KR20130141973A - Functional food composition containing decapentaplegic protein originated from silkworm in order to prevention and improvement of bone-related diseases - Google Patents

Abstract

The present invention relates to a food composition containing Dpp protein originated from silkworm as an active ingredient for the prevention and treatment of bone-related diseases. The present invention provides a food composition for the prevention and treatment of bone-related diseases containing Dpp protein originated from silkworm which contains the amino acid sequence with the sequence number 2 or a base sequence with the sequence number 2 for coding the amino acid sequence. The food composition for the prevention and treatment of bone-related diseases contains the Dpp protein originated from silkworm which is separated from silkworm and induces the production of bones by improving the differentiation of osteoblast necessary for the bone production.

Description

FUNCTIONAL FOOD COMPOSITION CONTAINING DECAPENTAPLEGIC PROTEIN ORIGINATED FROM SILKWORM IN ORDER TO PREVENTION AND IMPROVEMENT OF BONE-RELATED DISEASES}

The present invention relates to a food composition for preventing and improving bone-related diseases including silkworm-derived Dpp protein, and more particularly to a silkworm-derived Dpp (Decapentaplegice) protein that induces bone formation by increasing osteoblast differentiation required for bone formation. It relates to a food composition for the prevention and improvement of bone-related diseases comprising.

Bone morphogenetic protein (BMP) belongs to the TGF-β super family and is responsible for inducing osteoblast differentiation.

In particular, BMP-2, BMP-4 and BMP-7 proteins are known to directly induce osteoblast differentiation. Among them, BMP-2 is developed as a recombinant protein and is approved and sold in the US and Europe. have.

In addition, as patents expire recently, several companies, including Daewoong Pharmaceutical, Cowell Medi, and Korea Bonn Bank, are developing and marketing them as medical devices.

Several studies have reported that human BMP genes are homologous to Dpp (Decapentaplegic) genes in fruit flies.

The Drosophila Dpp gene plays an important role in the musculoskeletal formation in the Drosophila developmental stage, and its signaling mechanism has been reported to be similar to the BMP protein signaling mechanism related to human bone formation.

Related prior arts include Republic of Korea Patent No. 1135779 (Registration date: 04/04/2012, Name: Composition for promoting osteoblast differentiation including diosgenin and its use) and Republic of Korea Patent No. 686685 (Registration date: 2007 February 16, name: composition for inducing bone differentiation of mesenchymal stem cells containing a histone deacetylase inhibitor and a method for increasing bone differentiation using the same).

An object of the present invention is to remove the Dpp gene homology with the Drosophila Dpp and human BMP genes from silkworms to improve human bone disease, thereby increasing the osteoblast differentiation required for bone formation to induce bone formation It is to provide a food composition for the prevention and improvement of bone-related diseases comprising the derived Dpp protein.

The present invention for achieving the above object is characterized in that the food composition for preventing and improving bone-related diseases contains silkworm-derived Dpp protein as an active ingredient.

The silkworm silkworm pupa and the silkworm is characterized by containing more from male silkworm.

The Dpp protein is characterized by consisting of the amino acid sequence of SEQ ID NO: 2.

The amino acid of SEQ ID NO: 2 is characterized in that encoded from the nucleotide sequence of SEQ ID NO: 1.

The Dpp protein is characterized by promoting osteoblast differentiation or activity.

The bone-related diseases include osteoporosis, bone metastatic lesions, primary tumors produced in the bone, rheumatoid or degenerative arthritis, periodontal disease, inflammatory alveolar bone resorption disease, inflammatory bone resorption disease and paguet disease (Paget's disease) is characterized in that it is selected from the group consisting of.

Health functional food of the present invention is characterized by containing a food composition for the prevention and improvement of the bone-related diseases.

The food additive of the present invention is characterized by containing a food composition for preventing and improving the bone-related diseases.

According to the present invention, by providing a basis for the prevention and improvement effect on bone-related diseases, it is possible to expect a therapeutic value increase of silkworms, which is licensed as a functional food and relatively undervalued, and is economically developed in food and pharmaceutical materials. This is secured.

It is expected that the expression of the Dpp gene will be expanded in the pupal stage than in the larvae and in male silkworms as compared to the female silkworms. .

1 is a photograph showing cDNA amplification of the Dpp gene from baekjajam (5 year old silkworm larvae) fat tissue.
Figure 2 is a diagram showing the nucleotide sequence change between the Dpp gene reported in GeneBank (FJ572058.1) and Baekjam-derived Dpp gene of the present invention.
Figure 3 is a view showing the amino acid sequence changes between the Dpp gene reported in GeneBank (FJ572058.1) and Dak gene derived from jadeite of the present invention.
Figure 4 shows the 'pCEP4-Bm Dpp' plasmid DNA.
Figure 5 shows the introduction of silkworm Dpp gene through genomic PCR.
(Top) internal control gene (Mouse GAPDH)
(P) "pCEP4" plasmid DNA vector
(P) "pCEP4-Bm Dpp" plasmid DNA vector (invention)
1. C3H10T1 / 2 Cells
2. C3H10T1 / 2 cells with pCEP4 plasmid DNA
3. C3H10T1 / 2 Cells Incorporated with pCEP4-Bm Dpp plasmid DNA
Figure 6 shows the introduction of silkworm Dpp gene via RT-PCR.
(Top) internal control gene (Mouse GAPDH)
(Bottom) "pCEP4-Bm Dpp" plasmid DNA (invention)
1. C3H10T1 / 2 Cell cDNA
2. C3H10T1 / 2 Cell NO-RT Conditions
3. pCEP4 cDNA Conditions C3H10T1 / 2 Cells
4. C3H10T1 / 2 Cells Introduced with pCEP4 NO-RT
5. C3H10T1 / 2 Cells Incorporated with pCEP4-Bm Dpp cDNA
6. pCEP4-Bm Dpp NO-RT Condition C3H10T1 / 2 Cells
7 is a diagram showing the expression level of four specific genes (Runx2, Osterix, Osteocalcin, ALP) that is osteoblast differentiation after introduction of the "pCEP4-Bm Dpp" plasmid DNA of the present invention.
C3H10T-1 / 2 Normal Cells: C3H10T-1 / 2 Cell Expression
C3H10T-1 / 2 Cells-pCEP4: C3H10T-1 / 2 Cell Expression with pCEP4
C3H10T-1 / 2 Cells-pCEP4 Bm Dpp: C3H10T-1 / 2 Cell Expression Introduced with 'pCEP4-Bm Dpp'
8 is a diagram showing the RT-PCR results of cDNA synthesized from RNA isolated from tissue specific.
hemocyte TR: blood cell tissue total RNA
hemocyte cDNA: blood cell tissue cDNA
Midgut TR: Colonic Tissue Total RNA
Midgut cDNA: mid-tissue cDNA
Fat body TR: fat tissue total RNA
Fat body cDNA: fat tissue cDNA
9 shows tissue specific expression of silkworm Dpp gene.
silk gland: silk gland tissue
hemocyte: blood cell tissue
Midgut: Medium Tissue
Fat body: fat tissue
Figure 10a is a diagram showing the expression rate according to sex of silkworm larvae derived Dpp gene.
larvae male: larvae
larvae female: female larvae
Figure 10b is a diagram showing the expression rate according to sex of silkworm pupa derived Dpp gene.
pupae male: pupa on male
pupae female: female pupa
11 is a diagram showing the expression rate of silkworm-derived Dpp gene by developmental stage.
pupae male: pupa on male
larvae male: larvae
pupae female: female pupa
larvae female: female larvae

Hereinafter, the present invention will be described in detail, and detailed descriptions of well-known functions and configurations that may unnecessarily obscure the subject matter of the present invention will be omitted.

In general, human bone formation promoting protein (BMP) is used as an agent which plays a role of inducing differentiation of osteoblasts.

However, such a BMP requires a process for the expression and production of the protein using animal cells and human-derived genes at the drug manufacturing level for safer and more efficient use, and thus, it is difficult to continuously supply economically.

To compensate for this, the present invention was to seek the present invention by quoting that the human BMP gene has homology with the Decapentaplegic (Dpp) gene in Drosophila.

In other words, the present invention provides a food composition for the prevention and improvement of bone-related diseases containing the silkworm-derived Dpp protein consisting of the amino acid sequence of SEQ ID NO: 2 shown in Figure 3 as an active ingredient.

In addition, the present invention is characterized in that the amino acid of SEQ ID NO: 2 is encoded from the nucleotide sequence of SEQ ID NO: 1 shown in FIG.

In Example 1 of the present invention, the sequence information of the Dpp gene, which is a gene for inducing bone cell formation for the improvement of human bone disease, is confirmed, and total RNA is isolated and cDNA is synthesized from baekjap fat tissue.

The cDNA was confirmed amplification degree through gene amplification using a specific primer (see Fig. 1)

In order to confirm the nucleotide sequence of the gene, the amplified gene was inserted into the cloning vector pGEM-T Easy vector by TA cloning, and the nucleotide sequence and amino acid sequence of the amplified gene were confirmed by sequencing. 2 and 3)

In addition, in Example 2, the Dpp gene was inserted into a gene vector (pCEP4 plasmid DNA) that can be expressed in animal cells in order to confirm the bone cell formation promoting function of the Dpp protein to construct pCEP4-Bm Dpp plasmid DNA. 4)

The pCEP4-Bm Dpp plasmid DNA thus produced was introduced into stem cells to confirm that differentiation was induced into osteoblasts (see FIGS. 5 and 6).

Here, "stem cells" are cells that can easily proliferate in vitro and can be differentiated into various cell types (fat cells, chondrocytes, bone cells), preferably mesenchymal stem cells, which are adult stem cells.

As described above, if differentiation was induced into osteoblasts by introducing pCEP4-Bm Dpp plasmid DNA into stem cells, the expression levels of four proteins (Runx2, Osterix, Osteocalcin, ALP) were also increased (see FIG. 7).

In other words, the four proteins, Runx2, Osterix, Osteocalcin, and ALP, are osteoblast differentiation markers as proteins identified during osteoblast differentiation by BMP.

Thus, it was confirmed that silkworm-derived Dpp protein plays a role in promoting bone formation.

In addition, the Dpp gene has an expression rate regardless of the sex or developmental stage of the silkworm, but in order to identify a gender or developmental stage showing a higher expression rate, in Example 3, the Dpp gene is determined by sex and developmental stage of the silkworm. Each tissue was isolated from silkworm larvae and the tissue DPP gene expression rate was confirmed (see FIGS. 8 to 11).

As a result, male silkworms showed higher expression rate than female silkworms, and in the developmental stage, the expression rate was higher in pupa than in larva, and finally it was confirmed that the highest expression rate in male pupa.

The silkworm-derived Dpp protein can be used as a food composition for the prevention and improvement of bone-related diseases. The bone-related diseases include osteoporosis, bone metastatic lesions, primary tumors produced in the bone, rheumatoid or degenerative arthritis, periodontal disease, inflammatory alveolar bone resorption disease, inflammatory bone resorption disease and paguet Paget's disease.

In addition, the food composition for preventing and improving the bone-related diseases is to be used as a health functional food or food additives, in this case other effects that can give a synergistic effect of the main effect within the range that does not impair the main effect of the present invention. It may contain a component or the like.

For example, fragrances, pigments, fungicides, antioxidants, preservatives, moisturizers, thickeners, inorganic salts, emulsifiers, synthetic polymers, etc. may be further included to improve physical properties, as well as water-soluble vitamins, oil-soluble vitamins, and polymer peptides. It may further include auxiliary components such as polymer polysaccharide and seaweed extract.

The above ingredients may be appropriately selected and blended by those skilled in the art according to the dosage form or purpose of use, and the amount may be selected within a range that does not impair the object and effect of the present invention.

In addition, the formulation of the composition may also be in various forms such as solutions, emulsions, viscous mixtures, tablets, powders, etc., which may be ingested by various methods such as simple drinking.

As described above, the application field of the food composition for preventing and improving bone-related diseases of the present invention is not shown in the following specific examples, but the health functional food or food having the effects of preventing and improving bone-related diseases with the above-mentioned efficacy. It will be apparent to those skilled in the art that it can be used as an additive.

In this way, high-quality protein can be produced at low cost, and economical efficiency can be secured when developing it as a food and pharmaceutical material. Also, by providing a basis for the prevention and improvement effect on bone-related diseases, it is only approved as a functional food and relatively The value added of undervalued silkworms can be expected.

Hereinafter, the present invention will be described in detail with reference to examples, but the scope of the present invention is not limited thereto.

<Example 1> Gene cloning for securing Dpp gene derived from silkworms

1) Experiment and Results

Ⓛ Genetic information acquisition and cDNA synthesis

From the web page of NCBI, silkworm-specific Dpp gene (GeneBank: FJ572058.1) sequencing information (total 1,285 bp, gene coding region up to 164-1,273 bp) was confirmed.

Total RNA isolation and cDNA synthesis were performed from baekjajam (5 day 3 silkworm larvae) adipose tissue, and gene amplification was performed using specific primers. As a result, the amplified Dpp gene target (square) could be confirmed as shown in FIG. 1.

② Gene base sequence confirmation

The gene amplified by the PCR reaction was inserted by TA cloning into pGEM-T Easy vector, which is a cloning vector, to confirm Dpp gene sequencing information derived from Baekjam.

As a result, as shown in Figure 2 when comparing the Dpp gene reported in GeneBank (FJ572058.1) and Dpp gene derived from Baekjamjam it was confirmed that a total of seven nucleotide sequence changes appear.

Thus, it was confirmed that there is a slight difference between the nucleotide sequence and amino acid sequence of the Dpp gene of the known Dpp gene (GeneBank: FJ572058.1) and the white jade silkworm used in the present invention (Fig. 3).

Example 2 Confirmation of Bone Cell Formation Promoting Function of Silkworm-derived Dpp Gene

1) Promote bone cell formation of silkworm-derived Dpp gene In order to confirm the functionalities, pCEP4-Bm Dpp plasmid DNA is constructed by inserting the Dpp gene into a gene vector (pCEP4 plasmid DNA) that can be expressed in animal cells.

1. Experiment

pCEP4-Bm Dpp plasmid DNA was introduced into C3H10T-1 / 2 cells to confirm that differentiation was induced into osteoblasts.

Here, C3H10T-1 / 2 cells are pluripotent embryonic mesenchymal stem cell lines derived from rodents and are capable of differentiating into osteoblasts or adipocytes depending on conditions.

C3H10T-1 / 2 cells into which pCEP4-Bm Dpp plasmid DNA is introduced through the introduction process as shown in FIG. 4 express the silkworm Dpp gene, and if the expressed silkworm Dpp protein plays the same role as human BMP protein, C3H10T- results. It can be seen that 1/2 cells are differentiated into osteoblasts.

2. Results

Ⓛ Confirmation of silkworm Dpp gene introduction possibility through genomic PCR

The genomic DNA of C3H10T-1 / 2 cells into which pCEP4-Bm Dpp plasmid DNA was introduced is isolated, and as shown in FIG. 5, the internal control gene (upper), pCEP4 plasmid DNA vector (middle), and pCEP4-Bm Dpp plasmid DNA ( Expression level was confirmed using specific primers for ha).

As a result, the C3H10T-1 / 2 cells to which the pCEP4-Bm Dpp plasmid DNA was delivered were confirmed by genomic PCR that the silkworm Dpp gene was introduced.

② Confirmation of intracellular expression potential of silkworm Dpp gene introduced through RT-PCR

Total RNA from C3H10T-1 / 2 cells into which pCEP4-Bm Dpp plasmid DNA was introduced was isolated. As shown in FIG. 6, cDNA was synthesized and internal control gene (upper phase) and pCEP4-Bm Dpp plasmid DNA ( Expression level was confirmed using specific primers for ha).

 As a result, it was confirmed by RT-PCR that the C3H10T-1 / 2 cells to which the pCEP4-Bm Dpp plasmid DNA was delivered were correctly expressed in the silkworm Dpp gene.

In addition, as shown in Figure 6, unlike the normal cells, cells containing the Dpp gene was confirmed that the transcript of the Dpp gene is expressed.

2) Confirmation of functionality for promoting bone cell formation of Dpp gene

Stem cells or progenitor cells (cells prior to differentiation into bone cells; C3H10T-1 / 2 cells) were used to confirm the functionalities for promoting bone cell formation of the Dpp gene.

1. Experiment and Results

PCEP4-Bm Dpp plasmid DNA containing the Dpp gene was infected with stem cells or progenitor cells to deliver the target gene, Dpp gene, into the cell.

After transduction, bone cell differentiation was measured using a quantitative Real-Time RT-PCR method.

Schematic of the process by which stem cells or progenitor cells are differentiated into osteoblasts by BMP protein. Expression of four proteins (Runx2, Osterix, Osteocalcin, ALP) increases specifically in the signaling process for differentiation into osteoblasts, and these are indicative substances that can be differentiated into osteoblasts.

Therefore, if stem or progenitor cells are differentiated into bone cells after introduction of pCEP4-Bm Dpp plasmid DNA, gene expression of Runx2, Osterix, Osteocalcin, ALP, etc. is increased, and thus the degree of differentiation can be measured and functional confirmation is possible.

As shown in FIG. 7, the expression of four specific genes (Runx2, Osterix, Osteocalcin, ALP), which are osteoblast differentiation markers, was increased, and pCEP4-Bm Dpp plasmid was compared to normal cells or cells into which only plasmid vector DNA was introduced. It was confirmed that the expression of differentiation marker genes increased significantly in the cells into which DNA was introduced.

The silkworm Dpp gene can be confirmed to play a role in promoting bone formation.

<Example 3> Expression of Dpp gene expression rate from silkworms on the osteoporosis improvement effect of male silkworm bunde

1) Confirmation of Dpp gene expression rate by silkworm tissue

Total RNA isolation and cDNA synthesis were performed from four different tissues of silkworm silkworms at 5 days and 5 days (eg, silk glands, midgut, fat body, blood cells, etc.).

As a result, as shown in the RT-PCR results of Figure 8, genomic DNA was completely removed and confirmed that the cDNA was made correctly.

In addition, as a result of RT-PCR for confirming the tissue specificity of cDNA isolated and synthesized from each tissue, it was possible to confirm the expression rate of Dpp inheritance from each tissue.

That is, as shown in FIG. 9, when the expression rate of the Dpp gene expressed in the silk gland is 100, it was confirmed that the expression of the Dpp gene in the blood cells and the mid-tissue was more than two times, and the expression rate of the Dpp gene for each tissue was fat body <dogline (Silk gland) <Midgut (Midgut) <Hemocytes were found in the order of expression.

2) silkworm gender Dpp gene expression rate

Currently, silkworms are mainly used for male silkworms, and as shown in the quantitative PCR results of FIGS. 10A and 10B, as a result of confirming the Dpp gene expression rates of female silkworms and male silkworms in larvae and pupa, both larvae and pupa are derived from male silkworms. The expression rate of the Dpp gene was confirmed to be high.

3) Confirmation of Dpp gene expression rate by silkworm developmental stage

Currently, silkworms are mainly used for male pupa, and as shown in the quantitative PCR results of FIG. 11, the Dpp gene expression rate was confirmed in larvae and pupae.

As shown in the above quantitative PCR results, it was confirmed that the Dpp expression of female chrysalis was significantly less than that of male chrysalis, and the expression of female chrysalis was much higher than that of male larvae. Confirmed to show. As a result, it was confirmed that the Dpp gene expression of the male chrysalis showed the highest result.

In this regard, food and medicine materialization using silkworm pupa can be an important indicator of the effects on the improvement and treatment of human bone diseases such as osteoporosis. Since protein can be produced at low cost, it is expected to secure economic feasibility when developing it as a food and medicine material.

In addition, by providing evidence on the improvement and treatment effect for bone-related diseases, it is possible to expect a therapeutic value-added increase in silkworms, which are licensed as functional foods and relatively undervalued.

The expression of the Dpp gene is more expressed in pupa than in larvae and in male silkworms than in female larvae.

<110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> FUNCTIONAL FOOD COMPOSITION CONTAINING DECAPENTAPLEGIC PROTEIN          ORIGINATED FROM SILKWORM IN ORDER TO PREVENTION AND IMPROVEMENT          OF BONE-RELATED DISEASES <130> 2012-0083-10-A <160> 2 <170> Kopatentin 2.0 <210> 1 <211> 1285 <212> DNA <213> DECAPENTAPLEGIC GENE FROM SILKWORM <400> 1 agagattcgt tgttatgtgc cgttgcttcg ttcgtgaact agtgcgagtc aatctcttcc 60 cgaagcgttg accctgcgtg cgaatccctc gcgaggatat cgcggatttt gcgtacccgt 120 ctacgttgga tgcgttaaaa ttttgggtgg gtcgaggaga attatgcgtg gggcgtgcgc 180 atgcgcggtg gtgtgcgcgt tggtggcgct gtgcgcagcc gcgggcctcg acgaagcgac 240 gcgtgttgct gcagagaagc agttattggc gttgttgggt ctgccgaaga gaccgtcgcg 300 tcgatccgct ccagtaccgc ctataccacg cgccatgcga atgctttacg aggcaagcgg 360 agccataccg gccgctgcgg caaacacggc ccgttcatat cagcacgtac cgacggagct 420 cgatgcgagg ttcccaggcg aacatcgttt tcgcgtattc ttcaacataa gtggagtacc 480 ctctgatgaa gtagctcgtg gtgctgatct caaatttcat cgcgcgactg aagagacggg 540 tcctcagcgc ctattactat atgacgttgt acgtcctggt cgtcgaggga aaacgactcc 600 aattctaaga cttctcgatt ccgtgacatt attgccaggc gagggcacag tgacagcaga 660 cgccattgac gcagtgcgac ggtggctcct tgaaactgat caaaaccatg gactattagt 720 gcgtgttatt aaagaaggcc aacacaacgt tgatgcaaaa cggccacacg taagagttcg 780 aagacgagcg accgaagacg aagaagaatg gcgctctcag caacccttgc tgttgctgta 840 cactgaagat gcgcgagcca gagaagcacg cgagaatggg gagtcgcgtc taactcgaag 900 taaaagagca acacaacggc gtggtcatcg acctcaccac cgtcgtaaag aagctcgtga 960 aatctgccag cggcggccac tgtttgtcga cttcgcggaa gtcggctgga gcgactggat 1020 tgtcgcgcct cctggttacg aagcttattt ctgtcagggt gactgcccgt tcccgctcgc 1080 agatcatcta aatggcacta atcacgcaat agtgcaaact ttagtgaatt cagtggaccc 1140 agccttagtg cctaaagcgt gttgtatacc aacacaacta tcgcctattt ctatgttata 1200 tatggacgaa cataatcaag tggtgcttaa aaactatcag gatatgatgg tgatgggctg 1260 cggttgccga tgacgaaggc acgcc 1285 <210> 2 <211> 369 <212> PRT <213> DECAPENTAPLEGIC PROTEIN FROM SILKWORM <400> 2 Met Arg Gly Ala Cys Ala Cys Ala Val Val Cys Ala Leu Val Ala Leu   1 5 10 15 Cys Ala Ala Ala Gly Leu Asp Glu Ala Thr Arg Val Ala Ala Glu Lys              20 25 30 Gln Leu Leu Ala Leu Leu Gly Leu Pro Lys Arg Pro Ser Arg Arg Ser          35 40 45 Ala Pro Val Pro Pro Ile Pro Arg Ala Met Arg Met Leu Tyr Glu Ala      50 55 60 Ser Gly Ala Ile Pro Ala Ala Ala Ala Asn Thr Ala Arg Ser Tyr Gln  65 70 75 80 His Val Pro Thr Glu Leu Asp Ala Arg Phe Pro Gly Glu His Arg Phe                  85 90 95 Arg Val Phe Phe Asn Ile Ser Gly Val Pro Ser Asp Glu Val Ala Arg             100 105 110 Gly Ala Asp Leu Lys Phe His Arg Ala Thr Glu Glu Thr Gly Pro Gln         115 120 125 Arg Leu Leu Leu Tyr Asp Val Val Arg Pro Gly Arg Arg Gly Lys Thr     130 135 140 Thr Pro Ile Leu Arg Leu Leu Asp Ser Val Thr Leu Leu Pro Gly Glu 145 150 155 160 Gly Thr Val Thr Ala Asp Ala Ile Asp Ala Val Arg Arg Trp Leu Leu                 165 170 175 Glu Thr Asp Gln Asn His Gly Leu Leu Val Arg Val Ile Lys Glu Gly             180 185 190 Gln His Asn Val Asp Ala Lys Arg Pro His Val Arg Val Arg Arg Arg         195 200 205 Ala Thr Glu Asp Glu Glu Glu Trp Arg Ser Gln Gln Pro Leu Leu Leu     210 215 220 Leu Tyr Thr Glu Asp Ala Arg Ala Arg Glu Ala Arg Glu Asn Gly Glu 225 230 235 240 Ser Arg Leu Thr Arg Ser Lys Arg Ala Thr Gln Arg Arg Gly His Arg                 245 250 255 Pro His His Arg Arg Lys Glu Ala Arg Glu Ile Cys Gln Arg Arg Pro             260 265 270 Leu Phe Val Asp Phe Ala Glu Val Gly Trp Ser Asp Trp Ile Val Ala         275 280 285 Pro Pro Gly Tyr Glu Ala Tyr Phe Cys Gln Gly Asp Cys Pro Phe Pro     290 295 300 Leu Ala Asp His Leu Asn Gly Thr Asn His Ala Ile Val Gln Thr Leu 305 310 315 320 Val Asn Ser Val Asp Pro Ala Leu Val Pro Lys Ala Cys Cys Ile Pro                 325 330 335 Thr Gln Leu Ser Pro Ile Ser Met Leu Tyr Met Asp Glu His Asn Gln             340 345 350 Val Ala Leu Lys Asn Tyr Gln Asp Met Met Val Met Gly Cys Gly Cys         355 360 365 Arg    

Claims (9)

Contains silkworm-derived Dpp (Decapentaplegice) protein as an active ingredient,
Food composition for preventing and improving bone-related diseases.
The method of claim 1,
The silkworm is characterized by silkworm pupa,
Food composition for preventing and improving bone-related diseases.
3. The method of claim 2,
The silkworm is characterized by a male silkworm,
Food composition for preventing and improving bone-related diseases.
The method of claim 1,
The Dpp protein is composed of the amino acid sequence of SEQ ID NO: 2,
Food composition for preventing and improving bone-related diseases.
5. The method of claim 4,
The amino acid of SEQ ID NO: 2 is encoded from the nucleotide sequence of SEQ ID NO: 1,
Food composition for preventing and improving bone-related diseases.
The method according to any one of claims 1 to 5,
The Dpp protein is characterized in that to promote osteoblast differentiation or activity,
Food composition for preventing and improving bone-related diseases.
The method according to any one of claims 1 to 5,
The bone-related diseases are osteoporosis, bone metastatic lesions, primary tumors produced in the bone, rheumatoid or degenerative arthritis, periodontal disease, inflammatory alveolar bone resorption disease, inflammatory bone resorption disease and paguet disease (Paget's disease), characterized in that selected from the group consisting of,
Food composition for preventing and improving bone-related diseases.
A health functional food containing a food composition for preventing or ameliorating a bone-related disease of any one of claims 1 to 7.
A food additive containing a food composition for preventing or ameliorating a bone-related disease of any one of claims 1 to 7.

Images