KR19980031143A - Growth Promotion Method of Lactic Acid Bacteria by Hemp Addition - Google Patents

Abstract

Lactic acid bacteria are used to produce various lactic acid bacteria beverages. In this case, if the lactic acid bacteria proliferate per unit time, the efficiency is increased and the productivity is increased.

In the present invention, the growth of lactic acid bacteria is promoted by mixing 0.5-2.0% using hemp produced in Korea, as well as the unique components of the hemp remain in the lactic acid bacteria fermented food, thereby improving nutritionally. In addition, new consumption laws for villages are developed and the value of their use increases, contributing to the increase of farm household income.

Description

Growth Promotion Method of Lactic Acid Bacteria by Hemp Addition

The present invention relates to the production of fermented foods by a shortened fermentation process by promoting the growth of lactic acid bacteria related to yogurt production and kimchi production by adding a specific amount of natural hemp.

Lactobacillus is a bacterium or bacillus that is inactive and breathable gram-like, mostly catarase-negative, difficult to grow on simple media, and grows only on media with complex nutrients. In other words, depending on the species, various vitamins, minerals and nucleic acids are required, and amino acids or peptides are needed as nitrogen sources and carbohydrates can be fermented as energy sources. Therefore, the fermentation ability of lactic acid bacteria depends a lot on the selection of the medium, and researches on this have been conducted since 1924 when Mickle added vegetable extracts to artificial media of lactic acid bacteria and thereby promoted the growth of lactic acid bacteria. In particular, Kuiken reported that tomato juice promoted lactic acid bacteria in synthetic media, Metcalf and Colio studied lactic acid bacteria growth factors in cabbage, red pepper, and spinach. The effects of phosphoric acid and ganoderma lucidum extract on the growth of lactic acid bacteria were studied. Ginseng extract and ganoderma lucidum extract showed distinct growth promoting effects in case of L. Casei and S. thermophilus. The effect was also reported to increase as the increase, and evenly, Chlorera cytoplasm of Ecido pyrus (L. acidophil) lus) has been reported to improve growth. On the other hand. Eugene reported the growth inhibition effect of lactic acid bacteria growth effect of the addition of leek, garlic, red pepper and ginger. Chung et al. Reported that the higher the concentration of the extract, the higher the concentration of the extract, the lower the acidity.

Yam is a perennial vine herb belonging to the family Dioscorea, and the circumferential enlarged tubers of circumferential tubers used for edible or medicinal color are opaque or yellowish brown. It contains ingredients such as cosamine, tyrosine, glutamic acid, diastases, and vitamins (A, B, C). It is a plant known as a rejuvenating medicine since ancient times. It is also called as a medicine for herbal medicine, and it is used as a medicine that goes into licorice after licorice. Recently, it is added to health food and its usage is increasing. Established a factory to produce and sell 14 kinds of hemp processed foods including powders, teas and beverages. In addition, hemp is known as one of the most edible starch corrosion in Japan, and recently, in the United States, it has a high digestibility using horse starch, and is a baking source, biscuits, sauces, soups, etc. for children and the sick as a calorie source or health food. It is used as a raw material. As such, demand for special applications using hemp is increasing.

The present invention aims to reduce the fermentation time of lactic acid bacteria using foods such as yogurt and yogurt containing useful ingredients of hemp by promoting the growth of lactic acid bacteria by mixing a certain amount of raw horse and dry horse during fermentation of lactic acid bacteria.

Although hemp is used for various purposes, research on the use of hemp in fermented foods is very insignificant. Therefore, pH, total acidity, and viable cell count were evaluated for the effects on the growth and fermentation of lactic acid bacteria by adding hemp to milk, the lactic acid bacterium medium, and the functionalities of these products after the production of yogurt using the most effective lactic acid bacteria. Through the test, the effect of the addition on the lactic acid bacteria fermented food was evaluated. Through this research, the nutritional improvement and the shortening of the manufacturing process of fermented foods added with hemp can be expected by using it as a growth-promoting substance of lactic acid bacteria in the production of Marl fermented foods (yogurt and kimchi), one of the special products in Gyeongbuk region. It is believed that this will help the local economy as well as food processing.

1 is a production process diagram of the present invention.

2 is a (a) (b) (c) is a graph of the pH, acidity, viable count according to the concentration of the present invention.

3 is a (a) (b) (c) is a graph of the measurement of pH, acidity, viable cell number by time of the present invention.

4 is a (a) (b) (c) is a bar graph of pH, acidity, viable count according to the addition of the heat treatment hemp of the present invention.

5 is a (a) (b) (c) is a bar graph for measuring the pH, acidity, viable count according to the concentration of the horseshoe dry horse of the present invention.

Figure 6 is a (a) (b) (c) is a bar graph of the comparison of pH, acidity, viable count of fermented milk according to the addition of dry horse.

Figure 7 is a (a) (b) (c) is a graph of pH, acidity, viable cell comparison according to the shape of hemp when yogurt fermentation by L.casei bacteria.

The present invention is inoculated with milk starter cultured in a medium of MRS such as skim milk, such as skim milk in the yogurt fermentation experiment time at 37 ℃ or 30 ℃ depending on the type of blended dry horse and raw horse 0.5 to 2.0% based on the dry weight It is related with the provision of the yoghurt manufacturing method characterized by the above-mentioned. It will be described through specific experimental examples below.

Experimental Materials and Methods

1. Sample

Ma (yam) was purchased from Nonghyup cooperative in Yeongju, Gyeongbuk, raw yam (4) in the refrigerator and dry yam (dry yam) was stored at room temperature. General components of hemp are shown in Table 1.

Table 1. Composition analysis table of hemp (Unit:%, dry base)

The moisture content of dry horse and raw horse used in the experiment was 10% and 75%, respectively.

In addition to general ingredients, hemp contains 15 kinds of amino acids including saponins, tannins, polyphenols, allantoin, aminobutyric acid, sugars such as arabinose, mannose, and rhamnose, as well as euonic acid, keridone acid, and nitosterol. Phosphorus, calcium, sodium, iron, potassium, magnesium, manganese, zinc, selenium and the like.

Specific examples according to the manufacturing method of the present invention are as follows.

Example 1

Pyeongju, Yeongju, Gyeongbuk, prepared raw horses (95% water content) and dry horses (10% water content) purchased from the Agricultural Cooperatives.

1) Starter Manufacturing

10 ml of skim milk (Skim milk, Seoul milk) solution was dispensed into 10 ml each of 15 ml test tubes, and the strains were cultured in a medium sterilized in Autoclave (Sanyo electric Co.) for 15 minutes at 121 ° C. In order to maintain the vitality of bacteria, two passages were used as a motherstarter, and the incubators were refrigerated (4 ° C). L. casei YIT9081 and L. acidophillus were isolated from Leuc. Mesenteroides were cultured in 10 ml MRS medium and used as mother starters.

2) Fermented medium production

The yogurt production method including the type of hemp and the processing conditions added to the milk medium (pasteur milk) is collectively shown in FIG. Measure the solid content on the basis of% (W / V) building content and add to milk twice to homogenize to make 200ml. (The solid content was calculated based on water content of 10% dry horses and 75% dry horses. .)

Put the 200ml sample in the Erlenmeyer flask and heat it for 30 minutes at 85 ℃ in shaking water bath. After cooling, 0.5% starter was inoculated and fermented in a 37 ° C. incubator for 19 hours and then refrigerated at 4 ° C.

As another example of the present invention, after measuring the raw horse according to%, homogenized by dividing twice in distilled water and passed through gauze, centrifuged at 4800 rpm for 20 minutes to remove the remaining starch in the filtrate (supernatant) Solution). This viscous supernatant was heat-treated only with 200 ml of milk, and then added to the cooled medium at a level of 0.03% or 0.08%, followed by inoculation of the starter, followed by the same method as the first.

Another example of the present invention is similar to the first example, but only 1% of raw horse meat was added after only heat treatment of milk.

3. pH and total acidity measurement

Yogurt aged in the refrigerator was analyzed after homogenizing at 5,000 rpm for 20 seconds. The pH was measured using a pH meter (88 ATC, Seolim Science) .Total acidity was taken by taking 10 ml of yoghurt sample and diluting it with 10 ml of distilled water, and then 4-5 drops of 0.1% -phenolphthalein solution. 0.1N NaOH solution was added thereto.

The end point of the titration was calculated from the amount of 0.1 N-NaOH consumed (mL) in the titration based on pH 8.9 when it was scarlet.

Calculation

F = titer of 0.1N-NaOH solution (1.02)

A = titration (ml) of 0.1N-NaOH solution

Specific gravity of milk = 1.030

4. Viable cell count

The viable cell count was evaluated by diluting the fermentation broth stored at 4 ° C. with sterile distilled water. The curd of the fermentation broth was homogenized for 20 seconds at 5000 rpm, then 1 ml was taken and diluted in 9 ml sterilized distilled water. 0.1 ml sample solution was taken from this and diluted 10 ⁷ successively using 0.9 ml distilled water. Take 40 ul from the final diluted samples (10 ⁷ ) and inoculate three in MRS agar plate (MRS 3% (24g) / Agar 1.5% (12g) / 800ml DW) and incubate for 3 days at 37 ° C. The mean value of the numbers was determined.

5. Curd formation and whey separation measurements

The curd formation ability of yogurt was observed or photographed visually after storing the 20 ml milk medium incubated for 19 hours in a 37 ℃ incubator in a 4 ℃ refrigerator.

6. Sensory test (ranking test)

The sensory test was conducted by a panel agent consisting of 10 graduates from the Department of Food Science and Technology and Keimyung University. Evaluation criteria were evaluated by evaluating taste and aroma, and then ranking products as overall evaluation scores. Analysis of the evaluation data was performed by analysis of variance.

1, Effect of Hemp Addition on Growth of L. casei YIT9018

end. Effect of concentration on hemp

The effect of horses on the growth of L. casei YIT9018 was examined by adding the concentrations of dry horses and horses at levels of 0.5%, 1%, 2%, and 3%, followed by pH, total acidity, viable cell count and sensory test of fermentation broth. It was. As shown in FIGS. 2-1, 2, and 3, the milk medium containing the hemp was fermented for 19 hours compared to the non-added milk medium, and showed a remarkable effect on pH, total acidity, and growth of bacteria. In particular, the difference on the growth of lactic acid bacteria was also observed depending on the concentration of hemp added.

The pH change was lower as the amount of hemp added increased compared to the milk without hemp. In FIG. 2-1, the milk sample had the highest pH of 5.65, and when the raw yam was added to 2% level, the pH was lowered to 4.39, and the pH value was almost constant when 3% was added. When dry yam was added, the pH tended to decrease by 3%. The change in total acidity tended to increase with the amount of hemp added rather than the acidity of milk samples. In FIG. 2-2, the acidity of the milk sample was the lowest at 0.375%, the raw horse continued to increase to 0.87% up to the 3% addition level, and the dry horse continued to increase to 0.89% up to the 2% addition level and then 0.80% at the 3% addition level. Fell slightly. This phenomenon was almost consistent with the pH change of yogurt.

The number of viable cells showed that the number of bacteria increased when hemp was added rather than milk medium. The number of bacteria in milk medium is 1.68 × 10 ⁹ cell / ml, whereas when 1% dry horse is added, it is 8.5 × 10 ⁹ cell / ml and when raw horse is added, it is 5.9 × 10 ⁹ cell / ml. Was increased (Fig. 2 (c)). On the other hand, when more than 2% of hemp was added, a decrease in viable cell count was observed. In addition, there was no difference in taste and flavor with milk fermented products until the 1% addition level of hemp in the sensory evaluation of these fermented products.

However, at the level of 2% or higher, the sensory evaluation was worse than that of the fermented milk product. Therefore, when assessing the growth effect of lactic acid bacteria by the addition of hemp, the concentration of hemp was determined based on the highest 1% level.

I. Effect of incubation time

3- (a) (b) (c) is the result of measuring the change of pH, total acidity and viable cell number according to incubation time after adding 1% hemp. In the fermentation process, yoghurt showed full curd formation after 19 hours, and thus 19 hours were determined as the culture standard. Samples were taken at 7 hours, 14 hours and 19 hours.

The pH change with the incubation time was lowered by the difference between the addition of hemp and the milk medium. 3- (a) showed a slight difference in pH change at 7 hours, but pH at 19 hours was 5.57 in milk medium, 4.58 in dry horse and 4.43 in raw horse. Indicated.

The change in total acidity showed that the acidity was significantly increased compared to the milk medium at the time of the addition of hemp. In FIG. 3-2, the milk medium at 19 hours was not changed as much as in the pH change. In contrast to 0.43%, dry horses were 0.79% and raw horses were 0.84%, which was about twice as high.

In addition, when the fermentation time was increased, the number of cells was increased, and the number of bacteria was increased. In FIG. Yogurt fermented only with milk showed a viable cell count of 3.0 × 10 ⁹ cells / ml after 19 hours of fermentation, whereas the number of bacteria was 1.2 × 10 ¹⁰ cell / ml when fresh horse was added and 1.0 × 10 ¹⁰ cell / ml when dry horse was added. As a result, both dry horses and live horses showed the effect of lactic acid bacteria growth. In particular, to evaluate the difference between the effects of raw and dry horses, fermentation experiments were repeated five times with milk added to the 1% level. In the case of total acidity, live horses were 0.773 and dry horses were 0.748, and live bacteria and dry horses were 8.3 × 10 ⁹ and 7.7 × 10 ⁹ , respectively. From these mean values, it was found that raw horses were more effective in promoting lactic acid bacteria (L, casei) growth than dry horses.

(Figure 7)

In the comparison of curd formation and whey separation with the addition of hemp, the whey separation of yogurt was significantly reduced when heat-dried dry horse and raw horse were added at 1% level. Less separation

The whey separation phenomena of yogurt may depend on the number of inoculated bacteria and their acid production rate. However, in the case of the added horseshoe, the whey separation phenomena due to the viscous radish present is expected.

2. Effect of Hemp Addition on Growth of L. casei YIT9018

end. Heat treatment effect of hemp

In order to compare the effects on the growth of L. casei YIT9018 during the addition of heat-treated and unheated horses, samples were prepared and analyzed in 1 and 3.

4-1, 2, and 3 showed a slight decrease in pH than the heat treatment at the 1% level of raw horses, and the total acidity was about 2 times higher than in yogurt without hemp. Indicated. Showed the highest value as 1.3 × ¹⁰ 10 in the case of using the In heat treatment anhan saengma the number of viable cells, in the case of the heat-treated 1% saengma can live yoghurt bacteria number using a 1.0 × ¹⁰ 10, only milk 2.2 × 10 ⁹ It was about 5 times higher than that. Therefore, when yogurt was added to milk at 1% level, it was very effective in pH, total acidity and viable cell number regardless of heat treatment. However, heat treatment at 85 ° C. for 30 minutes resulted in complete curd formation and low whey separation. The precipitation of casein curd together with severe whey separation in the case of untreated heat when 1% of raw horse was added was thought to be due to proteolytic degradation of protease among enzymes present in raw horse. This is similar to the addition of the coagulant rennin for the production of cheese curd. Therefore, it is absolutely necessary to heat treatment to minimize whey separation in order to add 1% level of raw yam to produce a solid yogurt.

I. Effect of Raw Viscous Matter on L. casei YIT9018

The process of preparing viscous solution from starch and insoluble matters removed from raw horses through filtration and centrifugation is shown in Figure 1 (Method 2) and their effects on the growth of L. casei YIT9018 were investigated. . The addition of 0.08% (w / v) of the viscous liquid, which was not heat-treated, resulted in a pH and total acidity when compared to the case of fermenting 1% level of horses (raw horses, dry horses) with milk (30 minutes at 85 ° C). Similar trends were observed in viable cell numbers. In Fig. 5-3, the number of viable cells of lactic acid bacteria was 2 × 10 ⁹ cell / ml in the case of milk medium, and 1% of the raw horse was added at 8.3 × 10 ⁹ cell / ml in dry horse 1%, which was 1.1 × 10 ¹⁰ cell / ml. The addition of the viscosities of 0.08% was high as 1.1 x 10 ¹⁰ cells / ml. Therefore, a small amount (0.03-0.08%) of the viscous liquor of the raw horse was added without heat treatment, and the result was almost the same or improved than that of the 1% level of heat-treated horse. Therefore, it was evaluated that adding a small amount of viscous liquid of raw horse rather than adding heat-treated hemp further contributed to promoting the growth of lactic acid bacteria. However, the addition of untreated heat of 1% or more adversely affected the quality of yogurt by enzymatic action.

3. Comparison of viscosity of heat treated 1% viscous materials by heat treatment

Raw and dry horses were added to water and milk at a level of 1% to make 200 ml, and then filtration and centrifugation gave viscous solution from which insoluble materials were removed. These solutions were heat treated at 85 ° C. for 30 minutes, and samples without heat treatment were obtained by comparison using a Kenon capillary viscometer (NO. 125) in a 20 ° C. constant temperature water bath. At 20 ° C., a value of 1.006 cP (no heating), which is a water density of 0.998 g / cm 3, was shown. In comparison with dry horses, horses showed higher comparative viscosity, which means more viscous material than dry horses. Therefore, in the manufacturing process of dry horse, the viscous material in raw horse is judged to be partially destroyed or reduced in functionality due to physical change. The heat treated horses also showed a slightly increased comparative viscosity than before heat treatment, which may be attributed to the increase in molecular weight through partial coagulation of the viscous polysaccharides. In the case of heat treatment of raw horse viscous, the main viscous manna was found to be sensitive to heat. That is, when heat treatment of more than 1% of the concentration of hemp at 85 ℃ 30 minutes it was found that insoluble precipitates are formed remarkably, this precipitation phenomenon was more severe as the concentration of hemp increased. Therefore, if the raw horses were heat-treated at a concentration of 1% or more, the functionality (viscosity) of the viscous material would be greatly reduced.

4. Growth growth effect of various lactic acid bacteria by adding 1% of dry horse

Lactic acid bacteria require different nutritionally demanding media depending on the bacteria. The growth promoting effect of lactic acid bacteria of raw horse and dry horse was compared with milk medium through 5 replicates. Overall, raw horses showed better results in pH, total acidity, and viable cell count than dry horses (Fig. 7). However, raw horses are not more storageable than dry horses, and seasonal problems arise in securing raw materials. Therefore, the effect of promoting the growth of lactic acid bacteria involved in fermented milk and kimchi fermentation was compared by using dry horse which is advantageous in terms of shelf life and securing of raw materials. The growth promoting effect of adding 1% dry horse was the highest in L. casei YIT9018 based on the pH, acetate, and viable cell counts of FIGS. 8-1, 2, and 3. In comparison of pH and total acidity, L. acidophilluis and L. casei YIT9018 were similar to pH 4.48, pH 4.41, and total acidity 0.94% and 0.95%, respectively. In viable cell count, L. casei YIT9018 was 6.9 × 10 ⁹ cell / ml, L. acidophillus was 1.5 × 10 ⁹ cells / ml, Leuc. mesenteroides ATCC 27258 had the highest viable cell count in L. casei 9018 at 1.6 × 10 ⁹ . In particular, the increase rate of viable cell number by the addition of hemp was 3, 2.5 and 1.7 times in L. casei YIT9018, L. acidophillus, Leuc and mesenteroides, respectively. Thus, L. casei YIT9018 and L. acidophillus, Leuc. It was found that the growth promoting effect of lactic acid bacteria was increased by the addition of hemp in order of mesenteroides.

5. Sensory test comparison

Yogurt samples for sensory evaluation were used yogurt samples aged and fermented after adding raw horsetail, viscous material of dead horse and dry horse to milk and milk.

The ranking check results are as follows.

① The milk added 0.03% of viscous liquid.

② Add 1% of dry horse heat treated to milk.

③ 0.08% of viscous liquor solution was added to milk.

④ 1% of raw horse meat heat-treated in milk.

⑤ milk

In the sensory test, yogurt made from milk added with hemp rather than milk was evaluated as good. The addition of 0.03% of raw horse viscous solution showed the best results, and 0.08% of viscous solution was obtained. When only milk was fermented for 19 hours, complete curd formation was not achieved, and the taste of milk itself was strong, with little sourness peculiar to yogurt. On the other hand, when hemp was added, high value was obtained in the sensory test because of the soft texture due to the sourness and viscosity of yogurt as well as the production of curd.

The growth promoting effect of lactic acid bacteria with the addition of raw horse and dry horse was evaluated by pH, total acidity, viable cell count and sensory test of fermented yogurt.

The growth of lactic acid bacteria (L. casei 9018) was significantly increased when fresh and dry horses were added to the milk medium, and were more effective at pH and total acidity including viable cell count under the same fermentation conditions. The hemp added milk medium showed complete curd formation by fermentation at 37 ° C. for 19 hours, whereas milk showed incomplete curd formation.

Raw and dry horses showed the maximum fermentation promoting effect at the 1% addition level, and at the concentration of 1% or higher, the pH increased, the total acidity and the viable cell number were decreased, and the organoleptic effect of the fermented product was particularly bad.

The addition of heat-treated milk medium to the viscous material extracted from the raw horse also showed significant effects on pH, total acidity and viable cell count. The addition of 0.08% showed almost the same effect as the addition of heat treated 1% hemp, and the sensory evaluation of fermented products was evaluated as very good. Therefore, the use of viscous material from which starch and insoluble materials have been removed from raw horses is more effective than the addition of heat treated 1% hemp. However, when 1% of raw horse was added without heat treatment, whey separation was very severe in pH, total acidity, and viable cell count, but was similar to that of heat-treated horse. The comparative viscosity of viscous solution of 1% (W / V) hemp showed that the raw horse was higher than dry horse regardless of heat treatment.

In milk medium supplemented with 1% dry horse, L. acidophillus, and Leuc. The growth promoting effect of mesenteroides strains can be seen, and the increase rate of viable cell number by the addition of hemp was based on milk medium. L. casei YIT9018, L. acidophillus, Leuc. In mensenteroide, 3, 2.5 and 1.7 times respectively.

Claims (4)

In the yogurt production method in which a starter incubated with lactic acid bacteria in a medium such as skim milk is sufficiently inoculated into milk and fermented sufficiently at 37 ° C., the milk medium contains 10% dry horses and 75% raw horses at 0.5 to 2.0%, respectively. Lactic acid bacteria growth promoting method characterized in that the formulation. The method of claim 1 Heated raw horse and dry horse 1% in milk, homogenized twice, heat treated in 85 ℃ water bath for 30 minutes, inoculated with lactic acid bacteria starter about 0.5% and fermented in 37 ℃ incubator for 19 hours to form curd. Lactic acid bacteria growth promotion method characterized by. The method of claim 1, After homogenizing the raw horses twice in distilled water and centrifuging the filtrate obtained by passing through gauze, lactic acid bacteria were grown by adding about 0.08% (W / V, dry matter) to the milk medium without heat treatment. Promotion method. The method of claim 1, Lactic acid bacteria L.casei YIT9018, L.acidophillus, Leuc. How to promote mesenteroides growth.