KR0127899B1 - Salt tolerant microorgan and making method of doinjang - Google Patents

Abstract

Halotolerant variant sacchavomyces cereisiae ver. cerevisiae N-2 is KFCC-10823 cultivated at 18% salt concentration for producing the septic soybean paste. The above variant produces the soybean paste by using the above variant as a preservative.

Description

Manufacturing method of preservative-free doenjang using saline and mutant strains

1 is a general manufacturing process of doenjang.

Figure 2 is a manufacturing process diagram of the flame-resistant mutant strain for the preparation of the present invention preservative-free miso.

Figure 3 is a manufacturing process of preservative-free miso using salt-resistant mutants of the present invention.

Figure 4 is a graph showing the change in volume of miso wrapping paper by treatment in 20 ℃ temperature storage.

5 is a graph showing the change in volume of doenjang wrapping paper by treatment in 30 ℃ temperature storage.

Figure 6 is a graph showing the changes of yeast and mold water in doenjang by treatment at 20 ℃ temperature storage.

FIG. 7 is a graph showing changes in yeast and mold water in doenjang at 30 ° C temperature storage.

The present invention relates to a method for preparing an unpreservative miso, and in particular, to manufacture and manufacture a salt-resistant mutant that can produce a large amount of alcohol instead of the existing potassium sorbate or sorbic acid, which is used as a preservative of miso. The present invention relates to a method for preparing an antiseptic miso using an salt-resistant mutant strain.

Doenjang, which is currently manufactured by a doenjang manufacturer, is manufactured through a general manufacturing process as shown in FIG.

However, doenjang prepared in this way has a shelf life of 6 months or more depending on the type of product, but changes in consumers' perception about chemical synthetic preservatives such as potassium sorbate (potassium sorbata), which are used as preservatives for doenjang Because of the increasing interest in avoiding chemical preservatives among consumers, some manufacturers are using alcohol instead of synthetic preservatives, but this is also an increase in manufacturing cost due to artificial addition. There is an urgent need for manufacturing techniques that can be substituted for this.

The present invention has been invented in view of the above-described setting, using an antimicrobial agent to produce a large amount of ethanol having a very high storage effect of miso, using microorganisms related to manufacturing soy sauce instead of a chemical synthetic preservative such as potassium sorbate, and using an antiseptic agent using the same. Its purpose is to provide a method of making doenjang.

The method for producing miso of the present invention for achieving the above object is a large amount of ethanol, which is effective for storage properties, in place of a chemical synthetic preservative such as potassium sorbate, which is used as a preservative of miso in the fermentation process in the existing doenjang manufacturing method. It is characterized by adding a flame resistant strain.

Hereinafter, with reference to the accompanying drawings will be described in detail the manufacturing method of the present invention preservative-free miso as an example and a test example.

Example 1

Salt-resistant yeasts were bred in the process steps as shown in FIG. That is, in order to prepare a salt-resistant mutant for preparing an unpreservative miso, ATCC 42940 (saccharomyces cerevisiae ATCC 42940) was inoculated into 5 ml of YEPD medium having the chemical composition shown in Table 1 to produce an alcohol-producing strain saccharomyces cerevisiae. After shaking culture for 1 day, the cells were recovered by centrifugation (3,000 rpm, 5 minutes), and the mixture was clouded in 1 ml of 0.1 M phosphate buffer solution at pH 8.0. 0.15 ml of ethylmethanesulfonate was added thereto, followed by centrifugation (3,000 rpm, 5 minutes), followed by washing three times with 1 ml of mulberry water, and then, in a selective medium having the chemical composition shown in Table 2 below. Mutant strain N-2 with excellent flame resistance among the colonies (colony) shown in the selective medium after incubation for 5 days at ℃ (Saccharomyces cere visiae var, cererisiae N-2) Selected as.

TABLE 1 Chemical Composition of YEPD Medium

[Table 2] Chemical composition of selective medium

Example 2

Saccharomyces cere visiae var, Saccharomyces cere visiae var, deposited as the accession number (repair number) KFCC-10823 to the Korean Bacillus Association, the salt-resistant mutant injection method of the mutant strain N-2 selected in Example 1 cerevisiae N-2) was cultured in a medium for measuring alcohol production ability having the chemical composition shown in Table 3, and the alcohol production capacity and growth were compared with ATCC 42940 in the parent strain Sakaromyces cerevisiae and the results are shown in Table 4. Indicated.

[Table 3] Chemical composition of the medium for alcohol production capacity measurement

[Table 4] Growth and Alcohol Production Capacity of Improved Strains and Parents

As shown in Table 4, the parent strain was able to grow up to about 8% salt concentration, while the improved salt-resistant mutant strain was able to grow up to about 18% salt concentration, and the alcohol production was about 0.5% in 4 days culture. The mutant strain, an improved strain, was about 4%.

These results can be seen that the production of doenjang using improved strains considering that the salt concentration of the existing doenjang around 10%.

Example 3

Saccharomyces cerevisiae mutant obtained in Example 1 was prepared by using the N-2 (Saccharomyces cerevisiae var. Cerevisiae N-2) in the cerevisiae to prepare the doenjang as shown in FIG. After sterilization for 30 minutes at a temperature of 80 ℃ and filled in 100g each to a polyethylene film (PE film).

In order to investigate the shelf life of the filled product, the volume of the film packaging was investigated for 48 days in the incubator at 20 ° C. and 30 ° C., and the volume of the film was shown as 4 and 5, and the number of yeasts and molds was determined as shown in Table 5 below. Irradiation was carried out using a medium having a composition, and the results are shown as FIG. 6 and FIG.

[Table 5] Chemical Composition of Yeast and Mold Count Measurement Medium

Test Example 1. (Packaging Paper Volume Measurement)

Changes in the packaging volume of doenjang at 20 ° C. and 30 ° C. of Example 3 are shown in FIGS. 4 and 5, respectively. As is apparent from FIG. 4 and FIG. 5, only treatments without preservative added at 20 ℃ and 30 ℃ treatments showed a drastic change in the volume of the wrapping paper, and a slight change in treatment at 30 ℃ treatment in potassium sorbate added. However, in the treatment added with the improved strain (N-2 in the mutated cerevisiae to Saccharomyces cerevisiae), there was no significant change in packaging volume at the temperature treatment at 20 ℃ and 30 ℃.

Test Example 2 (Microbial Trend)

The changes of yeast and mold water in miso at 20 ° C. and 30 ° C. temperature treatment apparatus of Example 3 are the same as those in FIGS. 6 and 7, respectively. As shown in FIG. 6 and FIG. 7, the wrapping paper rapidly expanded after about 16 days in the treatment zone where the preservative was not added. On the other hand, the treatment with potassium sorbate was gradually decreased at 20 ℃, whereas it increased slightly after 8 days at 30 ℃.

In addition, the treatment group to which the improved strain (N-2 in mutated cerevisiae added to Saccharomyces cerevisiae) can see a sharp decrease in the number of yeasts and molds at 20 ℃ and 30 ℃, improving the shelf life of miso. It is believed to be able.

Test Example 3 (Expected Shelf Life)

A rare analysis of the packaging volume of the doenjang for the 0th order reaction to obtain the reaction equation and the correlation coefficient are shown in Table 6 below.

In Table 6, X is the storage days, Y is the packaging volume. Sample No. 1 is a preservative-free control sample No. 2 is treated with potassium sorbate added, and sample No. 3 is an antiseptic miso added with N-2 (mutant strain), an improved strain.

[Table 6] Reaction formula and correlation coefficient for packaging volume during miso storage

Using the reaction rate constant for the zero-order reaction, the Q ₁₀ value of the doenjang's packaging volume was determined using the reaction rate constant for the zero-order reaction. The volume of the complete expansion phenomenon was determined to be 90ml and the results of predicting the shelf life of doenjang using Q ₁₀ values are shown in Table 7 below.

[Table 7] Estimated shelf life at each temperature of doenjang

※ In the case of 30 ℃, the time until the packaging volume becomes 90ml, and room temperature is the value measured based on the annual average temperature of Korea (12.5 ℃).

As shown in Table 7, the additive-free group without preservatives was about 16.6 days at 12.5 ° C., Korea's annual average temperature, about 360.1 days when the potassium sorbate was added, and potassium sorbate was added when the modified strain was added. An estimated 803.7 days was increased by 2.2 times over the treated treatment.

As is clear from the above test examples, the improved strain had a much higher storage effect than potassium sorbate, a chemical synthetic preservative, and since it can replace the chemical synthetic preservative, it is possible to manufacture an antiseptic miso using an improved strain. have.

Claims (2)

Saccharomyces mutated cerevisiae N-2 (Saccharomyces, a saline-infected mutant, Saccharomyces cerevisiae, deposited as the accession number (repair number) KFCC-10823 to the Korean spawn association, which can be fed at a salt concentration of 18%, for the production of mubang-free miso). cererisiae var.cerevisiae N-2). Saccharomyces cerevisiae var. Cerevisiae N-2 (Saccharomyces cerevisiae var. Cerevisiae N-2) in the conventional method of preparing doenjang using potassium sorbate as a preservative, instead of potassium sorbate, a variant of saccharomyces cerevisiae var. Method for producing an antiseptic miso using salt-resistant mutant strains, characterized in that using as a preservative.