KR101034467B1 - Microorganism cultivating medium containing enzymatic hydrolysate of wheat bran and konjak - Google Patents

Abstract

PURPOSE: A medium containing hydrolysate of wheat bran and konjac for culturing microorganism is provided to ensure excellent productivity of cellulase, xylanase, and mannanase. CONSTITUTION: A medium for culturing microorganism of Bacillus sp., Aspergillus sp., or Lactobacillus sp. contains 0.1-5 weight% of wheat bran hyrolysate and 0.1-5 wight% of konjac hydrolysate. The medium is LB, PD, or PY medium. A feed additive for livestock contains microorganism or culture thereof.

Description

Microorganism cultivating medium containing enzymatic hydrolysate of wheat bran and konjak}

The present invention relates to a culture medium for culturing microorganisms, and more specifically, a high-temperature concentrated and decomposed product obtained by treating wheat bran or konjac with metase and cellulase or xylanase and cellulase, respectively, and then prepared for powder growth. Regarding a medium for culturing microorganisms containing bran and konjac enzymatic hydrolyzate of bran which have an effect of enhancing the production and viable cell count of enzymes such as metase, cellulase and xylanase of microorganisms in culture of microorganisms in addition to the medium will be.

As domestic and international interest in food safety increases, efforts are being made to produce high quality agricultural products in an eco-friendly manner. In reality, the issue of the persistence of the antimicrobial feed additives and the appearance of resistant bacteria added for the purpose of promoting livestock growth and improving feed efficiency has been the subject of debate, and the use of antibiotics has been realized. For this reason, it is urgent to develop useful resources to replace these antimicrobial feed additives, and for this, research on probiotics, prebiotics, enzymes, etc., is being actively conducted.

The intestinal flora of livestock is closely related to the productivity of livestock, and the maintenance of the intestinal flora is directly related to the prevention of livestock disease, the reduction of antibiotic use, and the improvement of productivity. Is done.

Probiotic means a microbial agent that functions usefully in the digestive tract of livestock and attracts much attention because microorganisms inhibit harmful microorganisms, maintain microbial balance, improve health and contribute to animal nutrition in the intestines of animals. . Basically, probiotics eliminate the harmful microbes that reach the digestive tract and grow in the digestive tract, and propagate useful bacteria such as lactic acid bacteria to restore and maintain the balance of the normal intestinal flora.

Currently, Bacillus, lactic acid bacteria, and yellow bacteria are commercially developed as a probiotic with yeast. Among these probiotic strains, Bacillus and Rhesus bacteria produce enzymes that can degrade feed-ingradable components in the intestine and culture of livestock.

In particular, Bacillus subtilis and Kookgi bacteria produce degrading enzymes of fibrous and semifibrous materials, which are contained in large amounts in the feed, and by hydrolyzing the cellulose and hemicellulose layers that surround the grains, the livestock easily nutrients in the grains. In addition to making it available, it has the effect of improving the intestinal digest of livestock. Feeding with high hemicellulose content tends to lead to stool or swelling in livestock. Bacillus subtilis and erythrocytes, which produce xylanase, mannase and cellulase, can prevent this and increase the value of feed. As the quality of wheat products is improved by improving the quality variation of wheat products, they can be used as a stable feedstock, so the value of these enzymes is high. It also has the effect of normalizing microorganisms in general, suppressing digestive diseases of harmful bacteria through the production of soft organic acids and replacing some antibiotics.

Lactobacillus is a safe bacterium that does not have a big problem to use as a live bacterium except in special cases because it does not give an unusual pathogenicity to animals or humans. Lactobacillus is known as a pronoun of probiotics, among which Lactobacillus is a gram-positive lactobacillus, which is a representative dominant bacteria in the intestines of animals. It is estimated that Lactobacillus is a microorganism that plays a very important role in the health of animals because it is detected in high numbers in healthy animals and low in sick animals.

Therefore, various studies have been made on the probiotics produced using the microorganisms having the above-mentioned efficacy, and in particular, Korean Patent Registration No. 439635 is a cell culture mixture obtained by culturing Bacillus, Lactobacillus, Aspergillus, etc. Microbial agent for the addition of livestock feed, which is effective in reducing the odor of livestock manure and the increase of livestock composed of a mixture of a mixture and an excipient (bran, soybean meal, barley stone), Korean Patent Registration No. 306386, Lactobacillus, Aspergillus Microbial preparation consisting of a culture cultured by mixing and formulating vermiculite and rice bran in a culture medium, Korean Patent Registration No. 396022, Koji containing crab or shrimp shell after first fermentation by mixing a liquid microbial culture Method for producing livestock probiotics prepared by secondary fermentation and drying in medium, Korean people Patent registration No. 944757 is a livestock probiotic fermented by adding functional foods such as jujube, wolfberry and garlic, and microbial substrates such as wood vinegar liquor and chitosan into water and administering fibrin-degrading microorganisms, acid-resistant filamentous bacteria and lactic acid bacteria, and Korean Patent Registration No. No. 936015 discloses a feed additive for livestock including a probiotic which survives after culturing in a medium containing garlic components prepared by adding garlic extract to MRS medium and garlic by-products consisting of stems, roots and epidermis of garlic. have.

However, the probiotic as described above has a problem in that the productivity of the useful enzyme is reduced or the productivity of the useful enzyme is increased, but the cell growth is lowered while increasing the bacterial growth of Bacillus or fungi depending on the composition of the culture medium.

Therefore, the present inventors have studied the medium to increase the useful enzyme productivity and viable cell number of microorganisms, microorganisms of wheat bran and konjac enzyme hydrolysate obtained by using bran or konjac as xylanase and cellulase or metase and cellulase The present invention was completed by confirming that the effect of increasing the useful enzyme productivity and the number of microorganisms when culturing microorganisms in the culture medium for microorganisms added to the medium for growth was excellent.

Accordingly, an object of the present invention is to provide a medium for culturing microorganisms which can increase the enzyme productivity and the number of microorganisms by additionally adding an enzyme hydrolyzate of bran or konjac to the medium for microbial growth.

Furthermore, another object of the present invention is to provide a method for culturing microorganisms using the microorganism culture medium.

The medium for culturing the microorganism according to the present invention for solving the above problems is a medium in which one or more of 0.1 to 5% by weight of the bran enzyme hydrolyzate and 0.1 to 5% by weight of the konjac enzyme hydrolyzate are added to the medium for microbial growth. Include.

The bran enzyme hydrolyzate suspends the bran in a 0.01 to 0.10 weight ratio compared to 10 mM sodium phosphate buffer (pH 6.0) and simultaneously adds cellulase and xylanase at 800 to 1000 U and leaves it at 40 to 60 ° C. for 2 to 4 hours. After carrying out the hydrolysis reaction, vacuum dried and powdered at 70 ~ 90 ℃ to use the prepared, konjac enzyme hydrolyzate suspended konjac in a 0.01 ~ 0.10 weight ratio to 10mM sodium phosphate buffer (pH 6.0) and met with nanase The cellulase is added at a time of 800 ~ 1000U at the same time and the hydrolysis reaction of bran while leaving for 2 to 4 hours at 40 ~ 60 ℃, vacuum dried and powdered at 70 ~ 90 ℃ is used.

When the amount of added bran and konjac hydrolyzate is less than 0.1% by weight, respectively, the enzyme productivity and viable cell number of microorganisms are low, and when it is more than 5%, there is no significant difference in enzyme productivity and viable cell number.

The production of polysaccharide degrading enzymes of microorganisms is often affected by the carbon source in the culture. Rather than inducing direct enzyme production of polysaccharides, enzyme production can be induced by their hydrolyzed low sugars. However, in the presence of monosaccharides or disaccharides such as glucose, fructose and sucrose, which are favorably used by microorganisms, polysaccharide degrading enzyme production by polysaccharide addition is not effectively induced.

Therefore, in order to induce the hydrolase production of fibrous or semifibrous polysaccharides, it is advantageous not to add monosaccharides and disaccharides which are readily available to microorganisms to the medium, but the growth of microorganisms does not increase unless these are added. Occurs. Therefore, in order to increase the growth of microbial cells and increase the production of enzymes, polysaccharides are treated with enzymes and converted to low saccharides, and when used together as an additional carbon source, enzyme production and viable cell number can be increased.

Wheat bran contains a large amount of fibrin and semifibrous polysaccharide xylan, and konjac is a meetable polysaccharide, in which mannose and glucose are linked by beta 1,4 bonds, which are usually insoluble and randomly form beta 1,4 bonds. When hydrolyzed with one or more enzymes selected from endo enzymes such as xylanase, cellulase or mannanase enzymes, it is possible to convert to highly soluble low sugars, but very small amounts of glucose, mannose and xylose Since only the production of the enzyme hydrolyzate of bran and konjac by the production method as described above when added to the medium for the growth of microorganisms can increase the amount of enzyme production and viable bacteria.

The medium for the growth of the microorganism may be at least one selected from the group consisting of LB medium, PD medium and PY medium, in addition, it may be a medium commonly used in culture of microorganisms.

Microbial culture medium containing bran and konjac enzymatic hydrolyzate according to the present invention is sterilized for 15 minutes in a high pressure steam sterilizer at 121 ℃ in a flask or fermenter and inoculated with the microorganisms after inoculating the microorganisms in a sufficient amount of air for 24 to 72 hours Can be cultured with shaking or aeration so that is supplied.

The microorganism may be at least one selected from the group consisting of Bacillus, fungi and lactic acid bacteria.

Bacillus subtilis ( Bacillus subtilis ), Bacillus lentus ( Bacillus lentus ), Bacillus licheniformis ( Bacillus licheniformis ), Bacillus coagulans ( Bacillus coagulans ), Bacillus pumilus ( Bacillus pumilus ) and Bacillus sere At least one selected from the group consisting of Bacillus cereus .

The fungus may be at least one selected from the group consisting of Aspergillus oryzae , Aspergillus sojae , and Aspergillus niger .

The lactic acid bacteria is Lactobacillus casei (Lactobacillus casei), Lactobacillus Planta room (Lactobacillus plantarum), Lactobacillus Bulgaria kusu (Lactobacillus bulgaricus), Lactobacillus ramno Versus (L actobacillus rhamnosus), Lactobacillus know also required Ruth (Lactobacillus acidophilus ), Lactobacillus reuteri , Lactobacillus fermentum , Lactobacillus gasseri , Lactobacillus brevis and Lactobacillus pentosus ( Lactobacillus pentosus ) It may be one or more kinds.

In the medium for culturing the microorganism of the present invention, yeast and photosynthetic bacteria may be cultured in addition to Bacillus, fungi and lactic acid bacteria.

The yeast may be at least one selected from the group consisting of Saccaromyces cerevisiae , Saccharomyces ellipsoidus and Saccaromyces bayanus .

The photosynthetic bacteria may be one or more selected from the group consisting of red sulfur bacteria, purple non-sulfur bacteria, and green sulfur bacteria.

Since the culture medium for microbial culture of the present invention does not contain animal-derived components, the cultured microbial culture using the same is prepared by a method of preparing a conventional livestock feed and then administered to the animals alone or mixed as an additive for livestock feed. Depending on the characteristics of the culture, can be fed to livestock such as cattle, horses, pigs, poultry such as ducks, chickens, turkeys, etc. In addition, it is prepared by the conventional method for improving soil, lettuce, cabbage, cabbage, radish It can also be used as soil improvement agent for crop cultivation such as pepper and pepper.

In the culture medium for microbial culture according to the present invention, by adding an enzyme hydrolyzate of bran or konjac to the medium for growing microorganisms, cellulase, xylanase and mannanase which can decompose fibrous and semifibrous materials during microbial culture. It is excellent in productivity and cell growth, and it is a medium prepared by adding bran or konjac hydrolyzate of enzyme and hydrolyzate of konjac. It is easy.

In addition, when the microbial cells cultured in the microorganism culture medium according to the present invention and its culture solution as animal feed additives are used as the main components of the feed, it is possible to efficiently decompose the fibrous and semi-fibrous components that are difficult to degrade in the body of the livestock. It improves feed efficiency of livestock animals, maintains the intestinal flora of livestock and prevents digestive diseases. It also helps to grow soil microorganisms by decomposing fibrous and semi-fibrous carbohydrates in soil. The effect is to improve the soil.

Hereinafter, preferred embodiments of the present invention will be described. However, the present invention is not limited to the embodiments described herein and may be embodied in other forms.

Example 1 Preparation of Bran Enzyme Hydrolysate

5 g of bran was suspended in 100 ml of 10 mM sodium phosphate buffer (pH 6.0), and cellulase and xylanase were added simultaneously at 1000 U and left for 3 hours at 50 ° C. to carry out hydrolysis of bran, followed by vacuum drying at 80 ° C. and powdering. The bran enzyme hydrolyzate was prepared.

Example 2 Preparation of Konjac Enzyme Hydrolysate

Suspend 5g konjac in 100ml of 10mM sodium phosphate buffer (pH 6.0), add 1000M each of mannanase and cellulase, and leave it at 50 ℃ for 3 hours to perform hydrolysis of konjac and vacuum-dry the reaction solution at 80 ℃. Powdered to prepare konjac hydrolyzate.

Example 3 Preparation of Medium for Culture of Microorganisms Containing 0.1% Bran Enzyme Hydrolysate

 After adding 0.1% of the bran enzyme hydrolyzate prepared in Example 1 to the LB medium and sterilizing for 15 minutes in a 121 ℃ autoclave to prepare a medium for culturing microorganisms containing bran enzyme hydrolyzate.

<Examples 4 to 47> Preparation of microorganism culture medium

In the same manner as in Example 3, except that the medium for culturing the microorganisms of Examples 4 to 47 under the same conditions as in Table 1.

<Comparative Examples 1 to 21> Preparation of culture medium for microbial culture

In the same manner as in Example 3, but was prepared in the culture medium for the culture of Comparative Examples 1 to 21 under the same conditions as in Table 1. In Comparative Examples 5, 12 and 19, bran and konnyaku which were not enzymatically treated were used, respectively.

division
Badge type Addition amount (% by weight) Bran Enzyme Hydrolysate Konjac Enzyme Hydrolysate Other additives Example 3 LB badge 0.1 - - Example 4 LB badge 0.3 - - Example 5 LB badge One - - Example 6 LB badge 3 - - Example 7 LB badge 5 - - Example 8 LB badge - 0.1 - Comparative Example 9 LB badge - 0.3 - Example 10 LB badge - One - Example 11 LB badge - 3 - Example 12 LB badge - 5 - Example 13 LB badge 0.1 0.1 - Example 14 LB badge 0.15 0.15 - Example 15 LB badge 0.5 0.5 - Example 16 LB badge 1.5 1.5 - Example 17 LB badge 2.5 2.5 - Example 18 PD badge 0.1 - - Example 19 PD badge 0.3 - - Example 20 PD badge One - - Example 21 PD badge 3 - - Example 22 PD badge 5 - - Example 23 PD badge - 0.1 - Example 24 PD badge - 0.3 - Example 25 PD badge - One - Example 26 PD badge - 3 - Example 27 PD badge - 5 - Example 28 PD badge 0.1 0.1 - Example 29 PD badge 0.15 0.15 - Example 30 PD badge 0.5 0.5 - Example 31 PD badge 1.5 1.5 - Example 32 PD badge 2.5 2.5 - Example 33 PY Badge 0.1 - - Example 34 PY Badge 0.3 - - 35 on real PY Badge One - - Example 36 PY Badge 3 - - Example 37 PY Badge 5 - - Example 38 PY Badge - 0.1 - Example 39 PY Badge - 0.3 - Example 40 PY Badge - One - Example 41 PY Badge - 3 - Example 42 PY Badge - 5 - Example 43 PY Badge 0.1 0.1 - Example 44 PY Badge 0.15 0.15 - Example 45 PY Badge 0.5 0.5 - Example 46 PY Badge 1.5 1.5 - Example 47 PY Badge 2.5 2.5 - Comparative Example 1 LB badge - - - Comparative Example 2 LB badge - - Glucose 0.3 Comparative Example 3 LB badge - - Mannose 0.3 Comparative Example 4 LB badge - - Xylose 0.3 Comparative Example 5 LB badge Bran 0.1 Konjac 0.1 - Comparative Example 6 LB badge 0.05 0.05 - Comparative Example 7 LB badge 8 8 - Comparative Example 8 PD badge - - - Comparative Example 9 PD badge - - Glucose 0.3 Comparative Example 10 PD badge - - Mannose 0.3 Comparative Example 11 PD badge - - Xylose 0.3 Comparative Example 12 PD badge Bran 0.1 Konjac 0.1 - Comparative Example 13 PD badge 0.05 0.05 - Comparative Example 14 PD badge 8 8 - Comparative Example 15 PY Badge - - - Comparative Example 16 PY Badge - - Glucose 0.3 Comparative Example 17 PY Badge - - Mannose 0.3 Comparative Example 18 PY Badge - - Xylose 0.3 Comparative Example 19 PY Badge Bran 0.1 Konjac 0.1 - Comparative Example 20 PY Badge 0.05 0.05 - Comparative Example 21 PY Badge 8 8 -

Test Example 1 Evaluation of Enzyme Productivity and Bioburden of Microorganisms According to Enzymatic Hydrolysates of Bran and Konjac

Bacillus bacteria

Comparison of enzyme productivity and viable cell count of Bacillus bacteria according to Examples 3 to 17 (medium hydrolyzate of bran and konnyaku are added or mixed to LB medium) and microorganism culture medium prepared in Comparative Examples 1 to 7 Evaluation was made.

After inoculating the microbial culture medium prepared in Examples 3 to 17 and Comparative Examples 1 to 7 in a flask and inoculated Bacillus subtilis in each medium and incubated for 24 hours at 35 ℃, centrifugal Separated (10 minutes at 13000rpm) and the supernatant was taken to measure the production of cellulase, mannanase and xylanase and compared and evaluated, and the number of viable cells cultured after culturing Bacillus subtilis in the same manner as described above Comparative evaluation was made. Enzyme production and viable cell counts are shown in Table 2 below.

division
Enzyme Production (unit / mL) Viable count
(CFU / mL) Cellulase Met meet Xylanase Example 3
(0.1% bran enzyme hydrolyzate) 1.3 2.8 1.8 6.5 × 10 ⁸ Example 4
(0.3% bran enzyme hydrolyzate) 2.5 3.7 2.1 1.7 × 10 ⁹ Example 5
(1% of bran enzyme hydrolyzate) 5.9 6.0 5.5 2.6 × 10 ⁹ Example 6
(3% of bran enzyme hydrolyzate) 6.7 6.5 5.8 3.2 × 10 ⁹ Example 7
(5% of bran enzyme hydrolyzate) 6.9 6.4 5.7 3.5 × 10 ⁹ Example 8
(Konjac enzyme hydrolyzate 0.1%) 1.3 28.7 1.7 6.6 × 10 ⁸ Example 9
(0.3% of konjac enzyme hydrolyzate) 1.7 21.5 2.2 1.6 × 10 ⁹ Example 10
(1% Konjac Enzyme Hydrolyzate) 2.5 35.2 3.3 2.5 × 10 ⁹ Example 11
(3% Konjac Enzyme Hydrolyzate) 2.9 38.3 3.7 3.2 × 10 ⁹ Example 12
(5% Konjac Enzyme Hydrolyzate) 3.3 39.7 3.5 3.4 × 10 ⁹ Example 13
(Of bran and konjac
0.1% of each enzyme hydrolyzate)
3.0
36.2
3.9
1.1 × 10 ⁹ Example 14
(Of bran and konjac
0.15% of each enzyme hydrolyzate)
3.5
48.2
4.8
1.7 × 10 ⁹ Example 15
(Of bran and konjac
0.5% of each enzyme hydrolyzate)
7.8
55.1
7.5
2.6 × 10 ⁹ Example 16
(Of bran and konjac
1.5% of enzyme hydrolyzate)
8.5
59.3
8.6
3.1 × 10 ⁹ Example 17
(Of bran and konjac
2.5% of each enzyme hydrolyzate)
8.7
60.5
8.9
4.6 × 10 ⁹ Comparative Example 1 (no addition) 0.2 1.1 0.3 2.0 × 10 ⁸ Comparative Example 2 (0.3% glucose) 0.3 1.2 0.3 1.8 × 10 ⁹ Comparative Example 3 (0.3% mannose) 0.3 1.2 0.3 1.7 × 10 ⁹ Comparative Example 4 (0.3% xylose) 0.2 1.1 0.3 6.1 × 10 ⁸ Comparative Example 5
(0.1% bran and konjac each) 0.6 1.7 0.7 2.8 × 10 ⁸ Comparative Example 6
(Of bran and konjac
0.05% of each enzyme hydrolyzate)
0.4
1.3
0.7
4.3 × 10 ⁸ Comparative Example 7
(Of bran and konjac
8% each enzyme hydrolyzate)
8.5
59.1
8.5
3.7 × 10 ⁹

As shown in Table 2, 0.3% bran enzyme hydrolyzate (Example 4), konjac enzyme hydrolyzate 0.3% (Example 9) and 0.15% (Example 14) of enzyme hydrolyzate of bran and konjac, respectively, were added. Bacillus cultivated in bran and konjac hydrolyzate free (Comparative Example 1) Cellulase, metase and xylanase produced 9-18 times, 3-44 times and 7-16 times higher than Bacillus cultured in LB medium. Found to be high.

In addition, Bacillus bacteria cultured in LB medium containing 0.3% of glucose (Comparative Example 2), mannose (Comparative Example 3) and xylose (Comparative Example 4), respectively, produced cellulase, mannanase and xylanase, No addition of konjac enzyme hydrolyzate (Comparative Example 1) The results were similar to those of Bacillus cultured in LB medium.

Therefore, microbial culture medium prepared by adding one or more of 0.1 to 5% by weight of wheat bran and 0.1 to 5% by weight of konjac hydrolyzate according to the present invention together with cellulase, mannanase and xylanase productivity in Bacillus culture. The viable cell count was confirmed to be excellent.

On the other hand, when konjac hydrolyzate is added, the amount of nanase produced by Bacillus subtilis is relatively higher than that of cellulase and xylanase. This is because the mannase produced by Bacillus subtilis is the enzyme hydrolyzate of konjac (low sugar). ) Is expected to induce production more effectively.

2. mildew

Comparison of enzyme productivity and viable cell count of Bacillus bacteria according to Examples 18 to 32 (medium added with bran and konjac hydrolyzate to PD medium, respectively, or mixed) and microbial culture medium prepared in Comparative Examples 8-14 Evaluation was made.

After inoculating the microbial culture medium prepared in Examples 18 to 32 and Comparative Examples 8 to 14 in a flask and inoculated each medium with Aspergillus oryzae incubated at 35 ℃ for 72 hours, centrifugation cellulase and taking the supernatant (10 minutes at 13000rpm), met kinase and xylene Rana was measured Comparative evaluation by the dehydratase production, and then culturing the Aspergillus duck material (Aspergillus oryzae) in the same manner as the number of cultured bacteria Was measured and compared. Enzyme production and viable cell counts are shown in Table 3 below.

division
Enzyme Production (unit / mL) Viable count
(CFU / mL) Cellulase Met meet Xylanase Example 18
(0.1% bran enzyme hydrolyzate) 3.5 0.3 0.7 1.3 × 10 ⁸ Example 19
(0.3% bran enzyme hydrolyzate) 4.7 1.4 2.3 3.6 × 10 ⁸ Example 20
(1% of bran enzyme hydrolyzate) 6.3 2.5 3.7 4.4 × 10 ⁸ Example 21
(3% of bran enzyme hydrolyzate) 7.8 4.7 3.3 4.9 × 10 ⁸ Example 22
(5% of bran enzyme hydrolyzate) 7.9 4.9 3.6 5.3 × 10 ⁸ Example 23
(Konjac enzyme hydrolyzate 0.1%) 1.8 1.5 0.9 1.4 × 10 ⁸ Example 24
(0.3% enzyme hydrolyzate of konjac) 3.1 3.5 2.4 3.7 × 10 ⁸ Example 25
(1% Konjac Enzyme Hydrolyzate) 4.7 5.1 3.1 4.5 × 10 ⁸ Example 26
(3% Konjac Enzyme Hydrolyzate) 5.1 6.2 3.8 4.9 × 10 ⁸ Example 27
(5% Konjac Enzyme Hydrolyzate) 5.2 6.5 4.0 5.4 × 10 ⁸ Example 28
(Of bran and konjac
0.1% of each enzyme hydrolyzate)
6.7
3.5
2.5
2.5 × 10 ⁸ Example 29
(Of bran and konjac
0.15% of each enzyme hydrolyzate)
9.0
7.1
4.8
3.7 × 10 ⁸ Example 30
(Of bran and konjac
0.5% of each enzyme hydrolyzate)
9.9
7.3
5.6
4.5 × 10 ⁸ Example 31
(Of bran and konjac
1.5% of enzyme hydrolyzate)
11.1
8.7
6.8
5.0 × 10 ⁸ Example 32
(Of bran and konjac
2.5% of each enzyme hydrolyzate)
10.3
9.0
7.0
5.4 × 10 ⁸ Comparative Example 8 (no addition) 0.5 0.2 0.1 4.1 × 10 ⁷ Comparative Example 9 (0.3% glucose) 0.5 0.2 0.1 3.6 × 10 ⁸ Comparative Example 10 (Monose 0.3%) 0.5 0.2 0.1 3.5 × 10 ⁸ Comparative Example 11 (0.3% xylose) 0.5 0.2 0.1 3.2 × 10 ⁸ Comparative Example 12
(0.1% bran and konjac each) 1.4 0.5 0.4 4.8 × 10 ⁷ Comparative Example 13
(Of bran and konjac
0.05% of each enzyme hydrolyzate)
2.1
1.7
1.5
8.1 × 10 ⁸ Comparative Example 14
(Of bran and konjac
8% each enzyme hydrolyzate)
9.5
7.8
4.4
5.5 × 10 ⁸

As shown in Table 3, PD medium to which 0.3% bran enzyme hydrolyzate (Example 19), konjac enzyme hydrolyzate 0.3% (Example 24) and 0.15% of bran and konjac enzyme hydrolyzate (Example 29) were added, respectively. The fungus cultured on the bran was 6-18 times higher, 7-35 times higher and 23-48 times higher in cellulase, metase and xylanase production than molds cultured in wheat bran and konjac hydrolyzate (Comparative Example 8). appear.

In addition, the fungi cultured in PD medium containing 0.3% of glucose (Comparative Example 9), mannose (Comparative Example 10), and xylose (Comparative Example 11), respectively, produced cellulase, mannanase, and xylanase production from bran and konjac. No enzyme hydrolyzate (Comparative Example 8) The result was similar to that of the fungus cultured in PD medium.

Therefore, microbial culture medium prepared by adding one or more of 0.1 to 5% by weight of wheat bran and 0.1 to 5% by weight of konjac enzyme hydrolyzate according to the present invention is viable bacteria with cellulase, mannanase and xylanase productivity in mold culture. The number was confirmed to be excellent.

3. Lactobacillus

Examples 33 to 47 (medium added with bran and konjac hydrolyzate to PY medium, respectively, were added or mixed) and the degree of growth (lactic acid bacteria) of lactic acid bacteria according to the culture medium for microbial culture prepared in Comparative Examples 15 to 21 Comparative evaluation was made.

After injecting the microbial culture medium prepared in Examples 33 to 47 and Comparative Examples 15 to 21 into the flask and inoculated each Lactobacillus casei in each medium and incubated at 35 ℃ for 24 hours, spectrophotometer The absorbance at 600nm was measured and compared. Culture absorbance measurement results are shown in Table 4 below.

division Media absorbance Example 33 (0.1% bran enzyme hydrolyzate) 1.8 Example 34 (0.3% bran enzyme hydrolyzate) 2.1 Example 35 (1% Bran Enzyme Hydrolysate) 2.7 Example 36 (3% wheat bran enzyme hydrolyzate) 4.9 Example 37 (5% Konjac Enzyme Hydrolysate) 4.7 Example 38 (0.1% Konjac Enzyme Hydrolyzate) 2.1 Example 39 (0.3% konjac enzyme hydrolyzate) 2.5 Example 40 (1% Konjac Enzyme Hydrolysate) 3.2 Example 41 (of bran and konjac
3% of each enzyme hydrolyzate) 4.3 Example 42 (of bran and konjac
5% of each enzyme hydrolyzate) 4.1 Example 43 (of bran and konjac
0.1% of each enzyme hydrolyzate) 3.1 Example 44 (of bran and konjac
0.15% of each enzyme hydrolyzate) 4.9 Example 45 (of bran and konjac
0.5% of each enzyme hydrolyzate) 8.1 Example 46 (of bran and konjac
1.5% of enzyme hydrolyzate) 9.5 Example 47 (of bran and konjac
2.5% of each enzyme hydrolyzate) 10.3 Comparative Example 15 (No Addition) 0.3 Comparative Example 16 (0.3% glucose) 2.0 Comparative Example 17 (0.3% mannose) 1.5 Comparative Example 18 (0.3% xylose) 0.3 Comparative Example 19 (0.1% of bran and konnyaku each) 0.5 Comparative Example 20 (of bran and konjac
0.05% of each enzyme hydrolyzate) 1.3 Comparative Example 21 (of bran and konjac
8% each enzyme hydrolyzate) 10.3

As shown in Table 4, to the PY medium to which 0.3% bran enzyme hydrolyzate (Example 34), konjac enzyme hydrolyzate 0.3% (Example 39) and 0.15% bran and konjac enzyme hydrolyzate (Example 44) were added. The culture absorbance of the cultured lactic acid bacteria was 7-16 times higher than that of the culture solution of the lactic acid bacteria cultured in PY medium without the addition of bran and enzyme hydrolyzate of konjac.

In addition, lactic acid bacteria cultured in PY medium to which 0.3% bran enzyme hydrolyzate (Example 34) and konjac enzyme hydrolyzate 0.3% (Example 39) were added, were used only with glucose (Comparative Example 16) used as a carbon source of lactic acid bacteria. Similar to the absorbance of the culture of lactic acid bacteria cultured in PY medium containing 0.3% of Nose (Comparative Example 17), it is estimated that oligosaccharides exist that promote the growth of lactic acid bacteria in the enzyme hydrolyzate of bran and konjac.

On the other hand, the lactic acid bacteria cultured in PY medium containing 0.3% xylose (Comparative Example 18) showed similar values to the lactic acid bacteria cultured in PY medium containing no bran and konjac hydrolyzate (Comparative Example 15). .

Therefore, it was confirmed that the growth of lactic acid bacteria is excellent in the culture medium for microbial culture prepared by adding one or more of 0.1 to 5% by weight of wheat bran and 0.1 to 5% by weight of konjac hydrolyzate according to the present invention.

Claims (11)

At least one microorganism culture medium selected from the group consisting of Bacillus, Aspergillus, and Lactobacillus bacteria, and 0.1 to 5% by weight of bran enzyme hydrolyzate and konjac based on the total weight of the microorganism culture medium. A microbial culture medium having an increase in the productivity of at least one enzyme selected from the group consisting of viable cells, or cellulase, xylanase and mannanase, characterized in that it comprises 0.1 to 5% by weight of enzyme hydrolyzate. The method of claim 1,
The enzyme used in the preparation of the enzyme hydrolyzate of bran and konjac is one or more medium selected from the group consisting of cellulase, xylanase and mannanase. The method of claim 1,
The medium for microbial growth is a medium for culturing microorganisms, characterized in that one selected from the group consisting of LB medium, PD medium and PY medium. delete The method of claim 1,
The genus Bacillus Bacillus subtilis ( Bacillus subtilis ), Bacillus lentus ( Bacillus lentus ), Bacillus licheniformis ( Bacillus licheniformis ), Bacillus coagulans ( Bacillus coagulans ), Bacillus pumilus and Bacillus Cereus ( Bacillus cereus ) medium for culturing microorganisms, characterized in that at least one selected from the group consisting of. The method of claim 1,
The genus Aspergillus is a microorganism culture medium characterized in that at least one selected from the group consisting of Aspergillus duck material ( Aspergillus oryzae ), Aspergillus sojae and Aspergillus niger ( Aspergillus niger ) . The method of claim 1,
The Lactobacillus genus is Lactobacillus casei , Lactobacillus plantarum , Lactobacillus plantarum , Lactobacillus bulgaricus , Lactobacillus rhamnosus ( L actobacillus rhamnosus), Lactobacillus acytocillus ( Lactobacillus acidophilus ), Lactobacillus reuteri , Lactobacillus fermentum , Lactobacillus gasseri , Lactobacillus brevis, Lactobacillus brevis and Lactobacillus pentosus Microbial culture medium, characterized in that at least one selected from the group. A method for culturing microorganisms using the microorganism culture medium according to any one of claims 1 to 3. A microbial cell or a culture thereof prepared by the microbial culture method of claim 8. An animal feed additive comprising the microbial cell of claim 9 or a culture thereof. The soil improvement agent for crop cultivation comprising the microbial cells of claim 9 or a culture thereof.