JP2002355004A - Lemon-fermented product and method for producing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a lemon fermented product exerting anti-oxidant action, and enabling, with ease, lemon to be extendedly and effectively used, and to provide a method for producing the lemon fermented product. SOLUTION: This lemon fermented product is obtained by subjecting at least one kind of fermented raw material selected from lemon peel, segment membrance and juice sac to microbial fermentation using Aspergillus saitoi; wherein this lemon fermentative product contains 8-hydroxyhesperetin having anti-oxidant action and exercise-oxidation stress-reducing action, and is obtained by subjecting Aspergillus saitoi-containing culture medium to pre-preparation of culture soil where shaking culture is carried out under the aerobic condition followed by inoculating the thus treated culture soil into the fermented raw material to subject it to microbial fermentation. Fermentation period for the microbial fermentation is desirable to be 2-14 days, and the fermentative raw material is preferable to comprise juice extraction residue obtained after extracting juice from lemons.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a fermented lemon product and a method for producing the same, which can be used for expanding and effectively using lemon. To be more specific, to add new added value by microbial fermentation treatment of pericarp, carbohydrate film, and sac, which are the components of lemon fruit, to use for beverages and pharmaceuticals with high antioxidant activity And a method for producing the same.

[0002]

2. Description of the Related Art Lemon fruit is composed of fruit juice, pericarp, carbohydrate membrane, sorghum and the like. Conventionally, lemon juice has been used by being added to beverages and the like, but use of other than juice has hardly been performed.

[0003]

However, in addition to the viewpoint of effective use of resources, the enforcement of the Food Recycling Law has increased the necessity of making effective use of components other than the juice of lemon fruit as much as possible. Was.

[0004] The present invention has been made by focusing on the problems existing in the prior art as described above. An object of the present invention is to provide a fermented lemon product and a method for producing the same, which exhibit a high antioxidant effect and can easily expand the use and effective use of a lemon.

[0005]

Means for Solving the Problems In order to achieve the above object, the fermented lemon product according to the first aspect of the present invention is obtained by using Aspergillus saitoi,
It is obtained by microbial fermentation of at least one fermentation raw material selected from lemon peel, carbohydrate membrane and canopy, and contains 8-hydroxyhesperetin.

[0006] The lemon fermented product of the invention according to claim 2 is
It is obtained by microbial fermentation of a fermentation raw material consisting of juice residue after juice is squeezed from lemon fruit using Aspergillus saitoi, and contains 8-hydroxyhesperetin. It is assumed that.

[0007] The fermented lemon product according to the third aspect of the present invention comprises:
The invention according to claim 1 or 2 has an antioxidant effect. The fermented lemon product according to the fourth aspect of the present invention is characterized in that, in the invention according to any one of the first to third aspects, it has a motor oxidative stress reducing effect.

According to a fifth aspect of the present invention, there is provided a method for producing a fermented lemon according to any one of the first to fourth aspects, wherein the fermented lemon is produced using Aspergillus saitoi. The fermentation material is microbial-fermented, whereby hesperidin contained in the fermentation material is microbial-converted to produce 8-hydroxyhesperetin.

In the method for producing a fermented lemon product according to the sixth aspect of the present invention, the medium containing Aspergillus cytoii is shaken under aerobic conditions prior to the microbial fermentation treatment. The method is characterized in that, after performing a preliminary culture treatment for culturing, the medium after the preliminary culture treatment is adhered to the fermentation raw material to carry out a microorganism fermentation treatment.

[0010] A method for producing a fermented lemon product according to the invention described in claim 7 is characterized in that, in the invention described in claim 5 or 6, the microorganism fermentation treatment is performed for 2 to 14 days. .

[0011]

Embodiments of the present invention will be described below in detail. The fermented lemon product is obtained by microbial fermentation of at least one fermentation raw material selected from lemon peel, carcinoma membrane, and sanoka using Aspergillus saitoi. This lemon fermented product contains 8-hydroxyhesperetin having a high antioxidant activity as an active ingredient, and has a health-promoting activity when added to, for example, beverages, foods, and pharmaceuticals. Used for health foods and health drinks.

As fermentation raw materials, peeled from lemon fruit,
At least one kind selected from the separated or isolated lemon peel (albedo and flavedo), the carcinoma membrane and the canopy is used. This fermented raw material may be refrigerated or frozen. Further, as the fermentation raw material, those separated from the lemon fruit by peeling or the like can be used, but since the labor required for production can be saved and the lemon fruit can be effectively used, the lemon fruit can be used. Most preferably, the juice residue after juice of the fruit juice is used. This squeezed residue contains, as main components, lemon peel (albedo and flavedo), a carcass membrane and a part of the sap, and a small amount of seeds and waste. It also contains a very small amount of lemon juice that could not be squeezed. In this fermentation material, hesperetin (3 ', 5,7-trihydr
Hesperidin (vitamin P), which is a glycoside of oxy-4′-methoxyflavanone (C ₁₆ H ₁₄ O ₆ ) and rutinose (L-rhamnosyl-D-glucose), is higher than the concentration in lemon juice. It is contained at a concentration.

[0013] The microbial fermentation treatment is a treatment in which Aspergillus cytoii is inoculated into the fermentation raw material, and then under predetermined fermentation conditions, Aspergillus cytoii transforms hesperidin in the fermentation raw material into 8-hydroxyhesperetin. The 8-hydroxyhesperetin has a structure represented by the following chemical formula 1.

[0014]

Embedded image This 8-hydroxyhesperetin has a chemical formula of C ₁₆ H ₁₄
In O _7, molecular weight of about 319 flavonoid compound of (3 ', 5,
7,8-tetrahydroxy-4'-methoxyflavanone or 2,3-dihyd
ro-5,7,8-trihydroxy-2- (3-hydroxy-4-methoxyphenyl)-
4H-1-benzopyran-4-one), which is an organic compound having a hydroxyl group at the 8-position of hesperetin. This 8-hydroxyhesperetin is soluble in methanol, ethanol, and dimethylsulfoxide (DMSO), and is slightly soluble but soluble in water. Furthermore, whereas the above-mentioned hesperetin has almost no antioxidant activity,
Hydroxyhesperetin has an extremely high antioxidant activity equivalent to that of α-tocopherol (vitamin E).

As a method of inoculating Aspergillus cytoii into a fermentation raw material, spores of Aspergillus cytoii can be directly sprinkled on the fermentation raw material and adhered.
Further, after a preliminary culture treatment in which a culture medium containing Aspergillus cytoii is cultured under shaking under aerobic conditions is performed in advance, the culture medium after the preliminary culture treatment is sprinkled so as to spread over the entire fermentation raw material, and the culture medium is adhered or adhered. It is also possible to inoculate by immersing the fermentation raw material in the subsequent medium. Among these inoculation methods, it is most preferable to sprinkle the medium after the preliminary culture treatment onto the fermentation raw material since the fermentation processing of the fermentation raw material proceeds relatively uniformly.

The pre-culture treatment is carried out in order to advance the microorganism fermentation process quickly and smoothly by sufficiently cultivating and activating Aspergillus cytoii used in the microorganism fermentation process in advance. This pre-culture treatment is carried out under aerobic conditions at 20 to 40 ° C. for at least 5 days, and preferably until the mycelium of Aspergillus cytoii covers the medium surface by about 1/3.

As the medium, a medium for filamentous fungi such as a potato dextrose-containing medium or a Zapec medium, or various liquid mediums containing organic substances such as okara are preferably used. Furthermore, in this pre-culture treatment, Aspergillus
In order to improve the growth of Cytoii, it is preferable to adjust the pH of the medium at the start of the culture to 3 to 7. The shaking speed during the shaking culture is preferably 50 rpm / min or more, more preferably 50 to 200 rpm.
pm / min. If the shaking speed is less than 50 rpm / min, the whole culture medium containing Aspergillus cytoii is not aerobic, so that hyphal growth cannot be sufficiently performed.
When the shaking speed exceeds 200 rpm / min,
There is a possibility that the shaking of the medium is severe and the formation of bacterial cells of Aspergillus cytoii is suppressed.

As the conditions for microbial fermentation when inoculating Aspergillus cytoii activated by the above-mentioned preculture treatment into the fermentation raw material, microbial fermentation can be easily performed under aerobic conditions. A culture vessel having a wide bottom and a shallow depth such as a culture dish formed in a bottom cylindrical shape is suitably used. Further, the fermentation material may be placed on the bottom of the culture vessel so as to spread the fermentation material evenly. The fermentation temperature is preferably 10 to 40 ° C, more preferably 20 to 40 ° C, as conditions suitable for growing Aspergillus cytoii. In addition, Aspergillus
As conditions suitable for the growth of Cytoii, it is preferable to carry out a microorganism fermentation treatment in a dark place.

The fermentation period of the microbial fermentation treatment is preferably 2 to 14 days, more preferably 2 to 10 days.
Days, more preferably 3 to 8 days. When the fermentation period is less than 2 days, a sufficient amount of 8-hydroxyhesperetin is not produced because microbial fermentation by Aspergillus cytoii has hardly progressed. On the other hand, when the period exceeds 14 days, the degradation of 8-hydroxyhesperetin produced by the microbial conversion proceeds, and the antioxidant effect decreases.

The fermented lemon product obtained by the above-mentioned microbial fermentation treatment is decomposed into fibers by fermentation, and becomes very brittle and easily collapsed. In the fermented lemon, there is a fermentation material with insufficient fibrous decomposition during fermentation and solids such as the mycelium of the inoculated strain formed during fermentation. Compared to fermentation raw materials, the volume is about 1/10. This fermented lemon product is preferably subjected to a heat sterilization treatment or the like, and then added as it is to materials such as beverages, foods, and pharmaceuticals, or can be added to the material after a mixer or homogenization treatment and used. It is. Moreover, after refine | purifying the active ingredient which has the antioxidant effect produced | generated in the said lemon fermentation product, you may comprise so that it may be added to the said raw material.

When purifying the active ingredient in the lemon fermented product, first, a solid content removing treatment for removing solid content mainly composed of the inoculum and the unfermented raw material is performed. In this solid content removal treatment, the fermented lemon containing solids is immersed in a polar solvent, the active ingredient is transferred into the solvent and extracted, and then filtered with gauze, a coarse mesh or the like, or 3000 rpm.
This is a process for removing solids by light centrifugation for about 30 minutes. The extraction conditions for extracting the fermented lemon product in a polar solvent are preferably extracted at room temperature for 2 hours or more from the viewpoint of extraction efficiency.

As the polar solvent, methanol, ethanol, their aqueous solutions or water are preferably used. Further, lower alcohols such as butanol and isopropanol or aqueous solutions thereof can also be used. Further, as the polar solvent, methanol, an aqueous solution thereof or water is preferably used from an economic viewpoint when a large amount of fermented lemon is treated, and water is most preferably used from the viewpoint of purification cost. When added directly to materials such as beverages, foods and pharmaceuticals, it is preferable to use ethanol, its aqueous solution or water. Also, Aspergillus
In order to kill microbes and stop microbial fermentation, it is necessary to use methanol, ethanol or a high concentration thereof (eg, 5
(0% by volume or more).

Furthermore, it is also possible to carry out a fiber content removal treatment for separating and removing water-soluble fiber components mainly composed of pectin contained in the lemon fermented product after the solid content removal treatment. This fiber content removal treatment is a treatment in which the fermented lemon product after the solid content removal treatment is centrifuged to remove water-soluble fiber components by settling at the bottom of the centrifuge tube. In addition, this fiber content removal processing is 8000 r when methanol or ethanol is used as the polar solvent.
It may be centrifuged with a centrifugal force of about 20 minutes at pm. When water or an aqueous solution is used as the polar solvent, the centrifugal force is centrifuged with a stronger centrifugal force.

The effects exerted by the above embodiment will be described below. -The fermented lemon product is obtained by microbial fermentation of at least one fermentation raw material selected from lemon peel, carcinoma membrane and canopy using Aspergillus cytoii. For this reason, the fermented lemon product contains 8-hydroxyhesperetin in which hesperidin contained in the fermentation raw material has been microbial converted by Aspergillus cytoii, and has an extremely high antimicrobial activity as compared with the fermentation raw material before the microbial fermentation treatment. It can exert an oxidizing effect. In particular, since the fermentation raw material contains hesperidin at a higher concentration than lemon juice, the production efficiency of 8-hydroxyhesperetin can be easily increased. This fermented lemon product eliminates active oxygen in vivo and suppresses the production of lipid peroxide, which can be useful for preventing lifestyle-related diseases such as cancer, arteriosclerosis, and diabetes complications caused by oxidative stress. In addition, it is possible to reduce oxidative stress caused by exercise to maintain health and recover fatigue.

Furthermore, since the fermented lemon product can be used in various products such as health foods and health drinks by adding it to materials such as beverages, foods and pharmaceuticals, the use of lemons is expanding. Can be easily achieved. In addition, since the fermentation raw material has been rarely used and has been disposed of so far, by using it as a raw material for producing the lemon fermented product of the present embodiment, its effective utilization is improved. In addition, the manufacturing cost can be easily reduced.

Furthermore, after the production of the lemon fermented product, only about one-tenth of the solid content of the fermented raw material is discarded, and the volume of the waste can be significantly reduced. For this reason, the transportation cost for disposal can be easily reduced, and it is extremely easy to comply with the Food Recycling Law. In addition, since this lemon fermented product uses lemon as a raw material and Aspergillus cytoii used for brewing alcoholic beverages such as shochu, there is no problem in ingestion into the human body.

The fermented lemon is contained in the fermented raw material by subjecting at least one fermented raw material selected from the peel of the lemon, the carcass membrane and the canopy to microbial fermentation using Aspergillus cytoii. Produced by microbial conversion of hesperidin to produce 8-hydroxyhesperetin. Therefore, it is possible to extremely easily produce a fermented lemon product that exhibits a high antioxidant effect and can easily expand the use and effective use of the lemon.

Further, by carrying out a pre-culture treatment of Aspergillus cytoii prior to the above-mentioned microbial fermentation, Aspergillus cytoii used for the microbial fermentation is sufficiently grown and activated in advance, and the microbial fermentation is carried out quickly and smoothly. Can proceed. In addition, by performing the microbial fermentation treatment for 2 to 14 days,
A fermented lemon product containing a large amount of 8-hydroxyhesperetin can be easily produced.

Further, by removing solids in the lemon fermented product by the solid content removing treatment, it is possible to easily produce a lemon fermented product in which the handleability and the concentration of the active ingredient are easily improved. Further, the removed solid content can be reused as a fermentation starter in the next microbial fermentation treatment, and in this case, waste can be further reduced.

[0030] Subsequent to the solid content removal treatment, the water-soluble fiber component in the lemon fermentation product is removed by the fiber content removal treatment, thereby easily producing a lemon fermentation product with further enhanced antioxidant action. it can. Further, the water-soluble fiber component thus removed can be added to a functional food or beverage having an intestinal function and reused, and the use of lemon and the effective use of lemon can be further effectively improved. be able to.

[0031]

EXAMPLES Examples and comparative examples which embody the above embodiment will be described below. <Manufacture of Lemon Fermented Product> (Example 1) In this experiment, Aspergillus cytoii strain (IAM No. 2210) obtained from the Applied Microbiology Research Promotion Association (commonly known as IAM) was used. The pre-culture treatment of this strain adjusted the pH to 5.0,
A potato dextrose-broth medium (manufactured by DIFCO) sterilized in an autoclave at 15 ° C. for 15 minutes was placed in a flask.
After the culture medium was sufficiently cooled, the obtained strain was inoculated with the strain, and cultured with shaking under aerobic conditions of 100 rpm. Seven days after the inoculation of the bacteria into the medium, the medium liquid after the preliminary culture treatment in which the cells were sufficiently grown in the flask was used as the inoculation bacteria used for the microorganism fermentation treatment.

As a fermentation raw material, juice residue remaining after juice was squeezed from lemon fruit was used. The squeezed residue is a lemon fruit in which the pericarp, the capsular membrane, and a portion of the sorghum remain. In this experiment, 4 kg of the residue with water remaining immediately after squeezing was used and spread evenly in a wide-bottomed container (bottomed cylindrical container) that had been autoclaved beforehand so that air permeability was improved. The inoculated bacteria were sprinkled so as to spread over the whole juice residue. Then, the squeezed residue after inoculation of the bacteria was subjected to a microbial fermentation treatment in a constant temperature room at 30 ° C. for 7 days under dark conditions.

Next, the squeezed residue (fermented lemon containing solids) after the microbial fermentation treatment was immersed in 12 L of methanol, and the extraction of the active ingredient was performed at 37 ° C. for 2 days in the dark. went. Subsequently, the obtained extract was subjected to an inverter high-speed cooling centrifuge 79 manufactured by Kubota Seisakusho.
After centrifugation at 3000 rpm for 30 minutes using the No. 30, only the supernatant was collected and concentrated under reduced pressure using an evaporator. Further, ethanol was added to the concentrate,
The supernatant obtained by centrifugation at 00 rpm for 20 minutes was concentrated under reduced pressure by an evaporator, and then dried by a vacuum freeze dryer for 24 hours to obtain 104.5 g of a powder of a fermented lemon product.

(Comparative Example 1) 4 kg of lemon juice residue was added to 12
The resultant was immersed in L of methanol and subjected to an extraction operation of the active ingredient under dark conditions at 37 ° C. for 2 days. Extract 3
After centrifugation at 000 rpm for 30 minutes, only the supernatant was collected and concentrated under reduced pressure using an evaporator. Further, ethanol was added to this concentrate, and the concentrate was added at 8000 rpm for 20 minutes.
The supernatant liquid obtained by centrifugation for 5 minutes was concentrated under reduced pressure by an evaporator, and then dried by a vacuum freeze dryer for 24 hours. As a result, 111.6 g of a lemon juice residue unfermented powder was obtained. .

<Confirmation of Active Ingredient in Lemon Fermented Product> It was confirmed as follows that the active ingredient contained in the lemon fermented product obtained in Example 1 was 8-hydroxyhesperetin. That is, after dissolving the powder of the fermented lemon product of Example 1 in methanol to a concentration of 1000 ppm, using a high-performance liquid chromatograph mass spectrometer (LCMS-QP8000α manufactured by Shimadzu Corporation), the following Table 1 was used. The components were analyzed and the molecular weight was confirmed under the analysis conditions shown in (1). The results are shown in FIG. Although data is not shown, 8-hydroxyhesperetin previously isolated by the present inventors (<production of hesperidin conversion product of Japanese Patent Application No. 2001-73577)) was dissolved in methanol to a concentration of 100 ppm. The same sample was analyzed under the same conditions.

[0036]

[Table 1] As a result, the isolated 8-hydroxyhesperetin (8
OH-HE), a single sharp peak appears 20.328 minutes after the start of elution, and its molecular weight (m / w) is 319.
It was shown to be. On the other hand, also in the fermented lemon product of Example 1, a peak appeared 20.385 minutes after the start of elution, and it was confirmed that the molecular weight of the peak was 319. Therefore, it was confirmed that 8-hydroxyhesperetin was contained in the fermented lemon product of Example 1. From these results, it is possible to produce 8-hydroxyhesperetin inexpensively by effectively utilizing unused resources that have been discarded by inoculating Aspergillus cytoii into lemon juice residue and performing microbial fermentation. It was shown to be.

<Measurement of Antioxidant Activity> DPPH (1,1-Diphenyl-2-picrylhydra) was prepared using the powder of the fermented lemon product of Example 1 and the powder of the unfermented lemon juice residue of Comparative Example 1.
zy) was measured.

As a preparation for the reagents used in this test, DP
After dissolving 20 mg of PH in 100 ml of ethanol,
By filtering through a 0.80 μm Millipore filter, a 500 μM DPPH solution was prepared. In addition, a Tris buffer (0.1 M, pH 7.4) was used as a buffer. As samples, the powder of the fermented lemon product of Example 1 and the powder of the unfermented lemon juice residue of Comparative Example 1 were each dissolved in methanol to give a concentration of 1000 ppm.
Three kinds of each sample liquid adjusted to 2500 ppm and 5000 ppm were used.

First, 2 ml of the DPPH solution and Tris
0.4 ml of each sample solution was added to a mixed solution obtained by mixing 1.6 ml of the buffer solution to prepare a total of 4 ml of a reaction solution. The mixture was vortexed well, placed in a dark room at room temperature and reacted for 20 minutes. In addition, as a control (blank test), the same reaction was performed using 0.4 ml of water instead of the sample solution.

Next, after each reaction solution was stirred again, 10 μl of each reaction solution was subjected to high performance liquid chromatography (HPLC) using a microsyringe (LC-10 manufactured by Shimadzu Corporation).
A) was injected. As the HPLC solvent, methanol / distilled water = 70/30, which had been sufficiently degassed, was used, and the measurement was carried out at a flow rate of 1 ml / min. The TSK-GEL OCTYL-80TS (15
0 × 4.6 mm D. ), Column temperature 35 ° C, UV
The measurement was performed at a detector wavelength of 517 nm.

Since a DPPH peak appeared approximately 9 minutes after the start of elution, the peak height (HEIGHT) was measured, and the DPPH radical scavenging ability (%) of each sample was determined by the following equation (1). Table 2 shows the results.

[0042]

(Equation 1)

[0043]

[Table 2] As a result, it was revealed that the lemon fermented product of Example 1 had a radical scavenging ability about three times stronger than that of Comparative Example 1 at all three concentrations. When the radical scavenging ability is close to 100%, it is highly probable that the increase in the numerical value does not represent an accurate value, and in fact, the increase may be more than three times. Therefore, it has been clarified that a fermented lemon product having extremely high antioxidant ability can be obtained by microbial fermentation of a lemon juice residue using Aspergillus cytoii.

<Study of optimal fermentation conditions> Aspergillus
We examined the optimal fermentation period in microbial fermentation treatment with Cytoii. Lemon juice residue immediately after juice extraction was used as a fermentation raw material. As a sample,
According to the method of Example 1, the microbial fermentation treatment was carried out on 0, 3, 5,
After fermenting the lemon for 7, 10, 12, 18 days, add 1
0 g each, and 100 ml of methanol was added for 37 g.
Extraction of the active ingredient for 2 days at
The supernatant obtained by centrifuging at 20 rpm for 20 minutes was used. Then, the DPPH radical scavenging ability was evaluated by the same method as in the above <Measurement of antioxidant activity>. The results are shown in FIG.

As a result, it was confirmed that the radical scavenging ability was maximized when the fermentation period was about 5 to 7 days. This radical scavenging ability is expected to increase until about the 7th day from the start of the microbial fermentation treatment, and then the activity is likely to decrease due to the progress of decomposition. In this experiment, it was also confirmed that the radical scavenging ability was increased about three times as compared with the time when the microorganism fermentation treatment was started (substantially unfermented).
When the radical scavenging ability is close to 100%, it is highly probable that the increase in the numerical value does not represent an accurate value, and in fact, the increase may be more than three times. Therefore, the fermentation period in the microbial fermentation process is 2 to
It was confirmed that by setting it to 10 days, a lemon fermented product having higher antioxidant activity than the conventional lemon juice residue could be obtained.

<Measurement of Exercise Oxidative Stress Marker> Powder of fermented lemon obtained by the method of Example 1 and Comparative Example 1
Using a powder of the unfermented lemon juice residue as a sample, the in vivo oxidative stress marker in rats was measured.

The experimental animals were Wistar rats (male, 1
0 weeks old), and after pre-breeding for one week, the rats were divided into three groups, and the fermented lemon product of Example 1, the unfermented lemon juice residue of Comparative Example 1, and physiological saline as a control sample Was orally administered to rats in each group. In addition, 1 ml of an aqueous solution whose concentration was adjusted to be 30% by weight was orally administered to rats. Furthermore, the group to which each sample was administered was further divided into an exercise group and a rest group, and the exercise group traveled downhill using a treadmill 4 hours after administration of the sample (downhill 16 hours).
°). The exercise time is 35 minutes (2 minutes at 15 m / min, 33 minutes at 20 m / min)
After the exercise, the rats were exsanguinated and their livers were removed, and TBA (Thiobarbituric) was used as an exercise oxidative stress marker.
acid) The amount of reaction product was measured.

The TBA method was used as a method for measuring a motor oxidative stress marker in rats. That is, pentobarbital was intraperitoneally administered to the rats to which the exercise load had been applied to anesthetize the rats, and then the rats were bled to death and the livers were removed and washed with physiological saline. Subsequently, the wet weight of the excised rat liver was measured, a KCl aqueous solution was added so as to be 30% by weight, and a sample homogenate was prepared using a Teflon (registered trademark) homogenizer. Next, capacity 12ml
0.10 ml of the sample homogenate and 0.02 m of the SDS aqueous solution were placed in a Pyrex (registered trademark) test tube with a screw stopper.
l, acetate buffer 1.50 ml, BHT acetic acid solution 50μ
1, TBA aqueous solution 1.50 ml and distilled water 0.70 ml
Were added in this order. After sealing the test tube and mixing well,
It was left as it was at 5 ° C. for 60 minutes. Thereafter, the mixture was heated in a boiling water bath for 60 minutes and then cooled, and extracted by adding 1.0 ml of distilled water and 5.0 ml of a butanol-pyridine mixed solution. After centrifugation at 3000 rpm for 10 minutes, the supernatant was recovered, and the absorbance at 532 nm was measured. The amount of TBARS in rat liver tissue (TBA reactive group mass: Thioba
rbituric acid reactive substance) was determined. Also,
Similarly, for the control sample to which physiological saline was orally administered, T
The BARS amount was determined. Table 3 shows the results.

[0049]

[Table 3] As a result, in rats to which physiological saline was given, a remarkable increase in the amount of TBARS production was observed when exercise load was given.
That is, the amount of TBARS produced by the rats in the rest group was 68.3 nmol / g, whereas the amount in the exercise group increased to 86.0 nmol / g. The rats in the exercise group to which the unfermented lemon juice residue of Comparative Example 1 was administered had 75.1 nmol / g and TBA.
The increase in RS production was reduced, and 68.2 nmo of the rats in the exercise group to which the fermented lemon of Example 1 was administered.
1 / g, which was almost the same as that of the rats in the rest group in the physiological saline administration group. Therefore, it was confirmed that the lemon fermented product of Example 1 had an extremely high TBARS production inhibitory effect, that is, a motor oxidative stress inhibitory effect.
In addition, the effect of suppressing exercise oxidative stress was confirmed, although slightly lower, in the unfermented lemon juice residue.

In general, when oxidative stress progresses, free radicals generated by the stress attack lipid components of the cell membrane to cause lipid peroxidation, and malondialdehyde (MDA) is liberated to the living body. It is said to have an adverse effect. In this experiment, this MDA was measured as the main TBARS and used as a marker for oxidative stress. The reason why the unfermented product of the lemon juice residue of Comparative Example 1 was observed to have the effect of suppressing motor oxidative stress was presumed to be due to the flavonoids (mainly eriocitrin and hesperidin) contained in the lemon juice residue. You. Further, Comparative Example 1 was added to the fermented lemon product of Example 1.
It is presumed that the higher radical scavenging ability was observed because the flavonoids in the lemon juice residue were structurally converted by the microbial fermentation treatment, and converted substances with increased antioxidant activity were produced. Then, it is considered that the elimination of free radicals (active oxygen) by the conversion substance suppressed TBARS generation due to exercise oxidative stress extremely effectively.

Further, the technical ideas that can be grasped from the above embodiment will be described below. The method for producing a fermented lemon product according to any one of claims 5 to 7, wherein a solid content removing process is further performed after the microbial fermentation process.

[0052] The method for producing a fermented lemon product according to any one of claims 5 to 7, wherein after the microbial fermentation process is performed, a solid content removal process and a fiber content removal process are further performed.

The fermented lemon product according to claim 1 or 2, wherein the fermented lemon product has a higher antioxidant effect than the fermentation raw material. The oxidative stress reducing action is higher than that of the fermentation raw material.
The fermented lemon product according to any one of the above.

A food comprising the fermented lemon product according to any one of claims 1 to 4. A beverage containing the fermented lemon product according to any one of claims 1 to 4. A sports drink comprising the fermented lemon product according to claim 4. A pharmaceutical product comprising the fermented lemon product according to any one of claims 1 to 4.

[0055]

As described above in detail, according to the present invention, the following effects can be obtained. According to the lemon fermented product of the invention as set forth in claims 1 to 4, while exhibiting a high antioxidant effect, the use expansion and effective use of lemon can be easily achieved.

According to the method for producing a fermented lemon product of the invention according to any one of the fifth to seventh aspects, a lemon fermentation product having a high antioxidant effect and capable of easily expanding and effectively utilizing the lemon can be obtained. Products can be easily manufactured.

[Brief description of the drawings]

FIG. 1 is a chromatogram obtained by analyzing the components of the fermented lemon product of Example 1.

FIG. 2 is a graph showing a relationship between a fermentation treatment period and a radical scavenging ability in Examples.

Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat II (Reference) A61P 39/06 A61P 39/06 // C12P 17/06 C12P 17/06 (C12P 17/06 C12R 1:66 C12R 1: 66) (72) Inventor Yoshiaki Miyake 45-2 Pokka Corporation R & D section, Kumanosho-shi, Nishi-Kasuga-gun, Nishi-Kasugai-gun, Aichi Pref. 45-2 Inventor Toshihiko Osawa 2615-409 72) Inventor Kenichiro Minato 163-203 F-term (reference) 4B016 LG02 LK18 LP13 4B018 LB03 MD08 MD52 MD91 ME06 4B064 AE46 CA05 CB02 CB07 DA01 DA10 4C086 AA01 AA02 BA08 MA01 MA04 NA14 AB88 CA25 NA14 ZC37

Claims (7)

[Claims] [1] Aspergillus cytoii
s saitoi), obtained by microbial fermentation of at least one fermentation raw material selected from lemon peel, carbohydrate membrane, and sanoka, and containing 8-hydroxyhesperetin. Fermented lemon. 2. An Aspergillus cytoii.
s saitoi), which is obtained by microbial fermentation of a fermented raw material consisting of juice residue after squeezing juice from lemon fruit, and containing 8-hydroxyhesperetin. object. 3. The fermented lemon product according to claim 1, which has an antioxidant effect. 4. The fermented lemon product according to claim 1, which has a motor oxidative stress reducing effect. 5. The method for producing a fermented lemon product according to claim 1, wherein the fermentation raw material is subjected to microbial fermentation using Aspergillus saitoi.
A method for producing a fermented lemon product, comprising converting 8-hydroxyhesperetin by microbial conversion of hesperidin contained in the fermentation raw material. 6. Prior to the microbial fermentation treatment, a preculture treatment in which a culture medium containing Aspergillus cytoii is subjected to shaking culture under aerobic conditions is performed, and the medium after the preculture treatment is attached to the fermentation raw material. The method for producing a fermented lemon according to claim 5, wherein the fermentation is performed by microbial fermentation. 7. The method for producing a fermented lemon according to claim 5, wherein the microorganism fermentation treatment is performed for 2 to 14 days.