KR100341796B1 - Lactic acid bacteria for the silage fermentation of low carbohydrate content forage and its using method - Google Patents

Abstract

An object of the present invention is to provide a novel Lactobacillus plantarum NLRI-101 strain suitable for plant silage fermentation with low sugar content.

This novel Lactobacillus plantarum NLRI-101 strain was isolated and identified from silage of good quality. Good quality can be obtained.

Description

Lactic acid bacteria for the silage fermentation of low carbohydrate content forage and its using method}

The present invention relates to a novel Lactobacillus plantarum NLRI-101 strain suitable for fermenting plant silage with low sugar content. In addition, the present invention also relates to a method for producing silage using the above strain.

The ruminant shaft is an important animal resource for humans who do not compete with humans through the use of forages. Forages overproduced during the growing season are dried or fermented and fed to the ruminants during the winter season. Fermented feedstocks are called silage. This method of manufacturing silage, like traditional kimchi, has a high possibility of abnormal fermentation and decay, and it is difficult to obtain a uniform product depending on natural fermentation. Thus, microorganisms are inoculated with plant silage to reduce the chance of growth of defective microorganisms and promote lactic fermentation. Such microorganisms are generally called inoculants, and the addition of this inoculant improves the quality of the silage. And livestock productivity (milk production and meat production) may be improved.

In general, feed crops have a low sugar content, and the silicic acid fermentation inocurant has a high sugar content and a high lactic acid production capacity and a wide range of temperature because the silicic acid temperature varies from 13 to 40 ° C. Growth must be possible under conditions. However, the conventional inocurant was developed to be suitable for fermentation of plants with high sugar content, but was not suitable for fermentation of plants with low sugar content. In particular, it was more difficult to manufacture silage in the case of plants having a lower sugar content than the general silage forage such as rice straw.

Accordingly, the present invention has been made to solve the problems of the prior art as described above, to provide a novel Lactobacillus plantarum NLRI-101 strain suitable for plant silage fermentation of low sugar content There is a purpose.

In addition, the present invention has another object to provide a method for producing silage using the novel Lactobacillus plantarum NLRI-101 strain as described above.

According to the present invention for achieving the above object, there is provided a novel Lactobacillus plantarum NLRI-101 strain (KFCC-11101) suitable for plant silage fermentation.

In addition, according to the present invention, there is provided a method for producing plant silage using the novel Lactobacillus plantarum NLRI-101 strain.

In addition, according to the present invention, the plant is preferably rice straw.

The present inventors have been studying the strain from the silage of high quality in order to isolate the strain suitable for fermentation of the fertilizer, the growth temperature range is similar to the production of silage, and even under low sugar content conditions, the growth rate is fast and lactic acid production ability It was successful to isolate good strains. Therefore, the present inventor named this strain Lactobacillus plantarum NLRI-101, and deposited it on September 9, 1999 with the Korean spawn association (accession number: KFCC-11101).

This strain was added to a high quality silage extract in a liquid medium containing 0.02% sodium azide (NaN ₂ ) (MRS) medium and incubated at 37 ° C. for 24 hours, followed by 0.02% using platinum (platinum wire). A novel Lactobacillus plantarum (Lactobacillus) obtained by smearing the MRS agar plate containing sodium azide (evenly inoculated with the strain) and incubating again at 37 ° C. for 48 hours. plantarum) NLRI-101 has the following characteristics.

(1) Morphology of bacteria when grown at 37 ° C for 48 hours in MRS agar plate medium

① Cell Type: Bacillus

② Mobility: None

③ Spore Formation Capacity: None

④ Gram Dyeing: Positive

(2) Morphology of mycelia when incubated at 37 ° C for 48 hours in MRS agar plate medium

① Shape: Round

② uplift: convex

③ Surface: Smooth

(3) physiological properties

① Growth temperature-Possible growth temperature: 13 ～ 43 ℃

Optimum growth temperature: 37∼39 ℃

② Growth pH-Possible growth pH: 3.5 ~ 7.5

Optimum pH: 6.5-6.8

③ effect on oxygen: anaerobic anaerobic

(4) Catalase:-

(5) Gas Formation:-

(6) Growth at 15 ℃: +

(7) Growth at 45 ℃:-

(8) Indole Production:-

(9) Lactic acid production: +

(10) sugar fermentation test

Amigdaline: +, D-arabinose:-, L-arabinose: +,

Cellobiose: +, esculin: +, fructose: +,

Galactose: +, glucose: +, gluconate: +,

Lactose: +, Maltose: +, Mannitol: +,

Mannose: +, melezitose: +, melibiose:-,

Rafinose:-, Ribose: +, Rhamnose: +,

Salysin: +, Sorbitol: +, Sucrose: +,

Trehalose: +, Xylose:-, Glycerol:-,

Erythritol:-, adonitol:-, beta methyl-xylside:-,

Sorboose:-, Doulsitol:-, Inositol:-,

Alpha methyl-di-mannoside: +,

Alpha methyl-di-glucoside:-,

Enacetyl Glucosamine: +,

Inulin:-, Amidone:-, Glycogen:-,

Xylitol:-, Beta Genthiobiose: +,

Turanos:-, Lysos:-, Tagatose:-,

Fucose:-, arabitol:-, 2 keto-gluconate:-,

5 Keto-Gluconate:-

+ And-to the above (4) to (9) indicate the presence or absence of production, respectively. For example, catalase-means not forming catalase. In addition, + and-in (10) indicate various listed sugars as '+' when using fermentation substrate and '-' when not used, respectively.

As described above, the NLRI-101 strain is Gram-positive, catalase-negative, grows at 15 ° C, fails to grow at 45 ° C, and is identified as Lactobacillus plantarum as a result of 45 sugar fermentation experiments without producing gas during glucose fermentation. It became.

Hereinafter, a preferred embodiment of a strain suitable for plant silage fermentation having a low sugar content according to the present invention and a method for producing silage using the same will be described in detail.

(1) Growth test at low sugar concentration

Novel Lactobacillus plantarum NLRI-101 (L. plantarum NLRI-101), Lactobacillus plantarum ATCC 10012 and Lactobacillus vulgaricus ATCC 33409 (L. bulgaricus ATCC 33409) ) Was incubated in MRS liquid medium for 24 hours, and then 1% was inoculated in nutrient limiting medium with glucose content adjusted to 0.1% and 0.5%, respectively, and samples were taken after 6 hours and 24 hours of culture. . The collected sample was serially diluted in 9 ml of 0.1% peptone solution, inoculated in MRS agar plate medium, incubated at 37 ° C for 48 hours, counted the number of cells formed, and the number of viable cells was measured. It was measured (see Table 4).

The composition of the medium used at this time is as follows.

As can be seen in Table 4, L. plantarum NLRI-101 (invention) according to the present invention is more viable than the L. plantarum ATCC 10012 (control 1) and L. bulgaricus ATCC 33409 (control 2) and low pH It can be seen. That is, it can be seen that the L. plantarum NLRI-101 strain according to the present invention is advantageous for producing silage compared to other strains.

(2) Silage Manufacturing Test

In general, the silage manufacturing method is to cut the plant, put it in a silo (silo) and seal after suppressing the fermentation about 60 days. In addition, the silage of raw straw was cut like a normal silage and put into a silo (since the mid 70's). However, in recent years, the development of agricultural machinery has made balers with large round balers without cutting materials, and then wrapped with vinyl. The storage method is called a technique for preparing raw straw straw silage by round bales. In other words, silage of fresh straw will add a new strain to the process of making balers.

By the above method, the microorganism distribution of rice straw silage (Table 5) and its characteristics (Table 6) two months after storage of the new Lactobacillus plantarum NLRI-101 strain after the addition of 10 ⁶ cfu / g of fresh straw straw silage preparation ) Was observed.

As can be seen from Table 5, when the L. plantarum NLRI-101 according to the present invention is added to raw rice straw, it is not added to inoccurant (no additives) or to other inoccurants (Company A and B). The number of lactic acid bacteria is high and the number of molds is low. That is, the L. plantarum NLRI-101 strain according to the present invention can be seen that it is advantageous for silage production because it increases the number of lactic acid bacteria to promote silage fermentation and lower the number of molds to promote corruption.

In addition, as can be seen in Table 6, L. plantarum NLRI-101 strain according to the present invention is higher in the lactic acid bacteria number than the case of adding no inocurant (no addition) or other inocurant (A company, B company) The Flieg score, which converts the mg equivalent of each acid into percentages, is also high, but the pH is low. Therefore, it can be seen that the L. plantarum NLRI-101 strain according to the present invention is advantageous for silage production.

Inocurant typically uses a strain complex containing Lactobacillus plantarum. In Tables 5 and 6, 'A' is a product prepared and sold by Medipharm, USA. It is composed of by-products and the total number of lactic acid bacteria is more than 5.0 × 10 ⁹ cfu / g. 'B' is a product of Pioneer, USA, consisting of Lactobacillus plantarum and Enterococcus faecium fermentation by-products, and the total number of lactic acid bacteria is 1.0 × 10 ¹⁰ cfu / g or more.

As described in detail above, the Lactobacillus plantarum NLRI-101 strain of the present invention is excellent in growth at low sugar content and excellent in lactic acid producing ability, so that when used as innocurant for silage production, good quality can be obtained.

In addition, the Lactobacillus plantarum NLRI-101 strain of the present invention can increase the utilization of rice straw because the silage can be produced using rice straw produced by 6.3 million tons per year.

The above describes the technical details of the strain suitable for the plant silage fermentation low sugar content of the present invention and the method for producing silage using the same with the accompanying drawings, which is illustrative of the best embodiments of the present invention by way of example It is not limited.

In addition, it is obvious that any person skilled in the art can make various modifications and imitations without departing from the scope of the technical idea of the present invention.

Claims (3)

Novel Lactobacillus plantarum NLRI-101 strain (KFCC-11101) suitable for plant silage fermentation. A method for producing plant silage using the novel strain of claim 1. The method of claim 2, wherein the plant is rice straw.