KR20220025679A - Method for producing fermented apple postbiotics with increased effective component content using lactic acid bacteria - Google Patents

Abstract

The present invention relates to a production method of apple fermented postbiotics with increased content of useful ingredients using lactic acid bacteria, wherein when using the production method according to the present invention, can provide the apple fermentation postbiotics with increased content of total polyphenols, total flavonoids, and L-ascorbic acid, and can usefully be used in the production of health functional foods for antioxidant, preventing obesity, enhancing immunity, and the like. The production method comprises: a step of processing; and a step of fermenting.

Description

Method for producing fermented apple postbiotics with increased effective component content using lactic acid bacteria

The present invention relates to a method for producing fermented apple postbiotics with an increased content of useful ingredients using lactic acid bacteria.

Lactobacillus is one of the most beneficial microorganisms available to humans, and is a non-pathogenic bacteria that live in the human intestine and perform intestinal and bacteriostatic actions. In addition, it has the characteristics of producing a large amount of lactic acid by fermenting sugars, growing well under low pH and anaerobic conditions, and producing various nutrients. When vegetables are fermented by lactic acid bacteria, their unique flavor and taste are improved, and the sensory properties are improved, and _vitamin C and various physiologically active substances are well preserved by organic acids. It is known to have the advantage of becoming a hygienic food by inhibiting the growth and proliferation of putrefactive and pathogenic bacteria.

Apples are fruits of apple trees, which are perennial plants belonging to the genus Malus sp. of the Rosaceae family, and are alkaline fruits with relatively little protein and fat, and high content of vitamin A, vitamin C and inorganic salts. Apples contain a large amount of pectin, a water-soluble dietary fiber, which lowers blood cholesterol, suppresses rapid rise in blood sugar, and inhibits fatty acid synthesis by secreting galacturonic acid in the intestine. In addition, it is known that the red skin of apples contains polyphenol-based antioxidants such as anthocyanins. As such, apples are not only effective in lowering blood pressure, preventing arteriosclerosis, preventing diabetes, antioxidant activity, anti-cancer activity and obesity prevention by dietary fiber, and enhancing immune function, but also show high consumption because they are good for health and skin care. . However, it is difficult to preserve the fresh flavor of apples for a long time due to the limitation of storage, and it is inconvenient to always consume it due to the seasonal limitation.

On the other hand, Korean Patent No. 1941967 discloses 'a fermented apple containing a large amount of calcium ions', and Korean Patent No. 1994172 discloses 'Apple fermented beverage using Weissella confusa SRCM100892 strain and jelly using the same. manufacturing method' is disclosed. However, the 'method for producing fermented apple postbiotics with an increased content of useful ingredients using lactic acid bacteria' of the present invention has not yet been disclosed.

The present invention was derived from the above needs, and the present inventors treated apple juice with cellulase and polygalacturonase, and then Lactobacillus plantarum CBG-C21 ( Lactobacillus plantarum CBG-C21) ) strain and freeze-drying to prepare apple fermented postbiotic powder, and compared to the enzyme-treated and unfermented apple powder, the total polyphenols, total flavonoids, and L of the apple fermented postbiotic powder - By confirming that the content of ascorbic acid significantly increased, the present invention was completed.

In order to solve the above problems, the present invention comprises the steps of: 1) treating an apple with an enzyme; and 2) inoculating lactic acid bacteria into the apples treated with the enzyme in step 1) and fermenting them.

In addition, the present invention provides an apple fermented postbiotics with an increased content of useful ingredients prepared by the above manufacturing method.

In addition, the present invention provides a health functional food comprising apple fermented postbiotics having an increased content of the useful ingredients.

By using the manufacturing method according to the present invention, it is possible to provide fermented apple postbiotics with an increased content of total polyphenols, total flavonoids and L-ascorbic acid, and it is used in the manufacture of health functional foods for antioxidant, obesity prevention, immunity enhancement, etc. It can be useful.

1 shows the total polyphenol content in the apple fermentation postbiotics of the present invention.
Figure 2 shows the total flavonoid content in the apple fermentation postbiotics of the present invention.
3 shows the L-ascorbic acid content in the apple fermentation postbiotics of the present invention.

In order to achieve the object of the present invention, the present invention

1) treating apples with enzymes; and

2) Inoculating the lactic acid bacteria in the enzyme-treated apple in step 1) and fermenting the apple.

As used herein, the term “postbiotics” refers to metabolic or fermentation products produced by probiotics, which are microorganisms beneficial to health, using prebiotics as a nutrient source. As a representative form of postbiotics, there are dead cells or fermented products processed by a specific method of probiotics.

In the present invention, the "apple fermentation postbiotics" means "apple lactic acid bacteria fermented product".

In the method for producing fermented apple postbiotics having an increased content of useful ingredients of the present invention, the apple may be in various forms such as apple extract, juice, concentrate or powder, and preferably prepared by mixing apple concentrate powder with water. It may be one apple juice, but is not limited thereto.

In addition, the enzyme may be cellulase (cellulase) and polygalacturonase (polygalacturonase), but is not limited thereto. The enzymes may be treated simultaneously or sequentially, but is not limited thereto.

In addition, the lactic acid bacteria may be a Lactobacillus plantarum strain, preferably a Lactobacillus plantarum CBG-C21 strain having an accession number of KACC92083P, but is not limited thereto.

In addition, the fermentation may be performed at 32-42° C. for 18-28 hours, preferably at 37° C. for 23 hours, but is not limited thereto.

In addition, the useful component may be total polyphenols (polyphenol), total flavonoids (flavonoid) and L-ascorbic acid (L-ascorbic acid), but is not limited thereto.

The manufacturing method according to the present invention is preferably,

1) treating 25-35% (w/v) apple juice with cellulase and polygalacturonase and reacting at 50-60° C. for 1.5-2.5 hours; and

2) Inoculate the Lactobacillus plantarum CBG-C21 strain at 3.0×10 ⁷ to 3.0×10 ⁹ CFU/ml into the apple juice treated with cellulase and polygalacturonase in step 1) at 32 to 42° C. Fermentation for 18 to 28 hours; may include, and most preferably,

1) 30% (w/v) apple juice is treated with 0.1% (w/v) cellulase and 0.1% (w/v) polygalacturonase and reacted at 55° C. for 2 hours; and

2) inoculating the Lactobacillus plantarum CBG-C21 strain at 3.0×10 ⁸ CFU/ml into the apple juice treated with cellulase and polygalacturonase in step 1) and fermenting at 37° C. for 23 hours; may include, but is not limited thereto.

In addition, the manufacturing method according to the present invention may further include the step of freeze-drying the fermented product of step 2), but is not limited thereto.

The present invention also provides an apple fermented postbiotics having an increased content of useful ingredients prepared by the above manufacturing method. The useful ingredients are the same as described above.

The postbiotics may be prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, effervescent tablet and beverage, but is not limited thereto.

The present invention also provides a health functional food comprising apple fermented postbiotics having an increased content of the useful ingredients. The useful ingredients are the same as described above.

There is no particular limitation on the type of the health functional food. Examples of health functional foods containing the apple fermented postbiotics include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, rice cakes, dairy products including ice cream, various soups, There are beverages, teas, drinks, alcoholic beverages, and vitamin complexes, and includes all health functional foods in the ordinary sense.

The apple fermented postbiotics according to the present invention has an increased content of total polyphenols, total flavonoids and L-ascorbic acid compared to apples without enzyme treatment and fermentation, and the antioxidant, obesity prevention, and immune function enhancement functions of the useful ingredients Various health functional foods can be manufactured using the

The health functional food may be prepared in any one formulation selected from powder, granule, pill, tablet, capsule, candy, syrup, effervescent tablet and beverage, but is not limited thereto.

When the fermented apple postbiotic of the present invention is used as a food additive, the fermented apple postbiotic may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The active ingredient may be appropriately used depending on the purpose of its use. In general, in the production of food or beverage, the apple fermented postbiotics of the present invention are added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health control, it can be used in an amount within a range that does not cause any problems in terms of safety.

The health functional food of the present invention includes ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when manufactured as a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.), disaccharides (eg, maltose, sucrose, etc.), oligosaccharides, polysaccharides (eg, dextrin, cyclodextrin, etc.) or sugar alcohols (eg, , xylitol, sorbitol, erythritol, etc.) is preferable. As the flavoring agent, natural flavoring agents (eg, taumartin, stevia extract, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) may be used. In addition to the above health functional food composition, various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonic acid It may further contain a carbonation agent and the like used in beverages.

Although the proportion of the added component is not very important, it is generally selected in the range of 0.01 to 0.1 parts by weight based on 100 parts by weight of the health functional food of the present invention.

Hereinafter, an embodiment of the present invention will be described in detail. However, the following examples are only illustrative of the present invention, and the content of the present invention is not limited to the following examples.

Preparation Example 1. Preparation of Apple Fermented Postbiotics

90.97 g of apple concentrate powder was weighed and mixed with 210 ml of purified water to prepare 30% (w/v) apple juice. Thereafter, the pH was corrected to 7.0 and dextrin was added, lyophilized and powdered, and used as a control (apple powder).

For the fermentation of apples, Lactobacillus plantarum CBG-C21 strain (Accession No.: KACC92083P) was cultured for 24 hours to prepare a strain culture solution of 6.0×10 ⁹ CFU/ml. The apple juice was treated with 0.1% (w/v) cellulase and 0.1% (w/v) polygalacturonase and reacted at 55° C. for 2 hours, followed by 2% ( w/v) glucose and 1% (w/v) casein peptone were added and autoclaved at 121° C. for 15 minutes. Thereafter, the enzyme-treated apple juice was inoculated with 5% (3.0×10 ⁸ CFU/ml) of the Lactobacillus plantarum CBG-C21 strain, and cultured at 37° C. for 23 hours to perform fermentation. After dextrin was added to the fermented apple juice after fermentation, it was freeze-dried and powdered to prepare an apple fermented postbiotic powder.

Example 1. Analysis of the content of useful ingredients

Total polyphenol, total flavonoid, and L-ascorbic acid content, which are useful antioxidant-related ingredients, were measured for the apple powder and apple fermented postbiotic powder of Preparation Example 1. The total polyphenol content was measured with some modifications of the Singleton and Rossi method (Singleton, V. and Rossi, J et al., 1965, American Journal of Enology and Viticulture. 16:144-158). Put 1 ml of the sample solution and gallic acid standard (20, 40, 60, 80, 100 mg/L) into a 25 ml quantitative flask containing 9 ml of tertiary distilled water, respectively, and phenol solution (Folin-ciocalteu's) phenol reagent) 1 ㎖ was added. After 5 minutes, 10 ml of a 7% Na ₂ CO ₃ solution was added and sufficiently stirred. Then, 25 ml of tertiary distilled water was filled, mixed, and reacted at 23° C. for 90 minutes. Absorbance was measured at 750 nm using a blank as a control. At this time, the standard calibration curve was obtained by measuring in the same manner as above using gallic acid as a standard material at a constant concentration, and the absorbance of the sample was substituted into the standard calibration curve to express the total phenol content in μg/g.

Total flavonoid content was determined by modifying Zhishen's method (Zhishen J. et al., 1999, Food Chemistry. 64:555-559). Add 1 ml of the sample solution and quercetin standard (20, 40, 60, 80, 100 mg/L) to a 10 ml quantitative flask containing 4 ml of tertiary distilled water, respectively, and at zero time, 5% NaNO ₂ 0.3 ml was added, and the mixture was sufficiently stirred. After 5 minutes, 0.3 mL of 10% AlCl ₃ was added, and at 6 minutes, 2 mL of 1M NaOH was added, and immediately, 2.4 mL of tertiary distilled water was added, mixed thoroughly, and reacted at room temperature for 1 minute, then using UV/Vis Spectrometer The absorbance of each solution was measured at 510 nm using the blank as a control. At this time, the total flavonoid content was obtained by measuring in the same way as above using a certain concentration of querycetin as a standard material, and substituting the absorbance of the sample into the standard calibration curve to express the total flavonoid content in μg/g.

The L-ascorbic acid content was measured using a Megazyme assay kit (Megazyme Ltd.). Specifically, 3% (w/v) metaphosphoric acid and 10 mM EDTA (ethylenediaminetetraacetic acid) were added to a 100 mg sample, homogenized at room temperature for 60 minutes, centrifuged for 10 minutes, and the obtained supernatant was filtered. was used. 152 μl of tertiary distilled water and buffer 1 were mixed with 10 μl of the filtered supernatant, and after 3 minutes, MTT[3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide] buffer 2 was added and reacted for 3 minutes The primary absorbance (A1) was measured at 578 nm using a spectrophotometer, and a PMS (5-methylphenazinium methyl sulfate) buffer was added and reacted for 8 minutes. Secondary absorbance (A2) was measured at 578 nm. .

As a result, as shown in Table 1 below, it was confirmed that the contents of total polyphenols, total flavonoids and L-ascorbic acid in the apple fermented postbiotic powder were significantly increased compared to the apple powder without enzyme treatment and fermentation (Fig. 1). to 3).

Measurement result of useful ingredient content in apple fermentation postbiotics useful ingredients apple powder apple fermentation
Postbiotics Powder Total polyphenols (μg/g) 375.2±2.44 453.8±2.09 Total flavonoids (μg/g) 19.99±0.15 26.68±0.23 L-Ascorbic Acid (g/100g) 1,867±19.55 1,960±12.17

Agricultural Biotechnology Research Institute KACC92083P 20150710

Claims (7)

1) treating apples with enzymes; and
2) Inoculating the lactic acid bacteria in the enzyme-treated apple in step 1) and fermenting the apple. The method of claim 1, wherein the enzyme is cellulase and polygalacturonase. The method of claim 1, wherein the lactic acid bacteria is a Lactobacillus plantarum CBG-C21 strain having an accession number of KACC92083P. The method of claim 1, wherein the useful ingredients are total polyphenols, total flavonoids, and L-ascorbic acid. manufacturing method. The method of claim 1,
1) treating apple juice with cellulase and polygalacturonase and reacting at 50 to 60° C. for 1.5 to 2.5 hours; and
2) Inoculating the Lactobacillus plantarum CBG-C21 strain having an accession number of KACC92083P into the apple juice treated with cellulase and polygalacturonase in step 1) and fermenting it at 32-42°C for 18-28 hours. ; A method for producing an apple fermentation postbiotic with an increased content of useful components of total polyphenols, total flavonoids, and L-ascorbic acid, including. An apple fermented postbiotic with an increased content of useful ingredients prepared by the method of any one of claims 1 to 5. A health functional food comprising apple fermented postbiotics with an increased content of useful ingredients of claim 6.

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