KR20160039394A - Novel microorganism separated from carnivorous plants and protease produced therefrom - Google Patents

Abstract

The present invention relates to a novel microorganism which digests feed by activity of digestive enzymes or commensal bacteria, is separated from carnivorous plants, and has protein decomposing activity, to a culture medium produced therefrom, having protein decomposing activity, and to a composition which generates low molecular weight collagen peptides by decomposing the protein and comprising the culture medium as an active ingredient. The novel microorganism Chryseobacterium sp. MK-8 produces a culture medium having stable protein decomposing activity at 15 to 50°C and in the pH range of 5.5 to 11.0 and low molecular weight collagen peptide generating activity.

Description

[0001] The present invention relates to a novel microorganism isolated from a carnivorous plant and a protease produced from the microorganism isolated from the carnivorous plant.

The present invention relates to a microorganism isolated from a carnivorous plant and a culture medium produced by the microorganism. More particularly, the present invention relates to a strain of Chrysobacterium sp. MK-8 derived from a carnivorous plant, a culture method thereof, And a composition for producing a low molecular weight collagen peptide by decomposing a protein containing the culture liquid as an active ingredient.

Carnivorous plant is an organ with incendiary and repellent functions. It is a plant that catches animals such as insects, daphnia, and microorganisms and obtains a part of nutrients through an organ having digestive and absorption functions. Collectively. As a synonym for carnivorous plants, insect plants, carnivorous plants, and insect plants are widely or partially used.

Carnivorous plants are largely green plants, all flowering plants, and seeds are all fruit seeds. The carnivorous plant is an Angiospermae in the Engler classification system and is a dicotyledoneae, and Lentibulariaceae belongs to the subcommunity (Sympetalae) In contrast, all other carnivorous plants belong to the subfamily Archichalamydeae.

The basic order of the ecosystem is that animals can not synthesize proteins, so they consist of food chains that feed on the plants that produce them, but the carnivorous plants have reversed the situation of these food chains and the plants have changed into eating insects. In other words, the carnivorous plant has an organ which functions to attract insects by changing parts of organs of the plant, and has an organ that functions to digest and absorb the insects that are caught. It was transformed into a survival system.

The carnivorous plant secretes secretions and digestive juices for incendiary and larvae through the glandular cells, and the absorption is carried out by the glandular cells. In addition to secretion of the digestive juices of the insecticidal plant itself, digestion may also be promoted by symbiotic fungi and the like.

On the other hand, proteolytic enzymes are enzymes that break down proteins into low molecular peptides and amino acids. They are produced from various microorganisms such as plants and animals, bacteria, fungi and yeast. Among them, protease derived from bacteria is widely distributed in animals, plants and microorganisms, and is also present both inside and outside of the cell. It is mainly mass produced outside the cells, has not only thermal stability, but also acts at a wide range of pH Therefore, it is recognized as a universal industrial enzyme. These bacterial proteases are used in various fields such as cosmetics, foods, pesticides, pharmaceuticals, detergents, leather, and cosmetics.

Proteolytic enzymes are divided into various types based on their substrate specificity, and their industrial applications vary depending on their characteristics. Endopeptidase cleaves randomly the protein that meets the specificity of the enzyme to form a peptide. Exopeptidase breaks down the protein into amino acid units along the peptide chain. Metallo protease, which contains a metal ion depending on the structure of the enzyme, and serine protease, which contains a serine residue in an amino acid, in the active site. In addition, acidity is divided into acidity, alkalinity, and neutral depending on acidity characteristics, and it is classified into heat resistance, mesophilic, and high temperature depending on temperature.

In the extreme conditions in which most carnivorous plants grow, such as wetlands, peatlands, sandy terrains, and rock walls, the nitrogen source (NO ₃ ^- , NH ₄ ⁺ (Components of nucleic acids (DNA, RNA) and ATP) are easily washed off into moisture and lack nutrients.

Therefore, it has been observed that the insect, daphnia, microorganisms and other nutrients are obtained through the insecticidal action of the carnivorous plant evolved by adaptation to the harsh environment of this survival, and the secretions and digestive juices of the carnivorous plant contain various enzymes Since these enzyme systems can be expressed by microorganisms that are symbiotic with the carnivorous plant, the microorganisms having proteolytic activity in the body organism of the carnivorous plant are searched for and isolated, and protease is produced therefrom It came to the following.

In general, collagen is also known as light protein or collagen, and is widely distributed in multicellular animals such as invertebrates and vertebrates, and is the most frequently found light protein. The main functions of collagen are known as firmness of skin, resistance and binding force of tissue, and support of cell adhesion. Such collagen has been known to have an effect of preventing the formation of wrinkles of the skin caused by thinning of the skin due to aging and photoaging due to ultraviolet irradiation, and thus it is used as a raw material for functional cosmetics. In addition, collagen is used as a raw material or an additive in various fields such as cosmetics, health drinks, health foods, medicines and cosmetics, and the number of industrial fields is increasing. Therefore, it is necessary to develop a method for efficiently producing collagen peptides.

Korean Patent Publication No. 10-2004-0101889 (published on December 03, 2004) Korean Registered Patent No. 10-0891975 (registered on Mar. 30, 2009) Korean Registered Patent No. 10-0358716 (registered on October 15, 2002)

Accordingly, an object of the present invention is to provide a novel microorganism MK-8 strain of Chrysosobacterium sp. Having excellent proteolytic activity, which is isolated from natural carnivorous plants, And a culture solution having the proteolytic activity generated therefrom.

It is another object of the present invention to provide a method for culturing the novel microorganism.

Another object of the present invention is to provide a composition for producing a low molecular weight collagen peptide by degrading a protein by containing the strain or a culture solution thereof as an active ingredient.

Another object of the present invention is to provide a method for producing a collagen peptide comprising treating the strain or a culture thereof with a protein.

In order to achieve the above object, the present invention provides a Chrysobacterium sp. MK-8 strain (accession number: KFCC11582P) derived from a carnivorous plant.

According to another aspect of the present invention, there is provided a culture medium having proteolytic activity produced from the above-mentioned Chrysobacterium sp. MK-8.

According to another aspect of the present invention, there is provided a method for culturing a strain of the present invention comprising culturing the strain in a medium containing a carbon source and a nitrogen source at a temperature of 4 to 37 DEG C and a pH of 5.5 to 8, Thereby providing a method for culturing Chryseobacterium sp. MK-8.

In order to accomplish still another object of the present invention, the present invention provides a composition for producing a low molecular weight collagen peptide by degrading a protein comprising the strain or a culture thereof as an active ingredient.

According to another aspect of the present invention, there is provided a method for producing a collagen peptide comprising treating a strain or a culture thereof with a protein.

As described above, the present invention provides a strain of Chrysobacterium sp. MK-8 derived from a carnivorous plant, a method of culturing the same, and a culture solution having proteolytic activity produced from the strain. The strain of the present invention produces a culture medium having proteolytic activity, and the culture medium of the present invention has a high proteolytic activity and is effective for decomposing protein-containing materials to produce low molecular weight collagen peptides. In addition, it is effective in the production of functional cosmetics because it decomposes keratin and produces low molecular weight collagen peptide by treating at the keratinous part of skin.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a micrograph of a microorganism Chrysobacterium sp. MK-8 isolated from a carnivorous plant according to the present invention. FIG.
FIG. 2 is a graph showing changes in proteolytic activity of proteolytic enzymes produced by the microorganism Chrysobacterium sp. MK-8 isolated from a carnivorous plant according to the present invention at various reaction temperatures.
FIG. 3 is a graph showing changes in proteolytic activity of proteolytic enzymes produced by the microorganism Chrysobacterium sp. MK-8 isolated from a carnivorous plant according to the present invention.
4 is a graph showing changes in storage stability of proteolytic enzymes produced by the microorganism Chrysobacterium sp. MK-8 isolated from a carnivorous plant according to the present invention at various storage temperatures.
FIG. 5 is a diagram showing a 16S rRNA base sequence of a microorganism Chrysobacterium sp. MK-8 isolated from a carnivorous plant according to the present invention.
Fig. 6 shows experimental results of the proteolytic activity of the proteolytic enzyme of the present invention.
FIG. 7 is a result of analyzing the amino acid components of the collagen peptide prepared by the method of the present invention and the existing collagen peptide (Test: collagen peptide produced by the production method of the present invention, Pig collagen: existing collagen peptide )

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, a detailed description of preferred embodiments of the present invention will be given with reference to the accompanying drawings. It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention. Accordingly, the invention in its broader aspects is not limited to the specific details and representative embodiments shown and described herein. Accordingly, It is to be understood that the invention includes equivalents and alternatives. In the following description of the present invention, detailed description of known functions and configurations incorporated herein will be omitted when it may make the subject matter of the present invention rather unclear.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Terms such as those defined in commonly used dictionaries should be interpreted as having a meaning consistent with the meaning in the context of the relevant art and are to be interpreted in an ideal or overly formal sense unless explicitly defined in the present application Do not.

The present invention provides a Chrysobacterium sp. MK-8 strain (accession number: KFCC11582P) derived from a carnivorous plant.

The microorganism of the present invention can be isolated from a naturally occurring carnivorous plant as follows. First, the palatability of the carnivorous plants, the supernatant, and the stems were separated and collected, and then each portion was homogenized with sterilized physiological saline and homogenized. Then, 1% non-fat milk powder or a solid medium supplemented with azocasein To select a strain that forms a transparent ring. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a photograph of a culture medium of a microorganism Chrysobacterium sp. MK-8 isolated from a carnivorous plant according to the present invention, wherein a strain in which a transparent ring is formed on a medium can be identified.

The above-mentioned carnivorous plant is a plant that survives by using an insect as a nutrient, and its kind is not particularly limited, but is preferably Nepenthes, Sarracenia, Drosera rotundifolia L., And Pinguicula. ≪ / RTI >

The microorganisms according to the present invention having the selected proteolytic activity as described above were found to be gram-negative, aerobic, and spore-forming when their strain characteristics, morphological, physiological, Bacteria of 0.5 - 1.0 mm in diameter without motility are shown. It is also positive for oxydase and catalase and can grow at 4-40 ℃, pH 4-10, NaCl 0-3.0% (W / V).

When the microorganism according to the present invention is based on the morphological, physiological and biochemical characteristics as described above, the ribosome subunit gene is isolated and its sequence is analyzed for identification of the microorganism of the present invention, As a result, the homology with Chrysobacterium taiwanense BCRC 17412 (T) was more than 98%. The microorganism according to the present invention was named as Chryseobacterium sp. MK-8, which is a new bacterium, based on the morphological, physiological and biochemical characteristics and the result of the nucleotide sequence analysis.

According to the present invention, a novel microorganism Chryseobacterium sp. MK-8 isolated from a natural carnivorous plant was deposited at Korean Society for Microbiological Research, KKB, Korea, March 17, 2014 No. 14-09 (Accession No: KFCC11582P).

The strain of the present invention is characterized by producing a culture solution having proteolytic activity.

Accordingly, the present invention provides a culture solution having proteolytic activity produced from Chrysobacterium sp. MK-8 of the present invention.

The culture broth produced by the strain of the present invention has proteolytic activity, and the culture broth has a proteolytic activity at a temperature of 15 to 50 ° C and a pH range of 5.5 to 11.0.

This effect of the present invention is well illustrated in the examples.

In one embodiment of the present invention, in order to isolate the proteolytic enzyme from the culture medium produced from Chrysobacterium sp. MK-8 of the present invention showing proteolytic activity, the microorganism is first cultured in a culture medium (yeast extract 0.3%, peptone 0.5%, azo casein 1.0% (or 1% skim milk powder), dextrose 0.5%, distilled water 1 L) was prepared and the surface was sterilized. The microorganisms were inoculated and cultured at 30 ° C for 24 hours , The culture was centrifuged to take up the supernatant, and the protein was concentrated to prepare a partially purified crude enzyme solution.

In another embodiment of the present invention, the stability of the separated protein coenzyme according to reaction temperature, reaction pH and storage period was examined.

As a result, as shown in FIG. 2, the culture broth of the present invention exhibited the most stable decomposing activity in the range of 30 to 40 ° C, exhibited activity of 70% or more at the maximum activity standard at 15 ° C, And showed very stable proteolytic activity at temperature. As shown in FIG. 3, the culture medium of the present invention was found to have optimal enzyme activity in the pH range of 5.5 to 11.0. As shown in FIG. 4, the culture medium of the present invention was cooled and stored at room temperature (25 ° C) To 50 ℃ showed no significant effect on the storage period and proteolytic activity was remarkably decreased at high temperature above 60 ℃.

Meanwhile, the present invention provides a method for culturing Chrysobacterium sp. MK-8 strain (accession number: KFCC11582P) derived from a carnivorous plant of the present invention.

The culture method of the present invention is a method for cultivating a chrysanthemum sp. MK-8 derived from a carnivorous plant of the present invention into a culture medium containing a carbon source and a nitrogen source at a temperature of 4 to 37 DEG C and a pH of 5.5 to 8 And culturing the cells under conditions.

The medium used in the culturing method of the present invention may be any medium used for culture of known microorganisms belonging to the genus Chrysobacterium, and the culture conditions and the method may be selected so that a temperature of 4 to 37 DEG C and a pH of 5.5 to 8 are satisfied Unless otherwise specified.

According to the method of the present invention, it is possible to effectively cultivate the Chrysobacterium sp. Strain MK-8 derived from the carnivorous plant of the present invention, and to efficiently produce a culture having a high enzyme-degrading activity.

The culture medium of Chrysobacterium sp. MK-8 of the present invention is excellent in the decomposition activity of decomposing proteins to produce low molecular weight collagen peptides. This point is first disclosed in the present invention.

The activity of these cultivars of the genus Kriobotherium MK-8 is well illustrated in the examples of the present invention. In one embodiment of the present invention, the MK-8 culture broth is isolated and purified to separate the protein fraction having a high proteolytic activity, and the protein fraction is treated to have a tissue characteristic similar to that of a human skin surface layer to produce proteolytic activity and low molecular collagen peptide production activity Respectively. As a result, it was confirmed that the fraction of the culture broth of MK-8 strain was excellent in the proteolytic activity and the activity of producing low molecular weight collagen peptide.

Accordingly, the present invention provides a composition for producing proteolytic and low molecular weight collagen peptides comprising Chrysobacterium sp. MK-8 or a culture thereof as an active ingredient.

The composition of the present invention is characterized by containing Chrysobacterium sp. MK-8 or a culture thereof as an active ingredient. The culture medium is a cultured medium of Chrysobacterium sp. MK-8, which contains all of the culture medium or all of the culture medium excluding the microorganism or the culture medium.

The composition of the present invention not only removes keratin but also has excellent activity of decomposing keratin to produce low molecular weight collagen peptide useful for skin.

The present invention also provides a method for producing a low molecular weight collagen peptide comprising treating Chrysobacterium sp. MK-8 or a culture thereof with a protein-containing sample.

The protein may be any protein capable of degrading a peptide to produce a peptide, preferably a fibrous protein, more preferably collagen or keratin.

The collagen-containing sample may be any of those containing collagen, such as, for example, hide splits, pig skin, dried ossein, fish, and preferably pig skin , Pig skin).

The collagen peptide may be low molecular weight by hydrolyzing collagen with an enzyme, preferably having a molecular weight of 100,000 or less, and more preferably 10,000 or less. The collagen peptide is a kind of fibrous protein consisting of glycine, proline, hydroxyproline, glutamic acid and the like, which is small in amino acid containing sulfur in the amino acid. It is a light protein having a long and narrow band shape, It plays an important role as a constituent of the dermis layer in the skin, especially in the membranes covering the organs of the body, articular cartilage, eye cornea, bones and skin.

Keratin is a member of an intermediate fiber protein that makes intermediate fibers in animal cells, and is an important constituent protein that makes keratin fibers especially in epidermal cells such as skin. Keratin is the most abundant intercellular fibrous connective protein in the epidermis and about 20 types of keratin are found on the human epidermis.

Since keratin has a high molecular weight and is connected by cystine disulfide bonds, it is insoluble and does not decompose well and its molecular weight is large and can not be easily removed by detergent alone. These protein impurities can be effectively removed by small degradation. Keratin is a fibrous protein with collagen and pseudo-unit structure. When degraded to low molecular weight, keratin effectively produces low molecular weight collagen peptide.

Since the present invention uses an enzyme having an excellent proteolytic activity, the collagen peptide can be efficiently produced in a simple process from pork. The collagen peptide produced by the method of the present invention is much lower in molecular weight than normal collagen, and thus has excellent bioabsorbability and is suitable for food and cosmetic raw materials.

Hereinafter, the present invention will be described more specifically based on examples. It is to be understood, however, that these examples are for illustrative purposes only and are not intended to limit the scope of the invention.

Example 1 Isolation of Microorganisms from Carnivorous Plants

Four kinds of carnivorous plants such as Nepenthes, Sarracenia, Drosera rotundifolia L. and Pinguicula were prepared to isolate microorganisms having protease activity from the samples, A certain amount of specimen is collected from the larvae of the carnivorous plants and 9 ml of physiological saline solution sterilized in 1 g of the carnivorous plant sample, and then serially diluted with sterile physiological saline.

For the selection of the strains having the activity of degrading the protein, a medium (proteolytic microorganism separation medium) having a composition of 0.3% of enzyme extract, 0.5% of peptone, 0.5% of azo casein (or 1% of skim milk) and 1 L of distilled water was prepared, . Dissolve 100-200 μL of the diluted sample in the medium, and platemorize using a gonad rod.

Dissociated plates are cultured for 3 days in a 30 ° C incubator, and then microorganism populations with clear zones due to proteolysis are first screened. When a microorganism having a protease degrades the protein source in the azo casein or skim milk added as a substrate during microbial growth, a clear zone is formed around the colony. Using this, microorganisms having a proteolytic enzyme are selected.

<Example 2> Identification of Identified Microorganisms

The 16S rDNA sequence was analyzed to identify microorganisms with proteolytic activity. The nucleotide sequences of the isolated strains were determined by Solzent (Daejeon, Korea). The analyzed nucleotide sequences were analyzed using an EzTaxon data server (http://eztaxon-e.ezbiocloud.net/) And the species and the genus of the strain were identified and identified.

As a result of analysis, the sample No. SW-4 was 98.16% identical to Chrysobacterium taiwanense BCRC 17412 (T). Chryseobacterium taiwanense BCRC 17412 (T) strain is known to be mainly isolated from soil, and thus it can be seen that the relationship between the sample of the carnivorous plant and the soil is not related. Especially, these strains have been reported to have various enzyme activities. The microorganisms isolated through the present invention were 98.16% identical to Chryseobacterium taiwanense BCRC 17412 (T), which was considered to be a novel strain and was confirmed as Chrysobacterium sp. MK-8 And donated with the deposit number KFCC11582P on March 17, 2014 at the Korea Microorganism Conservation Center attached to the Korean Society of Bacterial Cultures.

<Example 3> Characterization of Chrysobacterium sp. MK-8

The morphological, physiological and fermentative characteristics of the named new microorganism Chrysobacterium sp. MK-8 were examined. Chryseobacterium sp. MK-8 showed Gram negative as a result of Gram stain and was well grown under aerobic conditions. It did not form a spore and had no mobility. It looks like a germ. In addition, it is possible to grow at 4 to 37 ° C (optimal 20 to 30 ° C), pH 5.5 to 8.0 (optimum 7.0 to 7.5) and 0 to 3.0% (w / v) NaCl positive for oxydase and catalase. The physiological characteristics of the Chryseobacterium sp. Strain MK-8 are shown in Tables 1 and 2 below. Table 2 shows the fatty acid composition of Chrysobacterium sp. MK-8, and tr (traces) is less than 1.0%.

Characteristics MK-8 characteristics  MK-8   Catalase +   Gluconate _   Oxidase +   Citrate _   Growth temperature (℃) 4 - 37   Histidine +   pH for growth 5.5 - 8   L-Rhamnose _   Growth with NaCl (%, w / v) 0 - 3   Sucrose +   Indole production +   Enzyme activity:   Nitrate reduction -   Esterase (C4) +   Hydolysis of:   Cystine arylamidase +   Urea -   Trypsin +   DNA +   α-Chymotrypsin +   Assimilation of:   α-Galactosidase -   Arginine dihydrolase -   β-Galactosidase w   Histidine +   β-Glucosidase +   D-Gluose +   N-Acetyl-β-glucosaminidase +   L-Arabinose +   DNA G + C content (mol%) 41.7   D-mannose -

fatty acid MK-8   Saturated C _{16 : 0} 1.7   Unsaturated C _{18 : 1} omega 5C tr iso-C _{17 : 1} omega 9C 7.84   Branched-chain iso-C _{13 : 0} 1.4 iso-C _{15 : 0} 48.7 iso-C _{16 : 0} tr iso-C _{17 : 0} tr anteiso-C _{15 : 0} 5 iso-C _{15: 0} 3OH 4 iso-C _{16 : 0} 3OH Tr iso-C _{17 : 0} 3OH 17.4   Hydroxy C _{16: 0} 3OH One C _{17: 0} 2OH One   * Summed feature 3 6.64

Example 4 Characterization of Protease Derived from Chrysobacterium sp. MK-8

In order to measure proteolytic activity, the culture broth was centrifuged, and the supernatant was used as a crude enzyme solution. The recovered crude enzyme solution was reacted with 300 mL of 0.5% casein and 200 mL of crude enzyme solution at 37 ° C for 10 minutes, and 500 mL of Tricholoacetic acid was added thereto to terminate the reaction. The absorbance was measured and the resolution was examined. The amount of protein was quantitated by BCA method to quantify the amount of protein in the cells. Bicinchoninic acid solution and copper sulfate solution were mixed at a ratio of 49: 1. After mixing 950 μL of the mixed solution and 50 μL of the sample (crude enzyme solution), the absorbance was measured at 37 ° C. for 20 minutes.

The stability of the enzyme according to the reaction temperature, the reaction pH and the storage period was examined and shown in FIGS. 2 to 4.

As a result of the investigation of the reaction temperature, it showed the most stable decomposition activity in the range of 30 to 40 ° C, showed activity of 70% or more at the maximum activity standard at 15 ° C and exhibited activity of 80% or more until 50 ° C, Lt; / RTI &gt; These results suggest that protease activity can be maintained at various temperature ranges depending on various temperature conditions exposed during the growth of carnivorous plants. The activity of the protease of the present invention showed a tendency to decrease sharply at a low temperature of 10 ° C or lower and a temperature of 60 ° C or higher, but it has an active temperature range suitable for use as a cosmetic raw material (see FIG. 2).

In addition, as a result of examining the proteolytic activity in the range of pH 3.5 to 13.0 during the reaction, the enzyme activity was not affected in the pH range of weak acidity and alkaline range (pH 5.5 to 11.0) , And the activity of degrading the enzyme by protein denaturation is markedly decreased at acidic and strong alkaline pH ranges (see FIG. 3).

As a result of examining the stability of the enzyme activity during the storage period, there was little decrease in the enzyme activity during one month of storage at 35 ° C, which strongly indicates the activity of the enzyme. No change was seen. On the other hand, the proteolytic activity was remarkably decreased at a high temperature of 60 ° C or higher, which was judged to be due to the thermal deformation of the enzyme itself (see FIG. 4).

Example 5 Preparation of Protease and Activity Measurement

<5-1> Preparation of protease

Chryseobacterium sp. In order to isolate the proteolytic enzyme from the culture medium produced from MK-8, the microorganism was cultured in a culture medium (yeast extract 0.3%, peptone 0.5%, azocasein 1.0% (or 1% skim milk), dextrose 0.5% 1 L) was prepared and the surface was sterilized. The microorganisms were inoculated and cultured at 30 DEG C for 24 hours. The culture supernatant was centrifuged to obtain a partially purified crude enzyme solution.

<5-2> Measurement of proteolytic activity

The sterilized pork slices were cut into 20 mm x 4 mm x 1 mm thick and placed in a falcon tube. The enzyme solution isolated in <5-1> was diluted to 50%, and 30 ml of the diluted enzyme solution was added to the enzyme solution ) Was confirmed.

As a control group, distilled water was used instead of crude enzyme solution.

As a result, as shown in FIG. 6, the control group showed no significant change in the pungency until the end of the reaction, but in the case of the group to which the crude enzyme solution was added, the pung was completely decomposed at the 6th day of the reaction. Thus, it was confirmed that the culture solution of the present invention had proteolytic activity.

<5-3> Measurement of collagen peptide composition

The results of the analysis of the amino acid composition of the collagen powder were compared with those of the commercial collagen powder.

As a result, as shown in Table 3 and FIG. 7, it was confirmed that the amino acid composition of the collagen peptide produced by the strain-derived collagen degradation composition of the present invention was similar to that of the existing collagen peptide.

Example Comparative Example 1
Pig collagen AA MOL % MOL % ASX 5.26 5.07 GLX 9.70 8.44 SER 2.68 4.50 GLY 36.56 36.59 HIS 2.01 0.21 ARG 0.92 0.56 THR 2.33 2.25 ALA 12.78 12.95 PRO 13.40 15.20 TYR 0.57 0.38 VAL 2.64 2.81 MET 0.73 0.79 ILE 1.45 9.01 LEU 3.07 1.13 PHE 1.79 0.10 LYS 4.10 0.00 TOTAL 100.00 100.00

As a result, the composition of the present invention comprising the culture medium having the proteolytic activity produced from the genus Chryseobacterium sp. MK-8 of the present invention as an active ingredient has excellent proteolytic activity, and the culture solution is treated to form collagen peptide Were produced.

As described above, the embodiments of the present invention have been described in detail with reference to the drawings. However, various modifications may be made without departing from the scope of the present invention. Therefore, the scope of the present invention should not be limited by the described embodiments, but should be determined by the scope of the appended claims and equivalents thereof.

Korea Microorganism Conservation Center (Domestic) KFCC11582P 20140317

<110> LEE, hyun gy <120> Novel microorganism separated from carnivorous plants and          protease produced therefrom <130> p2014 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 1412 <212> DNA <213> Chryseobacterium sp. MK-8 (KFCC11582P) <400> 1 ccagacggag ctaaatgcag ctgagcggag aggcccttcg gggtcttgag agcggcgtac 60 gggtgcggaa cacgtgtgca acctgccttt atcaggggga tagcctttcg aaaggaagat 120 taatacccca taatattttg gatggcatca tttaaaattg aaaactgagg tggataaaga 180 tgggcacgcg caagattaga tagttggtga ggtaacggct caccaagtcg atgatcttta 240 gggggcctga gagggtgatc ccccacactg gtactgagac acggaccaga ctcctacggg 300 aggcagcagt gaggaatatt ggacaatggg ttagcgcctg atccagccat cccgcgtgaa 360 ggacgacggc cctatgggtt gtaaacttct tttgtacagg gataaaccta tctacgtgta 420 gatagctgaa ggtactgtac gaataagcac cggctaactc cgtgccagca gccgcggtaa 480 tacggagggt gcaagcgtta tccggattta ttgggtttaa agggtccgta ggcggatgtg 540 taagtcagtg gtgaaatctc acagctcaac tgtgaaactg ccattgatac tgcatgtctt 600 gagtaaggta gaagtggctg gaataagtag tgtagcggtg aaatgcatag atattactta 660 gaacaccaat tgcgaaggca ggtcactatg tcttaactga cgctgatgga cgaaagcgtg 720 gggagcgaac aggattagat accctggtag tccacgccgt aaacgatgct aactcgtttt 780 tggggattta tcttcagaga ctaagcgaaa gtgataagtt agccacctgg ggagtacgaa 840 cgcaagtttg aaactcaaag gaattgacgg gggcccgcac aagcggtgga ttatgtggtt 900 taattcgatg atacgcgagg aaccttacca aggcttaaat gggaattgat cggtttagaa 960 atagaccttc cttcgggcaa ttttcaaggt gctgcatggt tgtcgtcagc tcgtgccgtg 1020 aggtgttagg ttaagtcctg caacgagcgc aacccctgtt actagttgct accattaagt 1080 tgaggactct agtaagactg cctacgcaag tagagaggaa ggtggggatg acgtcaaatc 1140 atcacggccc ttacgccttg ggccacacac gtaatacaat ggccggtaca gagggcagct 1200 acacagcgat gtgatgcaaa tctcgaaagc cggtctcagt tcggattgga gtctgcaact 1260 cgactctatg aagctggaat cgctagtaat cgcgcatcag ccatggcgcg gtgaatacgt 1320 tcccgggcct tgttacacac cgcccgtcaa gccatggaag tctggggtac cgaagtcggg 1380 accgaacaga gcgccaggta tcgctgttat tt 1412

Claims (6)

Chrysobacterium sp. Strain MK-8 derived from a carnivorous plant (accession number: KFCC11582P).
A culture medium having proteolytic activity produced from Chrysobacterium sp. MK-8 of claim 1.
3. The culture medium according to claim 2, wherein the culture liquid has a proteolytic activity at a temperature of 15 to 50 DEG C and a pH range of 5.5 to 11.0.
A method for producing a strain of Chrysobacterium sp. MK-8 comprising inoculating the strain of claim 1 in a medium containing a carbon source and a nitrogen source, and culturing at a temperature of 4 to 37 DEG C and a pH of 5.5 to 8 Lt; / RTI &gt;
A composition comprising the strain of claim 1 or a culture thereof as an active ingredient to decompose a protein to produce a low molecular weight collagen peptide.
A method for producing a low molecular weight collagen peptide comprising treating the strain of claim 1 or a culture thereof with a protein-containing sample.

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