KR20080040134A - Composition and method for promoting proliferation of beneficial microorganisms - Google Patents

Abstract

A composition comprising a fermented material prepared by inoculating lactic acid bacteria and/or yeast into a mixture of a carbohydrate and water is provided to be usefully used in order to produce a genetically engineered enzyme by a microorganism or other functional polysaccharides, lipids, peptides, amino acids and enzymes by promoting proliferation of effective microorganisms, thereby being used in order to prepare a fermented food, for producing an industrial enzyme, and for improving cattle feed. A composition for promoting proliferation of effective microorganisms comprises a fermented product obtained by inoculating at least one microorganism selected from the group consisting of Lactobacillus sp., Streptococcus sp., Pediococcus sp., Leuconostoc sp., Bifidobacterium sp., and Saccharomyces sp. yeast into a mixture of a carbohydrate and water and fermenting it. A method for promoting proliferation of effective microorganisms comprises a step of adding the composition to a culture medium of the effective microorganisms.

Description

Composition for promoting the growth of effective microorganisms and a method for promoting the growth of effective microorganisms {COMPOSITION AND METHOD FOR PROMOTING PROLIFERATION OF BENEFICIAL MICROORGANISMS}

1 is a graph showing the growth promoting activity of lactic acid bacteria of the composition for promoting the growth of effective microorganisms of the present invention.

Figure 2 is a graph showing the growth promoting activity of the yeast of the composition for promoting the growth of the effective microorganism of the present invention.

Figure 3 is a graph showing the growth promoting activity of Bacillus microorganisms of the composition for promoting the growth of effective microorganisms of the present invention.

The present invention relates to a composition for promoting growth or proliferation of effective microorganisms and a method for promoting growth or growth of effective microorganisms.

Microorganisms can be divided into beeficial microorganisms and noxious microorganisms, depending on whether they or their metabolites are effective or harmful to humans. For example, lactic acid bacteria, Saccharomyces yeast used for brewing or baking can be classified as effective microorganisms, and Vibrio microbes causing sepsis and Salmonella microorganisms causing typhoid diseases can be classified as harmful microorganisms. have.

The development of technology for microorganisms is directed to inhibiting the growth of harmful microorganisms, and to promoting the growth of effective microorganisms.

For example, International Publication (WO) 2004-014893, entitled "Antimicrobial Aza-bicyclic Derivatives, Compositions, and Uses thereof" or International Publication (WO), entitled "Antimicrobial Compositions," 2002-76209 and the like is a technology for inhibiting the growth of harmful microorganisms, and the name of the invention is Korean Patent No. 469989 or the invention "Method of cultivating and using useful microorganisms using marine humic colloid micelles and active minerals" Korean Patent Laid-Open Publication No. 2006-0016942, "A method for mass-producing a useful microbial culture liquid extracted from rice bran and a method for mass-producing a useful microbial active liquid using the useful microbial culture," is a technique for promoting the growth of effective microorganisms.

As interest in well-being increases recently, effective microorganisms, such as food fermentation, livestock feed improvement, compost manufacturing, purification of livestock waste, improvement of soil quality, and purification of wastewater, are continuously attracting attention.

The present invention also provides a technique capable of promoting the growth of effective microorganisms.

Accordingly, an object of the present invention is to provide a composition for promoting the growth of effective microorganisms and a method for promoting the growth of effective microorganisms using the same.

Specific embodiments of the present invention will be presented below.

In one aspect, the present invention can be understood as a composition for promoting the growth of effective microorganisms.

The inventors, as confirmed in the following Examples and Experimental Examples, mixed soybean powder and glucose powder with water, inoculated and fermented with lactic acid bacteria and / or yeast, and then the supernatant of the fermentation broth was lactic acid bacteria, yeast, mushrooms. When added to the microbial medium, such as mycelium, it was confirmed that the growth of the microorganism is significantly promoted compared with the case where the supernatant is not added.

The present invention is provided based on these experimental results.

Therefore, the composition for promoting the growth of the effective microorganism of the present invention is characterized in that it comprises a fermentation product obtained by incorporating water into carbohydrates that can be used as an energy source by lactic acid bacteria or yeast, and inoculating them with lactic acid bacteria and / or yeast.

In the present specification, including the claims, the "carbohydrate that can be used as an energy source by lactic acid bacteria or yeast" is meant to include all polysaccharides, oligosaccharides, disaccharides and monosaccharides, but preferably means to include oligosaccharides, disaccharides and monosaccharides Can be understood as It is well known that lactic acid bacteria and yeast use oligosaccharides, disaccharides and / or monosaccharides as energy sources. Recently, lactic acid bacteria and yeasts also have the ability to degrade polysaccharides (Kim Mi-jung et al., "Ozone-treated clean materials and L. acidophilus Ripening of Chinese Cabbage Kimchi Using Rice, 1993. pp.165-174, KHWA Kwon, 22 (2); Song, Hye-Kyung, "Degradation of Polysaccharides of Lactic Acid Bacteria Isolated from Kimchi," 1996.02, Master's Thesis, Korea University). Therefore, the "carbohydrates available to lactic acid bacteria or yeast as an energy source" should not be understood as excluding polysaccharides. However, when using a polysaccharide, it may be preferable to use a polysaccharide degrading enzyme, such as amylase, in order to facilitate the progress of fermentation by lactic acid bacteria and / or yeast.

Examples of the polysaccharides may include glycogen, amylose, amylopectin, and the like, and examples of the oligosaccharides (which can be understood as meaning including two to ten sugars or less per monosaccharide according to a conventional definition) include fructose, lactose, and the like. Examples of the disaccharides, polysaccharides, decomposition products obtained by the action of the decomposition of the polysaccharides can be exemplified. Examples of the disaccharides can be exemplified sugar, lactose, maltose and the like, and examples of the monosaccharides are fructose, glucose and the like Can be.

In addition, in the present specification, including the claims, the "lactic acid bacteria" is glucose, such as by decomposing a sugar to mean microbes that produce lactic acid, these lactic acid bacteria, the lactic acid bacteria genus Lactobacillus (Lactobacillus sp.), Streptococcus genus of lactic acid bacteria (Streptococcus sp .), Syracuse Phedi OKO in lactic acid bacteria (Pediococcus sp . ), Leuconostoc sp . ) And a Bifidobacterium lactic acid bacteria (Bifidobacterium sp . ), Specifically, Lactobacillus acidophilus , Lactobacillus plantarum , Lactobacillus fermentum , Bifidobacterium longgum, Bifidobacterium longum ), Bifidobacterium bifidum and the like.

In the present specification, which also includes claims, “yeast” is a genus of Saccharomyces Yeast, such as Saccharomyces rouxii , Saccharomyces cereviciae ), Saccharomyces oviformis ) and Saccharomyces steineri .

In addition, in the following, including the claims, "effective microorganism" refers to all microorganisms that are useful to humans or metabolites thereof, for example, Saccharomyces yeast, which has been used in brewing and baking, has a formal function and cholesterol. Lactobacillus with inhibitory activity, actinomycetes known to produce streptomycin, photosynthetic bacteria that function to fix nitrogen while coexisting with yeast or lactic acid bacteria, mycelium of edible mushrooms that can be used as health functional foods or medicines Bacillus microorganisms are involved in the production of koji, such as koji bacteria and Chunggukjang used for fermentation. From the point of view of use, it is used for food fermentation, livestock feed improvement, compost production, purification of livestock waste, improvement of soil quality, purification of waste water, etc. Defined as any microorganism that somehow produces useful results to humans Can. However, specifically may be understood as lactic acid bacteria, Saccharomyces yeast, mushroom mycelium, Bacillus microorganisms used in the experimental examples of the present invention, more preferably the specific examples used in the experimental examples of the present invention It can be understood to refer to a microorganism.

In addition, in the present specification including the claims, "fermented product" refers to a fermentation broth containing the fermentation microorganisms, lactic acid bacteria and / or yeast, or a fermentation broth in which the lactic acid bacteria and / or yeast is present in a cell lysis (cell lysis) state , Supernatant obtained by filtration or centrifugation of the fermentation broth. Although the embodiment of the present invention looks at the degree of proliferation activity of the microorganisms with the supernatant of the culture, the fermentation broth in which the fermentation broth itself or the lactic acid bacteria and / or yeast are in the state of cell lysis is also separated from the supernatant. It is natural that the microorganisms will have the proliferative activity as long as the microorganisms have the proliferative activity.

On the other hand, in a preferred embodiment of the present invention, it is preferable to mix soybean milk or milk with carbohydrates before inoculating carbohydrates with lactic acid bacteria and / or yeast, in order to activate the fermentation of lactic acid bacteria more. For this reason, in the following examples of the present invention, carbohydrates were used by mixing soybean powder. There is no particular restriction on the type of soybean powder or soy milk that can be used for the production of soybean powder. For example, peas, kidney beans (including the hinge kidney beans and red kidney bean), it is possible is to be used both seomoktae, cheongtae eastern bamkong, efflorescence (soybeans), herbs, bean, etc. bapmitkong, jakdukong, seoritae, putkong, in the viewpoint of taxonomic Glycine subgenus ( G. albicans Tind and Craven; G. arenaria Tind ; G. argyrea Tind ; G. canescens FJ Herm ; G. clandestina Wendl ; G. curvata Tind ; G. cyrtoloba Tind ; G. falcata Benth ; G. hirticaulis Tind . and Craven; G. lactovirens Tine. and Craven; G. latifolia ( Benth .) Newell & Hymowitz ; G. latrobeana ( Meissn .) Benth ; G. microphylla ( Benth .) Tind ; G. pindanica Tind . and Craven; G. tabacina (Labill) Benth ) and Soja Asoke ( G. soja Sieb . & Zucc ; G. max (L.) Merr . forma gracilis All beans belonging to Skvortz ) can be used.

In addition, in a preferred embodiment of the present invention, the fermentation of lactic acid bacteria and / or yeast is preferably progressed to the extent that the lactic acid bacteria and / or yeast is fully grown. “Enough” here can be understood as a condition in which lactic acid bacteria and / or yeast have already proliferated to the maximum and no longer grow. It is determined in relation to the fermentation microorganism and the fermentation temperature, and the person skilled in the art can easily determine the time that the fermentation is sufficiently fermented according to the type of the fermentation microorganism at a specific temperature within the range of its usual capacity. There will be. In the following examples of the present invention, fermentation was performed at 20 ° C. to 40 ° C. for about 15 days regardless of whether lactic acid bacteria or yeast were used as the fermentation microorganism.

In a preferred embodiment of the present invention, it is preferable that the mixture of carbohydrate and water is inoculated and fermented with lactic acid bacteria and yeast. It is shown by the following Examples and Experimental Examples of the present invention, because the fermentation product obtained by inoculating and fermenting the lactic acid bacteria and yeast was found to have higher proliferation activity of the effective microorganism.

In another aspect, the present invention can be understood as a method for producing a composition for promoting growth of effective microorganisms.

Method for producing a composition for promoting the growth of effective microorganisms of the present invention comprises the steps of mixing water with carbohydrates, inoculating lactic acid bacteria and / or yeast in the mixture of carbohydrates and water and proceeding the fermentation by lactic acid bacteria and / or yeast It is configured to include.

In the method for producing a composition for promoting growth of an effective microorganism of the present invention, the meanings of carbohydrate, lactic acid bacteria, yeast, effective microorganism, and the like are effective as they are.

In the preferred embodiment, the foregoing is effective as it is. Specifically, it is preferable to mix the carbohydrate with soybean powder or soymilk or milk before inoculating the carbohydrate with lactic acid bacteria and / or yeast, and further ferment the lactic acid bacteria and / or yeast. It is preferable to proceed to the extent that the lactic acid bacteria and / or yeast is sufficiently multiplied, and it is preferable to inoculate and ferment the mixed lactic acid bacteria and yeast rather than to inoculate each of the lactic acid bacteria or yeast in the mixture of carbohydrate and water.

In another aspect, the present invention can be identified as a method for promoting the growth of effective microorganisms, characterized in that the composition for promoting the growth of effective microorganisms as described above is added to the medium for effective microorganisms.

Also in the method for promoting the growth of the effective microorganism of the present invention, the meanings of the carbohydrate, the lactic acid bacteria, the yeast, the effective microorganism, and the like are effective as they are.

The amount of the composition for promoting the growth of effective microorganisms added to the medium may be added in the range of 1 part by weight to 15 parts by weight based on the weight of the medium.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.

< Example > Lactic acid bacteria and / or yeast Fermented product  Produce

Example 1 Lactic Acid Bacteria Fermentation Product Preparation Example 1

30 g of powder and 30 g of glucose obtained by pulverizing soybeans (medium soybeans) were put into 1 L of clean water and sterilized (120 ° C., 20 min). After sterilization, the mixture is cooled to room temperature, and the lactic acid bacteria ( Lactoba plantus Lactoba) cllus plantarum )) (ATCC8014) 10 ⁸ cells / ㎖ was inoculated at 3% (W / V).

After inoculating microorganisms, the cells were fermented by incubating for about 15 days in a temperature range of 20 ° C to 40 ° C. After 15 days of incubation, the pH of the culture solution was 4.0 ± 0.2, and it was confirmed that the lactic acid bacteria reached 1.0 × 10 ⁸ cells / ml.

After completion of the culture, the culture solution was sterilized (120 ° C., 20 min), cooled to room temperature, and the supernatant was separated by centrifugation (10,000 rpm, 20 min) or microfiltration (≦ 0.1 μm pore size). The separated supernatant was concentrated under reduced pressure until the total solid content was 5%. The concentrate thus obtained (solid content 5%) was refrigerated until used for the experiment.

Example 2 Yeast Fermented Product Preparation Example 2

30 g of powder and 30 g of glucose obtained by grinding beans were sterilized in 1 L of clean water (120 ° C., 20 min). After sterilization, it is cooled to room temperature, and the yeast ( Saccharomyces cerevisiae )) (IFO0203) 10 ⁸ cells / ml were inoculated at 3% (W / V).

After inoculating microorganisms, the cells were fermented by incubating for about 15 days in a temperature range of 20 ° C to 40 ° C. After 15 days of incubation, the pH of the culture medium was 3.0 ± 0.2 and the yeast reached 2.0 × 10 ⁸ cells / ml.

After completion of the culture, the culture solution was sterilized (120 ° C., 20 min), cooled to room temperature, and the supernatant was separated by centrifugation (10,000 rpm, 20 min) or microfiltration (≦ 0.1 μm pore size). The separated supernatant was concentrated under reduced pressure until the total solid content was 5%. The concentrate thus obtained (solid content 5%) was refrigerated until used for the experiment.

Example 3 Lactic Acid Bacteria and Yeast Fermentation Preparation Example 3

30 g of powder and 30 g of glucose obtained by grinding beans were sterilized in 1 L of clean water (120 ° C., 20 min). After sterilization, the mixture is cooled to room temperature, and the lactic acid bacteria ( Lactoba plantus Lactoba) cllus plantarum )) (ATCC8014) and yeast ( Saccharomyces cerevisiae ) (IFO0203) were adjusted to 10 ⁸ cells / ml, and the mixture was mixed at a weight ratio of 1: 1. 3% (W / V) Inoculated.

After inoculating microorganisms, the cells were fermented by incubating for about 15 days in a temperature range of 20 ° C to 40 ° C. After 15 days of culture, the pH of the culture medium was 3.5 ± 0.2, and the lactic acid bacteria reached 1.0 × 10 ⁷ cells / ml and yeast reached 4.0 × 10 ⁸ cells / ml.

After completion of the culture, the culture solution was sterilized (120 ° C., 20 min), cooled to room temperature, and the supernatant was separated by centrifugation (10,000 rpm, 20 min) or microfiltration (≦ 0.1 μm pore size). The separated supernatant was concentrated under reduced pressure until the total solid content was 5%. The concentrate thus obtained (solid content 5%) was refrigerated until used for the experiment.

Example 4 Lactic Acid Bacteria Fermentation Product Preparation Example 4

The same material as in <Example 1> was used to ferment in the same process under the same process conditions, except that Lactobacillus acidophilus ( Lactobacillus acidophilus ) (KCTC3171) was used as a fermentation microorganism.

After completion of the culture, the pH of the culture solution was 3.0 ± 0.1, and the lactic acid bacteria was found to be 1.0 × 10 ⁷ cells / ml.

Thereafter, the process of producing the fermented product is the same as in <Example 1>.

Example 5 Yeast Fermented Product Preparation Example 5

Saccharomyces rouxii (KCTC7880) was used as the fermentation microorganism in the same process using the same material as in Example 2, but the same process under the same process conditions.

After completion of the culture, the pH of the culture solution was 3.5 ± 0.1, and the yeast was found to be 1.0 × 10 ⁸ cells / ml.

Thereafter, the process of producing the fermented product is the same as in <Example 1>.

Example 6 Lactic Acid Bacteria and Yeast Fermented Product Preparation Example 6

Fermented by the same process under the same process conditions using the same material as in <Example 3>, except that the yeast as fermentation microorganism is Lactobacillus acidophilus ( Lactobacillus acidophilus ) (KCTC3171) and yeast is Saccharo Saccharomyces rouxii ) (KCTC7880) was used.

After completion of the culture, the pH of the culture medium was 3.8 ± 0.2, and the lactic acid bacteria were identified as 0.5 × 10 ⁷ cells / ml and the yeast was 2.0 × 10 ⁸ cells / ml.

Thereafter, the process of producing the fermented product is the same as in <Example 1>.

< Experimental Example > Lactic acid bacteria and / or yeast Fermented product  Microbial growth promotion activity experiment

Experimental Example 1 Lactic Acid Bacteria Growth Promoting Activity Experiment

The lactic acid bacteria Lactobacillus, Bacillus Planta room (Lactobacillus plantarum) (ATCC8014) and incubated in the thermostat value of 30 ℃ to GYP broth. The cultured by adding the fermented product of each of the above examples to the GYP liquid medium (1%, V / V), and the control was not added to the fermented product. The bacterial counts of Lactobacillus plantarum were measured every 24 hours after the start of the culture by dilution plate method using GYP agar medium. Each prepared plate was incubated in a thermostat at 30 ° C. for 24 to 48 hours to measure the number of colonies that appeared.

The results are shown in FIG. 1A (showing the growth promoting activity of the lactic acid bacteria of the fermented products obtained in Examples 1 to 3) and 1B (showing the growth promoting activity of the lactic acid bacteria of the fermented products obtained in Examples 4 to 6).

Referring to FIGS. 1A and 1B, fermented products obtained by fermenting only with lactic acid bacteria or yeast (fermented products of Examples 1, 2, 4, and 5) also have the effect of promoting the growth of lactic acid bacteria, but mixed bacteria of lactic acid bacteria and yeast It can be seen that the fermented product (fermented products of Examples 3 and 6) obtained by the fermentation with an excellent lactic acid bacteria growth promoting effect.

<Experimental Example 2> Experiment for yeast growth promoting activity

Yeast Saccaromyces Saccaromyces cerevisiae ) (IFO 0203) was incubated in YM liquid medium in a thermostat at 25 ° C. Incubated with the addition of the fermented product of each Example in a YM liquid medium (1%, V / V), was added as a control was not added to the fermentation. Saccharomyces cerevises every 24 hours after the start of culture The bacterial counts were measured by dilution plate method using YM agar medium. Each prepared plate was incubated in a thermostat at 25 ° C. for 5-7 days to measure the number of colonies that appeared.

The results are shown in Fig. 2A (showing the yeast growth promoting activity of the fermented products obtained in Examples 1 to 3) and Fig. 2B (showing the growth promoting activity of the yeast of the fermented products obtained in Examples 4 to 6).

2A and 2B, fermented products obtained by fermenting only with lactic acid bacteria or yeast (fermented products of Examples 1, 2, 4 and 5) also have a yeast growth promoting effect, but are mixed with lactic acid bacteria and yeast. It can be seen that the fermented product obtained by fermentation (fermented products of Examples 3 and 6) has an excellent yeast growth promoting effect.

<Experimental Example 3> Experiment on the promotion of the mushroom mycelium growth activity

Mushroom mycelia are Pellinus linteus ) mycelium (KCTC6190) was used. Mushroom mycelium was inoculated in MEB (Malt Extract Broth) medium, and cultured by adding the fermented product of each Example as an experimental group (1% added, V / V), and the fermented product was not added as a control. It was. After incubation, the entire sample was lyophilized to measure the weight of each sample.

The results are shown in Table 1 below.

Incubation Period (day) 0 One 2 3 4 5 6 Example 1 0 0.12 0.19 0.31 0.41 0.89 1.3 Example 2 0 0.11 0.23 0.35 0.42 0.92 1.2 Example 3 0 0.18 0.40 0.71 1.20 1.90 2.1 Example 4 0 0.13 0.17 0.32 0.51 0.97 1.5 Example 5 0 0.11 0.22 0.40 0.59 0.99 1.6 Example 6 0 0.16 0.37 0.68 1.31 1.79 1.99 Control 0 0.09 0.12 0.15 0.30 0.71 0.92 * Individual quantity indication by dry weight (g / 100mL)

Table 1 shows that the growth of the situation mushroom mycelium with the addition of the fermented product of each example is increased as a whole compared with the case without culture. In particular, it shows that the fermented product obtained by culturing with the mixed bacteria of lactic acid bacteria and yeast (microbial extract of Examples 3 and 6) has a very good growth promoting effect.

Experimental Example 4 Bacillus Experiment of proliferative activity of subtilus

Bacillus subtilis (KCTC1107) was inoculated in YM liquid medium and incubated at 40 ° C. and pH 7.0 for 7 days. The cultured by adding the fermented product of each of the above examples to the YM liquid medium (1%, V / V), and the control was not added to the fermented product. The growth curves under these two conditions are shown in Fig. 3a (showing the effect of promoting the growth of lactic acid bacteria of the fermented products obtained in Examples 1 to 3) and 3b (showing the effect of promoting the growth of lactic acid bacteria of the fermented products obtained in Examples 4 to 6) and same. The Y axis shows the Logarithmic exponential value of the measured number of individuals.

6A and 6B, similar to the above results, the growth of Bacillus subtilis also in the case of fermentation products obtained by fermentation with only lactic acid bacteria or yeast (fermentation products of Examples 1, 2, 4 and 5). It can be seen that the fermentation product (extracts of Examples 3 and 6) obtained by fermentation with a mixed bacterium of lactic acid bacteria and yeast, having a promoting effect, has an excellent growth promoting effect of Bacillus subtilis.

As described above, according to the present invention, carbohydrates (and soybean powder, milk and / or soy milk, etc.) is mixed with water, and inoculated therein and fermented by the inoculation of lactic acid bacteria and / or yeasts for promoting the growth of effective microorganisms A composition can be provided.

The composition can be used for the production of genetically engineered enzymes or other functional polysaccharides, lipids, peptides, amino acids and enzymes by microorganisms by promoting the proliferation of effective microorganisms, fermented food production using effective microorganisms, industrial It can be usefully used for enzyme production, livestock feed improvement, etc.

Claims (8)

A composition for promoting effective microbial growth comprising a fermentation product obtained by incorporating water into a carbohydrate that can be used as an energy source by lactic acid bacteria or yeast and inoculating one or more microorganisms selected from the group consisting of lactic acid bacteria and yeast. The method of claim 1, The carbohydrate is effective microorganism growth promoting composition, characterized in that it comprises one or more selected from the group consisting of oligosaccharides, disaccharides and monosaccharides. The method of claim 1, The lactobacillus is Lactobacillus sp . Lactobacillus sp . , Streptococcus sp . , Pediococcus spp . sp . ), Leuconostoc sp . ) And Bifidobacterium spp . ( Bifidobacterium sp. ) A composition for promoting effective microbial growth, characterized in that selected from the group consisting of. The method of claim 1, The yeast is a composition for promoting effective microbial growth, characterized in that Saccharomyces genus yeast. The method of claim 1, The composition for promoting the growth of effective microorganisms, characterized in that the mixture of soybean powder or soy milk or milk before inoculating the carbohydrate lactic acid bacteria or yeast. The method of claim 1, A composition for promoting the growth of effective microorganisms comprising inoculating both lactic acid bacteria and yeast. Promoting the growth of effective microorganisms comprising mixing water to carbohydrates, inoculating one or more microorganisms selected from the group consisting of lactic acid bacteria and yeast into the mixture of carbohydrates and water, and proceeding fermentation by the microorganisms Method for producing a composition for use. A method for promoting growth of effective microorganisms, comprising adding a composition for promoting growth of effective microorganisms according to any one of claims 1 to 6, to a medium of the effective microorganism.

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