JP2010248186A - Lipase inhibitor - Google Patents

Abstract

An object of the present invention is to provide a new material that is a surfactant having excellent biocompatibility and can be used in a wide range of fields such as pharmaceuticals, cosmetics, and foods and drinks, and that can inhibit the hydrolysis reaction of lipase.
A sugar-type biosurfactant produced by microorganisms using various biomass such as vegetable oil and sugar as a raw material is used as a lipase inhibitor. The lipase inhibitor is used as an active ingredient in pharmaceuticals, health foods or cosmetics that are effective in preventing, ameliorating, and treating various diseases involving lipase.
[Selection figure] None

Description

  The present invention relates to a lipase inhibitor useful for pharmaceuticals, cosmetics, foods and drinks, and the like, which is composed of a highly safe glycolipid (glycotype biosurfactant) produced by microorganisms.

  Lipase, a lipid-hydrolyzing enzyme, degrades the lipids contained in foods, beverages, cosmetics, etc. into glycerin and fatty acids, causing deterioration such as deteriorating odor and reducing the value of the product. . In addition, lipase produced by microorganisms resident in human skin breaks down lipids on the skin into glycerin and free fatty acids, and some of these free fatty acids have adverse effects on the skin. It is known that it causes flames, dandruff, etc., and free fatty acids are further decomposed to cause body odor. On the other hand, in the human body, lipase is a digestive enzyme secreted from the pancreas, and lipids taken orally are hydrolyzed into glycerin and fatty acids by the action of lipase to promote digestion and absorption in the body. Lipids are particularly high in energy, and fat intake by Japanese in recent years tends to increase, contributing to lifestyle-related diseases such as obesity and hyperlipidemia. In order to solve the problems caused by such lipases, lipase inhibitors that inhibit lipases have been studied.

  In particular, the lipase inhibitor is used in pharmaceuticals, foods and drinks, and is in direct contact with a human living body. As some of the most recent development examples are listed in the following patent documents, in recent years, development of lipase inhibitors comprising plant-derived components has been carried out extremely widely and actively. (Patent Documents 1 to 12).

JP 2006-056850 A JP2007-145753 JP 2007-161645 JP 2007-246429 A JP 2007-246471 A JP 2008-019180 JP 2008-074735 A JP 2008-273980 A JP 2009-029786 Japanese Patent No. 3689099 Japanese Patent No. 4044274 Japanese Patent No. 4233645

  Lipase acts in an environment where lipid and water coexist, and the use of a surfactant is essential for materials used in such situations. Therefore, an agent that inhibits or promotes the lipase reaction is highly useful when it exhibits an effect when it coexists with the surfactant, and the surfactant itself exhibits such an effect. However, many surfactants emulsify and disperse lipids to promote contact between lipase and lipid, promote hydrolysis reaction, keep the vicinity of the active center of lipase in a hydrophobic environment, and maintain enzyme activity itself. Shows the effect of increasing. On the other hand, in the case of an active agent having a high surface active effect, the enzyme protein is deactivated. Moreover, most of the surfactants currently used are synthetic chemicals made from petroleum, and only a few types can be applied to living organisms. As described above, at present, most surfactants cannot be applied as lipase inhibitors excellent in safety aimed at development in the present invention.

  In view of the above circumstances, the present invention aims to provide a new material which is a surfactant having excellent biocompatibility that can be used in a wide range of fields such as pharmaceuticals, cosmetics, and foods and drinks, and which can suppress the hydrolysis reaction of lipase. And

  As a result of intensive studies to solve the above problems, the present inventor has focused on biosurfactants, which are amphipathic lipids produced by microorganisms using various biomass such as vegetable oils and sugars as the target lipase inhibitors. In particular, we investigated the effect of adding glycoform biosurfactants that have high affinity to biomolecules such as proteins and have various physiological activities on the lipase reaction system, and showed a specific reaction inhibitory effect with very small amounts of addition. The inventors have found out that the present invention has been achieved and have made the present invention.

That is, the present invention is as follows.
(1) A lipase inhibitor comprising a microorganism-produced sugar-type biosurfactant as an active ingredient.

(2) The lipase inhibitor according to the above (1), wherein the microorganism-produced sugar type biosurfactant is a mannosyl alditol lipid described in the following chemical formula (1).
(Wherein R ¹ represents an aliphatic acyl group having 2 to 24 carbon atoms, R ² represents hydrogen or an acetyl group, and R ³ represents hydrogen or an aliphatic acyl group having 2 to 24 carbon atoms. R ² and R ³ may be the same or different, and n is an integer of 1 to 4.)

(3) The lipase inhibitor according to the above (1), wherein the microorganism-produced sugar type biosurfactant is a mannosyl erythritol lipid described in the following chemical formula (2).
(Wherein R ¹ represents an aliphatic acyl group having 2 to 24 carbon atoms, R ² represents hydrogen or an acetyl group, and R ³ represents hydrogen or an aliphatic acyl group having 2 to 24 carbon atoms. The R ² and R ³ at each position may be the same or different.)

(4) A pharmaceutical or health food for prevention, amelioration or treatment of adult diseases, characterized by containing the microbial-produced sugar-type biosurfactant according to any one of (1) to (3) as an active ingredient.

(5) A fat absorption inhibitor comprising the microorganism-produced sugar-type biosurfactant according to any one of (1) to (3) as an active ingredient.

(6) An obesity inhibitor comprising the microbial-produced sugar-type biosurfactant according to any one of (1) to (3) as an active ingredient.

(7) A dermatitis inhibitor comprising the microorganism-produced sugar-type biosurfactant according to any one of (1) to (3) as an active ingredient.

(8) An acne inhibitor comprising the microbial sugar-type biosurfactant according to any one of (1) to (3) as an active ingredient.

(9) A dandruff suppressant comprising the microorganism-produced sugar-type biosurfactant according to any one of (1) to (3) as an active ingredient.

The lipase inhibitor of the present invention inhibits, for example, pancreatic lipase, inhibits degradation of fat in the intestinal tract by pancreatic lipase, and suppresses its absorption. Therefore, it is effective in preventing, improving, and treating so-called adult diseases with various symptoms such as hyperlipidemia caused by excessive absorption of fat, obesity, or accompanying arteriosclerosis and ischemic heart disease. Used.
In addition, sebum degradation and free fatty acid production are suppressed by changing the activity of lipase, and as a result, it has the effect of preventing and improving various skin diseases such as acne and dandruff. Furthermore, since the lipase inhibitor of the present invention is a component produced by microorganisms from various biomass and is extremely excellent in safety, it can be used in a wide range of fields such as pharmaceuticals, cosmetics and health foods.

In Example 1, it is the graph which followed the time-dependent change of the hydrolysis reaction of Candida antarctica origin lipase B. In the comparative example 1, it is the graph which followed the addition effect of surfactant with respect to the hydrolysis reaction of Candida antarctica origin lipase B. In the comparative example 2, it is the graph which put together the addition effect of the sugar type surfactant with respect to the hydrolysis reaction of Candida antarctica origin lipase B. In Example 2, it is the graph which put together the addition effect of the sugar type surfactant with respect to the hydrolysis reaction of lipase derived from Rhizomucor miehei . In Example 3, it is the graph which put together the addition effect of the sugar type surfactant with respect to the hydrolysis reaction of Thermomyces lanuginosus origin lipase. In Example 4, it is the graph which evaluated the addition effect of MEL with respect to the hydrolysis reaction of porcine pancreas origin lipase. In Example 5, it is the graph which put together the addition effect of each different sugar type surfactant with respect to the hydrolysis reaction of Candida antarctica origin lipase B. In Example 6, it is the graph which put together the addition effect of each different sugar-type surfactant with respect to the hydrolysis reaction of lipase derived from Rhizomucor miehei . In Example 7, it is the graph which put together the addition effect of each different sugar-type surfactant with respect to the lipase hydrolysis reaction derived from Thermomyces lanuginosus . In Example 8, a graph summarizing the effect of adding each on the hydrolysis reaction of Rhizopus delemer lipase different sugar surfactants. In Example 9, it is the graph which put together the addition effect of each different sugar-type surfactant with respect to a porcine pancreas origin lipase hydrolysis reaction.

<Microbial-produced glycolipids / glycotype biosurfactants>
The microorganism-produced glycolipid used in the present invention (hereinafter referred to as “glyco-type biosurfactant” or “glyco-type BS”) is a kind of amphipathic lipid produced and secreted by microorganisms using various biomass such as vegetable oil and carbohydrate. The sugar type BS has a sugar chain structure as a hydrophilic group in the molecule, and various medium chain and long chain fatty acids (saturated, unsaturated, branched, hydroxy type, etc.) are bound to the hydrophilic group as a hydrophobic group. Has a molecular structure. Among various BSs, glycosylated BS is most frequently studied because of its excellent productivity, and many types of substances produced by bacteria and yeast have been reported.

Specific examples include rhamnolipid, sophorolipid, mannosyl alditol lipid, trehalose lipid, cellobiose lipid, and oligosaccharide lipid. These properties change depending on the molecular structure (sugar chain structure, lipid structure / composition), but if the structure is equivalent, the same effect can be achieved regardless of the production conditions (production microorganisms, types of raw materials). can get. Among the above, particularly mannosyl alditol lipid is widely distributed in familiar environment such as flowers and soil from being produced by Pseudozyma (Pseudozyma) or the genus Ustilago (Ustilago) genus yeasts such as, high safety is expected Above, a lipase inhibitory effect is shown with a very small amount of use.

<Mannosyl alditol lipid>
Mannosyl alditol lipid (hereinafter referred to as MAL) used in the present invention is obtained by culturing a MAL-producing microorganism, and a typical example of its chemical structure is represented by the following formula (1): 4-O-β-D -Mannopyranosyl-alditol as its basic structure. Here, alditol is a sugar alcohol in which both ends of a linear saccharide are reduced, and glycerin having 3 carbon atoms, erythritol having 4 carbon atoms, threitol, arabinitol having 5 carbon atoms, xylitol, ribitol. Examples thereof include allitol having 6 carbon atoms, sorbitol, mannitol, iditol, galactitol, and taritol.

However, in the above formula (1), R ¹ is an aliphatic acyl group having 2 to 24 carbon atoms, including straight-chain or branched, saturated or unsaturated aliphatic acyl group. R ² represents hydrogen or an acetyl group. R ³ is hydrogen or a C2-C24 aliphatic acyl group, and includes a linear or branched saturated or unsaturated aliphatic acyl group. Two R ² and R ³ may be the same or different. The type of aliphatic acyl group of the substituents R ¹ and R ³ and the number of carbons thereof basically depend on the fatty acid in the fats and oils contained in the MEL production medium, but the number of carbons is the MEL-producing bacterium used. Varies depending on the degree of utilization of fatty acids. Therefore, each obtained MEL is usually in the form of a mixture of compounds in which the fatty acid residue portion of the substituent R is different. Moreover, n in the said Formula (1) is an integer of 1-4.
Among the mannosyl alditol lipids, the mannosyl erythritol lipid represented by n = 2 in the formula (1) is most excellent in productivity, so that it is desirable to use it.

<Mannosyl erythritol lipid>
Mannosyl erythritol lipid (hereinafter referred to as MEL) used in the present invention is obtained by culturing a MEL-producing microorganism and is represented by n = 2 in the above formula (1). As typical examples of MEL, chemical structures of MEL-A, MEL-B, MEL-C, MEL-D, single-chain MEL, and three-chain MEL-A are shown below.

In these formulas, R ¹ is the same group as the above formula (1), and Ac represents an acetyl group.

<Characteristics of sugar-type BS>
The sugar type BS has a high surface activity, and can be used as various catalysts for surfactants and fine chemicals. The sugar-type BS can form various nanostructures in an aqueous solution. For example, in the case of MEL, vesicles, lamellar phases, cubic phases, sponge phases (coacervates), etc. can be formed in a wide range of concentrations and temperatures (T. Imura et al., Langmuir, 23, 1659-1663 (2007), W. Worakitkanchanakul et al, Colloid Surf. B, 65, 106-112 (2008)). Therefore, it can be used as cosmetics and external preparations for skin by taking a lamellar structure with excellent affinity to the skin, and it can be used for oral and parenteral administration by taking a capsule structure containing other drugs etc. It is possible to develop products with appropriate dosage forms.

  Furthermore, various physiological actions are known, and application to pharmaceutical uses is expected. For example, in the case of the above MEL, when MEL is allowed to act on human acute promyelocytic leukemia cell line HL60, there is an effect of inducing differentiation of leukemia cells that differentiates the granule system (H. Isoda et al., Lipids, 32, 263-271 (1997), H. Isoda et al., Biosci. Biotech. Biochem., 61, 609-614 (1997)), and when MEL acts on PC12 cells derived from rat adrenal medullary pheochromocytoma, neurite outgrowth It has a physiological activity such as an action of inducing differentiation of nervous system cells (Y. Wakamatsu et al., Eur. J. Biochem., 268, 374-383 (2001)). Furthermore, for the first time as a glycolipid produced by microorganisms, it becomes possible to induce apoptosis of melanoma cells (X. Zhao et al., Cancer Research, 59, 482-486 (1999)), and has a cancer cell proliferation inhibitory action. The sugar-type BS is considered to have excellent biodegradability and high safety in addition to these physiological actions, and is expected to be used for pharmaceuticals, cosmetics, foods and drinks, and the like.

  It has not been known at all that the above-mentioned sugar-type BS exhibits an inhibitory action on lipase activity, and is a new finding obtained by the present invention.

  The sugar type BS can be used as it is as a lipase inhibitor, as a single product purified by a conventional purification means such as silica gel column chromatography. Moreover, you may use as a mixture which mixed them. Furthermore, you may utilize in the state of the solution dissolved in the solvent which does not disturb enzyme activity, and aqueous solution.

  The above-mentioned sugar type BS has excellent lipase inhibitory activity, and as a medicine, cosmetics and food containing these, further as an anti-obesity agent, hyperlipidemia improving agent, acne, dandruff skin improving agent It can be used.

  As an application method as a medicine, either oral administration or parenteral administration can be adopted. In administration, the active ingredient can be mixed with a solid or liquid nontoxic pharmaceutical carrier suitable for administration methods such as oral administration, rectal administration, and injection, and administered in the form of a conventional pharmaceutical preparation. Examples of such preparations include solid preparations such as tablets, granules, powders and capsules, solutions such as solutions, suspensions and emulsions, freeze-dried preparations, and the like. It can be prepared by conventional means. Examples of the non-toxic pharmaceutical carrier include glucose, lactose, sucrose, starch, mannitol, dextrin, fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, amino acid, gelatin, Examples include albumin, water, and physiological saline. Further, if necessary, conventional additives such as a stabilizer, a wetting agent, an emulsifier, a binder, and an isotonic agent can be appropriately added.

  Further, the lipase inhibitor of the present invention includes, for example, moisturizing cosmetics, skin roughening cosmetics, makeup cosmetics, anti-dandruff cosmetics, hair growth cosmetics, itching cosmetics, cleaning cosmetics, sunscreen cosmetics, body odor cosmetics, fragrance cosmetics, etc. Can be added to cosmetics. The addition amount at this time is not particularly limited as long as the effect expected by the substance is effectively exhibited, and is 0.000001 to 50% by weight of the total amount, preferably 0.00001 to It is 10% by weight, more preferably 0.0001 to 1% by weight. In addition, when adding the lipase inhibitor of this invention to cosmetics, another active ingredient, an amino acid, a pharmaceutically acceptable excipient | filler, a pigment | dye, a fragrance | flavor, etc. can also be used in combination as appropriate. Further, the form of the product is also arbitrary, and for example, any of liquid, powder, cream and the like is possible. From the functional aspect, for example, milky lotion, cosmetic liquid, face cream, hand cream, lotion, essence, shampoo, rinse and the like are preferable.

  Moreover, the lipase inhibitor of this invention can be added to skin disease therapeutic agents, such as an acne therapeutic agent and a dermatitis therapeutic agent. The addition amount at this time is not particularly limited as long as the effect expected by the substance is effectively exhibited, and is 0.000001 to 50% by weight of the total amount, preferably 0.00001 to It is 10% by weight, more preferably 0.0001 to 1% by weight. In addition, when adding the lipase inhibitor of this invention to a skin disease therapeutic agent, another active ingredient, an amino acid, a pharmaceutically acceptable excipient | filler, a pigment | dye, a fragrance | flavor, etc. can also be used in combination as appropriate. Further, the form of the product is also arbitrary, and for example, any of liquid, powder, cream and the like is possible.

The sugar type BS used as an inhibitor in the present invention is not particularly limited, but as an example, Pseudozyma antarctica KM-34 (FERMP-20730) strain is used and cultured using soybean oil as a raw material. An experiment using MEL-A (the compound on the upper left of formula (3)) obtained by isolating and purifying the glycolipid component from the above as an inhibitor will be described. The present invention is not limited thereby.

Example 1
(Inhibitory effect on lipase B derived from Candida antarctica )
P-nitrophenyl acetate (pNPA) was selected as a substrate, and Candida antarctica- derived lipase B (CalB) (Novozym 435 CALB L manufactured by novozymes) was added for ester hydrolysis reaction. The progress of the reaction was evaluated by measuring UV light absorption of p-nitrophenol (pNP) produced by decomposition at a wavelength of 410 nm with an ultraviolet-visible spectrophotometer. The specific operation procedure is described below.

pNPA was dissolved in 30 mM Hepes buffer (pH 7.5) containing 6% DMSO to prepare 570 μL of a 2.5 mM substrate solution. Here, the reaction was started when 30 μL of the enzyme solution prepared at each concentration was added, and the change in absorbance at 410 nm was followed for 15 minutes, which was used as a control.
Next, a sample solution in which MEL-A was added to each concentration with respect to the substrate solution was prepared, and the enzyme solution was added thereto, and the change in absorbance with time was similarly traced. The results are shown in FIG.

  FIG. 1 is a graph when the lipase concentration is 3 U / mL. The production of pNP was significantly suppressed by adding MEL to the control (glycolipid non-addition system; indicated as 0% in the figure). The effect was seen with the addition of 0.001%, and no significant difference appeared even at higher concentrations. From this result, it was confirmed that the lipase inhibitory effect of MEL is sufficiently effective in a very small amount.

Comparative Example 1
(Effects of other surfactants on lipase)
Instead of MEL-A, Triton X-100 (t-octylphenoxypolyethoxyethanol: Triton), a nonionic surfactant that has been reported to promote lipase activity, was used in the same manner as in Example 1. Experiments were performed and the reaction was followed. FIG. 2 shows a summary of graphs that trace changes in absorbance with respect to the added amount of activator.

  As the amount of Triton added increased, the reaction was greatly accelerated, whereas when MEL-A was added, a certain reaction inhibitory effect was observed.

Comparative Example 2
(Effects of other sugar-type surfactants on lipase)
In place of MEL-A, a common sugar-type synthetic surfactant, span 20 (sorbitan monolaurate: Span), and sugar ester (sucrose fatty acid diester: SE) were used. Similar experiments were performed to track the reaction. The absorbance at 15 minutes after the start of the reaction in the control (non-surfactant added system) was defined as a reaction rate of 100%, and the reaction rate was calculated therefrom. A graph summarizing the results is shown in FIG. In the figure, the white bar represents the reaction rate when the activator addition amount is 0.001%, and the black bar represents the reaction rate when the addition amount is 0.01%.

  When other surfactants were added, the reaction of lipase was all promoted, and the reaction was greatly promoted as the amount added was increased. On the other hand, the progress of the reaction was suppressed only when MEL-A was added. The effectiveness of MEL as a lipase inhibitor was shown.

Example 2
(Inhibitory effect on lipase derived from Rhizomucor miehei )
Using Rhizomucor miehei lipase (Novozymes Corporation Novozym 388) lipase, an experiment was conducted in the same manner as in Example 1 and Comparative Examples 1 and 2. The results are summarized in FIG.

  Although a slight inhibitory effect was observed even with sugar type surfactants other than Triton, it was confirmed that the reaction was remarkably inhibited by adding MEL-A in particular.

Example 3
(Inhibitory effect on lipase from Thermomyces lanuginosus )
Experiments were performed in the same manner as in Example 1 and Comparative Examples 1 and 2, using Thermomyces lanuginosus- derived lipase (Lipozyme TL 100L manufactured by novozymes) as the lipase. The results are summarized in FIG.

  Although an inhibitory effect was observed even with sugar type surfactants other than Triton, it was confirmed that the reaction was significantly inhibited by adding MEL-A in particular.

Example 4
(Inhibitory effect on porcine pancreatic lipase)
The ester hydrolysis reaction was evaluated using p-nitrophenyl laurate (pNPL) as a substrate and porcine pancreas-derived lipase (Lipase from porcine pancreas, Type II, manufactured by SIGMA) as the lipase. 475 μL of a substrate solution prepared by dispersing pNPL to 2.5 mM in 50 mM sodium phosphate buffer (pH 7.0) containing 2% Triton as a substrate dispersant for 5 minutes in a warm bath (37 ° C.). Preheated, 25 μL of 30 mg / mL enzyme solution was added thereto, and allowed to react in a warm bath for 15 minutes. 1 mL of acetone was added to the reaction solution after the reaction to stop the reaction, and the insoluble part was precipitated by centrifugation. About the supernatant liquid, the progress of reaction was evaluated by measuring UV light absorption of wavelength 410nm of p-nitrophenol (pNP) produced | generated by decomposition | disassembly with an ultraviolet visible spectrophotometer.
Using the above results as a control, prepare a sample solution with MEL-A added to each concentration of the substrate solution, and add the enzyme solution to follow the change in absorbance over time. did. The absorbance ratio (%) obtained by adding each concentration of MEL when the absorbance in the control is 100 is shown in FIG.

  It was confirmed that the hydrolysis reaction of porcine pancreatic lipase can be suppressed by adding 1% of MEL-A even in the presence of Triton. Pancreatic lipase functions to break down lipids in the living body and promote digestion and absorption. By inhibiting this, obesity-suppressing effect is expected.

[Lipase inhibition test using other MEL]
In the following S Examples, the effects of MEL addition on the ester hydrolysis reactions of various lipases with different origins were evaluated for MELs other than MEL-A produced using the Pseudozyma antarctica KM-34 (FERMP-20730) strain. did. The used MEL is as follows.
1) Mel -A obtained by culturing with Pseudozyma antarctica KM-34 (FERMP-20730) strain using soybean oil as the main raw material, and recovering the glycolipid component from the culture solution, followed by isolation and purification, MEL-B, MEL-C (hereinafter abbreviated as A1, B1, and C1, respectively)
Fatty acid (R ¹ ) composition (%) ・ ・ ・ C8: 0 = 22.8, C10: 0 = 26.2, C10: 1 = 28.4, C10: 2 = 2.9, C12: 0 = 5.2, C12: 1 = 5.1, C14: 2 = 4.7, Unknown = 4.7
2) MEL-B (hereinafter abbreviated as B2) obtained by culturing with Pseudozyma tsukubaensis NMRC1940 strain using olive oil as the main raw material, recovering glycolipid components from the culture broth, and isolation and purification.
Fatty acid (R ¹ ) composition (%) ・ ・ ・ C8: 0 = 28.1, C10: 0 = 1.3, C10: 1 = 1.7, C10: 2 = 1.1, C12: 0 = 12.0, C12: 1 = 11.5, C14: 0 = 1.4, C14: 1 = 7.8, C14: 2 = 27.1, Unknown = 8.0
3) Mel -C (hereinafter abbreviated as C2) obtained by culturing with Pseudozyma siamensis strain CBS9960, using safflower oil as the main raw material, recovering the glycolipid component from the culture, and then isolating and purifying it. )
Fatty acid (R ¹ ) composition (%): Mannose 2nd: C2 = 18, C4 = 82, Mannose 3rd: C14: 0 = 2.8, C14: 1 = 4.3, C14: 2 = 32.6, C16: 0 = 23.5, C16: 1 = 12.5, C16: 2 = 16.6, C18: 2 = 7.6, Unknown = 0.1
4) Using Pseudozyma crassa strain CBS9959 , culturing oleic acid as the main raw material, collecting glycolipid components from the culture, and then isolating and purifying the resulting MEL-A, MEL-B, and MEL-C. (Hereafter, abbreviated as A3, B3, and C3, respectively)
Fatty acid (R ¹ ) composition (%): Mannose 2nd: C2 = 60, C4 = 40, Mannose 3rd: C14: 0 = 12.2, C14: 1 = 53.0, C16: 0 = 4.6, C16: 1 = 19.0, C18: 1 = 3.1, Unknown = 8.1
Each MEL basically has the molecular structure of MEL-A, B, and C described in Formula (3), but the R ¹ (fatty acid) in the structural formula is different due to the difference between the production microorganism and the oil component of the raw material. The molecular structure such as the composition of the site is different.

Example 5
Candida antarctica- derived lipase B (CalB) (Novozym manufactured by novozymes)
435 CALB L), the final MEL addition amount was 0.01%, and the lipase inhibitory activity of each MEL of A1, B1, C1, and B2 was measured in the same manner as in Example 1. As shown in the results of FIG. 7, each MEL clearly showed the inhibitory activity of the lipase.

Example 6
Rhizomucor miehei- derived lipase (Novozym 388 manufactured by novozymes) was used, and the final MEL addition amount was 0.01%. The others were the same as in Example 1, and A1, B1, C1, B2, C2, A3, B3 and C3 The lipase inhibitory activity of each MEL was measured. As shown in the results of FIG. 8, each MEL showed significant inhibitory activity of the lipase.

Example 7
Using a lipase derived from Thermomyces lanuginosus (Lipozyme TL 100L manufactured by novozymes), the final MEL addition amount was 0.01%, and the others were the same as in Example 1, and A1, B1, C1, B2, C2, A3, B3 and The lipase inhibitory activity of each CEL MEL was measured. As shown in the results of FIG. 9, each MEL showed significant lipase inhibitory activity.

Example 8
Rhizopus delemer- derived lipase (manufactured by Seikagaku Corporation) (Example 8) was used, and the final MEL addition amount was 0.01%, and the others were the same as in Example 1, and A1, B1, C1, B2, C2, The lipase inhibitory activity of each MEL of A3, B3 and C3 was measured. As shown in the results of FIG. 10, each MEL showed significant lipase inhibitory activity.

Example 9
Using lipase derived from porcine pancreas (Lipase from porcine pancreas, Type II manufactured by SIGMA), the final MEL addition amount was 0.01%, and the lipase inhibitory activity was examined in the same manner as in Example 4. The control was MEL-A (A1) derived from Pseudozyma antarctica KM-34 strain that showed porcine pancreatic lipase inhibitory activity at a concentration of 1% in Example 4. As is clear from the results of FIG. 11, B2 has almost the same inhibitory activity as A1, but B1, C1, C2, A3, B3 and C3 have pancreatic lipase inhibitory activity at lower concentrations, and pancreatic lipase inhibitory activity. The activity was found to be higher than A1.

Claims (9)

  A lipase inhibitor comprising a microbial-produced sugar-type biosurfactant as an active ingredient. The lipase inhibitor according to claim 1, wherein the microorganism-produced sugar type biosurfactant is a mannosyl alditol lipid represented by the following chemical formula (1).

(Wherein R ¹ represents an aliphatic acyl group having 2 to 24 carbon atoms, R ² represents hydrogen or an acetyl group, and R ³ represents hydrogen or an aliphatic acyl group having 2 to 24 carbon atoms. R ² and R ³ may be the same or different, and n is an integer of 1 to 4.) The lipase inhibitor according to claim 1, wherein the microorganism-produced sugar type biosurfactant is a mannosylerythritol lipid represented by the following chemical formula (2).

(Wherein R ¹ represents an aliphatic acyl group having 2 to 24 carbon atoms, R ² represents hydrogen or an acetyl group, and R ³ represents hydrogen or an aliphatic acyl group having 2 to 24 carbon atoms. The R ² and R ³ at each position may be the same or different.)   A pharmaceutical or health food for prevention, amelioration or treatment of adult diseases, characterized in that the microorganism-produced sugar-type biosurfactant according to any one of claims 1 to 3 is contained as an active ingredient.   The fat absorption inhibitor which contains the microorganisms production sugar-type biosurfactant in any one of Claims 1-3 as an active ingredient.   The obesity inhibitor which contains the microorganisms production sugar-type biosurfactant in any one of Claims 1-3 as an active ingredient.   A dermatitis inhibitor comprising the microbial sugar-type biosurfactant according to any one of claims 1 to 3 as an active ingredient.   An acne inhibitor comprising the microorganism-produced sugar-type biosurfactant according to any one of claims 1 to 3 as an active ingredient.   An antidandruff agent comprising the microbial-produced sugar-type biosurfactant according to any one of claims 1 to 3 as an active ingredient.