and oval.
As used herein, the terms "Saccharomyces cerevisiae", "Lactobacillus plantarum" and "Acetobacter aceti" are intended to encompass Saccharomyces cerevisiae, Lactobacillus plantarum and Acetobacter aceti, respectively, which are readily available to those skilled in the art (e.g., from domestic or foreign depositories), or those isolated and purified from natural sources using microbial isolation methods customary in the art.
In accordance with the present invention, the drug may be manufactured using techniques well known to those skilled in the art into a dosage form suitable for parenteral (parenteral), oral (oral) or topical (topically) administration, including, but not limited to: injections (injections) [ for example, sterile aqueous solution (sterile aqueous solution) or dispersion (dispersion) ], sterile powder (sterile powder), troche (tablet), tablet (troche), buccal tablet (dosage), pill (pill), capsule (capsule), dispersible powder (dispersible powder) or granule (granule), solution, suspension (suspension), emulsion (emulsion), syrup (syrup), elixir (elixir), syrup (slurry), external preparation (external preparation), and the like.
According to the present invention, the pharmaceutical may further comprise a pharmaceutically acceptable carrier (pharmaceutically acceptable carrier) which is widely used in pharmaceutical manufacturing technology. For example, the pharmaceutically acceptable carrier may comprise one or more agents selected from the group consisting of: solvents (solvent), buffers (buffer), emulsifiers (emulsifying), suspending agents (suspending agent), disintegrating agents (disintegrant), disintegrating agents (disintegrating agent), dispersing agents (dispersing agent), binding agents (binding agent), excipients (excipient), stabilizers (stabilizing agent), chelating agents (chelating agent), diluents (diluent), gelling agents (gelling agent), preservatives (preserving), wetting agents (wetting agent), lubricants (lubricating), absorption delaying agents (absorption delaying agent), liposomes (liposome) and the like. The selection and amounts of such agents are within the skill and routine skill of those skilled in the art.
According to the present invention, the pharmaceutically acceptable carrier comprises a solvent selected from the group consisting of: water, normal saline (normal saline), Phosphate Buffered Saline (PBS), aqueous alcohol-containing solutions (aqueous solution linking alcohol), and combinations thereof.
According to the invention, the medicament may be administered by a parenteral route (parenteral routes) selected from the group consisting of: intraperitoneal injection (intraperitoneal injection), subcutaneous injection (subcutaneous injection), intraepidermal injection (intraepithelial injection), intradermal injection (intraepithelial injection), intramuscular injection (intramucosal injection), intravenous injection (intravenous injection) and intralesional injection (intrafocal injection).
According to the present invention, pharmaceuticals can be manufactured into an external preparation (external preparation) suitable for topical application to the skin using techniques well known to those skilled in the art, including, but not limited to: creams (lotions), liniments (liniments), powders (powders), aerosols (aerogels), sprays (sprays), emulsions (positions), serums (serums), pastes (pastes), foams (foams), drops (drops), suspensions (suspensions), ointments (salves), and bandages (bandages).
According to the present invention, the external preparation is prepared by mixing the medicine of the present invention with a base (base) as well known to those skilled in the art.
According to the invention, the substrate may comprise one or more additives (additives) selected from the following group: water, alcohols, glycols, hydrocarbons such as petroleum jelly (jelly) and white petrolatum]Wax (wax) [ such as Paraffin and yellow wax (yellow wax)]Preserving agents (preserving agents), antioxidants (antioxidants), surfactants (surfactants), absorption enhancers (absorption enhancers), stabilisers (stabilizing agents), gelling agents (gelling agents) [ such as
Figure BDA0002360982460000071
974P(
Figure BDA0002360982460000072
974P), microcrystalline cellulose (microcrystalline cellulose) and carboxymethyl cellulose (carboxymethyl cellulose)]Active agents (active agents), humectants (humectants), odor absorbers (odor absorbers), fragrances (fragrans), pH adjusting agents (pH adjusting agents), chelating agents (chelating agents), emulsifiers (emulsifiers), occlusive agents (occluvium)e-agents), emollients (emulsifiers), thickeners (thinners), solubilizing agents (solvabilizing agents), penetration enhancers (penetration enhancers), anti-irritants (anti-irritants), colorants (colorants), and propellants (propellants), among others. The selection and amounts of such additives are within the skill and routine skill of those skilled in the art.
According to the present invention, the care product may further comprise an acceptable adjuvant (acceptable adjuvant) which is widely used in the art of care product manufacture. For example, the acceptable adjuvant may comprise one or more agents selected from the group consisting of: solvents, gelling agents, active agents, preservatives, antioxidants, screening agents, chelating agents, surfactants, colouring agents, thickening agents, fillers, fragrances and odour absorbers. The selection and amounts of such agents are within the skill and routine skill of those skilled in the art.
In accordance with the present invention, the cosmetic may be manufactured in a form suitable for skin care (skincare) or makeup (makeup) using techniques well known to those skilled in the art, including, but not limited to: aqueous solutions (aqueous solutions), aqueous-alcoholic solutions (aqueous-alcoholic solutions) or oily solutions (oil solutions), emulsions in the form of oil-in-water type, water-in-oil type or compound type, gels, ointments, creams, masks (masks), patches, wipes, powders, aerosols, sprays, lotions, serums, pastes, foams, dispersions, drops, mousses (mousses), sunblocks, lotions (toiletries), foundations (foundations), make-up removal products (make-up removal products), soaps (soaps) and other body cleansing products (body cleansing products), and the like.
In accordance with the present invention, the cosmetic may also be used in combination with one or more known active topical agents (external use agents) selected from the following: whitening agents (whitening agents) [ such as retinoic acid (tretinoin), catechins (catechin), kojic acid, arbutin and vitamin C ], moisturizers, anti-inflammatory agents (anti-inflammatory agents), bactericides (bacteriodes), ultraviolet absorbers (ultraviolets absorbers), plant extracts [ such as aloe vera extract (aloe vera extract) ], skin nutrients (skin nutrients), anesthetics (anesthesics), anti-acne agents (anti-acne agents), antipruritics (antipruritics), analgesics (analgesics), anti-dermatitis agents (antipermatitis agents), anti-hyperkeratotic agents (anti-hypercholesterolitic agents), anti-dry skin agents (anti-dry skin agents), anti-perspirants (anti-perspirant agents), anti-aging agents (anti-aging agents), anti-wrinkle agents (anti-rinking agents), anti-seborrheic agents (anti-anerrheic agents), wound healing agents (wound-healing agents), corticosteroids (corticosteriods), and hormones (hormones). The selection and amounts of such agents for external use are within the skill and routine skill of those skilled in the art.
According to the present invention, the food product may be used as a food additive (food additive) to be added during the preparation of the raw material or during the preparation of the food by conventional methods, and formulated with any edible material into a food product for ingestion by humans and non-human animals.
According to the present invention, the types of food products include, but are not limited to: beverages (leafages), fermented foods (fermented foods), bakery products (bakery products), health foods (health foods) and dietary supplements (dietary supplements).
Example 1 detection of growth Rate of microorganisms
First, rambutan (Nephelium lappacium) juice from roxburgh city in south thailand was obtained at a ratio of 1: 5-10 wt% of water, and adding glucose with a concentration of 10% (w/w) based on the total weight. Then, extracting for 0.5-1.5 hours at 50-100 ℃ to obtain a plant extract.
Then, after the culture medium of the plant extract is inoculated with Saccharomyces cerevisiae (Saccharomyces cerevisiae) BCRC 20271 to start fermentation, 1mL of bacterial liquid is taken out every 24 hours, and after sequence dilution, the bacterial liquid is coated on a Yeast Peptone Dextrose (YPD) plate, wherein red LED light sources with the wavelength of 620-750 nm (preferably 635nm) are irradiated in the whole fermentation process, and the plant fermentation product irradiating the red LED light sources is obtained. Then, the plant fermentation product irradiated with the red LED light source was used as an experimental group, and the preparation method of the plant fermentation product of the comparative group was substantially the same as that of the experimental group except that: replacing the red LED light source with blue light with the wavelength of 400-450 nm; the preparation method of the plant fermentation product for the control group was substantially the same as that of the experimental group except that: natural light replaces the red LED light source. Subsequently, YPD plates coated with the inoculum solution were placed in an incubator at 30 ℃ for 48 hours, and then colonies were counted to evaluate the growth rate of Saccharomyces cerevisiae. The results of the experiment are shown in FIG. 1.
FIG. 1 is a graph of data for the detection of growth rate of microorganisms. As can be seen from FIG. 1, the growth rate of Saccharomyces cerevisiae was significantly increased in the experimental group compared to the control group and the comparative group. Particularly, in the fermentation process, the red LED light source with the irradiation wavelength of 620-750 nm can increase the growth rate of the saccharomyces cerevisiae by 47% (compared with a control group), and on the contrary, the effect of increasing the growth rate is not achieved when natural light and blue light are irradiated. The results of this example show that illuminating red LED light source in the process of fermenting plants can effectively increase the growth rate of saccharomyces cerevisiae.
Example 2 analysis of content of effective component in plant
First, a standard solution was prepared, 10g of gallic acid (gallic acid) was dissolved in water and a volume of 10mL was added to the measuring flask. Next, 0. mu.L/mL, 20. mu.L/mL, 40. mu.L/mL, 60. mu.L/mL, 80. mu.L/mL, and 100. mu.L/mL of the standard solutions were prepared, and 100. mu.L of each standard solution was taken into a centrifuge tube having a volume of 10 mL. Thereafter, 500. mu.L of Folin-Ciocalteu's phenol reagent was added, mixed and allowed to stand for 3 minutes, and then 400. mu.L of 7.5% sodium carbonate (sodi. mu.M carbonate) was added, mixed and allowed to stand for 30 minutes. Next, 200. mu.L of each reaction solution was transferred to a 96-well plate, and the absorbance was measured at 750 nm.
In addition, the plant fermentation product irradiated with the red LED light source described in example 1 was used as an experimental group, and the preparation method of the plant fermentation product of the control group was substantially the same as the experimental group, except that: natural light replaces the red LED light source. The experimental and control groups were diluted five times with water and 100mL volumes were taken into microcentrifuge tubes. Thereafter, 500. mu.L of the foscarnet reagent was added, mixed and allowed to stand for 3 minutes, and then 400. mu.L of 7.5% sodium carbonate was added, mixed and allowed to stand for 30 minutes. Next, 200. mu.L of each set of the reaction solution was transferred to a 96-well plate, and the absorbance was measured at 750 nm. The results for total polyphenol content are shown in figure 2.
FIG. 2 is a data diagram of the analysis of the content of effective components in plants. As can be seen from fig. 2, the total polyphenol content in the experimental group was significantly increased (increased by 60%) compared to the control group. The results of this example show that irradiation of a red LED light source during plant fermentation can effectively increase the content of active ingredients (i.e., total polyphenols) in the plant (i.e., rambutan).
Example 3 procedure for the method of the invention for fermenting plants
Based on the results of example 1 and example 2, the present inventors decided to prepare a plant fermentation product using a red LED light source. First, rambutan (Nephelium lappacium) juice from roxburgh city in south thailand was obtained at a ratio of 1: 5-10 wt% of water, and adding glucose with a concentration of 10% (w/w) based on the total weight. Then, extracting for 0.5-1.5 hours at 50-100 ℃ to obtain a plant extract. Cooling the plant extract to room temperature for subsequent three-stage fermentation. The three-stage fermentation is to sequentially inoculate 0.01-0.5% (v/v) Saccharomyces cerevisiae BCRC 20271 with the plant extract, and ferment for 1-2.5 days; inoculating 0.01-0.25% (v/v) of Lactobacillus plantarum (TCI 028) (deposited in biological resource preservation and research center of food industry development research institute in 2017, 12 and 13 days, deposited with number BCRC910805) to ferment for 1-3 days; inoculating 3-10% (v/v) acetic acid bacteria (Acetobacter aceti) BCRC 11688 (all the strains are purchased from the Center for biological resource preservation and Research (BCRC) of Food Industry Development Institute (FIRDI) in Taiwan), fermenting for 3-10 days, and irradiating red LED light source with wavelength of 620-750 nm (preferably 635nm) in the whole fermentation process. Then, the mixture is concentrated under reduced pressure at 45 to 70 ℃ and filtered through a 200 to 400 mesh screen. And then, adding 40-70% of isomalto-oligosaccharide, and sterilizing to obtain the plant fermentation product.
Example 4 evaluation of the effectiveness of the fermented product of plants obtained by the method for fermenting plants according to the present invention in enhancing the antioxidant ability of skin
It is reported that after age 35, the antioxidant capacity of human body is greatly reduced, a large amount of free radicals are accumulated, and skin is rapidly aged. Therefore, this example discusses whether the plant fermentation product prepared by the method of example 3 can improve the antioxidant ability of the skin. Using a colorimetric method for reducing iron oxidation capacity (FRAP) to determine the oxidation resistance of the substance to be detected; wherein iron ions (Fe) can be converted by using a specimen having a reducing activity3+) Reduction to ferrous ion (Fe)2+) The color of the solution is changed from reddish brown to green, and the shade of the green compound is in direct proportion to the reduction capability, so that the method can be used for quantifying the reduction activity of the substance to be detected.
First, a calibration curve was prepared using ascorbic acid known to have reducing activity. Precisely weighing 10mL of L-Ascorbic acid (L-Ascorbic acid, Vit C, vitamin C available from Sigma, USA, No. A5960-25g), placing in a 10mL volumetric flask, adding ddH2O was quantified to 10mL, and 1mg/mL of L-ascorbic acid was prepared. Then, 1mL of the prepared L-ascorbic acid was placed in a 10mL volumetric flask, and ddH was added thereto2O was quantified to 10mL to prepare 1. mu.g/mL of L-ascorbic acid, and 100. mu.g/mL of L-ascorbic acid was serially diluted to 0, 10, 20, 40, 60, 80, and 100. mu.L/mL of L-ascorbic acid (as shown in Table 1), 250. mu.L of each concentration of standard solution was added to each test tube, and 250. mu.L of phosphate buffer solution (anhydrous sodium dihydrogen phosphate (NaH) was added thereto2PO4From J.T.Baker, No. 3828-01) and disodium hydrogen phosphate (Na)2HPO4Purchased from Sigma, No. 04270) at a ratio of 1: 1) was mixed well with a shaker (Vortex) and 250 μ L of 1% hematite (Potassium ferricyanide, K) was added3Fe (CN), purchased from Sigma, No. 244023), was mixed well with a shaker and then subjected to a 50 ℃ water bathThe cells were heated for 20 minutes and 250. mu.L of 10% Trichloroacetic acid (TCA, CCl) was added3COOH, from J.T. Baker, No. 0414-01) was mixed well with a shaker, centrifuged at 300g for 10 minutes, and taken out without shaking, then 300. mu.L of the supernatant of each tube of the reaction solution was taken out, and 300. mu.L of ddH was added2O and 120. mu.L of 1% ferric chloride (FeCl)3From Alfa Aesar, No. a16231) was mixed uniformly with a shaker and reacted for 10 minutes, and the absorbance at 700nm was measured to plot the regression curve equation of the standard solution.
TABLE 1 formulation of serially diluted L-ascorbic acid standards
Figure BDA0002360982460000111
Then, the plant fermentation product obtained in example 3 was used as an experimental group, and the plant fermentation product of the comparative group was prepared in a substantially same manner as that described in example 3 for fermenting plants, except that: natural light replaces the red LED light source. In addition, the plant extract of example 3 was used as a control group. The subsequent experiments were performed after 20-fold dilution of each group of samples. Then, 250. mu.L of each sample was put in 10mL glass test tubes, 250. mu.L of phosphate buffer solution was added and mixed uniformly by a shaker, 250. mu.L of 1% hematite was added and mixed uniformly by a shaker, then the mixture was heated in a 50 ℃ water bath for 20 minutes, 250. mu.L of 10% trichloroacetic acid was added and mixed uniformly by a shaker, and the mixture was centrifuged at 300g for 10 minutes with taking out the supernatant of 300. mu.L of each reaction solution, and 300. mu.L of ddH was added2O and 120 mu L of 1% ferric chloride are uniformly mixed by a shaker, then the mixture reacts for 10 minutes, the light absorption value of the mixture at 700nm is measured, the concentration is calculated by an interior difference method according to the regression curve formula of the standard solution, and then the reduction capacity of each group of samples is obtained by multiplying the dilution times. The results of this example are shown in FIG. 3.
FIG. 3 is a data diagram showing the effect of the plant fermentation product prepared by the method of the present invention on improving the antioxidant ability of skin. As can be seen from fig. 3, the reduction capacity of the experimental group was significantly improved compared to the control group and the comparative group. In particular, the reduction capacity of the experimental group was improved by 57% compared to the control group. The results of this example show that the plant fermentation product prepared by the method for fermenting plants of the present invention can effectively improve the oxidation resistance of skin.
Example 5 evaluation of the Effect of the plant fermentation product obtained by the method for fermenting plants according to the present invention on the enhancement of the anti-glycation Activity of skin
In this example, in order to test the anti-glycation activity of the fermented plant material obtained by the method for fermenting plants according to the present invention, the glycation activity was quantified by the efficiency of inhibiting the glycation of Collagen (Collagen) by D-fructose (D-fructose). Sugar molecules cross-link with proteins to form Advanced glycation end-products (AGEs), which induce intracellular oxidative stress and cause skin aging damage. First, the plant fermentation product prepared in example 3 was used as an experimental group, and the preparation method of the plant fermentation product of comparative group 1 was substantially the same as that for fermenting plants described in example 3, except that: the preparation method of comparative group 2, which replaces the red LED light source with natural light, differs from the method for fermenting plants described in example 3 in that: the plant extract obtained by water extraction was not subjected to three-stage fermentation and red LED light irradiation. The control method was not illuminated and no plant fermentate or extract was added. The subsequent experiments were performed after diluting each group of samples 10-fold. Thereafter, 0.2mL of a solution containing 0.06% NaN was added3And 0.2mL of a 1.5M D-fructose (200 mM sodium phosphate buffer, pH 7.4) solution, taking out 0.1mL of the mixed solution as an origin product, reacting the remaining mixed solution at 50 ℃ for 24 hours, taking out 0.1mL of the mixed solution as an end product, and measuring the fluorescence values of the origin product and the end product at an excitation wavelength of 360nm and an emission wavelength of 460nm, respectively, wherein Aminoguanidine (Aminoguadine, AG) is known to have an effect of inhibiting glycation. Finally, the elimination of high glycation end is calculated by the following formulaEfficiency of product (AGEs) capacity to represent its anti-glycation activity, wherein less production of highly glycated end products indicates higher anti-glycation activity.
Figure BDA0002360982460000121
The results of this example are shown in FIG. 4. FIG. 4 is a data graph showing the efficacy of the plant fermentation product prepared by the method of the present invention in enhancing the anti-glycation activity of skin. As can be seen from fig. 4, the percentage of glycation ratio in the experimental group was significantly reduced compared to the control group, comparative group 1 and comparative group 2. Wherein the percent glycation ratio of the experimental group is reduced by about 55% compared to the control group. The results of this example show that the plant fermentation product prepared by the method for fermenting plants according to the present invention can effectively improve the ability of removing high-degree glycation end products, and has excellent anti-glycation activity for improving skin.
Example 6 evaluation of the efficacy of the fermented product of plants obtained by the method for fermenting plants according to the present invention in promoting proliferation of elastin (elastin)
In the skin, the elastin is mutually matched with the collagen, which can help support the framework of the skin, improve the elasticity of the skin, strengthen the compactness of the skin and reduce the generation of wrinkles. Therefore, this example discusses whether the plant fermentation product prepared according to the method of example 3 can promote elastin proliferation. First, the plant fermentate prepared in example 3 (0.0625% concentration) was used as an experimental group, and the preparation method of the plant fermentate of the comparative group was substantially the same as the method for fermenting plants described in example 3, except that: natural light replaces the red LED light source. The control method was not illuminated and no plant fermentate or extract was added.
Next, the elastin (elastin) content analysis is performed, the process is as follows: use of FastinTMElastin Assay kit (Elastin Assay kit) inoculates 1X 10 with 2mL of medium in a 6-well plate5The cell line used is human dermal fibroblast CCD-966sk (
Figure BDA0002360982460000131
Number CRL-1881), Minimum Essential Medium (MEM) supplemented with 10% FBS, 1% Penicillin/streptomycin, and 1mM sodium pyruvate (Gibco), and incubated at 37 ℃ for 24 hours. Thereafter, each group of samples was treated with 2mL of the medium and then acted upon for 48 hours. Next, the medium was removed and washed once with 1X PBS, and then the cells were treated with trypsin (trypsin) for 3 minutes. Trypsin activity was stopped with medium and then transferred to a 1.5mL tube. Next, the cells were collected with trypsin and then transferred to a 1.5mL microcentrifuge tube, centrifuged at 300Xg for 5 minutes, and then washed with PBS (Gibco). After this time, the cell pellet was centrifuged at 300Xg for 5 minutes and then retained in about 300. mu.L of PBS. Next, 100. mu.L of 1.0M oxalic acid (oxalic acid) was added to 300. mu.L of the cell suspension and allowed to act in a heating block at 100 ℃ for 1 hour. An equal volume of Elastin precipitation Reagent (Elastin Precipitating Reagent) was added to each tube, the tubes were capped and vortexed briefly and allowed to sit for 15 minutes to complete the precipitation. Thereafter, the liquid contents of the tube were drained by centrifugation at 10,000Xg for 10 minutes, and then the majority of the fluid remaining in the tube was removed by gently tapping the inverted tube onto a paper towel. Next, 1mL of dye reagent was added, the contents were mixed by inverting the tube, and then the tube was placed on a mechanical shaker for 90 minutes. After emptying the unbound dye tube, the elastin-dye complex can be observed as a reddish brown deposit in the bottom of the test tube and in the inner lower wall. To each tube was added 250 μ L of Dye Dissociation Reagent (Dye Dissociation Reagent), the tube was capped and the Dye was released into solution by vortex mixer to ensure that all bound Dye had entered the solution. Thereafter, the contents of each tube were transferred to a 96-well plate, and the plate was placed in an ELISA reader (BioTek) and absorbance was measured at 513 nm.
Next, elastin immunofluorescence staining was performed, the procedure was as follows: inoculation of 5X 10 per well on chamber slides3Human dermal fibroblast CCD-966sk (BCRC code)60153), Minimum Essential Medium (MEM) (Eagle) (Eaglle in Erle's Balanced Salt Solution, Earle's BSS, GIBCO corporation, No. 41500-. Thereafter, cells were fixed with 4% paraformaldehyde (parafumaldehyde) for 10 minutes at room temperature, followed by 3 washes with 1 × PBS, and then 0.2% Triton X-100 (formulated in 1 × PBS (Gibco)) was added to penetrate the cells and allowed to act for 10 minutes at room temperature. Next, 1% BSA (in 1 XPBS) was added, blocked at 37 ℃ for 1 hour, and then washed 3 times with 1 XPBS. Thereafter, a primary antibody (elastin antibody (Merck; MAB2503), diluted with 1% BSA, acted at 37 ℃ for 1 hour) was added and washed 3 times with 1 XPBS. Subsequently, a secondary antibody (anti-mouse-alexa 488 antibody (Thermo), diluted with 1% BSA, acted at 37 ℃ for 1 hour) was added and washed 3 times with 1X PBS. Thereafter, Hoechst 33342(Thermo) was added and allowed to act at room temperature for 3 to 5 minutes, followed by washing 3 times with 1 XPBS. Next, the slide glass was fixed with a fixing gel (mounting gel) and observed under a fluorescent microscope. The results of this example are shown in fig. 5 and fig. 6A and 6B.
FIG. 5 is a data graph showing the efficacy of the plant fermentation product prepared by the method of the present invention in promoting elastin proliferation. FIGS. 6A and 6B are cell staining charts showing the efficacy of the plant fermentation product prepared by the method of the present invention in promoting elastin proliferation. As can be seen from fig. 5, the elastin content in the experimental group is significantly increased compared to the control group and the comparative group. Wherein the elastin content in the experimental group is increased by 13% compared with the control group. As can be seen from FIGS. 6A and 6B, the cell staining pattern of the efficacy of the plant fermentation product obtained by the method of the present invention in promoting elastin proliferation is shown, wherein the dots represent the nucleus and the rest represent elastin. Fig. 6A is the control group of fig. 5, and fig. 6B is the experimental group of fig. 5, and the observation result under the fluorescence microscope shows that the elastin content is more under the action of the plant fermentation product, and the elastin content is improved by 13% as shown in fig. 5 after the data. The results of this example show that the plant fermentation product prepared by the method for fermenting plants of the present invention can effectively promote the proliferation of elastin.
Example 7 evaluation of the effectiveness of the fermented product of plants obtained by the method for fermenting plants according to the present invention in promoting lipolysis
This example was conducted by using mouse bone marrow stromal cells to test the efficacy of the fermented plant material prepared by the method for fermenting plants of the present invention in promoting lipolysis. The mouse bone marrow stromal cells OP9 were purchased from American type culture Collection (American type culture Collection)
Figure BDA0002360982460000151
) Number CRL-2749TM. The cells were cultured in preadipocyte Expansion Medium (Pre-adipocyte Expansion Medium) prior to differentiation, containing 90% minimal essential Medium Alpha Medium (minimum essential Medium Alpha Medium, available from Gibco, usa, 12100-; and mouse bone marrow stromal cells were differentiated using Differentiation Medium comprising 90% minimal essential Medium Alpha Medium, 20% fetal bovine serum, and 1% penicillin/streptomycin.
To confirm that the fermented plant material obtained by the method for fermenting plants according to the present invention has the effect of promoting lipolysis, mouse bone marrow stromal cells were first differentiated into adipocytes (adipocyte) by differentiating 8X 104Mouse bone marrow stromal cells OP9 were seeded in 24-well culture plates with 500. mu.L of preadipocyte Expansion medium and cultured at 37 ℃ for 7 days with replacement of fresh differentiation medium every 3 days. Next, the formation of lipid droplets (lipid droplets) was observed using a microscope (ZEISS) to ensure that the cells had fully differentiated.
OP9 cells were then divided into 3 groups, including 1 control group, 1 comparative group, and 1 experimental group. The plant fermentate obtained according to the method described in example 3 was added to the cells of the experimental group at a concentration of 0.0625%. The preparation of the comparative cell-supplemented material was substantially the same as described in example 3 for the fermentation of plants, except that: natural light replaces the red LED light source. The cells of the control group were not treated. Next, each group of cells was cultured for 7 to 10 days, and the medium was changed every 3 days. Thereafter, Glycerol analysis (Glycerol assay) was performed on each group of cells, with reference to the user's instructions of the Glycerol cell-based assay kit of Cayman.
The decomposition rate of the lipolysis is examined by detecting the amount of glycerol produced. Cell culture supernatants were collected from each well plate, and 25 μ L of cell culture supernatant and standard (standard) were then transferred to a new 96 well plate. Thereafter, 100. mu.L of modified Free Glycerol Assay Reagent (recombinant Free Glycerol Assay Reagent) was added to each well, followed by reaction at room temperature for 15 minutes. Next, the OD of each group was read with ELISA reader (BioTek)540nmAnd (4) light absorption value.
The results of this example are shown in FIG. 7. FIG. 7 is a data graph showing the efficacy of the plant fermentation product prepared by the method of the present invention in promoting lipolysis. As can be seen from fig. 7, the oil droplet content of the experimental group was significantly reduced compared to the control group and the comparative group. The results of this example show that the fermented plant material obtained by the method for fermenting plants according to the present invention has the effect of promoting lipolysis.
Example 8 evaluation of the effectiveness of the fermentation product of plants obtained by the method for fermenting plants according to the invention in increasing leptin secretion
Leptin is an important factor for regulating body weight of human body, can suppress appetite and fat hyperplasia, increase basal metabolic rate and control body fat in a proper range. Many people with long-term obesity have the problem of insufficient secretion of lean voxels, are difficult to lose weight and are easy to regain weight. Thus, this example discusses whether the plant fermentation broth prepared according to the method of example 3 can increase leptin secretion. First, 200. mu.L of preadipocyte Expansion Medium (Pre-adipocyte Expansion Medium) was inoculated into a 96-well plate at 1X 104Mouse adipocytes (Mus musculus, mouse adipo)cyte cell)3T3-L1(
Figure BDA0002360982460000161
Number CL-173TM) Which comprises 90% Dulbecco's Modified Eagle's Medium (available from Gibco), 10% Bovine Serum (obtained from Gibco), and 1% Penicillin/streptomycin (available from Gibco) was added, followed by culturing at 37 ℃ for 48 hours. After 48 hours, the medium was removed and differentiation medium was added for 4 days (fresh differentiation medium was changed every two days) containing 90% Dulbecco's modified Egger's medium, 10% FBS, 1% Penicillin/streptomycin (penicilin-streptomycin), 1.0. mu.M/mL Dexamethasone (DEXA) (Sigma), 0.5mM/mL methyl isobutyl xanthine (Methysobutylxanthine, IBMX) (Sigma), and 1.0. mu.g/mL insulin (insulin) (Sigma). After 4 days, the differentiation Medium was changed to Adipocyte Maintenance Medium (Adipocyte Maintenance Medium) (fresh Maintenance Medium was changed every two days), which contained 90% Dulbecco's modified Egger's Medium, 10% FBS, 1% Penicillin/streptomycin (penicilin-streptomycin), and 1.0. mu.g/mL insulin. After 7 to 10 days after the initiation of the differentiation induction, the cells were completely differentiated, and then the formation of lipid droplets (lipid droplets) was observed with a microscope. After that, each group of samples (concentration 0.0625%) was added and the medium was changed every 48 hours for an additional 12 days. Next, the culture medium was collected and the Leptin content was determined using the Mouse LEP ELISA Kit (Mouse LEP (Leptin) ELISA Kit) (Elabscience). The results of this example are shown in FIG. 8.
FIG. 8 is a data plot of the utility of plant fermentations made by the present invention in increasing leptin secretion. As can be seen from fig. 8, the leptin content in the experimental group was significantly increased compared to the control group and the comparative group. In particular, the lean voxel content of the experimental group was increased by 30% compared to the control group. The results of this example show that the plant fermentation product produced by the method for fermenting plants of the present invention is effective in increasing leptin secretion.
EXAMPLE 9 Process of the method of the present invention for increasing the growth rate of microorganisms
First, rambutan (Nephelium lappacium) juice from roxburgh city in south thailand was obtained at a ratio of 1: 5-10 wt% of water, and adding glucose with a concentration of 10% (w/w) based on the total weight. Then, extracting for 0.5-1.5 hours at 50-100 ℃ to obtain a plant extract.
Then, 0.01-0.5% (v/v) of brewer's Yeast (Saccharomyces cerevisiae) BCRC 20271 is cultured in the culture medium of the plant extract to start fermentation for 1-3 days, 1mL of bacterial liquid is taken out every 24 hours, and after sequence dilution, the bacterial liquid is coated on a Yeast Peptone Dextrose (YPD) plate, wherein the red LED light source with the wavelength of 620-750 nm (preferably 635nm) is irradiated in the whole fermentation process, and the plant fermentation product irradiating the red LED light source is obtained. Then, the plant fermentation product irradiated with the red LED light source was used as an experimental group, and the preparation method of the plant fermentation product of the comparative group was substantially the same as that of the experimental group except that: replacing the red LED light source with blue light with the wavelength of 400-450 nm; the preparation method of the plant fermentation product for the control group is substantially the same as that of the experimental group except that: natural light replaces the red LED light source. Subsequently, YPD plates coated with the inoculum solution were placed in an incubator at 30 ℃ for 48 hours, and then colonies were counted to evaluate the growth rate of Saccharomyces cerevisiae. The results of the experiment are shown in fig. 9.
FIG. 9 is a data plot showing the efficacy of the method for increasing the growth rate of microorganisms of the present invention in increasing the growth rate of Saccharomyces cerevisiae. As can be seen from fig. 9, the growth rate of saccharomyces cerevisiae in the experimental group was significantly increased compared to the control group and the comparative group. Particularly, in the fermentation process, the red LED light source (i.e. experimental group) with the irradiation wavelength of 620-750 nm can increase the growth rate of the saccharomyces cerevisiae by 47% (compared with the control group), otherwise, the effect of increasing the growth rate is not achieved when natural light and blue light are irradiated. The results of this example show that the method for increasing the growth rate of microorganisms of the present invention can effectively increase the growth rate of saccharomyces cerevisiae.
Then, after 5% (v/v) acetic acid bacteria (Acetobacter aceti) BCRC 11688 is inoculated into a culture medium of the plant extract (AAB culture medium, to which 0.5% Yeast extract, 0.3% peptone (peptone) and 2.5% mannitol (mannitol)) and fermentation is started for 1-3 days, 1mL of a bacterial liquid is taken out every 24 hours, and after the bacterial liquid is sequentially diluted, the bacterial liquid is coated on a Yeast peptone dextrose (Yeast dextrose, YPD) plate, wherein a red LED light source with the wavelength of 620-750 nm (preferably 635nm) is irradiated in the whole fermentation process, so as to obtain the plant fermentation product irradiated by the red LED light source. Then, the plant fermentation product irradiated with the red LED light source was used as an experimental group, and the preparation method of the plant fermentation product of the comparative group was substantially the same as that of the experimental group except that: replacing the red LED light source with blue light with the wavelength of 400-450 nm; the preparation method of the plant fermentation product for the control group was substantially the same as that of the experimental group except that: natural light replaces the red LED light source. Subsequently, YPD plates coated with the inoculum solution were placed in an incubator at 30 ℃ for 48 hours, and then colonies were counted to evaluate the growth rate of acetic acid bacteria. The results of the experiment are shown in table 2.
TABLE 2
Group of Growth rate of acetic acid bacteria
Control group
100％
Comparison group 89％
Experimental group 122％
As can be seen from Table 2, the growth rate of acetic acid bacteria in the experimental group was significantly increased compared to the control group and the comparative group. Particularly, in the fermentation process, the red LED light source (i.e. experimental group) with the irradiation wavelength of 620-750 nm can increase the growth rate of acetic acid bacteria by 22% (compared with the control group), otherwise, the effect of increasing the growth rate is not achieved when natural light and blue light are irradiated. The results of this example show that the method for increasing the growth rate of microorganisms of the present invention can effectively increase the growth rate of acetic acid bacteria.
In summary, the method for fermenting plants and the plant fermentation product prepared by the method of the invention can increase the growth rate of microorganisms (such as saccharomyces cerevisiae and acetic acid bacteria) and the content of effective components in plants, and achieve the effects of skin care and skin beautification by increasing the oxidation resistance of skin, improving the anti-saccharification activity of skin and promoting the proliferation of elastin. On the other hand, the plant fermentation product can achieve the effect of reducing fat by promoting lipolysis and increasing leptin secretion.
The foregoing is by way of example only, and not limiting. It is intended that all equivalent modifications or variations without departing from the spirit and scope of the present invention shall be included in the claims.

Claims (20)

1. A method for fermenting a plant comprising the steps of:
extracting a plant with water to obtain a plant extract; and
sequentially fermenting the plant extract with Saccharomyces cerevisiae, Lactobacillus plantarum and Acetobacter xylinum to obtain a plant fermented product,
wherein the fermentation process comprises irradiating the plant extract, the brewer's yeast, the germ lactic acid bacteria, and the acetic acid bacteria with a red light source having a wavelength of 620-750 nm.
2. The method of claim 1, wherein the plant is rambutan.
3. The method of claim 1, wherein prior to said extracting, a concentration of 10% (w/w) glucose based on total weight is added.
4. The method of claim 1, wherein the fermentation time of the saccharomyces cerevisiae is 1 to 2.5 days, the fermentation time of the germ lactic acid bacteria is 1 to 3 days, and the fermentation time of the acetic acid bacteria is 3 to 10 days.
5. The method of claim 1, wherein the concentration of Saccharomyces cerevisiae is 0.01-0.5% (v/v), the concentration of Lactobacillus plantarum is 0.01-0.25% (v/v), and the concentration of Acetobacter xylinum is 3-10% (v/v).
6. The method according to claim 1, wherein the weight ratio of the plant to the water is 1: 5-10.
7. The method of claim 1, wherein the temperature of the extraction is between 50 and 100 ℃ and the time of the extraction is between 0.5 and 1.5 hours.
8. Use of the method of any one of claims 1 to 7 for increasing the growth rate of saccharomyces cerevisiae and the content of active ingredients in plants.
9. Use according to claim 8, characterized in that the plant is rambutan and the active ingredient is a polyphenol.
10. A plant fermentation obtained by the method according to any one of claims 1 to 7.
11. Use of the plant ferment of claim 10 for the preparation of a skin-care and fat-reducing composition.
12. The use of claim 11, wherein the skin care comprises enhancing the antioxidant capacity of the skin, enhancing the anti-glycation activity of the skin, and promoting elastin proliferation.
13. The use of claim 11, wherein the fat reduction comprises promoting lipolysis and increasing lean voxel secretion.
14. The use according to claim 11, wherein the composition is in the form of a medicament, a nutraceutical or a food product.
15. A method for increasing the growth rate of a microorganism, comprising the steps of:
extracting a plant with water to obtain a plant extract; and
subjecting the plant extract to a fermentation process with a microorganism to obtain a plant fermentation product,
wherein the microorganism is Saccharomyces cerevisiae or Acetobacter xylinum, and the fermentation process comprises irradiating the plant extract and the microorganism with a red light source with a wavelength of 620-750 nm.
16. The method of claim 15, wherein the plant is rambutan.
17. The method of claim 15, wherein prior to said extracting, a concentration of 10% (w/w) glucose is added based on total weight.
18. The method of claim 15, wherein the fermentation time of the microorganism is 1 to 3 days.
19. The method of claim 15, wherein the concentration of Saccharomyces cerevisiae is 0.01-0.5% (v/v), and the concentration of Acetobacter is 5% (v/v).
20. The method of claim 15, wherein the growth rate of the brewer's yeast is increased by at least 47% and the growth rate of the acetic acid bacteria is increased by at least 22%.
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