KR102121839B1 - Method for manufacturing functional fermented material of oriental melon with yeast and lactic acid bacteria - Google Patents

Abstract

The present invention relates to a method for producing a functional melon fermentation product through a complex fermentation of yeast and lactic acid bacteria, and more specifically, a melon extract fermentation for 7 days by adding 3% of the precursor MSG continuously after 1 day of yeast fermentation Was performed. Unlike GABA conversion experiments using several natural products, there was a big difference in GABA production of the final complex fermentation according to the heat treatment of the initial melon extract. In the case of heat treatment of the melon extract, after the first yeast fermentation, the second lactic acid fermentation shows some GABA production, whereas in the case of the non-heat treatment of the melon extract, the second lactic acid fermentation shows GABA production from day 1 and fermentation 4 days. In the results, all MSG precursors were converted to GABA. The fermentation material with enhanced functionality as described above is expected to be used as a material for functional foods and health foods.

Description

Method for manufacturing functional melon fermented product through complex fermentation of yeast and lactic acid bacteria {Method for manufacturing functional fermented material of oriental melon with yeast and lactic acid bacteria}

The present invention relates to a method for producing a functional melon fermentation product through a combination fermentation of yeast and lactic acid bacteria.

Melon is a plant belonging to the family Cucurbitaceae, and its origin is known as India, and it was developed by dividing it into oriental melon and western melon according to the spread. Currently, it is mainly grown in Korea, China, and Japan. In Korea, it is estimated that melons were cultivated before the Silla period, and various varieties of native melons were grown in each region. Melon is a kind of fruit and vegetable with a sweet taste and unique scent, and its cool taste is suitable for our taste, and it has been widely used as a fruit in summer since ancient times. Melon has a high sugar content, high carbohydrates of 7.3% to 7.4%, contains a lot of carotene and vitamin C, and is recognized as a diuretic action like watermelon. In addition, it is characterized by a high content of minerals such as calcium and phosphorus and vitamins A and C. The pharmacological action of melon contains a crystalline gourmet substance called elaterin in dried undercooked melon taps, and this ingredient has the best effect. In addition, melons have ingredients that act as Jinhae and Geodam, and they also help relieve constipation, so it is also effective for palliation, jaundice, and diuresis. It is known to relieve thirst, especially in summer when sweating a lot, and melon is a particularly good food that helps to recover from fatigue in the summer, when the constitution becomes acidic. As a way to use melon as a healthy food material and high value-added material, it is necessary to develop a material with enhanced functional substances through fermentation to produce useful substances and use it as a material for food and health food.

Lactic acid bacteria and yeast are representative fermentation microorganisms and are proven GRAS (generally recognized as safe) to produce the metabolites of lactic acid and alcohol through fermentation and yeast fermentation of vegetables, fruits, and animal raw materials. It acts to impart storability. In particular, lactic acid bacteria and yeast are probiotics, and when they suppress harmful bacteria in the human body, they are beneficial bacteria.

Korean Registered Patent No. 10-1324621 (Registered on October 28, 2013)

An object of the present invention is to provide a method for producing a melon fermentation product with enhanced gamma-aminobutyric acid (GABA) through yeast and lactic acid bacteria fermentation.

In addition, an object of the present invention is to provide a GABA enhanced melon fermentation product prepared by the above method.

In addition, an object of the present invention is to provide a GABA enhanced food composition comprising the melon fermentation product as an active ingredient.

In order to achieve the above object, the present invention is (1) cutting melon; (2) homogenizing the cut melon by adding water of 1 to 3 times the weight of the cut melon; (3) a primary yeast fermentation step of inoculating and incubating the homogenized melon extract with Saccharomyces cerevisiae starter; And (4) a secondary lactic acid fermentation step of inoculating and incubating the primary yeast fermentation product with a Lactobacillus plantarum EJ2014 (KCCM11545P) starter, gamma-aminobutyric acid (GABA). ) Provides an improved method for manufacturing melon fermentation products.

In addition, the present invention provides a GABA enhanced melon fermentation product prepared by the above method.

In addition, the present invention provides a GABA enhanced food composition comprising the melon fermentation product as an active ingredient.

The present invention relates to a method for producing a functional melon fermentation product through a complex fermentation of yeast and lactic acid bacteria, and more specifically, a melon extract fermentation for 7 days by adding 3% of the precursor MSG continuously after 1 day of yeast fermentation Was performed. Unlike GABA conversion experiments using several natural products, there was a big difference in GABA production of the final complex fermentation according to the heat treatment of the initial melon extract. In the case of heat treatment of the melon extract, after the first yeast fermentation, the second lactic acid fermentation shows some GABA production, whereas in the case of the non-heat treatment of the melon extract, the second lactic acid fermentation shows GABA production from day 1 and fermentation 4 days. In the results, all MSG precursors were converted to GABA. The fermentation material with enhanced functionality as described above is expected to be used as a material for functional foods and health foods.

Figure 1 shows the process of manufacturing the melon extract.
Figure 2 shows the Lactobacillus plantarum ( Lactobacillus plantarum ) single fermentation process of the melon extract.
Figure 3 shows the results of (A) pH, (B) acidity measurement of a single fermentation according to the heat treatment conditions of the melon extract. (a) non-heat treatment, (b) 85° C., 20 min heat treatment, (c) 121° C., 15 min heat treatment.
Figure 4 shows the results of the measurement of (A) live bacteria, (B) reducing sugars of a single fermentation according to the heat treatment conditions of the melon extract. (a) non-heat treatment, (b) 85° C., 20 min heat treatment, (c) 121° C., 15 min heat treatment.
Figure 5 shows the results of measuring the GABA content of a single fermentation according to the heat treatment conditions of the melon extract. (a) non-heat treatment, (b) 85° C., 20 min heat treatment, (c) 121° C., 15 min heat treatment.
6 shows a mixed fermentation process using Saccharomyces cerevisiae and Lactobacillus plantarum of melon extract.
7 shows the results of (A) pH, (B) acidity measurement of the mixed fermentation according to the heat treatment conditions of the melon extract. (a) non-heat treatment, (b) 85° C., 20 min heat treatment, (c) 121° C., 15 min heat treatment.
Figure 8 shows the results of (A) yeast group viable cell count, (B) lactobacillus cell viable cell count according to the heat treatment conditions of the melon extract. (a) non-heat treatment, (b) 85° C., 20 min heat treatment, (c) 121° C., 15 min heat treatment.
Figure 9 shows the results of measuring the GABA content of the mixed fermentation according to the heat treatment conditions of the melon extract. (a) non-heat treatment, (b) 85° C., 20 min heat treatment, (c) 121° C., 15 min heat treatment.

The present invention (1) cutting the melon; (2) homogenizing the cut melon by adding water of 1 to 3 times the weight of the cut melon; (3) a primary yeast fermentation step of inoculating and incubating the homogenized melon extract with Saccharomyces cerevisiae starter; And (4) a secondary lactic acid fermentation step of inoculating and incubating the primary yeast fermentation product with a Lactobacillus plantarum EJ2014 (KCCM11545P) starter, gamma-aminobutyric acid (GABA). ) Provides an improved method for manufacturing melon fermentation products.

Preferably, the homogenizing step of step (2) may be first homogenized with a hand blender, filtered, and then secondary homogenized with an electric blender, but is not limited thereto.

Preferably, the primary yeast fermentation in step (3) may be fermented at 25 to 30°C for 1 to 3 days, but is not limited thereto.

Preferably, during the culture of step (4), with respect to 100 parts by weight of the entire culture, MSG may include 1 to 10 parts by weight, but is not limited thereto.

Preferably, the secondary lactic acid fermentation in step (4) may be fermented at 25 to 30°C for 1 to 10 days, but is not limited thereto.

The Lactobacillus plantarum EJ2014 strain used in the Examples of the present invention was deposited as KCCM11545P at the Korea Microbial Conservation Center.

In addition, the present invention provides a GABA enhanced melon fermentation product prepared by the above method.

In the present invention, "gamma-aminobutyric acid (gamma-amino butyric acid; GABA)" is 4-aminobutyl acid, L-glutamate (L-glutamate) substrate is produced by a decarbonation reaction, as an enzyme involved in this There is glutamate decarboxylase (GAD) and is synthesized by a pyridoxid-5'-phosphate dependent pathway. GABA is a non-proteinaceous amino acid that is not found in proteins. It is a neurotransmitter present in the brain or spinal cord that improves blood flow and increases brain oxygenation, promoting brain metabolism and improving brain memory. GABA is known to inhibit neuronal activity, unlike glutamate, which activates nerves, and these functions have a profound effect on neuronal function and information processing. It is known to coordinate and refine sophisticated motor skills. GABA's effect of promoting brain blood flow and increasing oxygen supply have been shown to improve the effects of stroke, sequelae and cerebral arteriosclerosis by promoting metabolism of brain cells, and are used as medicines.

In the present invention, "starter (starter)" refers to a microbial culture medium used when preparing a fermentation product. Therefore, the type of starter microorganism determines the characteristics of the product and has an important effect on the quality of the product. Among microorganisms, bacteria, fungi, yeast, etc. are used as starters, and these can be used alone or in combination.

In addition, the present invention provides a GABA enhanced food composition comprising the melon fermentation product as an active ingredient.

In the case of the food composition of the present invention, there is no particular limitation on the type of the food. Examples of foods to which the mixed fermentation product can be added are meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, Drinks, alcoholic beverages and vitamin complexes.

Hereinafter, the present invention will be described in more detail through the following examples. However, the present invention is not limited by these examples.

< Example 1> According to heat treatment of melon extract Lactobacillus Planta rum( Lactobacillus plantarum ) Single fermentation

1. Preparation of melon extract

(1) Materials and methods

After washing Seongju melon in Gyeongbuk (2018 harvest), it was cut to about 1~2cm and used as a sample while being stored frozen. After mixing the same amount of frozen melon slices and distilled water, dilute it twice and then homogenize it first with a hand blender (HR1673, Koninklijke Philips NV Co., Ltd., Amsterdam, Nederland), and use an electronic blender (SMX-3610WS, Guandgdong Shunde Lefan Electric Appliance) Co., Ltd., Cheonan, Korea) to homogenize at a second high speed. The first homogenized melon solution was filtered to remove foreign substances such as seeds using a 70 μm 25 mesh and then homogenized at a second high speed. The yield of the final extract was 82.5% (w/w) and was used in a sterilized collection bottle and stored frozen at -18°C (FIG. 1).

(2) Results and discussion

As a result of physicochemical analysis of melon diluted twice, pH and acidity were 5.95 and 0.10%, respectively, and sugar content, reducing sugar, and solid content were 6.67 °Brix, 4.24%, and 5.75%, respectively (Table 1). These results were obtained from previous reports (Jo YJ et al. 2010, J. Korean Soc. Food Sci. Nutr. 39: 1359-1365), melons harvested in 2009 at pH 5.95, acidity 0.21%, sugar content 10.5 °Brix and solids content. There was a tendency similar to that reported as 10.8%, but the acidity, sugar content, and solid content were slightly lower, which was judged to be due to dilution.

Oriental melon pH 5.95±0.01 ¹⁾ Total acidity (%) 0.10±0.02 °Brix 6.67±0.06 Reducing sugar (%) 4.24±0.54 Soluble content (%) 5.75±0.23

¹⁾ Numerical values were expressed as mean±SD (n=3).

2. According to heat treatment of melon extract Lactobacillus Plantarum ( Lactobacillus plantarum ) Single fermentation

(1) Materials and methods

After diluting the melons harvested in Seongju-gun twice and dividing the two-stage homogenized melon samples under the conditions of no heat treatment and heat treatment at 85°C for 20 minutes and 121°C for 15 minutes, MSG 3% and yeast extract 0.5% were equal. Add. Later Lactobacillus plantarum EJ2014 was inoculated, and lactic acid fermentation was performed at 30°C for 7 days (FIG. 2). The fermentation composition is shown in Table 2.

Ingredient (a) (b) (c) Oriental melon
(x2 dilution, mL) 100 MSG ^{2 )} (g) 3 10% Yeast extract (mL) 5 Total (mL) 105

²⁾ Monosodium glutamate

(a) Non-heat treatment

(b) 85℃, 20min heat treatment

(c) 121℃, 15min heat treatment

(2) Results and discussion

1) pH, acidity result

As a result of the single fermentation characteristics according to the heat treatment conditions of melon, as the lactic acid fermentation progressed, the pH of all conditions was maintained after decreasing from pH 5.9-6.0 to pH 4 before fermentation (FIG. 3A).

In the case of acidity, the acid was generated at 0.06-0.10 before fermentation and showed 1.19-1.34% under all conditions on the first day of fermentation, but the condition without heat treatment decreased to 1.11% on the seventh day of fermentation (FIG. 3B).

2) Viable cell count, reduced sugar result

As a result of the viable cell count, as a result of fermentation, a high bactericidal activity of 8.5-8.6 log CFU/mL was observed on the first day of fermentation under all conditions, and the activity was maintained until the third day of fermentation. The condition without heat treatment was reduced to 8.1 log CFU/mL on the 7th day of fermentation (FIG. 4A).

The reducing sugar showed a slight difference from the beginning of fermentation. The reduced sugar of melon diluted twice was 4.24%, showing a difference of 2.68-3.25% and 1-1.5% of the reducing sugar before fermentation of a, b, and c samples. It is thought that the microorganisms originally possessed by melons are continuously fermenting at room temperature. It was found that lactic acid bacteria exhausted sugar in all conditions on the 4th day of fermentation, although the rate of exhausting sugar was slower than that of the unheated condition (FIG. 4B ).

3) GABA analysis results

To measure the change in the GABA content of the fermented product by lactic acid bacteria, the supernatant obtained after diluting the fermented product 3 times and centrifuging it was qualitatively analyzed using TLC. GABA was produced from day 1 of lactic fermentation, but MSG remained under all conditions until day 7 of fermentation (FIG. 5). However, as the MSG spot was much smaller on the 7th day of fermentation under the unheated condition, the condition without the heat treatment during fermentation with melon seems to help in the production of GABA.

< Example 2> According to heat treatment of melon extract Saccharomyces Ceremony Child( Saccharomyces cerevisiae ) And Lactobacillus Plantarum ( Lactobacillus plantarum ) Mixed fermentation

1. Materials and Methods

After diluting the melons harvested in Seongju-gun, Gyeongsangbuk-do twice, and then homogenizing the sample of the melon extract homogenized to the second time after dividing the sample into heat-treated conditions at 85°C for 20 minutes and 121°C for 15 minutes, Saccharomyces cerevisiae ( Saccharomyces cerevisiae ), and yeast fermentation was performed at 30°C for 1 day. Subsequently, 3% of MSG was added in the same manner, and Lactobacillus plantarum EJ2014 was inoculated to proceed with lactic acid fermentation at 30°C for 7 days (FIG. 6 ). The fermentation composition is shown in Table 3.

Ingredient (a) (b) (c) 1 ^st - Yeast Fermentation Oriental melon
(x2 dilution, mL) 100 2 ^nd - Lactic acid Fermentation 30% MSG (mL) 10 Total (mL) 110

(a) Non-heat treatment

(b) 85℃, 20min heat treatment

(c) 121℃, 15min heat treatment

2. Results and Discussion

(1) pH, acidity results

As a result of the pH, the pH was different depending on the presence or absence of sterilization at the beginning of fermentation. It seems that the microorganisms in the melon showed differences due to fermentation at room temperature. The conditions of heat treatment were pH 5.21-5.28, and the conditions without heat treatment were slightly decreased to pH 4.24, showing acidification, and then slightly increased as yeast fermentation progressed. On the first day of lactic acid fermentation, pH was significantly different depending on the presence or absence of heat treatment. The b and c conditions of the heat treatment decreased as lactic acid fermentation progressed, indicating 4.21-4.23 on the 7th day of lactic acid fermentation. In the a condition without heat treatment, the pH increased until day 1 of lactic fermentation, and then decreased to pH 6.44, and then decreased to pH 4.61 on day 7 of lactic fermentation (FIG. 7A).

Also in the case of acidity, there was a difference according to the presence or absence of heat treatment at the beginning of fermentation. The conditions a, 0.63%, b, and c showed 0.16% and 0.32%, respectively. The conditions b and c, which were heat treated on the 1st day of lactic acid fermentation, showed an increase in acidity of 0.88-0.90%, followed by 1.46-1.51% on the 7th day of lactic acid fermentation. A condition without heat treatment was maintained after showing 0.14% of lactic acid fermentation on day 1 and 2.15% of lactic acid fermentation on day 4 (FIG. 7B).

(2) Viable cell count measurement result

As a result of the viable cell count of yeast bacteria, as the fermentation started, it showed a high bactericidal activity of 8.8-9.0 log CFU/mL on the first day of fermentation under all conditions, but the activity decreased as the fermentation of lactic acid bacteria progressed. The condition without heat treatment was not observed after decreasing to 7.8 log CFU/mL on the 1st day of lactic acid fermentation, whereas the two conditions with heat treatment showed 6.2-6.5 log CFU/mL on the 7th day of lactic acid fermentation (FIG. 8A).

The results of the number of live bacteria of lactic acid bacteria showed the highest activity at 8.9-10.0 log CFU/mL on the first day of lactic acid fermentation, and slightly decreased or maintained. The condition without heat treatment was decreased until the 7th day of lactic fermentation, indicating 7.7 log CFU/mL (FIG. 8B).

(3) GABA analysis results

To measure the change in GABA content of the fermented mixture by yeast and lactic acid bacteria, the fermented product was diluted 3 times and centrifuged, and the obtained supernatant was qualitatively analyzed using TLC. Under the a condition without heat treatment, it was found that GABA was produced from the 1st day of lactic acid fermentation, and MSG was converted to GABA on the 4th day of lactic acid fermentation (FIG. 9).

In conclusion, while performing fermentation to enhance the functional substance GABA using melons grown in Seongju region, MSG, a precursor, was converted to some GABA through sole lactic fermentation using L. plantarum EJ2014 derived from rice bran. In particular, in the case of non-heat treatment of the melon extract (2 fold dilution), GABA material production increased.

The same melon extract was continuously fermented with lactic acid bacteria for 7 days immediately after 1 day of yeast fermentation in the presence of the precursor MSG 3%. Unlike GABA conversion experiments using several natural products, there was a big difference in the GABA production of the final complex fermentation according to the heat treatment of the initial melon extract. In the case of heat treatment of the melon extract, after the first yeast fermentation, the second lactic acid fermentation shows some GABA production, whereas in the case of the non-heat treatment of the melon extract, the second lactic acid fermentation shows GABA production from day 1 and fermentation 4 days. All MSG precursors were converted to GABA.

Therefore, it is possible to ferment by adding only the precursor MSG without adding fermentable sugar after preparing the melon extract through homogenization by diluting it twice with fresh water in the fermentation that strengthens GABA as a raw material for sugary fruits such as melon. Did. In particular, during the preparation of the melon extract, the first homogenized sample exhibited a phenomenon in which the growth of aerobic contaminating bacteria (such as archaea) was promoted while the solid content in the form of particles was exposed to the surface during the primary yeast fermentation, thereby generating an odor such as odor . Therefore, it was possible to produce high concentrations of GABA in a short period of time by primary yeast fermentation and secondary lactic acid fermentation, as conditions for minimizing destruction of nutrients through non-thermal treatment of the melon extract prepared through two-step homogenization.

Korea Microbial Conservation Center (Overseas) KCCM11545P 20140609

Claims (7)

(1) cutting the melon;
(2) adding 1 to 3 times the water weight of the cut melon to filter the melon cut by hand homogenizing with a hand blender (hand blender), followed by secondary homogenizing with an electric blender;
(3) the primary yeast fermentation step of inoculating and incubating the homogenized melon extract without heat treatment, and inoculating a Saccharomyces cerevisiae starter; And
(4) Gamma-aminobutyric acid (GABA) comprising a secondary lactic acid fermentation step of inoculating and incubating the primary yeast fermentation product with a Lactobacillus plantarum EJ2014 (KCCM11545P) starter. Enhanced melon fermentation method. delete The method according to claim 1, wherein the primary yeast fermentation in the step (3) is fermentation at 25 to 30° C. for 1 to 3 days. The method of claim 1, wherein in the step (4), GABA-enhanced melon fermentation method characterized in that 1 to 10 parts by weight of MSG is included with respect to 100 parts by weight of the entire culture. According to claim 1, wherein the secondary lactic acid fermentation of step (4) is 25 to 30 ℃, GABA enhanced melon fermentation method characterized in that the fermentation for 1 to 10 days. A GABA enhanced melon fermentation product prepared by the method of any one of claims 1, 3 to 5. A GABA-enhanced food composition comprising the melon fermented product of claim 6 as an active ingredient.

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