JP2010273598A - Method for manufacturing tyrosinase activity inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a tyrosinase activity inhibitor and a melaninogenesis suppressing agent by using hemicellulose content materials such as rice bran and wheat bran. <P>SOLUTION: Hemicellulose high content plants or its sections are treated by using enzyme or β-glucosidase or xylanase high production aspergillus oryzae having hemicellulose decomposition activity, and the tyrosinase activity inhibitor is manufactured, and used as a melanogenesis inhibitor, a cosmetic for bleaching, and a food browning inhibitor. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a method for producing a tyrosinase activity inhibitor characterized by treating a hemicellulose-containing material with an enzyme or koji mold, a tyrosinase activity inhibitor obtainable by the method, a melanin pigment production inhibitor comprising the same, The present invention relates to whitening cosmetics and food browning inhibitors.

  Tyrosinase is an oxidase of tyrosine, which is an amino acid. Tyrosine is oxidized by tyrosinase from tyrosine → dopa (L-DOPA) → dopaquinone. The dopaquinone produced here is chemically highly reactive, and in several stages of auto-oxidation, it passes through the red pigment, dopachrome, to become a melanin pigment, which is a dark brown polymer. Melanin is a deposited pigment that is the causative agent of stains, buckwheat and aza. For this reason, if the tyrosinase reaction can be specifically inhibited, it is considered possible to prevent abnormal melanin production, which is the cause of spots and freckles (Non-patent Document 1). Tyrosinase is also one of the causes of browning in foods such as fruits and vegetables. Therefore, it is considered possible to suppress browning of food by suppressing the tyrosinase reaction.

  On the other hand, rice bran and wheat bran, which are discharged as by-products when foods such as grains are processed and manufactured, contain abundant nutrients such as dietary fiber. However, it is only used for feeds for livestock and the like, soil improvers, etc., and has not been effectively utilized as it is (Non-patent Document 2). The dietary fiber of rice bran and wheat bran is mainly composed of three components of cellulose, hemicellulose, and lignin, which are constituents of plant cell walls. Among them, hemicellulose is a main component of dietary fiber in rice bran and wheat bran, and is a generic term for sugars other than cellulose. These sugars have various configurations, and the binding mode is also complicated. It forms hydrogen bonds with cellulose, which is the main component of the cell wall, and covalent bonds with lignin, and functions to reinforce the cell wall. It has a structure in which sugars in the side chain are bound to sugars in the main chain that is the backbone, and there are many types of hemicellulases that are enzymes that decompose them. So far, it has been known that arabinose and xylose produced by degradation by hemicellulase have an effect of suppressing blood glucose level and an antioxidant ability, but are not known to be involved in the inhibition of tyrosinase activity.

Aomori Prefectural Industrial Research Center Abstract of Research Results: Okara Useful Use, 3, (2005) Kenji Ozeki, reprint: The latest technology and functionality of fermented and brewed foods, published by CMC Publishing, 4,248, (2006)

  Therefore, the problem to be solved by the present invention is to use a hemicellulose-containing material such as rice bran and wheat bran that has been used as food waste and limited use, and a method for inhibiting tyrosinase activity and a drug that suppresses melanin production. Is to provide.

  The inventors of the present invention have intensively studied to solve the above problems. Surprisingly, it was found that a processed product of raw materials such as rice bran and wheat bran containing a lot of hemicellulose using hemicellulose-degrading enzyme or koji mold inhibits tyrosinase activity, and inhibits tyrosinase activity. The method and tyrosinase activity inhibitor were completed.

That is, the present invention is (1) a method for producing a tyrosinase activity inhibitor, characterized by treating a hemicellulose-containing material with an enzyme or koji mold,
(2) The method according to (1), wherein the hemicellulose-containing material is wheat bran or rice bran,
(3) The method according to (2), wherein the hemicellulose-containing material is wheat bran and the enzyme is selected from the group consisting of Sumiteam ARS, Maceroteam A, and OPTIMASE CX 72L.
(4) The method according to (2), wherein the hemicellulose-containing material is rice bran, and the enzyme is selected from the group consisting of pectinase G “Amano”, hemicellulase “Amano” 90, and maceroteam A.
(5) The method according to (1) or (2), wherein the koji mold is a β-glucosidase or xylanase high-producing koji mold,
(6) The method according to any one of (1) to (5), wherein the hemicellulose-containing material is heat-treated before being treated with an enzyme or koji mold.
(7) The method according to (6), wherein the heat treatment is performed using microwaves,
(8) A tyrosinase activity inhibitor obtainable by the method according to any one of (1) to (7),
(9) A melanin pigment production inhibitor containing the tyrosinase activity inhibitor according to (8),
(10) A whitening cosmetic comprising the tyrosinase activity inhibitor according to (8),
(11) A food browning inhibitor comprising the tyrosinase activity inhibitor according to (8),
I will provide a.

  ADVANTAGE OF THE INVENTION According to this invention, the manufacturing method of an effective tyrosinase activity inhibitor and the tyrosinase activity inhibitor obtained by it are provided. Since the production of melanin can be suppressed by using the inhibitor of the present invention, for example, skin spots, buckwheat and aza, melanin pigmentation of gums and gums, browning of foods and the like can be prevented.

FIG. 1 shows tyrosinase activity inhibition measurement in a treated product of rice bran (untreated microwave) with an enzyme agent or a koji mold extract. The vertical axis represents the inhibition rate (%) of tyrosinase activity. The horizontal axis represents the type of enzyme agent and koji mold extract treated with rice bran. In the table, “β-glu” is β-glucosidase high-producing gonococcal extract, “xyla” is xylanase high-producing gonococcal extract, “Pecti G” is pectinase G “Amano”, “Hemicell” is hemicellulase “Amano” 90 , “Sumichi PX” is Sumi Team PX, “Sumichi ARS” is Sumi Team ARS, “Sumichi ACH-L” is Sumi Team ACH-L, “72L” is OPTIMASE® CX 72L, “Macero A” is Macero Team A, “ "Pecti SS" represents pectinase SS (hereinafter abbreviations mean the same). FIG. 2 shows tyrosinase activity inhibition measurement in a treated product of rice bran (microwave-treated) with an enzyme agent or a koji mold extract. The vertical axis represents the inhibition rate (%) of tyrosinase activity. The horizontal axis represents the type of enzyme agent and koji mold extract treated with rice bran. FIG. 3 shows tyrosinase activity inhibition measurement in a processed product obtained by treating wheat bran (untreated with microwave) with an enzyme agent or koji mold extract. The vertical axis represents the inhibition rate (%) of tyrosinase activity. The horizontal axis represents the type of enzyme agent and koji mold extract treated with wheat bran. FIG. 4 shows tyrosinase activity inhibition measurement in a processed product obtained by treating wheat bran (microwave-treated) with an enzyme agent or a koji mold extract. The vertical axis represents the inhibition rate (%) of tyrosinase activity. The horizontal axis represents the type of enzyme agent and koji mold extract treated with wheat bran. FIG. 5 shows the measurement of antioxidant ability in a treated product of rice bran (untreated microwave) with an enzyme agent. The vertical axis represents the erasure rate (%), and the horizontal axis represents the type of enzyme agent treated with rice bran. The leftmost “rice bran” represents a control not treated with an enzyme agent. FIG. 6 shows the antioxidant ability measurement in the processed material which processed wheat bran (microwave unprocessed) with the enzyme agent. The vertical axis represents the elimination rate (%), and the horizontal axis represents the type of enzyme agent treated with wheat bran. The “wheat bran” at the left end represents a control not treated with an enzyme agent. FIG. 7 shows the measurement of SOD activity in a processed product obtained by treating rice bran (microwave-treated) with a combination of enzyme agents. The vertical axis represents the inhibition rate, and the horizontal axis represents the combination of enzyme agents. In the vertical axis, (untreated) indicates that rice bran has not been pretreated with microwaves, and (treated) indicates that rice bran has been pretreated with microwaves. FIG. 8 shows the measurement of SOD activity in a treated product obtained by treating wheat bran (microwave-treated) with a combination of enzyme agents. The vertical axis represents the inhibition rate, and the horizontal axis represents the combination of enzyme agents. In the vertical axis, (untreated) indicates that wheat bran has not been pretreated with microwave, and (treated) indicates that wheat bran has been pretreated with microwave.

  In one aspect, the present invention provides a method for producing a tyrosinase activity inhibitor, which comprises treating a hemicellulose-containing material with an enzyme or koji mold. The hemicellulose-containing material used in the method of the present invention may be any plant or part thereof as long as it contains hemicellulose. It goes without saying that the hemicellulose-containing material used in the present invention is preferably as the hemicellulose content increases. Preferred hemicellulose-containing materials include, for example, wheat bran, rice bran, soybean okara, bagasse, oat bran, corn bran, beet fiber and the like as food waste, and foods such as cereal hulls, green peas, and beans. Peas, seaweeds such as jellyfishes and hijiki, and processed products thereof, among which wheat bran and rice bran are particularly preferred. Here, wheat bran generally refers to the skin and germ of wheat grains, and contains a large amount of hemicellulose, cellulose, lignin and the like. In the method of the present invention, the hemicellulose-containing material may be solid or soluble. In general, in the method of the present invention, a hemicellulose-containing material can be treated in a solution containing an enzyme, a living organism, or an extract thereof. However, as described later, a living organism can be directly applied to a solid material. Good. The method of the present invention has an advantage that food waste that has been disposed of so far can be effectively used as a new application.

  The method of the present invention includes a treatment step using an enzyme or koji mold. The enzyme used for the treatment of the present invention may be of any kind as long as it has hemicellulose-degrading activity, and is typically hemicellulase. In this specification, hemicellulase refers to an enzyme that cleaves the main chain or side chain between constituent sugars of hemicellulose. For example, as commercially available hemicellulases, pectinase G “Amano” (Amanoenzyme Co., Ltd.), hemicellulase “Amano” 90 (Amanoenzyme Co., Ltd.), Sumiteam PX (Shin Nippon Chemical Industry Co., Ltd.), Sumiteam ARS (Shin Nihon Chemical Industry Co., Ltd.), Sumiteam ACH-L (Shin Nihon Chemical Industry Co., Ltd.), OPTIMASE (registered trademark) CX 72L (GENENCOR Co., Ltd.), Macero Team A (Yakult Pharmaceutical Co., Ltd.) and Examples include pectinase SS (Yakult Pharmaceutical Co., Ltd.). Hemicellulase may be extracted from an organism, and organisms as a source of hemicellulase are known. Among these organisms, those having high hemicellulase activity can be screened and used. Such screening methods are known to those skilled in the art. Examples of organisms having high hemicellulase activity include wood-rotting fungi and koji molds, and koji molds are particularly preferred. Furthermore, in the method of the present invention, not only one hemicellulase but also two or more hemicellulases may be used in combination, or an enzyme other than hemicellulase may be used in combination. In the method of the present invention, not only one species but also two or more species may be used in combination, and these organisms may be used in combination with hemicellulase. A preferred combination of the above enzyme agents is hemicellulase “Amano” 90, Macero Team A, OPTIMASE CX 72L, and Sumi Team PX, but is not limited thereto. By combining these enzymes, it is possible to obtain a tyrosinase activity inhibitor more effectively, such as being able to cleave not only the main chain but also the side chain of hemicellulose.

When an enzyme is used in the method of the present invention, the mixing ratio of the hemicellulase-containing material and the enzyme can be easily determined by those skilled in the art. When a living organism is used as a hemicellulase source in the method of the present invention, an extract of such an organism may act on the hemicellulase-containing material, or such an organism may be directly grown on the hemicellulase-containing material. For example, koji molds may be grown on heat-treated wheat bran and the extract may be used as a tyrosinase activity inhibitor. Even when such a biological extract is used in the method of the present invention, the mixing ratio of the hemicellulase-containing material and the extract can be easily determined by those skilled in the art. The ratio of these enzymes or organisms or extracts and hemicellulase-containing materials can vary depending on factors such as the type and form of the raw materials used, the type of enzymes and koji molds, and processing conditions. Moreover, the treatment conditions in the method of the present invention, for example, optimum pH, optimum temperature, treatment time, CO ₂ concentration, etc. generally exemplify pH 3.0 to 9.5, 40 ° C. to 70 ° C., etc. However, it can be appropriately determined by a person skilled in the art according to the situation.

  Furthermore, the method of the present invention may include a step in which the hemicellulose-containing material is pretreated. Hemicellulose has a structure for reinforcing cell walls in plants, and various constituent sugars form strong and complex bonds with main chains and side chains. Therefore, if the hemicellulose bond can be relaxed or cut as a pretreatment, the efficiency of the treatment can be increased. Examples of the pretreatment that can be used in the present invention include heat treatment using a microwave such as a microwave oven or an autoclave, acid-alkali treatment using an acid or alkaline solution, physical crushing treatment or pulverization treatment, drying treatment, pressure treatment, etc. Although processing etc. are effective, it is not limited only to these. Preprocessing by microwaves using a microwave oven can be performed more conveniently in the present invention. Such pretreatment in the method of the present invention is optional, but is an important factor for increasing the inhibition efficiency of tyrosinase activity.

  In a further aspect, the present invention provides a hemicellulase activity inhibitor (hereinafter sometimes abbreviated as “inhibitor”) obtainable by the above production method. Inhibitors obtained by the production method of the present invention mainly include molecules decomposed from hemicellulose. For example, polysaccharide molecules such as glucuronoxylan, xylan, xyloglucan, arabinan, glucomannan, mannan and galactan can be listed. Furthermore, the inhibitor obtained by the process of the present invention contains monosaccharides such as xylose, arabinose, glucose, fructose, and inositol produced by decomposing the above molecules, oligosaccharides to which they are bound, and the like. The inhibitor obtained by the method of the present invention preferably contains a large amount of xylose or arabinose. The inhibitor obtained by the method of the present invention may contain components other than components generated by the decomposition of hemicellulose, such as various peptides, proteins such as glutelin, globulin, and albumin, saccharides such as cellulose, lignin, etc. Other components may be added. The method of the present invention is characterized in that the hemicellulose-containing material is effectively decomposed by the enzyme, the koji mold or a combination thereof, and the greatest technical feature is to use the inhibitory effect of tyrosinase activity in the obtained inhibitor.

  The method of this invention can include the process of refine | purifying the said inhibitor as needed. The purification step may be performed by a general method among those skilled in the art, and can be performed using a method such as normal phase or reverse phase chromatography, ion exchange chromatography, gel filtration, and the like.

  The inhibitor of the present invention can be used by itself or in combination with a carrier or excipient. Furthermore, the inhibitor of the present invention can be used in combination with other drugs, for example, antioxidants, other enzyme inhibitors, for example, known tyrosinase activity inhibitors.

  It is known that tyrosinase promotes melanin synthesis by oxidizing tyrosine to produce dopa and further oxidizing dopa to induce dopaquinone. The inhibitor obtained by the method of the present invention inhibits the oxidation of such tyrosine and dopa by inhibiting the activity of tyrosinase, thereby suppressing the production of melanin. Therefore, this invention provides the melanin pigment production inhibitor containing the said inhibitor in the further aspect. Using the melanin production inhibitor of the present invention, for example, preventing, suppressing, improving or treating skin spots, buckwheat and aza, gingival gingival gingival melanin, etc. caused by excessive synthesis of melanin Can do. The melanin production inhibitor of the present invention can be applied directly to the affected area, for example, in the form of a lotion, emulsifier, cream, ointment, dietary supplement, drink, tablet, capsule, powder, granule, etc., or taken orally can do. The melanin pigment production inhibitor of the present invention may be a pharmaceutical composition, and the administration route can be easily selected by a doctor. When used as a pharmaceutical composition, suitable excipients or carriers can be used and these are known to those skilled in the art. A typical administration route as a pharmaceutical composition is transdermal administration or oral administration, but is not limited to these administration routes, and is appropriately selected according to conditions such as the type of disease, the state of the subject, and the target site. be able to.

  In another aspect, the present invention provides a whitening cosmetic containing the inhibitor. The whitening cosmetic composition of the present invention may be in a form that can be directly applied to the skin, for example, a lotion, cream, solution, emulsion, pasta, or the like, and can be taken orally, for example, a drink, a tablet, or a capsule. It may be in the form of an agent, powder, granule or the like. The whitening cosmetic composition of the present invention can be prepared by a general method, and includes excipients, fragrances, fats and oils, surfactants, preservatives, sequestering agents, water-soluble polymers, thickeners. , Pigments, UV protection agents, moisturizers, pH adjusters, detergents, desiccants, or emulsifiers, along with lotions, facial cleansers, whitening agents, UV care products, creams, emulsions, lotions, packs It may be in the form of an agent, a skin cleanser, or a gel, but is not limited thereto.

  In yet another aspect, the present invention provides a food browning inhibitor containing the inhibitor. The browning inhibitor of the present invention can be applied to a food or food material by a known method such as spraying, dipping or coating.

  The compounding quantity of the inhibitor of this invention in the said melanin pigment production inhibitor of this invention, cosmetics for whitening, or the browning prevention agent of foodstuffs can be suitably changed according to the objective, use, object, etc.

  Furthermore, the inhibitor of the present invention, or a melanin pigment formation inhibitor, a whitening cosmetic or a food browning inhibitor containing the same, other agents such as antioxidants, other enzyme inhibitors such as other May be used in combination with other tyrosinase activity inhibitors.

  EXAMPLES The present invention will be described specifically and in detail below with reference to examples, but the examples should not be construed as limiting the present invention.

A. Pretreatment of wheat bran and rice bran by microwave In the present invention, a sample of rice bran and wheat bran pretreated by microwave and a sample without pretreatment were used. Briefly, the microwave treatment is performed by putting 10 g of rice bran (Ozeki Co., Ltd.) or wheat bran (Nisshin Flour Milling Co., Ltd.) into an Erlenmeyer flask and setting the microwave oven to 500 W. In wheat bran, it was performed by heating for 50 to 60 seconds. Subsequently, each obtained powder was dried at 60 degreeC.

B. Preparation of extract with high yield of useful enzyme-producing koji mold In the present invention, β-glucosidase-producing bacteria (Ozeki et al., 7th Annual Meeting of Molecular Biology Conference, p. 78) and xylanase-producing bacteria (Takagi et al., 7th Annual Meeting of Molecular Biology Conference, p. 76) was used. The above useful enzyme-producing gonorrhea was inoculated into a bran medium (autoclave sterilized by adding 6 mL of distilled water to 10 g of bran in a 200 mL conical flask) and cultured at 30 ° C. for 3 days. To the culture broth, 50 mL of a bran medium extract buffer (10 mM acetate buffer, pH 5.0, 0.5% NaCl) was added and left for 3 hours. Then, it filtered using the nylon mesh filter and preserve | saved in a 50 mL falcon tube.

C. In the present invention, an enzyme agent (hemicellulase) for treating wheat bran and rice bran extract, and the following 8 enzyme agents as extract of koji mold and Two kinds of useful enzyme high-producing koji mold extracts were used.
・ Pectinase G “Amano” (Amanoenzyme Co., Ltd.)
・ Hemicellulase “Amano” 90 (Amanoenzyme Co., Ltd.)
・ Sumi Team PX (Nippon Chemical Industry Co., Ltd.)
・ Sumi Team ARS (Nippon Chemical Industry Co., Ltd.)
・ Sumi Team ACH-L (Shin Nihon Chemical Industry Co., Ltd.)
OPTIMASE (registered trademark) CX 72L (GENENCOR CORPORATION)
・ Macero Team A (Yakult Pharmaceutical Co., Ltd.)
・ Pectinase SS (Yakult Pharmaceutical Co., Ltd.)
・ Β-Glucosidase high production koji mold extract ・ Xylanase high production koji mold extract

First, 50 mM acetate buffer / Tris-HCl buffer was adjusted to the optimum pH of each enzyme agent described in Table 1 below. 1 g of wheat bran or rice bran (untreated and heat treated with microwave) is added to 10 mL of these buffers, and the above enzyme agent (10 mg for solid, 10 μL for liquid) or useful enzyme 300 μL of high-producing koji mold extract was added. Subsequently, it incubated, making it shake gently at the optimal temperature of each enzyme agent for 1 day. Then, it extracted with the filter paper and filtered using the membrane filter.
(Table 1. Optimal conditions for each enzyme agent)

D. Measurement of enzyme activity of prepared treatment (a) Inhibition of tyrosinase activity Rice bran and wheat bran (microwave treatment in order to select an enzyme agent or a koji mold extract that is highly efficient in degrading wheat bran and rice bran Inhibition of tyrosinase activity was measured in the treated products treated with 8 kinds of enzyme agents shown in Table 1 and extracts of high-producing gonococci with high useful enzymes shown in Table 1. The measurement method is described in Experimental Biomolecular Chemistry (Kyoritsu Shuppan) p. 151, according to the method described in 151. The microwave treatment was performed for 50 seconds with a microwave oven set to 500 W for rice bran and 50 to 60 seconds for a wheat bran, and with a microwave oven set to 500 W for wheat bran. As a result, both rice bran and wheat bran not pretreated with microwave, and rice bran and wheat bran treated with microwave, both treated with enzyme agent or high enzyme-producing gonococcus extract, have high tyrosinase activity inhibition. Although the rate was shown, the treatment pretreated with the microwave showed a higher inhibition rate than the non-pretreated treatment (FIGS. 1-4). In particular, when rice bran is used, it is high when treated with xylanase high-producing gonococcal extract, pectinase G “Amano”, Sumiteam PX or Maceroteam A. When wheat bran is used, xylanase high-producing gonococcal extract, hemicellulase “Amano” "90, found to be high when treated with Sumiteam PX or OPTIMASE® CX 72L. Moreover, when the difference with the presence or absence of the process by a microwave was seen about the same enzyme agent or a koji mold extraction, the processed material processed with the microwave showed the high tyrosinase activity inhibition rate compared with the thing which is not processed (FIG. 1). ~ 4). From the above, it was shown that a processed product obtained by treating a material containing a large amount of hemicellulose such as rice bran and wheat bran with an enzyme agent such as hemicellulase or a koji mold extract such as xylanase has an inhibitory effect on tyrosinase activity.

(B) D-xylose measurement In order to examine the relationship between the increase in tyrosinase activity inhibition and the release of D-xylose, three types of xylanase high-producing bacteria extract, OPTIMASE ( Registered trademark CX 72L (GENENCOR Co., Ltd.) and Macero Team A (Yakult Pharmaceutical Co., Ltd.) were selected, and the amount of D-xylose was measured using a D-xylose measurement kit (Nippon Biocon Co., Ltd.). Absorbance at 340 nm was measured according to the instructions in the instructions attached to the kit. The results are shown in Table 2 below. In the column of “Pretreatment” in the table, “◯” indicates that the microwave has been processed, and “X” indicates that the microwave has not been processed. From the column of “xylose content” in Table 2, treated products of rice bran with high xylanase-producing bacteria extract (described as xylanase bran extract in the table) and wheat bran are OPTIMASE (registered trademark). It was found that the treated product with CX 72L (described as OPTIMASE CX 72L in the table) contains a large amount of xylose. Moreover, it was shown that the processed material which performed the pre-processing by the microwave with all the 3 types which were tested has a higher xylose content than the thing which has not performed the pre-processing.
(Table 2. Measurement result of xylose content)

（表４．小麦フスマを酵素剤で処理した処理物におけるキシロース含有量）

（表５．米ヌカを酵素剤の組み合わせで処理した処理物におけるキシロース含有量）

（表６．小麦フスマを酵素剤の組み合わせで処理した処理物におけるキシロース含有量）

Furthermore, the amount of D-xylose was measured in order to examine whether the combination of enzyme agents increased the release of D-xylose as compared with the treated product treated alone. The results are shown in Tables 3-6. In Tables 5 and 6, hemicell is hemicellulase “Amano” 90, pectin G is pectinase G “Amano”, macero A is macero team A, sumi PX is sumi team PX, sumi ARS is sumi team ARS, 72L is OPTIMASE® CX 72L is meant. From Tables 3-6, the combination of the enzyme agent in which the xylose content was particularly increased compared to the processed product treated with the single enzyme agent was Hemicelle x Macero A, Rice Nuka (treated) when rice bran (untreated) was used. ), Pecti G x Sumitomo PX, Pecti G x Macero A, Wheat bran (untreated), Sumichi ARS x Macero A, Sumichi ARS x 72L, Macero A x 72L, wheat bran (treated) ) Was 72 L × Hemicell, 72 L × Sumichi PX, Hemicell × Sumichi PX.
(Table 3. Xylose content in processed product of rice bran treated with enzyme)

(Table 4. Xylose content in processed product of wheat bran treated with enzyme)

(Table 5. Xylose content in processed product of rice bran treated with enzyme combination)

(Table 6. Xylose content in processed products obtained by treating wheat bran with a combination of enzyme agents)

(C) Measurement of Antioxidant Capacity (Radical Catch) Next, in a processed product obtained by treating rice bran and wheat bran with each of the above enzyme agents, the antioxidant capacity was measured using an antioxidant capacity measurement kit (Aroka Co., Ltd.). Was measured. The measurement was performed according to the instructions attached to the kit. In the present invention, the sampling time is 5 seconds, the sampling point is 24 points, and the measurement time is 120 seconds. The activity inhibition rate was calculated from the following formula, and the elimination rate was calculated.
Activity inhibition rate (%) = 100− (A) / (B) × 100
[Where A is the amount of blank glucose produced and B is the amount of glucose produced by the extract. ]
Next, the IC ₅₀ value was determined as a sample concentration giving half of the control light emission amount or a sample concentration of 50% erasure rate (suppression rate). As a result, hemicellulase “Amano” 90 showed the highest erasure rate in the treated product of rice bran (untreated microwave) with the enzyme agent, and an erasure rate close to 60% was observed (FIG. 5). ). Also, a high erasure rate of 50% or more was observed for Sumiteam PX. In addition, a high erasure rate of about 50% was also observed for pectinase SS. On the other hand, hemicellulase “Amano” 90 showed the highest elimination rate in the processed product of wheat bran (untreated with microwave) with an enzyme agent, and a high elimination rate of 90% or more was observed (FIG. 5). 6). Further, an erasure rate of nearly 90% was also observed for pectinase SS. In addition, pectinase G “Amano” showed an erasure rate exceeding 85%, and Sumiteam PX showed an erasure rate close to 70%. In addition, wheat bran was generally observed to have a higher elimination rate than rice bran. From these results, it was found that the antioxidant ability was increased when materials such as wheat bran and rice bran were treated with an enzyme agent.

(D) SOD activity measurement (NBT reduction method)
SOD activity detection kit (Wako Pure Chemical Industries, Ltd.) was used to investigate whether processed products of rice bran and wheat bran with various enzyme preparations and useful enzyme high-producing gonococcal extracts can suppress active oxygen. SOD activity was measured using a strain). The measurement was performed according to the instructions attached to the kit. Next, by using the absorbance value obtained by this method, the inhibition rate of SOD activity was calculated by the following formula.
SOD activity value (% inhibition) = [(E1) − (E2)] / (E1) × 100
[In the _formula, a _{E1 = E EL -E BL-BL} , is _{E2 = E S -E S-BL} . ]

  As a result, the pre-treated rice bran processed product showed the highest inhibition rate when it was treated with a combination of pectinase G “Amano” and maceroteam A, and showed an inhibition rate of nearly 80% ( FIG. 7). In addition, in the processed product of rice bran that has not been treated with microwaves, the inhibition of hemicellulase “Amano” 90 and pectinase G “Amano”, and the combination of hemicellulase “Amano” 90 and maceroteam is 50% or more. A rate was observed (Figure 8). When attention was paid to the influence of pretreatment with microwaves, it was found that the combination of pectinase G “Amano” and maceroteam A resulted in an extremely high inhibition rate of the pretreatment. On the other hand, the wheat bran showed the highest inhibition rate when the wheat bran pretreated with microwave was treated with a combination of OPTIMASE (registered trademark) CX 72L × Sumiteam PX, and the inhibition rate was close to 60%. Indicated. In addition, a high inhibition rate of 50% or more was also observed when the wheat bran not treated with microwaves was treated with a combination of maceroteam A × OPTIMASE (registered trademark) CX 72L. From these results, it was shown that when each material was treated with an enzyme agent, the antioxidant ability was increased.

  According to the present invention, a method and an inhibitor for tyrosinase activity are provided, so that melanin pigmentation caused by tyrosinase activity can be prevented. Therefore, the present invention is related to skin whitening, darkening of gums and gums, and prevention of food browning. It can be used in fields such as cosmetics, medicine, and food.

Claims (11)

  A method for producing a tyrosinase activity inhibitor, which comprises treating a hemicellulose-containing material with an enzyme or koji mold.   The method according to claim 1, wherein the hemicellulose-containing material is wheat bran or rice bran.   The method of claim 2, wherein the hemicellulose-containing material is wheat bran and the enzyme is selected from the group consisting of Sumiteam ARS, Maceroteam A, and OPTIMASE CX 72L.   The method according to claim 2, wherein the hemicellulose-containing material is rice bran and the enzyme is selected from the group consisting of pectinase G “Amano”, hemicellulase “Amano” 90, and Macero Team A.   The method according to claim 1 or 2, wherein the koji mold is a β-glucosidase or xylanase high-producing koji mold.   The method according to any one of claims 1 to 5, wherein the hemicellulose-containing material is heat-treated before being treated with an enzyme or koji mold.   The method according to claim 6, wherein the heat treatment is performed using microwaves.   A tyrosinase activity inhibitor obtainable by the method according to any one of claims 1 to 7.   A melanin pigment production inhibitor comprising the tyrosinase activity inhibitor according to claim 8.   A whitening cosmetic comprising the tyrosinase activity inhibitor according to claim 8.   The browning prevention agent of the food containing the tyrosinase activity inhibitor of Claim 8.

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