KR101814550B1 - A Method for Preparing Bacterial Cellulose Using Scoria Extract and the Bacterial Cellulose Obtained Thereby - Google Patents
A Method for Preparing Bacterial Cellulose Using Scoria Extract and the Bacterial Cellulose Obtained Thereby Download PDFInfo
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- KR101814550B1 KR101814550B1 KR1020150137894A KR20150137894A KR101814550B1 KR 101814550 B1 KR101814550 B1 KR 101814550B1 KR 1020150137894 A KR1020150137894 A KR 1020150137894A KR 20150137894 A KR20150137894 A KR 20150137894A KR 101814550 B1 KR101814550 B1 KR 101814550B1
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- bacterial cellulose
- extract
- volcanic
- present
- bacterial
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- 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
- A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
- A61K8/73—Polysaccharides
- A61K8/731—Cellulose; Quaternized cellulose derivatives
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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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- 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
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- 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
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
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- 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
- A23V2250/00—Food ingredients
- A23V2250/50—Polysaccharides, gums
- A23V2250/51—Polysaccharide
- A23V2250/5108—Cellulose
Abstract
The present invention discloses a method for producing bacterial cellulose using a volcanic rock meal extract and bacterial cellulose obtained by the method. The production of the bacterial cellulose according to the present invention is increased and the bacterial cellulose obtained according to the present invention has excellent moisturizing activity and wrinkle improving activity, so that it can be usefully used as a cosmetic for the purpose.
Description
The present invention relates to a method for producing bacterial cellulose using a volcanic crust extract and bacterial cellulose obtained by the method.
Cellulose is a polysaccharide composed of β-1,4-glucose, which is the main component of cell walls of higher plants widely found in nature. Plant cellulose is composed of complex polysaccharides mixed with other polysaccharides such as pectin, hemicellulose and lignin, resulting in low crystallinity and low mechanical strength and adsorption properties (Proc Natl Acad Sci USA 1980 Nov; 77 (11): 6678-82.). On the other hand, cellulose produced by bacteria is a collection of pure cellulose that does not contain polysaccharides such as pectin, hemicellulose, and lignin, unlike cellulose produced by ordinary plants. A microfibril having a thickness of about 0.1 袖 m is formed by hydrogen bonding (Milda E et al., Food Hydrocoll 1994 8: 419-430.). ≪ / RTI > . As a result, it is easy to separate and purify, and has high crystallinity and excellent physical properties such as mechanical strength, adsorptivity, water retention, suspension stability, and tackiness and is widely used as a material for food, cosmetic and pharmaceutical industries (Vandamme FJ et al , Polym Degarad Stab, 1998 59: 93-99; Cho SH et al., J Korean Soc Food Sci Nutr 2002 31: 802-807; Sutherland IW., Tibtech 1998 16: 41-46).
Since acetic acid bacteria have been reported to produce bacterial cellulose by Brown in 1886, there are currently strains producing Acetobacter sp., Acrobacterium sp., Agrobacterium sp. Away Resorts in (Rhizobium sp.), Pseudomonas species (Pseudomonas sp.) and Sar Sina (Sarcina sp.) was known in the strain, especially bakteo in acetonitrile (Acetobacter sp.) in acetonitrile bakteo Xi Leeum (Acetobacter xylium , A. pasteurinanus and A. hansenii are most known. Recently, a strain of Gluconacetobacter sp. Has been isolated from cellulosic producing bacteria, which has been reclassified from Acetobacter genus by molecular systematic taxonomy using 16S rRNA sequence (Lee OS et al., J. Korean Soc. Food Sci. Nutr 2002 31: 572-577; Choi KH et al., J. Korean Soc. Food Sci. Nutr 2004 33: 170-175.).
It is reported that the strain producing celluloses having high purity is morphologically stomach bacterium and has a strong acid-fast bacterium and secretes cellulose from the cells. These strains, which produce bacterial celluloses, are used for the production of glucose, sucrose, molasses, fructose, mannitol, glycerol or naturally-occurring juice and persimmon juice, apple juice, Peptone, yeast extract and the like can be used as a carbon source (S. Masaoka et al., J. Ferment. Bioeng. 75, 1993, 18-22) 1998, 312-317) was used as a nitrogen source (Ramana KV et al., World J. Microbial Biotechnol., 16, 2000, 245-248, Yang YK et al., J. Ferment.
The manufacturing techniques of bacterial cellulose having various utility values can be roughly classified into a technique for improving the yield and a technique for improving the properties of the cellulose.
Techniques for improving the yield include techniques for selecting optimal one among various kinds of carbon source, auxiliary carbon source and nitrogen source and establishing culture conditions at an appropriate concentration (Korean Patent Publication No. 1998-0067009, Korean Patent Publication No. 2010 -0135455, Korean Patent No. 1435580, and Sung-Joon Kim, Chonnam National University, 2007), but the technology for improving physical properties has been developed for the purpose of improving elasticity, strength, decomposition and dispersibility, and improving water retention European Patent Publication No. 2002-013925, Japanese Patent Laid-Open No. 1991-228479, US Patent No. 10/162073, European Patent Publication No. 2002-022526, etc.).
The present invention discloses a method for producing a bacterial cellulase having an improved production yield and a moisturizing activity and a wrinkle-reducing activity by using an extract of a volcanic stone clay.
It is an object of the present invention to provide a method for producing bacterial cellulose using a volcanic rock meal extract.
It is still another object of the present invention to provide a composition for moisturizing or wrinkle-improving, which comprises the bacterial cellulose obtained by the above production method as an active ingredient.
Other and further objects of the present invention will be described below.
The inventors of the present invention confirmed that the production amount of bacterial cellulose was increased by using a culture medium supplemented with a volcanic stone extract, as confirmed in the following examples and experimental examples, and it was confirmed that the bacterial cellulase extract Cellulose showed excellent skin moisturizing activity (moisturizing activity) and excellent wrinkle reducing activity in human body test.
The present invention provides, on the basis of these experimental results, a method for producing a bacterial cellulase comprising the steps of (a) preparing a culture medium for bacterial cellulose production containing a carbon source and a volcanic rock extract, (b) inoculating the culture medium with a bacteria- (c) culturing in an aerobic condition after inoculation, and (d) obtaining bacterial cellulose from the culture.
As used herein, "Scoria" or "clay" refers to a porous basaltic rock mass having a volume of pore that is similar in volume to solids, including volcanic ash (less than 2 mm), volcanic ash These include volcanic rocks (gravel, lapilli, 2 to 64 mm), volcanic blocks (more than 64 mm), and volcanic bombs. Pine mushroom can be found in the topography formed by the volcano. In Korea, it is easily found in the volcanic island of Jeju Island. Jeju volcanic stone pine has various colors depending on its major constituents. The cluster with high silicon oxide content has dark gray color, the cluster with high oxidized aluminum content has yellowish brown or black color, and the cluster with high content of titanium oxide and iron oxide has reddish brown color . It is known that Jeju mushroom usually has a melting point of 1,120 ~ 1210 ℃, an absorption rate of 17.7 ~ 32.5% and a wear rate of 47.25 ~ 67.22%. The volcanic stone pine contains various minerals and is known to have excellent heavy metal adsorption, antimicrobial effect, and outward emission ability compared to jade, yellow loess, elvan, and loess, which are widely used in recent times. The volcanic stone clusters that can be used in the present invention are not particularly limited to Jeju pine clusters, but it is desirable to understand them as Jeju volcanic rocks which are known to have excellent functionality such as far infrared radiation and heavy metal adsorption effect.
The volcanic stone clusters of the present invention can be obtained by conventional methods in the art, and the volcanic stone clusters for the production of volcanic rock extract can be used in the form of raw stones or in the form of powders and preferably in the form of powders .
In this specification, "% (w / w)" means a concentration (w / w) expressed in terms of weight percents, Weight solute. For example, 10% (w / w) means that 10 g of the solute is dissolved in 100 g of the solution. Therefore, in the method of the present invention, when glucose is added to the medium in an amount of 10% (w / w), 90% by weight (w / w) of other medium components such as purified water and 10% Can be understood as meaning.
Terms not specifically defined in this specification refer to the national meaning of the word or the meaning commonly used in the art.
The present invention relates to a method for producing a bacterial cellulose having improved productivity using a volcanic stone meal extract.
In the method of the present invention, the carbon source of the culture medium for producing bacterial cellulose in step (a) includes glucose, sucrose, maltose, fructose, lactose, xylose, galactose, arabinose, glycerol, Starch, starch hydrolyzate, and the like. These carbon sources may be contained in the medium alone or in a mixture of two or more, and preferably 5% (w / w) to 20% (w / w) w) < / RTI > When using natural or natural products such as starch or starch hydrolyzate as a carbon source, these carbon sources can be sterilized and used to prevent unwanted fermentation by other microorganisms. Sterilization can be carried out using methods known in the art such as high temperature and high pressure sterilization, ultraviolet irradiation, and the like.
In the method of the present invention, the volcanic stone extract of the step (a) in the culture medium for the production of bacterial cellulose may be prepared by dissolving the volcanic stone clones, which are the object of extraction, irrespective of the extraction method, with water, anhydrous or lower alcohols having 1 to 4 carbon atoms Propanol, iso-propanol and n-butanol), acetone, ethyl acetate, methylene chloride, chloroform, 1,3-butylenelicolle, hexane, diethyl ether or a mixed solvent thereof Is understood as meaning that the obtained extract and the extract obtained by fractionation of the above-mentioned speckle in the extract. As long as it is extracted through the step of immersing the object to be extracted in the extraction solvent, it is understood that any of extraction methods such as cold beating, refluxing, heating, and ultrasonic wave can be applied. Nevertheless, the above-mentioned extract preferably means that the extract is obtained by extracting water with a solvent, and includes a concentrated liquid extract or a solid extract from which the extraction solvent has been removed. More preferably, it means an extract obtained by using water as an extraction solvent and heating. More preferably, it means an extract obtained by heating water to a concentration of 70 ° C to 80 ° C using water as an extraction solvent. In addition, the volcanic stone extract may further comprise a filtration step after solvent mixing. This filtration step is a step for removing the suspended solid particles of the volcanic stone clay from the solvent, and any method known in the art can be used such as gravity filtration, pressure filtration, vacuum filtration, centrifugal filtration and the like.
In addition, the method of the present invention may further include a solvent extraction and a post-filtration sterilization step for the production of a volcanic rock meal extract in a medium for bacterial cellulose production in the step (a). This sterilization step is intended to prevent contamination by the microorganisms present in the volcanic rock meal extract, and as a result, to prevent contamination of the culture medium for bacterial cellulose production. As the sterilization method, any method known in the art such as high temperature and high pressure sterilization method, ultraviolet irradiation, filtration and the like can be used.
In addition, in the method of the present invention, the extract of the volcanic rocks in the culture medium for the production of bacterial cellulose in the step (a) may contain the bacterial cellulose production rate, moisturizing activity and wrinkle- And the like. Excessive inclusion of components of the volcanic rock extract such as minerals may inhibit the growth of bacteria having cellulosic production ability, which may lead to a decrease in the production of cellulose. On the other hand, when the volcanic rock meal extract is less contained, the cellulose-producing ability by the bacteria may not be higher than that of the medium containing no volcanic rock extract, and the moisturizing and wrinkle-improving functional activity by the volcanic stone meal extract may not be sufficiently exhibited . (W / w) to 90% (w / w), preferably 60% (w / w) to 90% (w / w), more preferably 70% w) to 85% (w / w).
In addition, in the method of the present invention, the culture medium for producing bacterial cellulose in step (a) may further comprise an auxiliary carbon source. The auxiliary carbon source is not essential for the growth of microorganisms having the ability to produce bacterial cellulose or for the production of bacterial cellulose by such microorganisms but for the addition of bacterial cellulose to promote the growth of microorganisms having bactericidal cellulogenicity and / or the production of bacterial celluloses by such microorganisms Such as lactic acid, citric acid, succinic acid, citric acid, acetic acid, ethanol, formic acid, malic acid, skin acid and the like, and furthermore, (Kim SY, et al., Appl Biochem Biotechnol., 129-132, 705-15, 2006), apple juice Et al., J. Appl. Microbiol. Iotechnol., ≪ RTI ID = 0.0 > 12 (5), 722-728, (Jonas R, et al., Polym Degrad Stab., 59, 101-92), and beer waste (Park JK, et al., Kor Chem Eng Res., 44 (1), 52-57, 2006) 106, 1998), pineapple juice (see Japanese Patent Application No. 1999-299034), citrus juice, and the like. It is preferable that the medium for the production of bacterial cellulose of the present invention further uses citrus juice (Citrus juice).
In the method of the present invention, the culture medium for producing bacterial cellulose in step (a) may further comprise a nitrogen source. Examples of the nitrogen source include natural nitrogen sources such as yeast extract, soytone, peptone, tryptone, malt extract, soybean powder, soybean bean meal, chongkukjang powder and doenjang powder, Or inorganic nitrogen sources such as ammonium salts such as ammonium sulfate, ammonium chloride and ammonium phosphate, nitrate salts and urea can be used. These nitrogen sources may also be used in admixture of one or more species, and may be contained in the medium preferably in the range of 0.1% (w / w) to 8.0% (w / w). When a natural nitrogen source is used, it is preferable to sterilize the same for the same reason as described in connection with the case of using a natural carbon source.
In the method of the present invention, the culture medium for bacterial cellulose production in step (a) may further comprise inorganic salts and trace elements. Examples of the inorganic salts that can be used include sodium hydrogen phosphate, magnesium sulfate, iron chloride, calcium salt, manganese salt, cobalt salt, molybdate salt and chelate metal salt. Examples of trace salts include amino acids, vitamins, fatty acids, Sulfite pulp waste liquid, and the like. These inorganic salts and trace elements may be used in admixture of one or more species and may be included in the medium in the range of 0.001% (w / w) to 3.0% (w / w).
In the method of the present invention, after the culture medium for producing bacterial cellulose is prepared in the step (a), a sterilization step may be further included. This sterilization step is intended to prevent contamination of other microorganisms, and as a result, to increase bacterial cellulose production yield. As the sterilization method, any method known in the art can be used such as a high-temperature and high-pressure sterilization method, ultraviolet irradiation, and the like.
In the method of the present invention, any microorganism known in the art can be used as the microorganism having the ability to produce bacterial cellulose in the step (b). Examples of such microorganisms include Acetobacter sp., Acacobacter sp., Gluconacetobacter sp., Agrobacterium sp., Rhizobium sp., Pseudomonas sp. Pseudomonas sp. And Sarcina sp. May be used, but Gluconacetobacter sp. Is preferably used. It is furthermore preferable that Gluconacetobacter sp. Gel_SEA623-2 is used.
In the method of the present invention, it is preferable that the microorganism having the ability to produce a bacterial cellulase in step (b) is inoculated after pre-culture for activation thereof before inoculation. In the pre-culture, the medium components such as the carbon source and the auxiliary carbon source, the composition of the components, the pre-culture conditions, the pre-culture period, and the like can be referred to as described above.
In the method of the present invention, the culturing temperature in step (c) is preferably in the range of 10 to 40 ° C, more preferably 25 to 35 ° C, and more preferably 28 to 30 ° C.
In the step (c) of the present invention, the culture period is preferably 5 to 20 days, more preferably 7 to 14 days.
In the culture of step (c) of the present invention, the oxygen concentration is preferably in the range of 1 to 100% (w / w), particularly 20 to 80% (w / w) for making aerobic conditions.
In the culturing in the step (c) of the present invention, the culture method may be either a static culture method or a continuous culture method, and preferably a static culture method may be used.
The present invention relates to a bacterial cellulose obtained by the above-mentioned process for producing a bacterial cellulase.
The bacterial cellulose of the present invention can be understood as a bacterial cellulose obtained by using an extract of a volcanic stone clay, thereby to be understood as a bacterial cellulose improved in skin moisturizing and skin wrinkle improving activity.
In view of the above, the present invention can be understood as a skin moisturizing composition comprising, as an effective ingredient, the bacterial cellulose obtained by the above production method.
In another aspect of the present invention, the bacterial cellulose obtained by the above-described method can be regarded as a composition for improving skin wrinkles containing an extract as an active ingredient.
As used herein, the term "active ingredient" alone means an ingredient which exhibits the desired activity or which can exhibit activity together with a carrier which itself is not active.
In the meantime, the "composition for skin moisturizing" or "composition for improving skin wrinkles" of the present invention may contain any amount (effective amount) of bacterial cellulose using a volcanic stone as an effective ingredient thereof as long as it can exhibit skin moisturizing activity or wrinkle- , A typical effective amount will be determined within the range of from 0.001 wt% to 99.900 wt% based on the total weight of the composition. The term "effective amount" as used herein refers to an amount of an effective ingredient capable of preventing, ameliorating, treating, or delaying the onset of symptoms of a symptom. Such effective amounts can be determined experimentally within the ordinary skill of those skilled in the art.
The skin moisturizing or skin wrinkle improving composition of the present invention can be identified as a cosmetic composition in a specific embodiment.
When the composition of the present invention is identified as a cosmetic composition, the cosmetic composition can be prepared in various forms, for example, emulsion, lotion, cream (underwater type, water type, multiphase), solution, suspension (Water and water), anhydrous products (oil and glycol), gel, mask, pack, powder and the like.
The composition of the present invention may contain an acceptable carrier in cosmetic preparations other than those containing the active ingredient.
As used herein, the term " acceptable carrier for cosmetic preparation "refers to a compound or composition which is already known and used in cosmetics, or which is a compound or composition to be developed in the future, and which has no toxicity that the human body can adapt to when contacted with skin.
The carrier may be included in the composition of the present invention in an amount of from about 1% by weight to about 99.99% by weight, preferably from about 50% by weight to about 99% by weight of the composition, based on the total weight thereof.
However, since the ratio depends on the above-mentioned formulation of the cosmetic product and its specific application site (face or hands) or the desired amount of application thereof, the ratio is to limit the scope of the present invention in any aspect It should not be.
Examples of the carrier include alcohols, oils, fatty acids, silicone oils, wetting agents, moisturizing agents, viscosifying agents, emulsifiers, stabilizers, sunscreens, coloring agents and flavoring agents.
Because the compounds / compositions which can be used as the carrier and which can be used as alcohols, oils, fatty acids, silicone oils, humectants, moisturizers, viscosifiers, emulsions, stabilizers, sunscreens, Those skilled in the art will be able to select and use the appropriate substance / composition.
In another specific embodiment, the composition of the present invention can be identified as a food composition.
When the composition of the present invention is identified as a food composition, it can be used as a food composition such as beverage such as juice, carbonated beverage, ionic drink, processed oil such as milk and request route, gum, rice cake, Korean food, bread, , Capsules, and the like.
The food composition of the present invention can be manufactured as a health functional food, a health supplement food, a special nutrition supplement food, a functional beverage and the like.
The food composition of the present invention may contain sweetening agents, flavoring agents, physiologically active ingredients, minerals and the like in addition to the active ingredients thereof.
Sweetening agents may be used in an amount that sweetens the food in a suitable manner, and may be natural or synthetic. Preferably, natural sweeteners are used. Examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.
Flavors may be used to enhance taste or flavor, both natural and synthetic. Preferably, a natural one is used. When using natural ones, the purpose of nutritional fortification can be performed in addition to the flavor. Examples of natural flavoring agents include those obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or those obtained from green tea leaves, Asiatica, Daegu, Cinnamon, Chrysanthemum leaves and Jasmine. Also, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, banks and the like can be used. The natural flavoring agent may be a liquid concentrate or a solid form of extract. Synthetic flavors may be used depending on the case, and synthetic flavors such as esters, alcohols, aldehydes, terpenes and the like may be used.
Examples of the physiologically active substance include catechins such as catechin, epicatechin, gallocatechin and epigallocatechin, and vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine and riboflavin.
As the mineral, calcium, magnesium, chromium, cobalt, copper, fluoride, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium and zinc can be used.
In addition, the food composition of the present invention may contain preservatives, emulsifiers, acidifiers, thickeners and the like as needed in addition to the above sweeteners.
Such preservatives, emulsifiers and the like are preferably added in a very small amount as long as they can attain an application to which they are added. The term " trace amount " means, when expressed numerically, in the range of 0.0005% by weight to about 0.5% by weight based on the total weight of the food composition.
Examples of the preservative which can be used include calcium sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate and EDTA (ethylenediaminetetraacetic acid).
Examples of the emulsifier which can be used include acacia gum, carboxymethyl cellulose, xanthan gum, pectin and the like.
Examples of the acidulant that can be used include acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, and phosphoric acid. Such an acidulant may be added so that the food composition has a proper acidity for the purpose of inhibiting the growth of microorganisms other than the purpose of enhancing the taste.
Agents that may be used include suspending agents, sedimentation agents, gel formers, bulking agents and the like.
As described above, according to the present invention, it is possible to provide a method for producing bacterial cellulose in which the production yield is improved using a volcanic stone seedlings, and the bacterial cellulose obtained by the method for producing the bacterial cellulose. Further, the bacterial cellulose of the present invention can be usefully used for cosmetics or foods for moisturizing and wrinkle-improving purposes.
Fig. 1 schematically shows a process for producing a volcanic stone meal extract.
Figure 2 is a schematic representation of a method for producing bacterial cellulose using a volcanic rock meal extract.
FIG. 3 shows the result of confirming skin moisture change with time in application of bacterial cellulose using a volcanic stone meal extract.
Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.
<Examples> Bacterial cellulose using a volcanic ash extract
A culture medium was prepared using a volcanic stone meal extract and the bacterial celluloses were obtained by cultivating Gluconacetobacter genus gel_SEA623-2 (KACC 91526P) on the corresponding media.
Example 1 Preparation of Bacterial Bacteria for Producing Bacterial Cellulose
For the experiment, Gluconacetobacter sp. Gel_SEA623-2 strain was purchased from the National Institute of Horticultural Science, Citrus Experiment Station (Lee SY et al., Kor. J. Hort. Sci. Technol.
Gluconacetobacter sp. The gel_SEA623-2 was inoculated on a G10 agar medium (glucose 100 g / L, yeast extract 10 g / L, calcium carbonate 20 g / L, agar 15 g / L) and cultured at 30 ° C for 4 days, The colonies were cultured in a pre-culture medium (10% w / w, 8.8% w / w sucrose, 1% w / w ethanol, , and the culture bottle inlet was covered with sterilized filter paper (HYUNDAI Micro) and cultured at 30 ° C for 5 days to obtain seeds for culturing (Seed) ).
<Example 2> Production of volcanic stone meal extract
The volcanic stone scoria was collected from the uplift of Jeju Island in order to produce the volcanic stone pine mushroom extract, and then it was ground with a ball mill and standardized by shaking less than 40 mesh. The volcanic stone clusters were mixed with 10 times the weight of purified water, heated to 75 ± 5 ° C, and kept at the temperature for 24 hours. After 24 hours, the extract was filtered through filter paper (Advantec No. 2, 300 mm) and sterilized at 121 ° C for 15 minutes.
Example 3 Production of Bacterial Cellulose Using a Volcanic Stone Pine Mushroom Extract
The volcanic stone extract of Example 2 was used to prepare bacterial cellulase culture medium of strain Gluconacetobacter gel_SEA623-2.
Bacterial cellulose culture medium consisted of volcanic rock meal extract (79.2% w / w), citrus juice (wenzhou citric juice) 10 (% w / w), sugar (8.8% w / w), acetic acid (1% w / , Ethanol (1% w / w), and a balance of purified water. The medium was sterilized at 121 ° C for 15 minutes.
To the sterilized bacterial cell culture medium was added Gluconacetobacter sp. The cells were inoculated with 1% (w / w) of the culture medium containing the gel_SEA623-2 seeds and cultured under aerobic conditions at 30 ° C for 14 days to produce bacterial cellulose.
< Comparative Example > Comparison of bacterial cellulose Manufacturing example
≪ Comparative Example 1 & gt ; Comparative Production Example 1 of Bacterial Cellulose
The bacterial cellulose was produced through the same manufacturing process as that of Example 3 except that the volcanic rock extract was removed from the bacterial cellulase culture medium and the remaining amount was adjusted to the purified water.
≪ Comparative Example 2 & gt ; Comparative Production Example 2 of Bacterial Cellulose
The bacterial cellulose was produced through the same manufacturing process as in Example 3, except that the volcanic rock clusters in the bacterial cellulase culture medium were in the form of powders Respectively. The volcanic ash powder is prepared by preparing a powder having a particle size ranging from 0.05 to 5 μm through a pulverizer, adding supercritical carbon dioxide to the powder at a temperature of 100 ° C. and a pressure of 600 bar to remove foreign matter and microorganisms, w). All samples were prepared using 100 ml of medium and compared with the cross-sectional thickness (mm) and weight (g) of cellulose after incubation.
< Experimental Example 1> Yield analysis of bacterial cellulose
The amount of bacterial cellulose produced through the Examples and Comparative Examples was checked and the results are shown in Table 1.
The amount of bacterial cellulose was evaluated based on the thickness of the white gel formed on the upper layer of the cultured medium. The thickness and thickness of the thickest part and the thinnest part were measured and measured in mm.
The dried bacterial cellulose was then washed and dried in a drier (OF-22, Palmswell Bio) at a temperature of 60 ° C for 24 hours to measure the dry weight of the resulting bacterial cellulose. The microbalance FX -300i, AND) and recorded. All experiments were repeated 3 times and mean values are shown in Table 1.
As shown in Table 1, when the bacterial cellulose was produced by adding the volcanic rock meal extract as in the example, it can be confirmed that the thickness and the weight were increased. This suggests that the extract of the volcanic rocks has an effect of increasing the productivity of the bacterial cellulose.
< Experimental Example 2> Volcanic stone Identification of the function of bacterial cellulosic extracts using Pine Mushroom Extract
≪ Experimental Example 2-1 >
The human body test was conducted to confirm the moisturizing activity of the bacterial cellulose prepared using the volcanic stone meal extract of Example.
The bacterial cellulose obtained through the above <Examples> and <Comparative Example> was washed several times with purified water to remove the medium components, and then treated with 1% NaOH aqueous solution at 80 ° C for one day to remove the cells. Bacterial cellulose was immersed in 1% acetic acid solution to neutralize and then thoroughly rinsed with purified water to prepare a sample at a concentration of 10 mg / ml (1%, w / v).
Twenty adult women aged 30 to 55 without skin abnormalities such as scars, dots, acne, erythema, and capillary dilatation were selected as subjects. The subjects were kept clean and dry in order to make the measurement conditions the same, and the skin was stabilized at the place where the constant temperature and humidity (22 ± 2 ℃, R.H. 40 ~ 60%) was maintained for at least 30 minutes. If the test product has abnormal reactions such as erythema, edema, scaling, itching, stinging, burning, tightness, prickling or other adverse reactions, Respectively.
Four specimens were applied to the selected test site (1.5 cm x 1.5 cm) of the forearm using a micropipette in an amount of 2 μl /
The skin moisture was measured using a moisture meter (CM825, Courage and Khazaka Electronic Co., Germany). The moisture meter is made by measuring the static load capacitance of the current transmitted through the probe contacting the skin. The water content and the static load capacity are proportional to each other, and the higher the water content, the higher the measured value, and the measurement factor is the arbitrary unit (A.U.).
The amount of water change in the non-cannulated area was calculated by the following equation.
≪ Amount of moisture change to the non-coated area >
(%) = ((Result by product measurement time - Before product application) - (Result by non-canonical measurement time -
The statistical analysis was performed to measure the moisture change with time in the non-coated area and the measurement of the moisture change amount with time in the bacterial cellulose used site prepared through the above <Examples>, <Comparative Example 1> and <Comparative Example 2> Independent t-test method. All statistical results were considered to be statistically significant at 5% (p <0.05), which is the most frequently used biometric statistical analysis, and statistical analysis was performed using SPSS 18.0 software. The test results are shown in [Table 2], [Table 3] and [Figure 1].
As can be seen from the results of [Table 2], [Table 3] and [Figure 1], the bacterial cellulose using the volcanic stone meal extract of Example had a water content higher than that of the <Comparative Example> Is high. In particular, it was confirmed that when the bacterial cellulose was produced by using the volcanic stone clusters in the form of an extract, the moisturizing activity was superior to that of the <Comparative Example 2> using the volcanic stone meal powder.
<Experimental Example 2-2> Wrinkle improving activity
A human body test was conducted to confirm the wrinkle-improving activity of bacterial cellulase using the volcanic stone meal extract of Example.
Twenty adult women aged 30 to 55 years without any skin abnormalities such as scars, dots, acne, erythema, and capillary vasodilation were selected as subjects. On the right and left sides were applied twice daily for 12 weeks, morning and evening. <Example>, <Comparative Example>, or a control sample was applied to the right side and the left side of the face of the subject. The degree of relaxation of the wrinkles was measured using a skin image analyzer (Visiometer). The degree of wrinkles was evaluated with no improvement, slight improvement, moderate improvement, and high improvement, and the results are shown in Table 4. The wrinkle depth (탆) was measured by an image analysis method on the wrinkle replica before and after the action, and the results are shown in Table 5 below. Distilled water was used as a control.
As shown in [Table 4] and [Table 5], in the group to which the bacterial cellulose of the <Example> was applied using the volcanic stone meal extract, the bacterial cellulase using the medium of <Comparative Example 1> or the cluster of the volcanic stone of <Comparative Example 2> Wrinkle improving activity was shown to be superior to the group to which the bacterial cellulose using the powder as the medium was applied. As described above, the component of the volcanic stone extract contained in the culture medium increased the yield of bacterial cellulose and increased the content of the functional ingredient, thereby producing a bacterial cellulose having excellent properties.
Claims (10)
(b) inoculating the medium with bacteria having bactericidal cellulase production ability,
(c) culturing in an aerobic condition after inoculation, and
(d) obtaining bacterial cellulose from the culture,
The bacterium having the bacterial cellulase-producing ability is Gluconacetobacter sp. Gel_SEA623-2 in the genus Gluconacetabacter,
Characterized in that the volcanic ash-bearing hot-water extraction water is contained in the medium in an amount of 70% (w / w) to 85% (w / w)
A method for producing bacterial cellulose using a volcanic rock meal extract.
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| KR20200002498A (en) * | 2018-06-29 | 2020-01-08 | 주식회사 송이산업 | Antioxidant and anti-inflammatory composition comprising mixed extracts of ecklonia cava extract and scoria |
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| KR102560988B1 (en) * | 2020-12-04 | 2023-07-28 | 주식회사 송이산업 | cosmetic compound extract suitable for preventing wrinkles |
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| KR20200002498A (en) * | 2018-06-29 | 2020-01-08 | 주식회사 송이산업 | Antioxidant and anti-inflammatory composition comprising mixed extracts of ecklonia cava extract and scoria |
| KR102121048B1 (en) | 2018-06-29 | 2020-06-09 | 주식회사 송이산업 | Antioxidant and anti-inflammatory composition comprising mixed extracts of ecklonia cava extract and scoria |
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