KR20230146494A - A Natural Complex Preservative Agent for a Grain or a Grain Product - Google Patents

Abstract

The present invention relates to natural complex preservatives for grains or grain products. A preservative for grains or processed grain products comprising more than 50 wt% of Cordyceps sinensis extract and broccoli extract, wherein the Cordyceps sinensis extract and broccoli extract are bio-converted.

Description

A Natural Complex Preservative Agent for a Grain or a Grain Product}

The present invention relates to natural complex preservatives for grains or grain products, and in particular to natural complex preservatives for grains or grain products based on extracts of Cordyceps sinensis and broccoli.

Due to changes in dietary patterns due to industrial development, demand for alternative foods that can be easily eaten by children and adults is increasing. Grain processed foods that are substitute foods include bread, rice cake, noodles, cereals, or similar products produced using grains. These processed foods have been used as snacks, but as living environments change, they are gradually becoming convenient products that can replace staple foods. As the consumption of processed grain products increases, various products that can actively reflect consumer tastes are being developed and released. However, most processed grain products have a high moisture content, which makes it easy for microorganisms to grow, making it difficult to ensure safety against microorganisms. . For example, microorganisms such as E.coli Bacillus cereus and Salmonella spp., which are widely distributed in processed grain products, are bacteria that cause food poisoning and can be introduced through various routes such as raw materials, production process, packaging process, or distribution process. Although synthetic preservatives have been used since the 1930s to address microbial risks, there may always be controversy and concerns about safety. To solve this problem, microbial control such as process improvement is being implemented, but quality problems or disease accidents in processed grain products caused by microorganisms still occur. Research is being conducted on chitosan, natural extracts, microbial metabolites, organic acids, or similar antibacterial materials to replace synthetic preservatives. However, despite such research and development, there are many difficulties in producing preservatives with natural extracts that have effects such as ensuring the safety of processed grain products against microorganisms and extending the storage period. Patent Publication No. 10-2016-0141605 discloses a method for producing a natural preservative containing a rosemary composition. In addition, International Publication No. WO 2017/083363 describes natural preservatives derived from extracts of Albizia amara seeds and Magnolia officinalis bark that can be used for the preservation of food, cosmetics, nutritional supplements, oral hygiene products, and pharmaceuticals. Begin. Natural preservatives need to have an antibacterial effect against various microorganisms and have properties that can be added to grains or grain processed products to maintain or improve the unique characteristics of grains or grain processed products. However, the prior art does not disclose natural preservatives with such properties.

The present invention is intended to solve the problems of the prior art and has the following purposes.

Prior art 1: Patent Publication No. 10-2016-0141605 (Dain Materials Co., Ltd., published on December 9, 2016) Method for producing natural preservative containing rosemary composition Prior Art 2: International Publication No. WO 2017/083363 (Unigen, Inc., 2017.05.18. Natural preservatives and antimicrobial agents and compositions thereof

The purpose of the present invention is to provide a natural complex preservative for grains or processed grain products that is composed of Cordyceps sinensis extract and broccoli extract and has improved antibacterial and preservation properties.

According to a preferred embodiment of the present invention, the preservative for cereals or processed cereal products comprises at least 50 wt% of Cordyceps sinensis extract and broccoli extract, and the Cordyceps sinensis extract and broccoli extract are biotransformed.

According to another suitable embodiment of the present invention, it comprises at least one ingredient selected from the group consisting of garlic extract, green tea extract, citron extract, fennel extract, dandelion extract, lactic acid bacteria culture, yeast culture, licorice extract, goldenrod extract and turmeric extract. do.

According to another suitable embodiment of the present invention, the microorganism for bioconversion is Saccharomyces sp. Or Lactobacillus Plantarum.

The natural complex preservative for grains or processed grain products according to the present invention is composed of Cordyceps sinensis extract and broccoli extract and can have high antibacterial properties against various types of microorganisms or molds. The preservative according to the present invention can improve antioxidant activity by including garlic or green tea extract, thereby improving the effect of inhibiting the growth of bacteria, yeast, or mold. In the present invention, grains or processed grain products are treated with an antibacterial agent to maintain their original properties and at the same time extend their shelf life. The antibacterial agent according to the present invention can be applied to various foods.

Figure 1 shows an example of the production process of a natural complex preservative for grains or processed grain products according to the present invention.
Figure 2 shows changes in components due to biological conversion during the manufacturing process of the preservative according to the present invention.
Figure 3 shows the results of the antioxidant effect of the preservative according to the present invention applied to processed grain products.

Below, the present invention will be described in detail with reference to the embodiments shown in the attached drawings, but the examples are for a clear understanding of the present invention and the present invention is not limited thereto. In the description below, components having the same reference numerals in different drawings have similar functions, so unless necessary for understanding the invention, the description will not be repeated, and well-known components will be briefly described or omitted, but the present invention It should not be understood as being excluded from the embodiments.

Figure 1 shows an example of the production process of a natural complex preservative for grains or processed grain products according to the present invention.

The natural complex preservative for grains or processed grain products according to the present invention may contain more than 50 wt% of Cordyceps sinensis extract and broccoli extract. For the production of such a complex preservative, Cordyceps sinensis and broccoli can be prepared (P11, P12). Cordyceps can be Cordyceps militaris, C.sobolifera, C.sphecocephala, or similar Cordyceps. Cordyceps sinensis can be cultured under defined conditions for the preservative according to the invention, for example in liquid media, solid batches or worms (P11). Cordyceps sinensis can be cultured in a medium containing brown rice, potatoes, corn, or similar grains, and culture conditions can be set to increase the content of Cordycepin. In media containing brown rice, the content of cordycepin changed depending on growth conditions, and the content differences were shown in Table 1 below.

<Comparison of growth rate and content according to cultivation temperature>

<Comparison of growth rate and content according to cultivation humidity>

<Comparison of growth rate and content according to cultivation light intensity>

As can be seen from Tables 11, 12 and 13, when the culture temperature was 10 to 18°C, the growth rate improved and the content of cordycepin increased as the culture temperature increased. And when the temperature was higher than 18°C, the growth rate decreased and the content of cordycepin decreased along with it. Additionally, when the humidity was gradually increased to 90%, the growth rate improved and the cordycepin content increased accordingly. However, it has been shown that humidity exceeding 90% has a disadvantageous effect. In the case of light intensity, the growth rate remained similar in the range of 200 to 2,000 Lux, but the content of cordycepin was found to increase as the light intensity increased. Therefore, it is advantageous for Cordyceps to be cultured at a temperature of 10 to 18 ° C, a relative humidity of 90% or less, and a light intensity of 200 to 2,000 Lux. Broccoli can be dried and then crushed to form a powder (P12). For example, broccoli can be dried to have a moisture content of 10 wt% or less and crushed to a size of 200 mesh or less. Yes (P12). When Cordyceps sinensis and broccoli are prepared in this way, the active ingredients can be extracted, respectively (P21, P22). Extraction of Cordyceps sinensis and broccoli can be done in various ways, for example, the active ingredients can be extracted by hot water extraction, microwave extraction, or supercritical extraction. The extraction yield and antibacterial activity of broccoli may vary depending on the extraction conditions, and therefore, it is advantageous to be extracted according to appropriate extraction conditions. Table 2 shows the extraction conditions of broccoli and the resulting extraction yield and antibacterial activity.

<Yield and antibacterial activity according to broccoli extraction conditions>

++++: 10 ㎜ or more, +++: 7 ㎜ or more, ++: 3 ㎜ or more, ±: Less than ㎜, -: Unable to confirm

1) Extraction yield: weight loss on drying after extraction/weight of raw material used for extraction *100

2) Antibacterial activity: Comparison of clear zone through bio assay (㎜)

As can be seen from Table 2, the yield or antibacterial activity increases depending on the extraction temperature or extraction time, and it can be seen that the effect of temperature is greater than that of time. However, the antibacterial activity was found to decrease 24 hours after crushing. In addition, it is advantageous to store it for 6 hours after crushing and then extract it, and it is believed that the extraction efficiency is increased due to tissue softening due to the enzymatic reaction within the broccoli after crushing.

Cordyceps sinensis extract and broccoli extract can be mixed (P30), and then Saccharomyces cerevisiae (KCTC 7296), Lactobacillus plantarum subsp. Alternatively, it can be conjugated by microorganisms such as plantarum (KCTC 3108) and Weissella cibaria (KCTC 3807) (P40). And the antibacterial activity according to the mixing ratio and type of microorganism is presented in Table 3.

<Antibacterial activity according to mixing ratio and bioconversion>

1) The more + there are, the better the antibacterial activity (++++: clear zone 10 ㎜ or more and MIC Value 1.0 % or less)

2) S ; Saccharomyces sp., L: Lactobacillus Plantarum, W: Weissella cibaria

3) Bio; each bioconversion → Mix

4) Con; Mix → bioconversion = Conjugation

As can be seen from Table 3, it can be seen that the antibacterial activity increases when the Cordyceps sinensis extract exceeds 50 wt%, and also when bioconversion is performed by mixing Saccharomyces sp. and Lactobacillus Plantarum, the antibacterial activity increases. there is. In order to confirm that the cause of this improvement in antibacterial activity was due to biological conjugation, changes in ingredients were confirmed using high-performance liquid chromatography (HPLC), and are presented in Figure 2. The left side of Figure 2 shows the contents of Sulforaphane and Cordycepin, respectively, and the right side shows the changes in components after bioconversion, showing the components before and after bioconversion. As can be seen from Figure 2, it can be seen that new components appear as the amount of cosedipine and Sulfora phane increases.

Bioconverted preservatives can be applied alone or mixed with other ingredients as natural preservatives or freshness maintainers. According to one embodiment of the present invention, the preservative may include garlic extract, green tea extract, citron extract, fennel extract, and dandelion extract. Such extracts may be extracted by methods such as hot water extraction, microwave extraction, or supercritical extraction. Additionally, one or more of these extracts may be added to the preservative and may be added at a stage before or after bioconversion. At least one ingredient selected from the group consisting of garlic extract, green tea extract, citron extract, fennel extract, dandelion extract, lactic acid bacteria culture, yeast culture, licorice extract, golden extract, and turmeric extract in a ratio of 1 to 50 wt% based on the total preservative. can be added.

Antioxidant activity was measured for the preservative according to the invention.

Antioxidant activity measurement

To measure antioxidant activity, radical scavenging ability was confirmed, and the free radical method was modified. For accurate comparison, a preservative containing green tea extract was freeze-dried and made into powder. 0.2 mL of sample dissolved in each concentration of freeze-dried preservative using MeOH was mixed with 1.8 mL of 0.2 mM 2,2-diphenyl-1-picrylhydrazyl (DPPH) solution and left in the dark for 30 minutes. Absorbance was measured at 517 nm with a UV/Vis spectrophotometer, and the sample concentration SC50 (concentration for 50% scavenging of radical) required to reduce DPPH radicals by 50% was confirmed using the measured absorbance value. L-ascorbic acid and BHA (Butylated Hydroxy Anisole) were used as positive controls.

Processed grain products with preservatives

Raw noodles and white bread were selected as processed grain products for measurement, and were made with the ingredient composition shown in Table 4. The raw noodles were made by putting salt-dissolved water and all-purpose flour in a kneader, kneading for 8 minutes in step 3, and then refrigerating for 15 minutes. It was put to rest. The noodles after the toilet paper were molded using a noodle making machine to have a thickness of 2.5 mm and a width of 2 mm, and then divided into 100 g portions. A mixed product was used for the bread. After thoroughly mixing water and yeast powder, it was placed in a kneader bowl and the mixed powder was added in small portions several times. The dough, which underwent secondary fermentation, first at 25°C for 30 minutes and secondly at 25°C for 120 minutes, was baked in an oven preheated to 180°C for 30 minutes and cooled at room temperature for 30 minutes. The cooled bread was cut into 15 mm pieces and divided into 200 g pieces. Subdivided raw noodles and bread were sprayed with preservatives of different concentrations, and the spray amount was adjusted to be less than 10% (w/w). The spray-treated food was left to stand at room temperature for 10 minutes and then packaged in a sealed pack. Raw noodles were stored at 12°C and bread was stored at 25°C, and changes were observed.

<Processed grain products with preservatives applied>

Microbiological characteristics of processed grain products

For microbial analysis of processed grain products, 10 g of sample and 100 mL of physiological saline were placed in a sterilized sample bag and homogenized at 100 rpm for 120 seconds using a Stomacher. 1 mL of the homogenized sample solution was diluted in steps of 10-1, and the total number of bacteria was determined by smearing on an Aerobic Count Plate and culturing at 37°C for 24 to 72 hours to determine the number of colonies. The total number of yeast molds was determined by smearing the Yeast and Mold Count plate and culturing it at 25°C for 72 to 120 hours to determine the number of colonies. The confirmed colony number was converted to the dilution factor and expressed as log cfu/g.

Statistical analysis

The significance of the measurement results was expressed as mean ± standard deviation using SPSS ver.20 software (IBM, Armonk, USA) . Statistical significance was tested using Duncan's multiple range test ( P ≤0.05).

Results and Discussion

Establishment of complex extract mixing ratio and fermentation conditions

By checking the mixing ratio and fermentation conditions, antibacterial activity was evaluated based on clear zone and MIC (minimum inhibitory concentration) values, and the best conditions were selected as shown in Table 5. The antibacterial activity of the complex extract before fermentation showed a relatively excellent effect when more than 50% of Cordyceps sinensis extract was added, and 5.7 mm (clear zone) and 1.5% (MIC vale) in the Cordyceps sinensis 75%/broccoli 25% complex extract (d). It was confirmed. The strains used for fermentation were Saccharomyces cerevisiae, Lactobacillus plantarum, and Weisella cibaria, and a mixed strain fermentation method was used. It was confirmed that the antibacterial activity increased significantly during fermentation using S/L (Saccharomyses cerevisiae & Lactobacillus plantarum) at all mixing ratios, and the antibacterial activity of 10.1 mm and 0.89% in the Cordyceps 75%/broccoli 25% composite extract (e) Activity was confirmed. In addition, the antibacterial activity of Cordyceps sinensis extract and broccoli extract was confirmed by fermentation before/after mixing, and it was confirmed that the antibacterial activity tended to increase when the mixed extract was fermented. The final selected conditions were extracts (g) fermented using Saccharomyses cerevisiae and Lactobacillus plantarum after mixing 75% Cordyceps sinensis extract/25% broccoli, which showed the best effect with a clear zone of 14.3 mm and MIC of 0.54%. The measurement effect is presented in Figure 3.

<Measurement of the effect of preservatives on processed grain products>

1) ++++ : clear zone more than 10mm & MIC value less than 10%, +++ : clear zone more

than 10mm, ++ : clear zone more than 7mm & MIC value less than 15% + : clear zone less than 7mm, - : not detected of antibacterial activity

2) S: Saccharomyces cerevisiae , L: Lactobacillus plantarum subsp . plantarum , W: Weissella cibaria

3) F-M: Mix after fermentation, M-F: Fermentation after mix

Referring to the DPPH radical scavenging activity results in Figure 2, both the positive control and the sample showed a concentration-dependent trend. In the positive control, the added concentration of extract required to scavenge 50% of DPPH radicals was 49.95 μg/mL and 58.83 μg/mL for L-ascorbic acid and BHA, respectively. Preservatives including green tea were found to be 592.52 μg/mL. The preservatives containing green tea were found to be lower than the positive control group, and were found to have greater antioxidant activity than 1,000 μg/mL, which corresponds to the value of the preservative before bioconversion.

Microbiological characteristics of processed grain products

Microbiological characteristics during storage of bread treated with green tea-containing preservatives are presented in Table 6.

<Microbiological characteristics of raw noodles treated with green tea-containing preservative (BGC)>

ac) Small letter refer difference significants( P 005) in the same row by storage duration through Duncan TEST

AD) Capital letter refer difference significants( P 005) in the same column by different concentration treatment through Duncan TEST

Referring to Table 6, the initial total bacteria and yeast/mold counts in the 90% EtOH treatment group were 2.01 log cfu/g and 1.34 log cfu/g, respectively, and on the 10th day of storage, they were 8.18 log cfu/g and 5.69 log cfu/g. increased rapidly. In the case of treatment with 0.1% preservative containing green tea, the total bacterial count was confirmed to be 3.71 log cfu/g on the 5th day of storage, which increased to 6.22 log cfu/g on the 10th day of storage. Additionally, it was confirmed that yeast/mold growth control showed no significant difference with 90% EtOH. In the case of treatment with 0.3% preservative containing green tea, the total bacterial count was 3.75 log cfu/g on the 10th day of storage, and the growth of yeast/mold was suppressed until the 5th day of storage, but it was 4.30 log on the 10th day of storage. It was confirmed that there was a rapid increase in cfu/g. In the case of the group treated with 0.5% preservative containing green tea, the total number of bacteria was 2.13 cfu/g and yeast/mold was 0.96 log cfu/g on the 10th day of storage, which confirmed that the growth of microorganisms was extremely suppressed. In particular, in the case of yeast/mold, it was confirmed that the number of microorganisms decreased from the initial 1.73 log cfu/g to 0.96 log cfu/g.

<Microbiological characteristics of bread treated with green tea-containing preservative (BCG)>

ac) Small letter refer difference significants( P 005) in the same row by storage duration through Duncan TEST

AC) Capital letter refer difference significants( P 005) in the same column by different concentration treatment through Duncan TEST

Table 7 shows the microbial characteristics of bread treated with preservatives containing green tea. In the case of 90% EtOH treatment, the initial total bacteria and yeast/mold counts were found to be 0.32 log cfu/g and 0.22 log cfu/g, respectively. However, on the 10th day of storage, it increased to 5.32 log cfu/g and 4.62 log cfu/g. In the case of the group treated with preservatives including green tea, the initial bacteria were similar to the 90% EtOH treatment group, and in the group treated with 0.3% preservative, the number of bacteria was 4.72 log cfu/g and 4.37 log cfu/g on the 10th day of storage, and in the group treated with 0.5% BGC, on the 10th day of storage. It was confirmed that the increase in microorganisms decreased to 3.01 log cfu/g and 3.37 log cfu/g. In particular, in the case of the preservative 0.8% treatment group, the total number of bacteria was 1.32 log cfu/g and the yeast/mold was 0.81 log cfu/g, and it was confirmed that the growth of microorganisms was inhibited by more than 99.9% compared to the 90% EtOH treatment group.

Referring to the above measurement results, the Cordyceps sinensis and broccoli extract used as a natural complex antibacterial agent were mixed in a ratio of 75:25, and Saccharomyces cerevisiae and Lactobacillus Plantarum subsp. It can be seen that bioconversion to a mixed strain of plantarum shows the highest antibacterial activity. The antibacterial activity of this preservative is Clear zone 14.3 mm and MIC value 0.54%. In addition, in the case of a preservative mixed with garlic and green tea extract, the antioxidant activity (RC50) required to scavenge 50% of DPPH radicals was 592.52 μg/mL, which was slightly higher than the positive control L-Ascorbic acid and BHA, but the antioxidant activity was reduced by biotransformation. It was found that activity could be improved. When treated with such a preservative, the growth of total bacteria and yeast/mold is suppressed. For example, in the case of raw noodles, growth inhibitory activity of microorganisms can be confirmed at over 0.3% of preservative. In particular, when treated with 0.5% BGC, growth inhibitory activity was shown for total bacteria 2.13 log cfu/g and yeast/mold 1.01 log cfu/g. gave. In the case of white bread, when treated with more than 0.5% of preservative, growth was suppressed with significant differences in total bacteria and yeast/mold. In particular, when treated with 0.8% preservative, total bacteria were 1.32 log cfu/g and yeast/mold was 0.81 log on the 10th day of storage. It was found to be inhibited by more than 99.9% in cfu/g compared to the 90% EtOH treatment group. Various ingredients can be mixed with Cordyceps sinensis extract and broccoli extract to bioconvert and be applied as a preservative for various types of grains or processed grain products, and are not limited to the presented examples.

The natural complex preservative for grains or processed grain products according to the present invention is composed of Cordyceps sinensis extract and broccoli extract and can have high antibacterial properties against various types of microorganisms or molds. The preservative according to the present invention can improve antioxidant activity by including garlic or green tea extract, thereby improving the effect of inhibiting the growth of bacteria, yeast, or mold. In the present invention, grains or processed grain products are treated with an antibacterial agent to maintain their original properties and at the same time extend their shelf life. The antibacterial agent according to the present invention can be applied to various foods.

Although the present invention has been described in detail above with reference to the presented embodiments, those skilled in the art will be able to make various variations and modifications without departing from the technical spirit of the present invention by referring to the presented embodiments. . The present invention is not limited by such variations and modifications, but is limited by the claims appended below.

P11: Cordyceps growing P12: Shredded broccoli
P21, P22: Ingredient extraction P30: Mixing
P40: Biological Conversion

Claims (2)

In preservatives for cereals or processed cereal products,
At least one ingredient selected from the group consisting of garlic extract, green tea extract, citron extract, fennel extract, dandelion extract, lactic acid bacteria culture, yeast culture, licorice extract, golden extract, and turmeric extract;
50 wt% or more of Cordyceps sinensis extract; and
Contains broccoli extract,
Broccoli is dried to have a moisture content of 10 wt% or less, then crushed to a size of 200 mesh or less, and the broccoli made into powder is crushed at an extraction temperature of 60 to 120 ℃ and stored for 6 to 24 hours. extracted,
A preservative characterized in that Cordyceps sinensis extract and broccoli extract are mixed and bio-converted by microorganisms, and then at least one ingredient is added. The preservative according to claim 1, wherein the microorganism is Saccharomyces sp. or Lactobacillus Plantarum.