KR102166789B1 - Mutant lactic acid bacterium strain having acid and heat resistance, and culturing composition comprising this strain as effective gredient - Google Patents

Abstract

The present invention relates to Lactobacillus fermentum JS 1 (KCCM-80171), which is a lactic acid bacterium mutant strain capable of survival at high temperature and low pH, and to a culture composition thereof. In more detail, the present invention relates to Lactobacillus fermentum JS1, which is a mutant strain of Lactobacillus fermentum having blood sugar lowering, cholesterol lowering, antioxidant, and lipid peroxidation inhibitory effects, to a culture composition containing the strain as an active ingredient, and to the use of the composition.

Description

Acid-resistant, heat-resistant lactic acid bacteria mutant strain and a culture composition comprising the strain as an active ingredient {Mutant lactic acid bacterium strain having acid and heat resistance, and culturing composition comprising this strain as effective gredient}

The present invention relates to a lactic acid bacteria mutant strain capable of viability at high temperature and low pH and a lactic acid bacteria culture composition comprising the strain as an active ingredient, and more particularly, it is possible to survive at high temperature and low pH, lowering blood sugar, lowering cholesterol, It relates to a Lactobacillus fermentum (Lactobacillus fermentum) JS1 lactic acid bacteria mutant strain (Accession No. KCCM-80171) having an antioxidant, lipid peroxidation inhibitory effect, and a lactic acid bacteria culture composition comprising the strain as an active ingredient.

Lactobacillus is a bacterium that has been usefully used by humans for the longest time among bacteria, and is particularly commonly used in fermented dairy products. Lactobacillus is reported to improve absorption of nutrients, relieve lactose intolerance, improve intestinal motility, and have anticancer effects.

Since Mechnikov's longevity theory, various studies have been conducted on the physiological effects of foods using lactic acid bacteria or lactic acid bacteria.

As the action of lactic acid bacteria contained in fermented milk such as yogurt, it is known that the intestinal microbial total improvement effect and intestinal action are known. Recently, it has been reported that lactic acid bacteria have various functions, for example, immune activity, antibacterial activity and anti-tumor activity. In addition, it has been reported that it has various functions of lactic acid bacteria, for example, immune activity, antibacterial activity, and antitumor activity. As described above, since various health effects of lactic acid bacteria are expected, strains such as Lactobacillus acidophilus, Streptoccocuss thermophilus, and Bifidobacterium genus detected in human intestine Fermented milk and lactic acid bacteria beverages are commercially available.

However, since these lactic acid bacteria are weak in acid resistance or heat resistance, they are almost killed by gastric acid secreted from the stomach when humans ingest them, and thus have a disadvantage that they cannot reach the intestines.

To solve this problem, a method of capturing and ingesting lactic acid bacteria in a capsule made of an acid-resistant and enteric-soluble coating was invented, but even if the lactic acid bacteria safely pass through the stomach and reach the intestine, the capsule in the intestine It has a problem in that it is not properly decomposed or dissolved by the pH of the intestine, or the capsule is chewed in the mouth so that the lactic acid bacteria inside are leaked out of the capsule.

Thus, the present inventors were devising a way to pass the stomach without trapping the lactic acid bacteria in the capsule, as a result of researching lactic acid bacteria having acid resistance at low pH, Lactobacillus fermentum that can survive at high temperature and low pH (Lactobacillus fermentum) JS1 Lactobacillus mutant strain was discovered and the present invention was completed by confirming its function.

An object of the present invention is to provide a Lactobacillus fermentum (Lactobacillus fermentum ) JS1 lactic acid bacteria mutant strain capable of survival at high temperature and low pH.

Another object of the present invention is to provide a culture composition comprising the Lactobacillus fermentum JS1 lactic acid bacteria mutant strain as an active ingredient.

Another object of the present invention is that the culture composition of the Lactobacillus fermentum JS1 lactic acid bacteria mutant strain has an antioxidant, cholesterol lowering, antioxidant, and lipid peroxidation inhibitory function, so it is a dietary supplement, functional food, animal and fish. It is to provide a use for auxiliary feed and fermented feed, microbial fertilizer for soil improvement, and water quality improvement.

Another object of the present invention is to provide a method for producing lactic acid bacteria rice by spraying and spraying the plant lactic acid bacteria, which is the Lactobacillus fermentum JS1 lactic acid bacteria mutant strain, onto rice, and the rice prepared by the method.

In order to achieve the above object, the present invention provides a Lactobacillus fermentum JS1 lactic acid bacteria mutant strain (accession number KCCM-80171) having heat resistance and acid resistance having 16S rRNA encoded by the DNA of SEQ ID NO: 1.

In addition, the present invention provides a lactic acid bacteria culture composition comprising the Lactobacillus fermentum JS1 lactic acid bacteria mutant strain having a function of lowering blood sugar, antioxidant, inhibiting production of lipid peroxidation and lowering cholesterol as an active ingredient.

In addition, the present invention provides an auxiliary feed composition that can be used for livestock or fish using the lactic acid bacteria culture composition, a fermented feed production method using a mushroom waste medium, and a water quality improvement method.

In addition, the present invention provides an organic fertilization, characterized in that the lactic acid bacteria culture composition is added to the livestock meal, and a soil improvement method, characterized in that the organic fertilization is mixed into the soil.

In addition, the present invention is preferably spraying and spraying the lactic acid bacteria mutant strain on rice, and provides a lactic acid bacteria-containing rice prepared by the above method.

Hereinafter, the present invention will be described in detail.

The present invention provides a Lactobacillus fermentum JS1 lactic acid mutant strain (accession number KCCM-80171) having heat resistance and acid resistance having 16S rRNA encoded by the DNA of SEQ ID NO: 1.

The present inventors obtained the Lactobacillus fermentum wild strain from Hyupjeon Corporation in Japan on February 11, 2016 (see FIG. 1), and after successful artificial culture, the medium produced by mixing with various grains was the worst. Lactobacillus mutant strains capable of viability at high temperatures and low pH were prepared by culturing under the conditions of culture. Specifically, during the acid resistance and heat resistance experiments conducted in the process of preparing the lactic acid strain culture composition, a mutant strain having excellent viability was found at pH 2 to 3, and the mutant strain was isolated and cultured by a conventional method. Mentum (Lactobacillus fermentum) was identified as a different strain from the strain, and was named'Lactobacillus fermentum JS1 bacteria'. The Lactobacillus fermentum JS1 strain according to the present invention has the 16S rRNA encoded by the DNA of SEQ ID NO: 1, and was deposited with the Korea Microbiological Conservation Center on October 26, 2017, and was given accession number KCCM-80171.

In addition, the present invention provides a lactic acid bacteria culture composition comprising the Lactobacillus fermentum JS1 lactic acid bacteria mutant strain having a function of lowering blood sugar, antioxidant, inhibiting production of lipid peroxidation and lowering cholesterol as an active ingredient.

In the lactic acid bacteria culture composition of the present invention, the composition is a Lactobacillus fermentum JS1 lactic acid bacteria mutant strain per 1 liter of water, glucose 20 to 30 g, yeast extract 10 to 20 g, soypeptone 5 to 8 g, casein 3 to 5 g, MgSO ₄ .7H ₂ O 0.1~0.15g, K ₂ HPO ₄ 1~2g, MnSO ₄ .5H ₂ O 0.05~0.1 g, sodium acetate 3~5g, ammonium citrate 2~3g, Tween 80 1 ~ 2g, neoline (neoline) is preferably prepared by culturing in a medium mixed with 0.25 ~ 0.3g, the composition is a yeast Saccharomyces cerevisiae (Saccharomyces cerevisiae) strain per 1 liter of water Protia Protease peptone No3 5~10g, malt extract 3~5g, glucose 10~20g, yeast extract 10~15g, sodium acetate 5~10g, ammonium citrate 3~5g It is more preferable to additionally include a yeast culture solution.

In addition, the present invention provides an auxiliary feed composition that can be used for livestock or fish using the lactic acid bacteria culture composition, a fermented feed production method using a mushroom waste medium, and a water quality improvement method.

In addition, the present invention provides an organic fertilization, characterized in that the lactic acid bacteria culture composition is added to the livestock meal, and a soil improvement method, characterized in that the organic fertilization is mixed into the soil.

In addition, the present invention is preferably spraying and spraying the lactic acid bacteria mutant strain on rice, and provides a lactic acid bacteria-containing rice prepared by the above method.

As described above, Lactobacillus fermentum according to the present invention Unlike conventional lactic acid bacteria, the lactic acid bacteria culture composition containing JS1 was resistant to heat and acid, had a completely new property of decomposing ammonia, which is theoretically impossible, and showed excellent efficacy in lowering blood sugar. Based on the experimental results that have been shown to be very effective in the development of these bacteria and in lowering blood sugar levels of these bacteria, it is possible to eliminate the side effects of drugs and the discomfort of regular insulin supply in diabetics, as well as new treatments as a result of this study. By providing methods, it will be able to contribute greatly to the promotion of public health.

1 is a guidebook of lactic acid bacteria for wild-type Lactobacillus fermentum before mutation obtained from Hyupjeon Corporation in Japan.
Figure 2 is a graph showing the blood glucose level of ob/ob mice (n=5) ingested with a lactic acid bacteria-containing product ( ^a p <0.01: ^b p <0.001 (compared with the control group))
Figure 3 is a plant lactic acid bacteria Lactobacillus fermentum of the present invention (Lactobacillus fermentum) JS1 Raw horse and yeast Saccharomyces cerevisiae This is a design of a method for improving the water quality of a small river channel equipped with a plastic box containing raw horses.
4 and 5 are the vegetable lactic acid bacteria Lactobacillus fermentum of the present invention (Lactobacillus fermentum) JS1 Raw horse and yeast Saccharomyces cerevisiae This is a test report showing the content of each strain contained in rice containing raw malt.

Hereinafter, preferred embodiments according to the present invention will be described in more detail by presenting them in more detail. However, the following embodiments are provided so that the present invention may be sufficiently understood by those of ordinary skill in the art, and may be modified in various other forms, and the present invention is limited by the above embodiments. It is not.

<Example 1> Mutant Lactobacillus fermentum Manufacturing of JS1

The present inventors were guided by wild-type Lactobacillus fermentum lactic acid bacteria progenitor bacteria before mutation from Hyupjeon Corporation in Japan (FIG. 1).

In order to prepare a mutation from the wild-type Lactobacillus fermentum, it was sequentially subcultured in the following three-step medium composition. Specifically, first, after sterilization of Petri dish MRS agar at 121° C. for 15 minutes, 20 ml of MRS agar was added to Petri dish in a clean bench, and 24 hours later, the protobacteria were inoculated in this medium and cultured at 37° C. for 48 hours. Thereafter, 250cc of MRS liquid medium was sterilized at 121°C for 15 minutes, cooled in a clean bench, colonies cultured on the MRS agar medium were inoculated with platinum, and cultured at 42°C for 15 to 18 hours, and then in a 2°C refrigerator. Stored frozen. Finally, if you mix 92% bean curd, 5% rice bran, 1% glutinous rice powder, and 2% black soybean powder, sterilize a medium containing 78% moisture at 121℃ for 15 minutes, cool it to 42℃, and then lactic acid bacteria in an Erlenmeyer flask. After adding 1% of the medium amount and incubating for 5 hours, 160 ml of 99% sodium hydroxide was diluted in 1 liter of water to prepare 4 mol, and the pH of the medium was adjusted to 2.5. After raising the culture temperature to 98°C and incubating for 36 hours, the cycle was repeated 200 times for 3 years. In other media whose pH was adjusted to 1.5, they were cultivated 200 times at 98°C, and the final DNA was examined to prepare a natural mutant strain, JS1 strain, at the end of Cheonshin Mango, and this was deposited as of October 26, 2017. Deposited with.

<Example 2> Lactobacillus Fermentum Preparation of JS1 lactic acid bacteria culture solution composition

A method of preparing the lactic acid bacteria culture medium composition is as follows.

Put 200cc distilled water into a 250cc Erlenmeyer flask, add 11g of MRS broth medium, and mix the medium thoroughly by boiling water. Put it in an autoclave, sterilize at 121℃ for 15~30 minutes, take it out, cool it down to 35℃ in a clean bench, and then use PET. After inoculation of the Lactobacillus fermentum JS1 progenitor obtained in Example 1 cultured on dish MRS agar medium, the inoculum was stored in a refrigerator after incubation at 37 to 40° C. for 20 to 30 hours.

Then, this culture is carried out with glucose 20~30g per 1 liter of water, yeast extract 10~20g, soypeptone 5~8g, casein 3~5g, MgSO ₄ .7H ₂ O 0.1~0.15g, K ₂ HPO ₄ 1~ 2g, MnSO ₄ .5H ₂ O 0.05~0.1g, sodium acetate 3~5g, ammonium citrate 2~3g, Tween 80 1~2g, neoline 0.25~0.3g Put in a tank, dilute, sterilize at 121℃ for 1 hour, cool to 37℃, inoculate 1~3% of the inoculum, incubate at 39~42℃ for 18~24 hours, and then use a centrifuge to separate lactic acid bacteria. After mixing 10 to 15% of skim milk powder and 5 to 10% of trehalose to the amount, it was taken out after vacuum freeze drying at -80°C for 72 hours in a vacuum freezer and pulverized with a crusher to prepare a lactic acid bacteria raw powder, put in an aluminum bag, and stored in a refrigerator.

The number of lactic acid bacteria in the original end was 3.5 × 10 ¹¹ cfu/g to 4.5 × 10 ¹¹ cfu/g.

The raw powder was diluted with glucose according to the intended use and mixed at 1.5×10 ⁷ cfu/g, 1.5×10 ⁸ cfu/g, and 1.5×10 ⁹ cfu/g.

Using the lactic acid bacteria raw powder, sterilized water was added and diluted to 6.8 × 10 ⁷ ml, which was used in the following experiments.

<Example 3> Yeast ( Saccharomyces cerevisiae ) Of the culture solution

YM liquid medium was put in a 250cc Erlenmeyer flask, 200ml of distilled water was added and diluted, and the medium was thoroughly mixed by boiling water, put in an autoclave, sterilized at 121℃ for 15-30 minutes, and then removed and cooled in a clean bench. After inoculating the yeast progenitors cultured on Petri dish YM agar medium, the inoculum was incubated at 37° C. for 72 hours in an incubator, and then stored in a refrigerator.

Then, the main culture is 5-10 g of protease peptone No3 per 1 liter of water, 3-5 g of malt extract, 10-20 g of glucose, 10-15 g of yeast extract, 5-10 g of sodium acetate, and ammonium citrate. Put in a culture tank at a ratio of 3~5g, dilute, sterilize at 121℃, cool to 37℃, inoculate 1~3% of the inoculum, incubate at 37℃ for 2-3 days, and then use a centrifuge. Add 10 to 15% skim milk powder and 5 to 10% trehalose to the amount of separated sludge, mix with yeast sludge, and then vacuum freeze dry at -80℃ for 72 hours with a vacuum freezer, take out, crush with a crusher, and place in an aluminum bag. Then, the raw yeast was refrigerated.

The number of yeast in the raw yeast is 2 × 10 ⁹ cfu/g ~ 1.5 × 10 ¹⁰ cfu/g, and the raw yeast is mixed with the lactic acid bacteria culture composition when used as an auxiliary feed or microbial fertilizer. In addition, when the raw yeast was used as an auxiliary feed or microbial fertilizer, corn flour or glucose was mixed, and the lactic acid bacteria were 1.5 × 10 ⁷ cfu/g, and the yeast was 1.5 × 10 ⁶ cfu/g.

<Comparative Example 1>

Lactobacillus acidophilus ( Lactobacillus acidophilus) was used as a lactic acid bacteria commercially available beverages from other companies.

<Comparative Example 2>

Lactobacillus casei ( Lactobacillus casei) was used as a lactic acid bacteria commercially available beverages from other companies.

<Test Example 1> Acid resistance test

Lactobacillus permentum according to the present invention The acid resistance test was performed to confirm the viability of JS1 bacteria even at low pH. At this time, a BCP-added medium was used as the medium, and the culture conditions were cultured at 37-39°C for 72 hours, and the pH was adjusted to pH 2 and pH 3 using citric acid and HCl, respectively. The yellow colonies generated after culturing under the above conditions were measured as lactic acid bacteria colonies, and the results are shown in Table 1.

pH before pH adjustment 1 hour later 2 hours later Example 2 pH 3.0 6.8 × 10 ⁷ ml 5 × 10 ⁷ ml 6.5 × 10 ⁶ ml pH 2.0 6.8 × 10 ⁷ ml 6.2 × 10 ⁶ ml 4.8 × 10 ⁵ ㎖ Comparative Example 1 pH 3.0 6.8 × 10 ⁷ ml 3.0 × 10 ⁴ ml 6.3 × 10 ² ml pH 2,0 6.8 × 10 ⁷ ml 4.1 × 10 ³ ml 5.5 × 10 ² ml

<Test Example 2>

<2-1> Experimental animals and breeding conditions

C57BL/6J-Leb ^ob (ob mouse) is an animal used in obesity-type diabetes experiments. It was purchased from The Jacson Laboratory (JAX) in the United States, and the breeding facility was carried out in the SPF barrier system, and the breeding conditions were automatically controlled (23℃, humidity 52 +). 10%, noise 60 phone or less, odor 20 ppm or less, lighting 200 to 250 Lux), and the contrast was automatically adjusted in a 12 hour cycle (18:00 to 06:00). A polycarbonate cage (180W × 240D 125H mm) was used for the breeding box, and the crushed cottonwood sawdust was used as the litter, and the animal breeding box exchange was carried out twice a week. All equipment and litter used in the breeding room were autoclaved (121°C for 30 minutes sterilization, 30 minutes drying). As for the feed, mouse experimental animal feed was purchased and allowed to be freely ingested and reared for 3 weeks.

The composition of the feed is water 12.5% by weight, crude protein 22% by weight, crude fat 4.3% by weight, crude fiber 4.1% by weight, ash content 6.3% by weight, calcium 0.8% by weight.The negative water of the control group is filtered with water and then UV water sterilizer (Dynamics, M600 , USA) passed through the water was ingested.

<2-2> Selection of animals and preparation of samples

In the case of the test animals used in this experiment, at the age of 3 to 4 weeks, animals showing obesity were selected and used, and the age of the test animals was 9 weeks old male. Immediately before the test, body weight and basal blood sugar were measured, and the average level was similarly adjusted for each group, and the average blood sugar value at this time was around 380-420 mg/dl.

1 kg of the lactic acid bacteria raw powders of Examples 2 to 3 and Comparative Examples 1 to 2 were mixed in a ratio of 1.5 × 10 ⁸ cfu/g using glucose, dissolved in 10 liters of drinking water, and put in a water bottle so as to be ingested for 3 weeks. .

<2-3> Dietary intake and dietary efficiency

During the experiment, they were fed freely (ad libitum), and water was supplied with a water bottle at a certain time every day to freely intake. Dietary intake was measured for a certain time every day, and body weight was measured on days 2, 4, 8, 12, 16 and 21, respectively, during the experiment.

Dietary efficiency was calculated by dividing the amount of increase in the total by the amount of dietary intake during the same period. The effects on dietary intake and dietary efficiency are shown in Table 2 below. Dietary intake indicated the amount of feed (daily intake) consumed by the control group and the experimental group by diluting the lactic acid bacteria powder in drinking water, and there was no significant difference. In addition, when comparing the feed efficiency ratio, which is used as an indicator of weight gain (g) to feed intake (g), it was almost consistent with the weight change.

Comparison of dietary intake, weight gain and dietary efficiency ratio (FER) group Dietary intake (g/day) Weight gain
(g/21days) FER Control 5.10 ±0.70 7.26 ±1.20 0.07 ±0.01 Example 2 5.30 ±0.50 7.15 ±0.56 0.06 ±0.01 Example 3 5.30 ±0.60 7.52 ±0.57 0.07 ±0.01

FER = weight gain (g)/diet intake (g) ratio

All figures are mean ±S.E. (n=5).

<Test Example 3> Measurement of active concentration

Blood collection was started at 14:00 using the orbital vein blood collection method, so that it did not take more than 30 minutes to collect all animals. The concentration of blood glucose in all subjects was measured using one touch basic (Lifescan Co. USA). Blood glucose was measured by the same method after 2, 4, 8, 12, 16, and 21 days.

In Examples 2 to 3 and Comparative Examples 1 to 2, changes in blood glucose levels according to intake of dissolved water of lactic acid bacteria and yeast bacteria were compared and observed. Drinking water was used as a control group, and changes in blood glucose levels according to intake of dissolved water for lactic acid bacteria and yeast bacteria in Examples 2 to 3 and Comparative Examples 1 to 2 were compared and observed. As a control group, drinking water was used, and the results of comparing the blood sugar inhibitory effect for 3 weeks while freely ingesting the lactic acid bacteria and yeast dissolved water of Examples 2 to 3 and Comparative Examples 1 and 2 are shown in FIG. 2. As shown in Figure 2, the lactic acid bacteria and yeast lysed water of Examples 2 to 3 began to show a significant blood sugar lowering effect after 4 days compared to the control group, and when comparing the blood sugar level on the last 21 days, about 45% and 35%, respectively, compared to the control group. Showed a remarkable hypoglycemic effect. In addition, Comparative Examples 1 to 2 also began to drop blood sugar levels from the 4th day, and the blood sugar level on the last day of the 21st day showed a blood sugar lowering effect of about 17% to 21% respectively compared to the control group.

<Test Example 4>

<4-1> Blood collection and separation

Anesthetize the experimental animal with ether, collect blood from the heart, leave it at low temperature for 2 hours, and then centrifuge at 700 g for 10 minutes to obtain the supernatant serum obtained by adding sodium heparin (100 Units, Sima), an anticoagulant, per 1.0 ml of blood. It was put into a CBC bottle (complete blood cell count: green cross) treated with ml and used for analysis while cryopreserving at -70°C.

<4-2> Measurement of neutral lipids and proteins

The content of triglyceride (TG) as a neutral lipid in serum was treated with a TG kit reagent (Sigma, USA) for measuring serum triglycerides, and the content of triglycerides in serum was quantified based on a standard calibration curve. The protein content in the serum was measured according to Lowry's method (1951) using BSA (standard serum albumin) as a standard substance.

<4-3> Measurement of cholesterol content

The total cholesterol content was measured by the o-phthalaldhyde method according to the method of Rude1 et al. (Rude1 LL, 1973). First, 0.1 ml of samples were taken, heated in a water bath at 60° C. for 15 minutes, and then cooled. 5.0 ml of nucleic acid was added thereto, mixed, and 3.0 ml of distilled water was added again, followed by well-mixing for the next 1 minute, and the layers were separated to obtain 1.0 ml of a hexane layer. The hexane layer was concentrated to dryness with nitrogen, 2.0 ml of o-phthalaldhyde reagent was added and mixed well. After 10 minutes, 1.0 ml of concentrated sulfuric acid as a color developing reagent was added and mixed. The absorbance was measured at 550 nm using a spectrophotometer within 10 to 90 minutes after sulfuric acid addition, and the content of total cholesterol in the blood was quantified by a standard calibration curve.

<4-4> Measurement of lipoprotein-cholesterol content

To measure the content of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol in serum, HDL-cholesterol (HDL-C555, Eiken Japan), LDL-cholesterol (BLF, Eiken Japan) kit reagent was used.

<4-4-1> Measurement of HDL cholesterol content

0.3 ml of serum was put into a test tube, 0.3 ml of a precipitation reagent was added thereto, mixed well, and allowed to stand at room temperature for 10 minutes, followed by centrifugation at 700 g for 10 minutes. After that, 50 µl of the supernatant, 50 µl of the standard solution (100 ml/dl), and 3.0 ml of the HDL color developing reagent were added to 50 µl of distilled water as a blank, mixed well, and heated in a water bath at 37°C for 5 minutes. Using the blank as a control, absorbance was measured at 555 nm to quantify the content of HDL-cholesterol (mg/dl serum).

<4-4-2> Measurement of LDL cholesterol content

Add 0.1 ml of serum and 0.1 ml of standard serum to a test tube, add 4.0 ml of BLF kit reagents I and II to each, mix well for 5 seconds, and then leave for 25 minutes at room temperature (25 ±3℃) and distilled water within 10 minutes. As a control group, absorbance was measured at 650 nm to quantify the LDL-cholesterol content (mg/dl serum).

<4-4-3> Calculation of atherosclerosis index (Atherogenic index)

The atherosclerosis index (AI), which is used as an indicator of the onset of atherosclerosis, known as an early symptom of adult diseases, was calculated by subtracting the total cholesterol content according to the method of Haglund et al. (1991) and dividing it by HDL-cholesterol. .

To investigate the effect of plant lactic acid bacteria (Lac, F-100, 50) and animal lactic acid bacteria fermented beverages (HYW-100, 50) on the content of triglyceride (TG), cholesterol and lipoprotein cholesterol, which are known to cause adult diseases. Table 3 shows the effect of administration for 3 weeks while allowing the obese mouse to freely ingest the lactic acid bacteria dissolved water of Examples 2 to 3 and Comparative Examples 1 to 2, respectively.

In the lipid composition content of ob/ob mice, Lac. F effect of beverage administration parameter Control Example 2 Example 3 Comparative Example 1 Comparative Example 2 protein
(Mg/dl serum) 25.510±0.58
(100%) 25.90±0.64
(101.6%) 29.940±31
(105.6%) 27.150±39
(106.5%) 27.210±32
(106.7%) Triglycerides
(Mg/dl serum) 137.51±1.35
(100%) 119.88±0.39
(87.2%) 125.63±7.14
(91.4%) 124.57±4.80
(90.6%) 129.69±2.32
(94.3%) Total cholesterol
(Mg/dl serum) 123.956±92 ^C
(100%) 104.33±2.23 ^C
(84.2%) 115.72±3.23 ^a
(93.4%) 112.60±5.45 ^a
(90.8%) 121.81±4.55 ^a
(98.3%) LDL-cholesterol
(Mg/dl serum) 112.03±5.78 ^c
(100%) 83.630±0.47 ^c
(74.7%) 90.92±1.77 ^c
(81.2%) 91.87±2.27 ^c
(82.0%) 95.46±3.44 ^c
(85.2%) HDL-cholesterol
(Mg/dl serum) 56.02±2.90 ^c
(100%) 68.070±0.96 ^c
(121.5%) 65.47±0.38 ^c
(116.9%) 59.17±2.12 ^c
(105.6%) 58.21±1.15 ^c
(103.9%) Arteriosclerosis Index (AI) 1.20±0.12 ^c
(100%) 0.520±0.09 ^c
(43.8%) 0.75±0.07 ^c
(62.8%) 0.91±0.17 ^a
(74.4%) 1.08±0.16 ^c
(90.1%)

＊ All figures are mean ±S.E (n=5)

＊＊ ^a p <0.05: ^b p <0.01: ^c p <0.001 (compared with control)

It was found that the content of neutral lipids and total cholesterol in the group to which the fermented vegetable lactic acid bacteria fermented beverages of Examples 2 to 3 were administered were significantly suppressed to 8.2 to 12.5% and 6.4 to 15.6%, respectively, compared to the control group. When comparing the lowering effect of LDL-cholesterol, which is directly related to the onset of actual adult diseases, about 17.8 to 24.5% or more of the inhibitory effect was found compared to the control group. In addition, it was found that the atherosclerosis index (AI), which is known as an early onset index of adult diseases, was significantly suppressed by almost 37.5 to 55.6% in the group administered with the fermented vegetable lactic acid beverage of Examples 2 to 3. It is interesting to note that this decrease in the arteriosclerosis index almost coincides with the decrease in the content of LDL-cholesterol. This is because LDL-cholesterol content and an increase in atherosclerosis index are directly related to the onset of adult diseases.

It can be judged that the level of HDL-cholesterol, which is widely known as an anti-cholesterol factor, and is highly associated with exercise and longevity, is also very effective in preventing adult diseases that the plant lactobacillus administration group increased by 15.6-21.2%.

Animal lactic acid bacteria fermented beverages of Comparative Examples 1 to 2 also showed that the levels of neutral lipids and LDL-cholesterol also decreased compared to the control group. However, compared to the plant lactic acid bacteria of Examples 2 to 3, the degree of inhibition was found to be weak.

The vegetable lactic acid bacteria of Examples 2 to 3 according to the present invention significantly lowers the content of cholesterol in blood, which plays an important role in lipid metabolism as well as lowering blood sugar, so it is expected to be of great help in preventing adult diseases.

<Test Example 5> Measurement of active oxygen and lipid peroxide (LP0)

<5-1> Measurement of active oxygen

Measurement of hydroxyl radicals, known as the most powerful free radicals among free radicals, is a method of measuring the degree of destruction of deoxyribose, and is a method of measuring the degree of formation of hydroxyl radicals. Ribose is destroyed to produce aldehyde, and this aldehyde was measured according to a method (Halliwell, B, 1981) in which color was developed by reacting with thiobabituric acid in an acidic solution.

The content of lipid peroxide (LP0) in serum, known to be produced by these oxygen radicals, was measured according to the experimental method of Choi et al. (Choi, J, H, 1994).

<5-2> Determination of enzyme activity

The measurement of superoxide dismutase (SOD) activity, which is important as a scavenging enzyme for reactive oxygen species, was quantified according to the method of Oyanagui et al. (Oyanagui, Y, 1984). In addition, the activity of glutathione peroxidase (GSHPx) in serum was measured as a method to measure the decrease of NADPH at 340 nm as an important scavenging enzyme in vivo (Lawrence, R, A, 1978).

<Test Example 6> Statistical processing of analysis results

All experimental results of the present invention were statistically processed to calculate the mean value and standard deviation, and the significance test between each experimental group was performed by Student's t-test (Steel, R, G, D, 1960).

Superoxide radicals reported as active oxygen generated in the initial stage among free radicals among free radicals among free radicals as the effect of administration of plant lactic acid bacteria dissolved water in Examples 2 to 3 on lipid peroxidation in the blood of obese mice. radical) and the formation of a hydroxyl radical known as the most powerful free radical and the content of lipid proxide (LPO) produced at this time are shown in Table 4 below. In addition, biodefensive enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) are powerful antioxidant enzymes that exist in the body. It was found that it not only inhibits the generation of free radicals such as radicals and superoxide radicals, but also very effectively inhibits the generation of lipid peroxide, which is known to be closely related to the onset of adult diseases and aging.

Effects of Lac and F Beverages on Reactive Oxygen Species and Scavenger Enzyme Activities in Ob/ob Mice parameter Control Example 2 Example 3 Comparative Example 1 Comparative Example 2 Lipid peroxide (LPO)
(nmol/ml serum) 5.10±0.22 ^c
(100%) 4.36±0.14 ^c
(85.7%) 4.55±0.27 ^b
(89.4%) 4.48±0.38 ^a
(88.0%) 4.51±0.32 ^a
(88.6%) Hydroxy radical
(nmol/mg protein) 4.26±0.41 ^a
(100%) 3.67±0.26 ^a
(86.2%) 3.90±0.12 ^b
(91.6%) 3.79±0.12 ^b
(89.0%) 3.91±0.09 ^c
(91.8%) Superoxide radical (nmol/mg protein) 86.59±3.01 ^a
(100%) 71.74±3.10 ^C
(82.9%)
77.83±4.85 ^b
(89.9%) 78.29±4.10 ^b
(90.4%) 82.23±0.80 ^a
(95.0%) Super Oxide Dismutase (SOD)
(unit/mg protein) 212.82±7.96 ^c
(100%) 258.96±5.4 ^c
(121.7%) 245.26±3.04 ^c
(115.2%) 257.20±8.61 ^c
(120.9%) 242.43±5.32 ^c
(113.9%) Glutathione peroxidase (GSHPx)
IU/g protein 7.31±0.23 ^b
(100%) 8.63±0.56 ^b
(118.0%) 7.95±0.10 ^b
(108.7%) 7.90±0.51 ^b
(108.1%) 7.53±0.54 ^b
(103.0%)

* All figures are average ±S.E. (n=5)

^a p <0.05: ^b p <0.001 (compared to control)

When comparing the effect on the activity of SOD, an enzyme related to antioxidant, it can be seen that the activity is effectively increased to 14.8~21.5% compared to the control group. In addition, the activity of GSHPx was also 8.5 to 18.2% compared to the control, and it was found that ingestion of the dissolved water of plant lactic acid bacteria in Examples 2 to 3 increased the activity of antioxidant enzymes in vivo.

It is known that the production of reactive oxygen species (ROS) such as superoxide anion, hydroxyl radical, and hydrogen peroxide as free radicals in vivo not only causes various adult diseases, but also promotes the aging process. . It is present in the blood or all organs in the body to remove the superoxide anion generated by free radicals in the body or the harmful oxygen of hydrogen peroxide and the hydroxyl group. These defense systems include superoxide dismutase (SOD). , glutathione peroxidase (GSHPx), catalase, etc. are known. It has been reported that the activity of reactive oxygen species in milk and fermented milk products is lowered, and this mechanism is known to be closely related to antioxidant enzymes.

Therefore, the dissolved water of plant lactic acid bacteria of Examples 2 to 3 according to the present invention not only inhibits the generation of such reactive oxygen species, but also significantly increases antioxidant enzymes, and thus it is judged to affect not only adult diseases but also inhibition of the aging process.

In the present invention, in order to scientifically identify the therapeutic and preventive action of diabetes caused by ingestion of lactic acid bacteria, the dissolution water of plant lactic acid bacteria using Lactobacillus fermentum JS1 was used as a diabetes test animal, ob. Weight change while rearing for 3 weeks using a mouse, the effect of reducing the content of triglycerides and cholesterol in the blood, the effect of reducing LDL-cholesterol and atherosclerosis index (atherogenic radical), hydroxyl radical, superoxide radical (superoxide radical) and lipid peroxide (LPO) production inhibitory effect, and the effects of these active oxygen scavenging enzymes such as superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) I did.

Changes in blood glucose level showed remarkable hypoglycemic action of 35 to 45%, respectively, compared to the control group in the experimental group to which the plant lactic acid bacteria dissolved water of Examples 2 to 3 was administered. Long-term oral administration of the plant lactic acid bacteria dissolved water of Examples 2 to 3 was ob. It was found to have a significant hypoglycemic effect in mice. The content of triglycerides (TG), total cholesterol and LDL-cholesterol in the group administered with dissolved plant lactic acid bacteria of Examples 2 to 3 was not only found to have a lowering effect compared to the control group, but also had an arteriosclerosis index of 50% or more. There was a reduction effect. In the groups administered with dissolved water of plant lactic acid bacteria of Examples 2 to 3, not only the generation of superoxide radicals and hydroxyl radicals were significantly suppressed compared to the control group, but also the content of lipid peroxide (LPO) produced according to the active oxygen was significantly reduced. . In addition, it was confirmed that the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSHPx) as enzymes for removing active oxygen were increased compared to the control group. From the above results, as a result of administering the plant lactobacillus dissolved water of Examples 2 to 3 containing Lactobacillus fermentum JS1 to obese mice in a diabetes model for 3 weeks, it was experimentally demonstrated that it can prevent or delay adult diseases and aging in addition to the hypoglycemic effect. Proved to be.

<Test Example 7> Broiler breeding test

Vegetable lactic acid bacteria Lactobacillus fermentum JS1 raw powder and yeast Saccharomyces cerevisiae raw powder are mixed with glucose to make lactic acid bacteria 1 × 10 ⁷ cfu/g and yeast 1 × 10 ⁶ cfu/g, and then 0.2% of feed is mixed with broiler feed. And paid.

When breeding broilers, 4 repeated tests were performed on the control, antibiotics, and 0.2% lactobacillus-added groups of 50 per 3.3 m ² each. For the test breeding broilers, 50 count × 3 treatment groups × 4 repetitions = 600 numbers were tested.

Breeding test results (average of 4 repeated tests) division Breeder Breeding days Total feed Average weight Remark Control 200 number 35 days 680.4 kg 1.8kg Antibiotic sphere 200 number 35 days 650.2 kg 1.8kg 0.2% lactobacilli 200 number 32 days 620.4 kg 1.8kg 300g of feed savings per allowance

The average weight of the control group or antibiotic group was 1.8 kg on the 35 days of breeding, and the average weight of the group fed 0.2% lactobacillus reached 1.8 kg on the 3 days of breeding, which shortened the breeding period by 3 days, resulting in a saving of 300 g of average feed cost per animal. In addition, the odor of chicken manure was reduced by 60-70%, the occurrence of flies was suppressed, and thus eco-friendly broiler breeding was possible, and antibiotic-free chicken could be supplied. The breeding period of antibiotics was the same as that of the control, but the average feed cost was saved by 151g.

When 30,000 water is raised using 0.2% of lactic acid bacteria in general breeding farms, 9,000 kg of feed is saved, and antibiotic purchase costs are saved, which can greatly contribute to farm income, and by supplying antibiotic-free chicken meat, it contributes to the improvement of national health.

<Test Example 8> Fish breeding test

Plant lactic acid bacteria Lactobacillus fermentum JS1 raw powder and yeast Saccharomyces cerevisiae raw powder were mixed with glucose to make lactic acid bacteria 1 × 10 ⁷ cfu/g and yeast 1 × 10 ⁶ cfu/g. It was prepared and fed as aquaculture feed.

During breeding of the carp test, the control, antibiotic, and 0.2% lactic acid bacteria were subjected to four repeated tests of 100 animals each for 4 months. Breeding test carp number was tested in 3 treatments × 100 × 4 repetitions = 1,200.

Fish breeding test results (average of 4 repeated tests) Control Lactobacillus
fementum JS1 (0.2%) Flavomycine
4% mg/KG Liver pancreas weight 2.19±0.27 ^a 3.60±0.56 ^c 2.63±0.43 ^b HSI 2.38±0.50 ^a 3.21±0.53 ^c 2.78±0.21 ^b HPC 8.64±0.27 ^a 10.01±0.42 ^c 9.19±0.46 ^b chapter weight 2.11±0.27 ^a 2.91±0.38 ^c 2.61±0.26 ^b ISI 2.32±0.22 ^a 2.61±0.36 ^b 2.58±0.40 ^b IPC 6.39±0.32 ^a 8.31±0.38 ^d 6.82±0.23 ^b Length 21.63±0.67 ^a 28.44±1.70 ^c 23.29±1.44 ^b RGL 141.27±7.96 ^a 175.44±17.08 ^b 149.63±1.20 ^a

As a result of the carp breeding test, the length of the carp was increased by about 30% and the weight was increased by about 24% compared to the control group in the group fed 0.2% lactic acid bacteria. Compared to the antibiotic administration group, the 0.2% lactobacillus administration group increased the carp length by about 22%, but the weight increased by about 17%.

In addition, the water in the carp breeding tank had to be changed once a week for the control or antibiotic administration unit, but the lactic acid bacteria administration unit was replaced once every 8 weeks because the water in the tank was purified with lactic acid bacteria.

<Test Example 9> Manufacture of fermented feed from various mushroom waste media

If fermented feed is produced using mushroom waste media generated after cultivation of various mushrooms and used as livestock feed, it can contribute to farm household income by reducing feed costs.

Vegetable lactic acid bacteria Lactobacillus fermentum JS1 raw powder and yeast Saccharomyces cerevisiae raw powder are mixed with glucose to make lactic acid bacteria 1 × 10 ⁷ cfu/g, yeast 1 × 10 ⁶ cfu/g, and 0.2% of the mushroom waste medium is dissolved in water. And used. When making fermented feed, 10% corn flour, 10% bran, 80% mushroom waste medium, and 0.2% lactic acid bacteria were put in a fermenter and fermented at 40-50℃ for 48 hours to become fermented feed.

Another method is to put 80% of waste mushroom medium, 10% of bran, and 10% of corn flour into a fermenter, sterilize with high-pressure steam at 121℃ for 30 minutes, cool the medium to 40℃, and mix 0.2% of lactic acid bacteria and yeast to 40~ Fermentation at 50° C. for 15-18 hours completed fermentation feed.

When breeding broilers, you can feed 90% of broiler mixed feed and 10% of fermented mushroom waste medium feed, and for pig farming, 80% of pig mixed feed and 20% fermented mushroom feed may be fed. In (from 5 months to 13 months of age), 40% of Korean beef mixed feed and 60% of fermented mushroom waste medium are mixed and fed.In the pre-foliation period (14 months to 21 months of age), 60% of mixed feed and 40% of mushroom waste medium are provided. It was mixed and fed, and at the end of fattening (from 22 to 29 months of age), 80% of the mixed feed and 20% of the mushroom waste medium were mixed and fed.

The production of fermented mushroom waste medium fermented cattle and raising feedstock can significantly contribute to farm household income by reducing feed costs by more than 30%.

Feeding fermented mushroom waste medium reduces the odor of poultry by about 70% during poultry breeding, and reduces the odor of pig manure by about 80% in the case of pig farming, and reduces the odor of cattle by 90% when rearing cattle. The outbreak of flies in pig sheds and barns is suppressed, so eco-friendly livestock can be achieved, and eco-friendly chicken, pork, and beef can be supplied without the use of antibiotics, thus providing eco-friendly livestock products to the public, which can greatly contribute to the improvement of national health.

When broilers are bred using fermented feed, inosic acid in chicken is doubled, pig pigs are 5 times increased in pork, and fattening cattle are tripled in beef. In this case, 90% of first-class meat is produced, which increases farm household income.

Mushroom waste medium is a mushroom mycelium, so there is no need to use antibiotics because beta-clucan, a natural antibiotic, is contained in the mushroom mycelium.

Mushroom medium mainly uses solid wood sawdust, and the wood sawdust contains lignin, so animals should not consume lignin, but only mushroom mycelium can decompose the lignin in the sawdust, so lignin is completely decomposed in the mushroom waste medium. It does not exist, and the mushroom waste medium is made of sawdust and mushroom mycelium in which lignin is completely decomposed.

Mushroom waste medium fermented feed composition analysis table Ingredient name unit Analysis moisture % 56.11 Crude protein % 6.41 Crude Fat (F.E) % 2.76 Crude fiber % 12.58 Minutes % 3.17 Soluble Nitrogen % 19.76 calorie Kcal/kg 2,150 carbohydrate % 31.95 leaven cfu/g 2.2 × 10 ⁷ Lactobacillus cfu/g 5.4 × 10 ⁸

<Test Example 10> Microbial organic fertilizer manufacturing

Vegetable lactic acid bacteria Lactobacillus fermentum JS1 raw powder and yeast Saccharomyces cerevisiae raw powder are mixed with glucose to make lactic acid bacteria 1 × 10 ⁷ cfu/g, yeast 1 × 10 ⁶ cfu/g, and then 50% chicken meal and 50% sawdust are composted. After mixing in a fermentation plant, 0.2% of the amount of chicken meal and sawdust was dissolved in water, and then fermented for 30 days using a compost fermenter after dissolving 0.2% of the amount of chicken meal and sawdust in water. During fermentation of organic compost, the fermentation temperature was 85℃, but the lactic acid bacteria and yeast were not killed.

When growing crops, cultivate organic compost with lactic acid bacteria for growing vegetables such as spinach carrots, cabbages, radishes, and peppers per danbo, and 4,000kg when growing watermelons, melons, cucumbers, and eggplants, which are fruits and vegetables, and grind with a cultivator. It was cultivated by transplanting seeds or seedlings.

Then, since anthrax, fetopathy, sclerotic disease, plague, anti-arritic disease, blue-going death, etc. rarely occur, agricultural products can be produced without using pesticides. Lactobacillus secretes lactic acid from pathogens in the soil to kill pathogens, and lactic acid bacteria secrete lactic acid from the eggs of pests in the soil to dissolve the shells of pests, thereby inhibiting the occurrence of pests. And when aphids occur in crops, lactic acid bacteria and yeast powder are made 100 times and sprayed, and lactic acid bacteria secrete lactic acid and kill aphids.

Lactobacillus Organic Fertilizer Component Analysis Table Analysis item unit Analysis nitrogen % 0.40 Organic matter % 31.71 Phosphoric acid (P ₂ O ₅ ) % 0.78 Girly (K ₂ O) % 0.75 Number of lactic acid bacteria cfu/g 1.2 × 10 ⁶ Yeast bacteria count cfu/g 2.3 × 10 ⁵

<Test Example 11> Water quality improvement method

Vegetable lactic acid bacteria Lactobacillus fermentum JS1 raw powder and yeast Saccharomyces cerevisiae raw powder are mixed with glucose to make lactic acid bacteria 1 × 10 ⁷ cfu/g, yeast 1 × 10 ⁶ cfu/g, and then 90% cedar sawdust and 10% bran are mixed. After making the moisture to 50%, add 1% of the above-described lactic acid bacteria and yeast, and then ferment at a fermentation temperature of 40 to 45°C for 3 days, then lactic acid bacteria 1.5 × 10 ⁷ cfu/g, yeast 2 × 10 ⁶ It became a cfu/g composition.

80% of this composition, 10% of charcoal powder, and 10% of white clay are mixed, put in a mesh bag that does not leak sawdust, and put it in a plastic box that is open in all directions. It was installed so that water passed through the plastic box. It was installed in two places at 20m intervals as shown in FIG.

Since livestock wastewater is mixed and flowed from the upper stream in the small river, the water quality is contaminated by nitrogen and ammonia components in the flowing water. Because of the purification, the water quality has been improved, and moss does not occur on the bottom of the river, and the odor is removed from the water, and the water quality is purified.

In addition, if you put cedar sawdust fermented with lactic acid bacteria in a water tank under the air-conditioning water cooling tower on the roof of a large building and put it in a water-conducting mesh bag and immerse it in the cooling tower water tank, it kills germs in the water tank and does not produce odor. If you look at the water tank of the air conditioner water cooling tower of all large buildings, you can see that bacteria multiply, smell a lot, and the water is very turbid. If you use the above method, the water quality of the water tank is improved.

In addition, a lot of water stains are attached to the bathtub or floor tiles of a private home, but it is difficult to remove. Lactobacillus 1 × 10 ⁷ cfu/g and yeast 1 × 10 ⁶ cfu/g are dissolved in water with 100 times the amount of water and Spraying on the tile keeps it clean because it doesn't get scaled, but you can spray it 3-4 times a month.

Small river water quality analysis table Upper stream of small river 4km downstream from the upstream 8 km downstream from the upstream 1km upstream from the experimental waterway 200m downstream from the test channel 500m downstream from the test channel pH 6.3 7.2 7.6 7.9 7.2 7.1 TH 2 * 2 * 3.2＊ 4.8 ＊ 2.8 * 2 * NH ₃ 0.0ppm 0.2ppm 0.2ppm 0.23ppm 0.23ppm 0.0ppm NO ₃ 5mg/ℓ 11.5mg/ℓ 48.0mg/ℓ 48.0mg/ℓ 23.0mg/ℓ 23mg/ℓ NO ₂ 0.1mg or more 0.1mg or more 0.13mg or more Less than 0.1mg Less than 0.1mg Less than 0.1mg CO ₂ 3 mg 3 mg 5 mg 8 mg 5 mg 3 mg O ₂ 12mgO ₂ /ℓ 12mgO ₂ /ℓ 7mgO ₂ /ℓ 4mgO ₂ /ℓ 10mgO ₂ /ℓ 710mgO ₂ /ℓ COD
ppm 2mgO/ℓ 3mgO/ℓ 3mgO/ℓ 5mgO/ℓ 2mgO/ℓ 2mgO/ℓ

Water quality inspection items

pH (acidity) NH ₃ (ammonia concentration) NO ₂ (nitrogen acid) O ₂ (oxygen) TH (total hardness)

NO ₃ (acetate) CO ₂ (carbon dioxide) COD(ppm)

<Test Example 12> Method for producing lactic acid bacteria rice

Lactobacillus rice can be prepared by spray-spraying the plant-borne lactic acid bacteria mutant strain on rice, and when cooked using the lactic acid bacteria rice prepared in this way, the plant lactic acid bacteria did not die and grew to about 250 times the number of lactic acid bacteria in the lactic acid bacteria rice.

In the present invention, the culture medium of the vegetable lactic acid bacteria rice was cultured using soybean flour, rice bran, and glutinous rice in drinking water rather than using the above-described medium. Specifically, 100 to 130 g of soy flour, 40 to 50 g of rice bran, and 15 to 20 g of glutinous rice are added to 1 liter of water, sterilized at 121°C, cooled, and inoculated with lactic acid bacteria and yeast inoculum, and incubated at 39 to 42°C for 15 to 18 hours. After the medium was completely filtered, the lactic acid bacteria were used as lactic acid bacteria by spraying rice.

For example, in the case of 10 liters of lactic acid bacteria culture, add 10 liters of medium to 3 liters of drinking groundwater, sterilize at 121°C for 20 minutes, supplement 7 liters of drinking groundwater, and inoculate 100-130 ml of lactic acid bacteria/yeast bacteria inoculum. After incubation at ~42°C for 15-18 hours, cooling, and filtering the medium, it was used as a lactic acid bacteria rice spray.

The reason for using glutinous rice as a medium raw material is that glutinous rice contains a large amount of methionine, so that lactic acid bacteria use methionine, and the reason for supplementing drinking groundwater is that drinking groundwater contains a large amount of minerals. Since the mineral components are destroyed, the lactic acid bacteria cannot use the mineral components, so after sterilization, drinking groundwater is supplemented and cultured so that the lactic acid bacteria can sufficiently use the mineral components containing various components.

The manufacturing method of lactic acid bacteria rice is to be packaged in packaging after spraying on the polished rice with a spraying device just before the milling is completed and put in the packaging paper. The lactic acid bacteria sprayed rice has a long preservation period and maintains freshness.

The number of cultured lactic acid bacteria is 2 to 4 × 10 ⁹ cfu/g and the number of yeast bacteria is 1 to 1.4 × 10 ⁹ cfu/g. The packaging was finished after spraying 100 kg of rice with 1 liter of lactic acid bacteria. When cooking rice with lactic acid bacteria, put the rice in water and lightly filter the rice in water without rubbing it with your hands. Put it in a rice cooker and mix with water. When lactic acid bacteria are added to water and rubbed thinly, the number of lactic acid bacteria attached to the rice is washed away, reducing the number of lactic acid bacteria. When rice is cooked with lactic acid bacteria in a pressure cooker, the lactic acid bacteria multiply without dying even at high temperatures and grow to about 250 times the number of lactic acid bacteria contained in the rice.

4 and 5 of the present invention are the vegetable lactic acid bacteria Lactobacillus fermentum of the present invention JS1 Raw horse and yeast Saccharomyces cerevisiae This is a test report showing the content of each strain contained in rice containing raw malt.

Above, the present invention has been described in detail with reference to a preferred embodiment, but the present invention is not limited to the above embodiment, and various modifications by those of ordinary skill in the art within the scope of the technical idea of the present invention This is possible.

Korea Microorganism Conservation Center KCCM80171 20171026

<110> CHOI, Jung Sik CHOI, Byung Chul CHOI, Byung Sun <120> Mutant lactic acid bacterium strain having acid and heat resistance, and culturing composition comprising this strain as effective gredient <130> 2018TE-0012 <160> 1 <170> KopatentIn 2.0 <210> 1 <211> 1510 <212> DNA <213> Lactobacillus fermentum 16S rRNA <400> 1 ctgtcaggat gaacgccggc ggtgtgccta atacatgcaa gtcgaacgcg ttggtccaat 60 tgattgatgg tgcttgcacc tgattgattt tggtcgccaa cgagtggcgg acgggtgagt 120 aacacgtagg taacctgccc agaagcgggg gacaacattt ggaaacagat gctaataccg 180 cataacaacg ttgttcgcat gaacaacgct taaaagatgg cttctcgcta tcacttctgg 240 atggacctgc ggtgcattag cttgttggtg ggtaatggcc taccaaggcg atgatgcata 300 gccgagttga gagactgatc ggccacaatg ggactgagac acggcccata ctcctacggg 360 aggcagcagt agggaatctt ccacaatggg cgcaagcctg atggagcaac accgcgtgag 420 tgaagaaggg tttcggctcg taaagctctg ttgttaaaga agaacacgta tgagagtaac 480 tgttcatacg ttgacggtat ttaaccagaa agtcacggct aactacgtgc cagcagccgc 540 ggtaatacgt aggtggcaag cgttatccgg atttattggg cgtaaagaga gtgcaggcgg 600 ttttctaagt ctgatgtgaa agccttcggc ttaaccggag aagtgcatcg gaaactggat 660 aacttgagtg cagaagaggg tagtggaact ccatgtgtag cggtggaatg cgtagatata 720 tggaagaaca ccagtggcga aggcggctac ctggtctgca actgacgctg agactcgaaa 780 gcatgggtag cgaacaggat tagataccct ggtagtccat gccgtaaacg atgagtgcta 840 ggtgttggag ggtttccgcc cttcagtgcc ggagctaacg cattaagcac tccgcctggg 900 gagtacgacc gcaaggttga aactcaaagg aattgacggg ggcccgcaca agcggtggag 960 catgtggttt aattcgaagc tacgcgaaga accttaccag gtcttgacat cttgcgccaa 1020 ccctagagat agggcgtttc cttcgggaac gcaatgacag gtggtgcatg gtcgtcgtca 1080 gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc gcaacccttg ttactagttg 1140 ccagcattaa gttgggcact ctagtgagac tgccggtgac aaaccggagg aaggtgggga 1200 cgacgtcaga tcatcatgcc ccttatgacc tgggctacac acgtgctaca atggacggta 1260 caacgagtcg cgaactcgcg agggcaagca aatctcttaa aaccgttctc agttcggact 1320 gcaggctgca actcgcctgc acgaagtcgg aatcgctagt aatcgcggat cagcatgccg 1380 cggtgaatac gttcccgggc cttgtacaca ccgcccgtca caccatgaga gtttgtaaca 1440 cccaaagtcg gtggggtaac cttttaggag ccagccgcct aaggtgggac agatgattag 1500 ggtgaagtct 1510

Claims (11)

Lactobacillus fermentum ( Lactobacillus fermentum ) JS1 lactic acid bacteria mutant strain (Accession No. KCCM-80171) having heat resistance and acid resistance having 16S rRNA encoded by the DNA of SEQ ID NO: 1.
A lactic acid bacteria culture composition comprising the Lactobacillus fermentum JS1 mutant strain of claim 1 having a function of lowering blood sugar, antioxidant, lipid peroxidation, and lowering cholesterol as an active ingredient.
The method of claim 2, wherein the composition comprises a Lactobacillus fermentum JS1 lactic acid bacteria mutant strain of 20 to 30 g of glucose per 1 liter of water, 10 to 20 g of yeast extract, 5 to 8 g of soypeptone, 3 to 5 g of casein, MgSO ₄ .7H ₂ O 0.1~0.15g, K ₂ HPO ₄ 1~2g, MnSO ₄ .5H ₂ O 0.05~0.1 g, sodium acetate 3~5g, ammonium citrate 2~3g, Tween 80 1~ Lactic acid bacteria culture composition, characterized in that produced by culturing in a medium mixed with 2g, neoline (neoline) 0.25 ~ 0.3g.
According to claim 2, wherein the composition is a yeast Saccharomyces cerevisiae (Saccharomyces cerevisiae) strain per 1 liter of water protease peptone (protease peptone) No3 5 ~ 10g, malt extract (malt extract) 3 ~ 5g, Lactobacillus culture composition, characterized in that it further comprises a yeast culture solution cultured in a medium containing 10 to 20 g of glucose, 10 to 15 g of yeast extract, 5 to 10 g of sodium acetate, and 3 to 5 g of ammonium citrate.
An auxiliary feed composition usable for livestock or fish farming using the lactic acid bacteria culture composition of claim 3 or 4.
A method for producing fermented feed for mushroom waste medium using the lactic acid bacteria culture composition of claim 3 or 4.
A method for producing organic fertilization, characterized in that the lactic acid bacteria culture composition of claim 3 or 4 is added to livestock meal.
A soil improvement method, characterized in that the organic additional fertilization of claim 7 is mixed in the soil.
A method for improving water quality, characterized in that the lactic acid bacteria culture composition of claim 3 or 4 is placed in a mesh bag or plastic box through which water passes, and installed in flowing water or a water tank.
A method for producing lactic acid bacteria-containing rice, characterized in that spraying and spraying the lactic acid bacteria mutant strain of claim 1 on rice.
Rice containing lactic acid bacteria produced by the method of claim 10.

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