JP4036385B1 - Method for producing fermented papaya fruit - Google Patents

Abstract

An object of the present invention is to provide a method for producing a fermented papaya fruit having high hydroxyl radical scavenging activity.
[Solution] Glucose, lactic acid bacteria, amylase and miso enzyme are added to and mixed with wild immature papaya fruits, allowed to stand and fermented, and the water-soluble extract from the mixture is dried, and this is dried for at least 6 months. By aging as described above, a fermented papaya fruit having a hydroxyl radical scavenging activity of 2,000 μg or less / ml (IC50), or 0.2 μmol equivalent or more in terms of Trolox is produced.
[Selection figure] None

Description

The present invention relates to a method for producing a fermented papaya fruit having high hydroxyl radical scavenging activity.

  In general, the relationship between reactive oxygen species (ROS) and diseases is widely recognized, and it is also related to so-called lifestyle-related diseases such as cancer, cataracts, neurological diseases, liver diseases, and kidney diseases, as well as allergies and diabetes. . Antioxidants have been shown to be effective against ROS-related diseases.

  On the other hand, papaya (scientific name: Carica Papaya Linn) is not only used for food as a tropical fruit, but also contains a proteolytic enzyme known as papain, so it can be used as a compounding ingredient in foods, cosmetics, and pharmaceuticals. Widely used in the field.

  It has been suggested that the health functionality may be based on the ROS scavenging activity of the product, especially the hydroxyl radical scavenging activity.

  It is known that fermented papaya fruit products generally have an antioxidant action and are effective for various diseases. For example, the effectiveness against diseases such as epilepsy, amnesia and Alzheimer's disease, the immunity of the skin and colonic mucosa, and the possibility of suppressing allergic inflammation have been suggested.

  In addition, the use of processed papaya fermented products in foods (Patent Documents 1 to 4) and in the detergent / cleaning field (Patent Document 5) has been considered, and some of them have been put into practical use.

  However, the level of radical scavenging activity required for a fermented papaya fruit having high health functionality is not known, and a method for producing the fermented product has not been established.

  Under such circumstances, establishment of a fermented papaya fruit having high hydroxyl radical scavenging activity and its production technology has been expected.

JP 2001-120224 A

Japanese Patent Laid-Open No. 2003-24009

JP 2004-267026 A

JP 2006-75086 A

JP 2001-152188 A

The present invention has been made under the above background, and it is an object of the present invention to provide a method for producing a fermented papaya fruit having high hydroxyl radical scavenging activity and high health functionality.

  In order to solve the above-mentioned problems, the present inventors have made extensive studies, [1] establishment of a fermented papaya fruit having high hydroxyl radical scavenging activity and production technology thereof, and [2] papaya processing including the fermented papaya fruit. The present invention has been reached based on the development of food and [3] elucidation of the relationship between hydroxyl radical scavenging activity of fermented papaya fruit and disease.

The method for producing a fermented papaya fruit according to the present invention is a wild immature papaya having white seeds, which is mashed after slicing the seeds with the skin removed from the bottom and bottom of the flower. The first mixture obtained by adding glucose is left to age, and then the first mixture is filtered to be fractionated into a first filtrate and a first residue, and glucose is mixed into the first residue. The second mixture is allowed to stand, and the second filtrate is obtained by filtering the second mixture, and glucose is added to the mixture of the first filtrate and the second filtrate to obtain a glucose concentration. First, an intermediate raw material prepared at 40 to 60% is obtained, fermented after adding an enzyme for miso to this intermediate raw material, then further added with glucose, mixed and dried, and then crushed. Get the composition of Add the lactic acid bacteria and miso enzymes to the intermediate raw material, ferment and mix, and then add the glucose, mix, dry, and crush to obtain the second composition, and add amylase to the intermediate raw material Mixing and fermenting, then adding further glucose, mixing and drying, crushing to obtain a third composition, the first composition, the second composition and the third composition Is characterized by obtaining a fermented papaya fruit by mixing and homogenizing and aged for a long period of at least 6 months .

  Here, "wild immature papaya" is a wild papaya that is close to the original species, and the unripe hull is a blue papaya, and has been improved in varieties seen at stores, supermarkets, etc. Unlike papaya, it is about the size of a human fist and is packed with seeds.

  The lactic acid bacterium used in the present invention is not particularly limited, but Streptococcus, Pediococcus, Lactobacillus and the like are preferable, and Streptococcus lactic acid bacterium is particularly preferable. This is, for example, a raw material available as a food grade commercial product.

  The amylase used in the present invention is a general term for starch hydrolyzing enzymes, and is not particularly limited as long as it has quality conditions usable for food production. This is a raw material that can be obtained and used as, for example, a food grade commercial product, and the usage method depends on the quality of the enzyme, the quality of the active unit, etc., the quality condition of the target product, and the production conditions. Can be adjusted as appropriate.

  In the present invention, the enzyme for miso means fungus bodies, secretions, and the like of Aspergillus oryzae used in the production of sake, miso, soy sauce, and the like. In the above-mentioned “process for producing a fermented product by mixing lactic acid bacteria, amylase and miso enzyme with the intermediate raw material and then fermenting them,” an appropriate amount of the miso enzyme can be added and mixed. This is, for example, a raw material available as a food grade commercial product, and the usage method is appropriately adjusted according to the quality of the enzyme, the quality of the activity unit, etc., the quality condition of the target product, and the production conditions. it can.

  The amount of lactic acid bacteria, amylase, and miso enzyme used in the present invention is not particularly limited, but an appropriate amount can be determined each time depending on the quality and cost requirements. Moreover, the usage amount and usage method of these raw materials can be adjusted according to the quality of the target product.

  In the present invention, the hydroxyl radical scavenging activity is defined as follows.

  Evaluation of hydroxyl radical scavenging ability of fermented papaya fruit is performed by electron spin resonance spectroscopy (ESR method). The ESR system is an X-Band (RX type manufactured by JEOL Ltd.) with a digital high-speed sweep unit (Radical Research Co., Ltd.) incorporated into an improved radical detection system for the WIN-RAD system RDA-03W ESR data analyzer (Radical Research Co., Ltd.) ) Is connected.

  The ESR measurement conditions were as follows: sweep magnetic field region: 336.5 ± 5 mT, magnetic field modulation Field Mod; 0.079, Time Constant; 0.10 seconds, sweep time: 1 minute, output: 8.0 mW, and the result is CYPMPO ( 2- (5,5-Dimethyl-2-oxo-2-λ5- [1,3,2] dioxaphosphin-2-yl) -2-methyl-3,4-dihydro-2H-pyrrole-1-oxide, radical It is shown as the relative intensity of the radical adduct trapped by Research).

  In the present invention, the hydroxyl radical scavenging activity is the IC50 value (μg / ml, final concentration) of the concentration of the papaya fruit fermented product that reduces the signal relative intensity by 50% when measuring the control ESR without the sample (papaya fruit fermented product). ).

  Alternatively, the concentration of Trolox ((±) -Hydroxy-2,5,7,8-tetramethylchloro-2-carboxylic acid, Aldrich) corresponding to the hydroxyl radical scavenging activity, that is, exhibiting the same radical scavenging activity, is used. Obtained and shown as a unit (μmol Trolox equivalent / mg) corresponding to the IC50 value (μg / ml, final concentration) of the papaya fruit fermented product. In other words, Trolox is used as a preparation for evaluating hydroxyl radical scavenging activity, and evaluation is performed by determining the chemical equivalent of Trolox corresponding to the hydroxyl radical scavenging activity of the sample (fermented papaya fruit).

  This makes it possible to minimize data variation due to differences in ESR models, method differences, reagent purity differences, etc., and to provide data with objectivity.

  According to the present invention, since it has high hydroxyl radical scavenging activity, it contributes to maintaining and improving the health of the nation, and by providing a method for producing a fermented papaya fruit, a safe material can be supplied in large quantities and at low cost. Make it possible.

  The best mode for carrying out the present invention will be described below.

The present inventors continuously ingest, for example, an average of 3.0 g / day for 1.0 month or more of a papaya fruit fermented product produced by a mild type 2 diabetes patient using the method for producing a papaya fruit fermented product of the present invention. Assuming that the hydroxyl radical scavenging activity of the fermented papaya fruit is 2,000 μg or less / ml (IC50), or 0.2 μmol equivalent / mg in terms of Trolox, preferably 1,000 μg or less / Symptom improvement at ml (IC50) or when converted to Trolox 0.40 μmol equivalent / mg, more preferably 500 μg or less / ml (IC50), or Trolox converted 0.80 μmol equivalent or more / mg Found to be effective.

Manufacture of fermented papaya fruit product of the present invention for various lifestyle diseases including allergies, hypertension, cancer, cataracts, neurological diseases, liver diseases, and kidney diseases, as well as mild type 2 diabetes patients The effectiveness of fermented papaya fruit produced using the method was measured and examined under the above conditions. As in the case of mild type 2 diabetic patients, the hydroxyl radical scavenging activity of the fermented papaya fruit was not more than 2,000 μg. / Ml (IC50), or when converted to Trolox 0.20 μmol equivalent / mg, desirably 1,000 μg or less / ml (IC50), or when converted to Trolox 0.40 μmol equivalent / mg, More desirably, it is less than 500 μg / ml (IC50) or 0.80 μ in terms of Trolox. When ol equivalent or higher / mg, the symptoms of prevention, was found to be valid in improving.

Regardless of the effective intake under the above conditions, the intake method, and the intake period, the fermented papaya fruit produced using the method for producing a fermented papaya fruit of the present invention is a food, so in the present invention, the intake amount, the intake method, There are no special upper and lower limits or other conditions for the intake period. Therefore, various cases are envisaged, such as taking a large amount for a short period of time, taking a small amount for a long period of time, or taking with other health functional materials, but the present invention is not limited by this.

The fermented papaya fruit produced using the method for producing a fermented papaya fruit of the present invention can be ingested as it is, but can be ingested by mixing with other foods. The amount of the fermented papaya fruit added to other foods is 1.0% by weight or more, preferably 3.0% by weight or more, and more preferably 5.0% by weight or more based on the food. The food to be mixed can be in various states such as powder, liquid, and paste, but the present invention is not limited to these states.

  The amount and method of mixing the fermented papaya fruit product with other foods will depend on the cost, quality requirements including sensory evaluation, and the content of the required function.

The details of the intermediate raw material production of the fermented papaya fruit produced using the method for producing a fermented papaya fruit of the present invention are as follows.

  Harvest wild immature papaya, cut the flower part of the bottom and bottom, cut the whole seeds with the skin, crush and knead (mash) in a container. Glucose is added to this product and aged for 7 to 10 days. The aged product is filtered to obtain filtrate 1 and residue 1.

  Wild immature papaya fruit used as a raw material has a characteristic of having white seeds, and is different from mature papaya fruit having black seeds.

  In general, wild immature papaya fruits have higher hydroxyl radical scavenging activity and higher health functionality than cultivated immature papaya fruits, and therefore wild immature papaya fruits are used in the present invention. It is characterized by that.

  After leaving the mixture of residue 1 and glucose, filtrate 2 is obtained by filtration, glucose is added to the mixture of filtrate 1 and filtrate 2, and the final glucose concentration in the mixture is adjusted to 40 to 60% by weight. This product is used as an intermediate raw material for fermented papaya fruit.

  Details of the papaya fruit fermented product production are as follows.

  Lactic acid bacteria, miso enzyme and amylase are added to the intermediate material, mixed, fermented, glucose is added to the fermented product, dried, the dried product is pulverized into a powder, and left for 6 months or longer By aging, a fermented papaya fruit characterized in that the hydroxyl radical scavenging activity of the product is 2,000 μg or less / ml (IC50) or 0.20 μmol equivalent / mg in terms of Trolox can be obtained. . This product has excellent sensory evaluation quality and retains physiological activity useful as a health functional material.

  This product is milky white powder, but for easy intake, it is possible to change the dosage form such as granulation by granulation, pulverization by pulverization, tableting by tableting, addition of coloring agent, flavor etc. The addition of the mixture is not hindered.

  In the present invention, it is important to use wild immature papaya fruit as a raw material. If mature papaya fruit is used as a raw material, quality degradation is likely to occur during the manufacturing process, and the quality of the finally obtained fermented product is sensory-evaluated compared to when immature papaya fruit is used. Furthermore, it does not sufficiently retain the physiological activity expected as a health functional material.

  Hereinafter, the present invention will be further described by way of examples. However, the following examples are for illustrative purposes only and are not intended to limit the present invention in any way.

<Example>
1) Measurement of hydroxyl radical scavenging activity of papaya fruit fermented product and Trolox The evaluation of free radical and reactive oxygen species scavenging ability of the papaya fruit fermented product produced as described above was conducted by electron spin resonance spectroscopy (ESR) method. I went. The ESR device is a free radical detector that incorporates a digital high-speed sweep unit (Radical Research) into X-Band (JEOL Ltd. RX type), and an improved WIN-RAD system RDA-03W ESR data analyzer (radical research). System).

  The ESR setting conditions are as follows: sweep magnetic field region: 336.5 ± 5 mT, magnetic field modulation Field Mod: 0.079, Time Constant: 0.10 seconds, sweep time: 1 minute, output: 8.0 mW, and the result is DMPO or It is shown as the relative intensity of the radical signal trapped by CYPMPO. More specific description will be given below.

  100 μM ferrous sulfate, 100 μM MDETAPAC, 5 mM CYPMPO, various concentrations of sample (papaya fruit fermented product) or Trolox, 100 μM hydrogen peroxide mixed solution in a fully nitrogen-substituted 200 mM phosphate buffer (pH 7.8) Was subjected to ESR analysis. Of all 8 peaks due to CYPMPO-hydroxyl radical adduct generated by starting sweeping 1 minute after hydrogen peroxide addition, 4th peak (ESR signal) and 6 ESR signals due to Mn Among them, the peak of the second Mn from the low magnetic field side (ESR signal) was compared, and the hydroxyl radical was quantified by the ratio of each ESR signal, that is, the relative intensity of CYPMPO-hydroxyl radical / Mn. The relative intensity of the ESR signal of CYPMPO-hydroxyl radical / Mn observed in the control ESR measurement carried out without adding the test material or Trolox was taken as 100%. A sample (fermented papaya fruit product) was added to the ESR measurement solution, and the scavenging activity of the sample (fermented papaya fruit product) on hydroxyl radicals was examined by changing the relative intensity of the ESR signal of CYPMPO-hydroxyl radical / Mn. As a result, the concentration of the fermented papaya product that attenuates the relative signal intensity during the measurement of the control ESR by 50% was determined as an IC50 value (μg / ml), which was 704 μg / ml (final).

  Similarly, when an equivalent equivalent of Trolox corresponding to the above hydroxyl radical scavenging activity was measured, it was 0.6 μmol Trolox equivalent / mg. That is, it was confirmed that the activity is sufficient to exhibit health functionality.

2) Production Example of Intermediate Raw Material 100 kg of wild immature papaya fruit was collected, washed with water to remove foreign matters, and then the sliced portion and the bottom of the fruit were removed to obtain 85 kg of sliced product.

  20 kg of glucose was added to the sliced product, mixed well, allowed to stand for 4 days, and the mixture was filtered to obtain filtrate 1 and residue 1 as fruit juice.

  10 kg of glucose was added to 60 kg of the residue, mixed, allowed to stand for 4 days, and the mixture was filtered to obtain filtrate 2 as a fruit juice.

  Filtrate 1 and filtrate 2 were mixed, 2 kg of glucose was added and mixed well to obtain 40 kg of the desired intermediate material.

3) Example of production of final product Add 0.5 kg of miso enzyme to 20 kg of the intermediate raw material produced in 2) above, mix well, ferment at 40 ° C. for 48 hours, and then add 60 kg of glucose. After mixing, the mixture was dried at 18 ° C. for 1 month and pulverized to obtain Composition 1.

  After adding 5 kg of lactic acid bacteria and 0.1 kg of miso-enzyme to 15 kg of the intermediate raw material produced in the above 2), mixing well, fermenting at 40 ° C. for 48 hours, adding 30 kg of glucose and mixing well, It was dried at 18 ° C. for 1 month and pulverized to obtain Composition 2.

  Add 0.1 kg of amylase to 5 kg of the intermediate material produced in 2) above, mix well, ferment at 40 ° C for 48 hours, add 10 kg of glucose, mix well, then at 18 ° C for 1 month. It was dried for a while and pulverized to obtain a composition 3.

  Compositions 1, 2, and 3 were combined and homogenized, and aged for 6 months or more in a high aluminum barrier made of gas barrier to obtain 100 kg of the final product.

  This product was a milky white powder with a high fluidity and a moisture content of 5% by weight or less, and had an excellent taste taste sensory. The hydroxyl radical scavenging activity of this product is as described in 1) above.

4) Examples of use of final products Cookies and orange juice added with 0.1%, 1.0%, 10.0% by weight of the product obtained in 3) above were prototyped by 10 sensory evaluators. Tasting and tasting.

  As a result, it was confirmed that all of the cookies and orange juice added with 1.0% by weight and 10.0% by weight of the product can be ingested as food or beverages, and that this product has excellent sensory evaluation quality. It was done. However, the 0.1% by weight added product was evaluated to be inferior in sensory evaluation. In addition, in order to improve health functionality, it was evaluated that the use of a 0.1% by weight additive was not practical for the necessity of increasing the absolute intake of the product itself.

<Comparative Example 1> Product quality when matured papaya fruit is used as a raw material When mature papaya fruit is used as a raw material, it is produced in the same process as in 2) and 3) above. As a result, the final product had an unpleasant odor and became a quality that cannot be used sensuously as food.

<Comparative Example 2> Product quality when matured papaya fruit is used as a raw material A material obtained by sterilizing mature papaya fruit by heat treatment (100 ° C, 1 hour) is the same as in 2) and 3) above. When manufactured in the process, a product having the same functional properties as in the above 3) was obtained.

  Compared with the product obtained from the immature papaya fruit fermented product obtained in 3) above, the hydroxyl radical scavenging activity of this product was half or less, and the required quality could not be satisfied.

  As described above, as a result of comparing the examples of the present invention with Comparative Examples 1 and 2, the necessity and superiority of using immature papaya fruit as a starting material were shown.

Claims (1)

A first mixture of wild immature papaya with white seeds, with the addition of dextrose added to the mash after slicing the seeds with the skin removed from the bottom and the bottom of the flower, is removed. Aged,
Then, the first mixture is filtered to fractionate into a first filtrate and a first residue. Glucose is mixed with the first residue to form a second mixture, which is allowed to stand, and the second mixture is allowed to stand. A second filtrate is obtained by filtering the mixture, and an intermediate material prepared by adding glucose to the mixture of the first filtrate and the second filtrate to obtain a glucose concentration of 40 to 60% is obtained.
The first composition is obtained by adding the miso enzyme to this intermediate material and fermenting it, then fermenting, then adding further glucose, mixing and drying, and then crushing.
Add the lactic acid bacteria and miso enzyme to the intermediate raw material and ferment, and then add the glucose, mix and dry, and then crush and then obtain a second composition,
Add the amylase to the above intermediate raw material, ferment and ferment, then add further glucose, mix and dry, and then crush to obtain a third composition,
A fermented papaya fruit obtained by mixing and homogenizing the first composition, the second composition, and the third composition, and standing and aging for a long period of at least 6 months. Manufacturing method.