KR100232907B1 - Method for promoting growth of lactic acid bacteria by adding yam - Google Patents

Abstract

The present invention relates to a method for promoting the growth of lactic acid bacteria by the addition of horse mackerel. The combination of 0.5-2.0% of horse mackerel promotes the growth of lactic acid bacteria, as well as a unique component of the horse remaining in the fermented lactic acid bacteria, .

In addition, a new consumption law will be developed and the value of use will increase, which will also contribute to the increase in farm income.

Description

Method for promoting growth of lactic acid bacteria by addition of horse mackerel

The present invention relates to the production of fermented foods by a shortened fermentation process by adding specific amounts of natural margarines to promote the growth of lactic acid bacteria involved in yogurt production and kimchi production.

Lactic acid bacteria are staphylococci and bacillus. They are inactive, permeable to Gram-positive, mostly negative for catarrh. They are difficult to propagate in simple medium and grow only in medium containing multiple nutrients. In other words, various kinds of vitamins, minerals, and nucleic acids are required depending on the species, and amino acids and peptides as nitrogen sources and carbohydrates that can ferment as energy sources are needed. Thus, the efficacy of lactic acid bacteria depends greatly on the choice of medium. Studies have been conducted since 1924 when Mickle found that the addition of vegetable extract to the artificial medium of lactic acid bacteria promoted the growth of lactic acid bacteria. In particular, in the synthetic medium, Kuiken reported that tomato juice promotes the growth of lactic acid bacteria. Metcalf and Colio have studied lactic acid bacteria growth factors in cabbage, red pepper and spinach , Ganoderma lucidum and Ganoderma lucidum were investigated on the nutrition of lactic acid bacteria. The ginseng extract and Ganoderma lucidum extract showed distinct growth enhancement effect in each case of L. casei and S. thermophilus And the effect was also increased as the addition amount was increased. In addition, Yoon et al. Reported that the chlorella cytoplasm was found in Esophilus (L. acidophillus) on the growth of rice. On the other hand, it was reported that the inhibition effect of lactic acid bacteria on the growth of lactic acid bacterium by addition of corn, garlic, red pepper and ginger was higher than that of the extract, while the growth inhibition effect and acidity were decreased as the concentration of extract was higher .

Yam is a perennial vine plant belonging to the genus Dioscorea, and the inside of the circumferential bloom mass used for edible and medicinal purposes is milky white or yellowish brown and contains starch, glucose, fructose, sucrose, mucin, Amine, tyrosine, glutamic acid, diastase, and vitamins (A, B, C).

It is also known as an aphrodisiac plant. It is also known as a herb medicine in the oriental medicine. It is also used as a medicine to be followed by licorice, and recently it has been added to health food and its usage has been increasing. To produce and sell 14 types of processed foods such as powder, tea, and beverages. It is also known as one of the most commonly used starchy corroded foods in Japan. In recent years, it has been known that in the United States, baking, biscuits, sauces, soup, etc. for children and sick people As a raw material. As such, there is an increasing demand for special applications using hemp.

The present invention aims at promoting the growth of lactic acid bacteria by adding a certain amount of raw or dry germ during fermentation of lactic acid bacteria, thereby shortening fermentation time of foods using lactic acid bacteria such as yogurt and producing yogurt containing useful ingredients of hemp.

However, research on the use of hemp in fermented foods is extremely limited. The pH, total acidity and number of viable cells were measured and evaluated for the effect of lactic acid bacteria on the growth and fermentation of lactic acid bacteria by adding lactic acid bacteria to the lactic acid bacteria medium. After the yogurt was produced using the most effective lactic acid bacteria, The addition of fermented foods to lactic acid bacteria was evaluated by sensory evaluation. Through this study, it can be expected that the nutritional improvement and the shortening of the manufacturing process of the fermented food added with margarine can be expected by using marl as a growth promoting substance of lactic acid bacteria in the manufacture of fermented food (yogurt, kimchi) , Thereby contributing to the local economy as well as the food processing aspect.

FIG. 1 is a production process diagram of the present invention. FIG.

Figs. 2 (a), 2 (b) and 2 (c) are graphs showing the pH, acidity and viable cell count according to the concentration of the present invention.

FIGS. 3 (a), 3 (b) and 3 (c) are graphs showing the pH, acidity and viable cell counts of the present invention.

FIGS. 4 (a), 4 (b) and 4 (c) are measurement bar graphs showing the pH, acidity and viable cell count according to the addition of the heat-treatment step of the present invention.

5 (a), 5 (b) and 5 (c) are measurement bar graphs of pH, acidity and viable cell counts according to the concentrations of raw horses and horses in the present invention.

Figure 6 (a), (b), and (c) show comparative bar graphs of pH, acidity and viable cell counts of fermented milk according to the presence or absence of the addition of horse mackerel.

7 (a), (b), and (c) are graphs showing the comparative measurement of pH, acidity, and number of viable cells according to the morphology of yogurt fermentation by L. casei.

The present invention relates to a method for the production of yoghurt, comprising the step of inoculating a starter cultivated in a skim milk or MRS medium of a lactic acid bacterium into milk and adding 0.5-2.0% of dry matter or raw horses on a dry weight basis at 37 캜 or 30 캜, The present invention relates to a method for manufacturing a yogurt.

Hereinafter, the present invention will be described with reference to specific experimental examples.

□ Materials and Methods

1. Sample

The yam was purchased from Pyongyang Nonghyup cooperative association in Yeongju, Gyeongsangbuk - do, while the raw yam was stored in a refrigerator at 4 ℃ and the dry yam was stored at room temperature. The general components of the maize are listed in Table 1.

The water content of the raw horses and the raw horses used in the experiment were 10% and 75%, respectively.

In addition to the common components, there are also 15 kinds of amino acids including saponin, tannin, polyphenol, allantoin and aminobutyric acid, saccharides such as arabinose, mannose and rhamnose, and uronic acid, keridonic acid and nitosterol, Such as phosphorus, calcium, sodium, iron, potassium, magnesium, manganese, zinc, selenium and the like.

Specific examples of the production method of the present invention are as follows.

[Example]

Yeongju, Pyongyang, Gyeongsangbuk - do prepared seeds (75% moisture content) and gumma (10% moisture content) purchased from the Nonghyup.

1) Starter manufacturing

10 ml of skim milk (Seoul milk) was added to each 15 ml test tube, and the strain was cultured at 121 ° C for 15 minutes in an autoclave (Sanyo electric Co.) sterilized medium. In order to maintain the viability of the microorganism, 2 times subculture was used as a mother starter. The incubated starters were refrigerated (4 ℃).

L. casei YIT9018 and L. acidophillus were obtained from Leuc. mesenteroides were cultured in 10 ml MRS medium as the mother starter.

The strains used were Lactobacillus casei YIT9018 (Korean Yakult), Lactobacillus acidophilus CH5, Lactobacillus mesenteroides ATCC 27258, and Lactobacillus kefir ATCC 35411 from Korean Microorganism Conservation Center (KCCM).

2) Preparation of fermentation medium

Fig. 1 schematically shows a method for producing yogurt including the kind of yogurt added to the pasteur milk and the treatment conditions. (W / V) The solid content is measured on the basis of the content of the w / v, and the resultant is homogenized twice in the milk to make 200 ml (in this case, the solid content is calculated on the basis of the water content of 75% .)

The 200 ml sample thus prepared is placed in an Erlenmeyer flask and heat-treated at 85 ° C for 30 minutes in a shaking water bath. After cooling, 0.5% starter was inoculated and fermented in an incubator at 37 ° C for 19 hours and then stored at 4 ° C in a refrigerator.

As another example of the present invention, the raw horses were measured according to%, then homogenized by dividing twice in distilled water, passed through gauze, and then centrifuged at 4800 rpm for 20 minutes to remove residual starch in the filtrate. The supernatant Solution).

This impregnated supernatant was added to the cooled medium at a level of 0.03% or 0.08% after heat treatment of only 200 ml of milk and inoculation of the starter was carried out in the same manner as in the first step.

As another example of the present invention, it is similar to the first example, except that only 1% of raw milk was added after heat treatment of milk.

2. pH and total acidity measurement

The aged yogurt in the refrigerator was homogenized at 5,000 rpm for 20 seconds and analyzed. The pH was measured using a pH meter (88 ATC, Seimeung Scientific). The total acidity was determined by diluting 10 ml of yogurt with 10 ml of distilled water and adding 4-5 drops of 0.1% - phenolphthalein solution And titrated with 0.1 N NaOH solution.

The end point of titration was calculated from the amount of 0.1N NaOH consumed in the titration (ml) based on the pH 8.9 when it became subtle red.

<Calculation>

* F = Potency of 0.1N NaOH solution (1.02)

* a = 0.1 N NaOH solution (ml)

* Specific gravity of milk = 1.030

3. Measurement of viable cell count

The viable cell count was evaluated by diluting the fermentation medium stored at 4 ° C with sterilized distilled water. The curd of the fermentation medium was homogenized at 5000 rpm for 20 seconds, then 1 ml was taken and diluted with 9 ml of sterilized distilled water. From this, 0.1 ml of the sample solution was continuously diluted to 10 ⁷ with 0.9 ml of distilled water. 40ul of the final diluted samples (10 ⁷ ) were inoculated into MRS agar plate (MRS 3% (24g) / Agr 1.5% (12g) 800ml DW) and incubated at 37 ° C for 3 days. Was determined.

4. Curd formation and whey separation measurement

Curd formation of yogurt was carried out by visual observation or photographing after storing 20 ml milk medium cultured in a 37 ° C incubator for 19 hours in a refrigerator at 4 ° C.

5. Sensory evaluation (Ranking test)

Department of Food Science and Engineering, Keimyung University, and a panel of 10 male and female students. The evaluation criteria were evaluated by evaluating the taste and flavor and then ranking the products as the overall evaluation score. The analysis of the evaluation data was carried out by analysis of variance.

1. Effect on the Growth of L. casei YIT9018 by Addition

end. Effect of concentration on hemoglobin concentration

After the addition of 0.5%, 1%, 2%, and 3% of gamma and raw maize to the growth of L. casei YIT9018, the pH, total acidity, viable cell count and sensory evaluation of fermentation broth Respectively. As shown in Fig. 2 (a), (b), and (c), the marginally added milk medium showed significant effects on pH, total acidity and growth of bacteria after fermentation for 19 hours I could see. In particular, differences in the growth of lactic acid bacteria were observed depending on the concentration of added rice.

The change of pH was lowered as the amount of starch added was higher than that of milk without milk. In Figure 2 (a), the milk samples had the highest pH of 5.65 and the pH was lowered to 4.39 when the raw yam was added to the 2% level, and the pH value was almost constant at the 3% addition. The pH was decreased to 3% even with the addition of day yam.

Changes in total acidity showed a tendency to increase with the amount of hemp than with the acidity of milk samples. . The acidity of the milk samples was the lowest at 0.375% in the second step (b), and the raw horses continued to increase to 0.87% until the addition of 3%, and continued to increase to 0.89% . This phenomenon almost coincided with the pH change of yogurt.

When the number of viable cells was changed, it was found that the number of bacillus was increased by the addition of the mash medium. Whereas the number of bacteria in the milk medium inde 1.68 × 10 ⁹ cell / ㎖ case of adding 1% level of geonma is 8.5 × 10 ⁹ cell / ㎖, and saengma added and has at least three to five times with 5.9 × 10 ⁹ cell / ㎖ The number of bacteria was increased. On the other hand, as shown in Fig. 2 (c), a decrease in the viable cell count was observed at the addition of 2% or more of horseradish peroxidase. From the quality evaluation of these fermented products, there was no difference in taste and flavor of the milk fermented product until 1% addition of the fermented product.

However, at the level of 2% or more, the sensory evaluation was worse than that of milk fermented product. Therefore, the concentration of hemicellulase in the evaluation of the efficacy of lactic acid bacteria on the addition of hemp was set at a maximum of 1% level.

I. Effect of incubation time

3 (a), (b), and (c) show the results of measurement of changes in pH, total acidity and viable cell count according to the incubation time after adding 1% level of hemp. In the fermentation process, yogurt was incubated for 19 hours because the yogurt showed complete curd formation after 19 hours. Samples were taken at 7 hours, 14 hours, and 19 hours.

Changes in pH with incubation time decreased with time, and the addition of hemp was lower than that of milk medium. As shown in Fig. 3 (a), pH changes slightly at 7 hours, while the pH at 19 hours was 5.57 for milk medium, 4.58 for gymnospermum and 4.43 for raw horses. .

As shown in Figure 3 (b), there was no significant difference in the pH at 7 hours, but at 19 hours milk The medium was 0.43% compared to 0.79%, while the growth rate was 0.84%.

In addition, the number of viable cells increased with increasing fermentation time. In (c) of Fig. 3, the change of bacterial number with the addition of 1% Yogurt fermented with milk only showed 3.0 × 10 ⁹ cells / ㎖ viable cells after 19 hours of fermentation, while the number of viable cells was 1.2 × 10 ¹⁰ cells / ㎖ at the time of addition of raw cells and 1.0 × 10 ¹⁰ cells / ㎖, both ginseng and raw maize showed lactic acid bacterial growth promoting effect. In particular, to evaluate the difference between the effects of raw horses and horses, the fermentation of milk with 1% level of margarine was repeated 5 times. The total acidity was 0.773 for raw horses and 0.748 for dry horses. The number of viable cells was 8.3 × 10 ⁹ and 7.7 × 10 ⁹ , respectively. From these mean values, it was found that the growth promoting effect of L. casei was better than that of the raw horses.

As shown in FIG. 7, whey separation of yogurt was remarkably reduced when 1% of heat-treated horseradish and raw horses were added at a level of curd formation and whey separation according to the addition of hemp, Showed better curd formation and less whey separation.

The whey breakdown of yogurt can be influenced by the initial number of inoculated bacteria and their acid production rate. However, in the case of added raw horses, inhibition of whey breakdown by existing slime is predicted.

2. Effects on the growth of L. casei YIT9018 according to the addition of hemp

end. Heat treatment effect

In order to compare the effects of heat - treated and un - heat - treated raw rice on the growth of L. casei YIT9018, the samples were analyzed by the methods (1) and (3) in Fig.

4 (a), (b) and (c) show that the pH was slightly decreased and the total acidity was two times higher than that of the untreated yogurt Of the total. The number of viable cell counts was the highest at 1.3 × 10 ¹⁰ when the untreated raw horses were used, and the viable cell count was 1.0 × 10 ¹⁰ at the heat treated 1% raw horses. The number of viable cells of the yogurt using only milk was 2.2 × 10 ¹⁰ 5 times more. Therefore, when adding 1% of raw milk of yogurt to milk, it was very effective in pH, total acidity and viable count regardless of heat treatment.

However, the heat treatment at 85 占 폚 for 30 minutes resulted in the curd being completely formed, and the whey separation was small. It is thought that the addition of 1% of raw horses resulted in proteolytic degradation of proteases due to the precipitation of casein curd as well as severe whey segregation in the absence of heat treatment. This was similar to the addition of the coenzyme, Lenin, for the production of Cuzcurd. Therefore, in order to add 1% level of raw horses for the production of solid yogurt, it is absolutely necessary to perform heat treatment so as to minimize whey separation.

I. Effects on the L. casei YIT9018 in the Stomach of Rats

The preparation process of the viscous solution obtained by removing the starch and insoluble materials from the raw horses through filtration and centrifugation processes is shown in ② of Fig. 1 and the effect of these on the growth of L. casei YIT9018 was investigated. Compared with fermentation after heat treatment (at 85 ℃ for 30 min) with 1% level of tuna (raw or dried) with milk added with 0.08% (W / V) of heat - treated viscous solution, pH, total acidity , And the number of viable cells was similar. In Fig. 5 (c), the number of viable cells of the lactic acid bacteria was 2 × 10 ⁹ cells / ml for the milk medium, and 1.1 × 10 ¹⁰ cells / ml for the 1 ml of 8.3 × 10 ⁹ cells / , And 0.08% of raw viscous viscous substance was added as 1.1 × 10 ¹⁰ cells / ㎖. Therefore, the addition of a small amount (0.03 - 0.08%) of the viscous liquid of the raw horses was almost the same or better than the case of adding 1% level of hemp treated by heat treatment. Therefore, it was evaluated that adding a small amount of viscous liquid of raw horses rather than adding heat - treated horses would further contribute to the growth of lactic acid bacteria. However, the addition of 1% or more non-heat-treated raw rice had a bad influence on the quality of yogurt by enzymatic action.

3. Growth promotion effect of various lactic acid bacteria by addition of 1%

Lactic acid bacteria require different nutrient requirements depending on the bacteria. The growth promoting effect of Lactobacillus acidophilus was compared with that of milk medium through five repeated experiments. Overall, raw horses showed good results in pH, total acidity and viable cell counts rather than raw horses.

However, as shown in Fig. 7, the raw horses have no storage stability than the horseshoe horses, and seasonal problems arise in securing raw materials. Therefore, the growth promoting effect of lactic acid bacteria involved in the fermentation of fermented milk and kimchi fermentation was compared by using gamma which is advantageous in terms of storage and raw material securing. As shown in Figure 6 (a), (b), and (c), the growth promoting effect of 1% PH 4.48, pH 4.41 and total acidity were 0.94% and 0.95%, respectively. The viable cell counts of L. casei YIT9018 and L. acidophilus were 6.9 × 10 ⁹ cells / ㎖, 1.5 × 10 ⁹ cells / mesenteroides ATCC27258 was 1.6 × 10 ⁹ and showed the highest viable count in L. casei YIT9018. Especially, the increase rate of viable cell counts by the addition of hematocrit on the basis of milk medium was higher than that of L. casei YIT9018, L. acidophillus, Leuc. Mesenteroides showed 3 times, 2.5 times and 1.7 times, respectively. Thus, L. casei YIT9018 and L. acidophillus, Leuc. and mesenteroides in the order of growth of lactic acid bacteria.

4. Sensory test comparison

Yoghurt samples for sensory evaluation were fermented with milk, raw silk, raw horseradish, and yamma, and then aged yogurt samples were used.

The results of the rank test are as follows.

A. Milk contains 0.03% of vitreous viscous liquid.

N. Added 1% heat treated milk to milk.

C. Milk contains 0.08% of vitreous viscous liquid.

D. 1% added with heat-treated raw milk in milk.

ㅁ. milk

The sensory evaluation showed that yogurt was better in milk than in milk. The addition of 0.03% of raw mucilage solution showed the best result. The addition of 0.08% of viscous liquid resulted in good evaluation. When the milk was fermented for only 19 hours, no complete curd formation was made, and the sour taste such as the unique yogurt was small and the taste of the milk itself was strong. On the other hand, when the horse was added, it was considered that the sensory evaluation showed high value because of the soft texture due to the sour taste and the tannery of the yogurt, as well as the production of the perfect cupper.

Growth promoting effect of lactic acid bacteria on the addition of raw horseradish and horseradish were evaluated by pH, total acidity, viable cell count and sensory evaluation of fermented yogurt.

The growth of lactic acid bacteria (L. casei YIT9018) was remarkably increased when the raw horseradish and horseradish were added to the oil seedlings and it was more effective in pH and total acidity including viable cell count under the same fermentation condition. The marginalized medium exhibited complete curd formation by fermentation at 37 ° C for 19 hours, whereas milk showed incomplete curd formation.

At the concentration of 1% or more, the total acidity and the pH and number of viable cells were decreased, and especially, the sensory properties of the fermented product were adversely affected.

When added to the heat - treated milk medium, also the pH, total acidity and viable cell count were remarkable. The addition of 0.08% level of vitamins was almost the same as that of 1% level of hemicellulase. The sensory evaluation of the fermented product was very good. Therefore, it is considered more effective to use the viscous substance in which starch and insoluble matter are removed from the raw horses rather than the heat treated 1% level of horses. However, pH, total acidity, and viable cell counts were similar to those of heat - treated maize when 1% non - heat - treated raw rice was added.

L. acidophillus other than L. casei YIT9018, and Leuc. mesenteroides strains were investigated. The growth rate of viable cell counts by the addition of hematocrit on the basis of milk medium was investigated in L. casei YIT9018, L. acidophillus, Leuc. mesenteroides were 3 times, 2.5 times and 1.7 times, respectively.

As described above, according to the method of promoting the growth of lactic acid bacteria by the addition of the present invention, the growth of the lactic acid bacteria can be promoted by the addition of hemp, thereby shortening the manufacturing time of the fermented food, and the unique lactic acid bacteria fermented food And the nutritional improvement is greatly improved, so that the consumption preference of the fermented food is improved, and the farm income can be increased.

Claims (3)

A starter cultured in a medium such as skim milk is inoculated into milk and fermented sufficiently at 37 캜. The yogurt is produced by adding 0.5 to 2.0% by weight of hemp to the milk medium. A method of promoting the growth of lactic acid bacteria by. The method of promoting growth of lactic acid bacteria according to claim 1, wherein 1% of the heat-treated horses is homogenized twice in the milk culture medium, and then the mixture is heat-treated for 30 minutes in a water bath at 85 캜. A starter cultured in a medium such as skim milk is inoculated into milk to ferment lactic acid bacteria in a milk, and the yogurt is sufficiently fermented at 37 캜. The yoghurt having a moisture content of 75% is homogenized twice in distilled water and then passed through gauze. 0.03% to 0.08% (W / V, based on the dry matter content) of the supernatant obtained after the separation was added to the milk medium without heat treatment.