KR102214600B1 - Functional composition comprsing low molecular collagen peptide extract from skate skin and Functional food comprising the same - Google Patents

Abstract

[화학식 2]

The body fat reduction functional raw material composition according to the present invention is a body fat reduction functional raw material composition comprising a low molecular weight collagen peptide obtained by hydrolyzing skate skin as an active ingredient, and has a protein content of 90% by weight or more, and is represented by the following formula (1). 1 compound 58-89 mg/g and a low molecular weight collagen peptide containing 190-286 mg/g of the second compound represented by the following formula (2) as an active ingredient,
The low molecular weight collagen peptide is a step of collecting and freezing the raw materials of the skate skin (consisting of 100% skate skin) generated during the skate processing process; Putting the skin in a high-pressure reactor and extracting hot water at 121° C. for 90 minutes; Hydrolyzing the proteolytic enzymes Alcalase and Protamax by adding 0.5% of each of the raw materials for 2 hours; Deodorizing using activated carbon and ion exchange resin; Removing impurities through vacuum filtration; And concentrating and lyophilizing after cutting off based on a molecular weight of 1,000 Da. It characterized in that it is manufactured through.
[Formula 1]

[Formula 2]

Description

Functional composition comprsing low molecular collagen peptide extract from skate skin and Functional food comprising the same} through preclinical and human application experiments of low molecular collagen peptide derived from skate skin

The present invention relates to a body fat reduction functional raw material composition comprising a low molecular weight collagen peptide obtained by hydrolyzing skate skin as an active ingredient, and for the development and industrialization of functional raw materials for reducing body fat through preclinical and human application experiments of low molecular weight collagen peptide derived from skate skin. About.

Skate is a favorite food in Korea and has various pharmacological effects such as digestive function and appetite enhancement, immune function improvement and aging prevention, skin elasticity maintenance, and arthritis prevention. It is a form that is consumed without special processing, such as, scallop stew, and skatefish (liver).

Due to the functional characteristics and peculiar palatability of skates, the demand for skates has rapidly increased nationwide, from the form of being consumed only in small quantities in Jeollanam-do.

Fish skin and internal tissues contain a large amount of collagen, and it is reported that it is safe because it does not have the risk of human metastasis such as mad cow disease or foot-and-mouth disease, unlike collagen derived from the skin and bones of cattle and pigs.

Since the skate skin contains a large amount of collagen, it can be used as a functional food material with high added value. However, since collagen itself has a large molecular weight (known to have a molecular weight of 30 kDa or more), and is insoluble in water and has a low digestion and absorption rate, a low-molecular peptide is required to maximize absorption in the body.

Korean Patent Registration No. 10-1774701

The present invention has been conceived to solve the above problems, and an object of the present invention is to provide a functional raw material composition for reducing body fat comprising a low molecular weight collagen peptide obtained by hydrolyzing skate skin as an active ingredient.

In addition, the present invention is an experiment to extract collagen peptides by molecular weight by using the skin, a by-product generated during processing of skates, and to confirm the industrial applicability using the extract for the production of low molecular weight collagen peptides.General ingredients and molecular weight, amino acid composition, and deodorization It is to examine the possibility of industrial application through experiments such as.

The technical problems to be achieved by the present invention are not limited to the technical problems mentioned above.

In order to achieve this object, the body fat reduction functional raw material composition according to an embodiment of the present invention is a body fat reduction functional raw material composition comprising a low molecular weight collagen peptide obtained by hydrolysis of skate skin as an active ingredient, and has a protein content of 90 A low molecular weight collagen peptide containing 58 to 89 mg/g of the first compound represented by the following formula (1) and 190 to 286 mg/g of the second compound represented by the following formula (2) as an active ingredient,
The low molecular weight collagen peptide is a step of collecting and freezing the raw materials of the skate skin (consisting of 100% skate skin) generated during the skate processing process; Putting the skin in a high-pressure reactor and extracting hot water at 121° C. for 90 minutes; Hydrolyzing the proteolytic enzymes Alcalase and Protamax by adding 0.5% of each of the raw materials for 2 hours; Deodorizing using activated carbon and ion exchange resin; Removing impurities through vacuum filtration; And concentrating and lyophilizing after cutting off based on a molecular weight of 1,000 Da. It characterized in that it is manufactured through.

delete

[Formula 1]

[Formula 2]

In the body fat reduction functional raw material composition according to an embodiment of the present invention, the free amino acid of the low molecular weight collagen peptide is 6 to 8% by weight of the first compound and the second compound 19 based on 100% by weight of the total free amino acids. It may contain ~23% by weight.

As an example, the first compound may be Hydroxy proline, and the second compound may be Glycine.

In the body fat reduction functional raw material composition according to an embodiment of the present invention, the constituent amino acids of the low-molecular collagen peptide are 5.36 to 5.90 wt% of Aaspartic acid, 3.38 to 3.72 wt% of Threonine, and Serine based on 100 wt% of the total of the constituent amino acids. 9.77~10.71 wt%, Glutamic acid 9.77~10.71 wt%, Glycine 21.27~23.64 wt%, Alanine 7.65~8.51 wt%, Valine 2.88~3.17 wt%, Methionine 1.72~2.16 wt%, Isoleucine 2.34~2.58 wt%, Leucine 3.50 to 3.87 wt%, Tyrosine 0.73 to 0.86 wt%, Phenylalanine 2.07 to 2.28 wt%, Lysine 3.54 to 3.88 wt%, Ammonia 0.84 to 0.92 wt%, Histidine 1.49 to 1.62 wt% and Arginine 7.11 to 7.93 wt% I can.

In the functional raw material composition for reducing body fat according to an embodiment of the present invention, the free amino acid of the low molecular weight collagen peptide is 0.03 to 0.05% by weight of Phosphoserine, 0.01 to 0.02% by weight of Taurine, and Aspartic acid based on 100% by weight of the total free amino acid. 6.46~7.11 wt%, Hydroxy proline 6.10~6.85 wt%, Threonine 3.29~3.42 wt%, Serine 5.38~5.71 wt%, Glutamic acid 9.93~10.78 wt%, Proline 8.28~9.02 wt%, Glycine 19.97~22.09 wt%, Alanine 6.92~7.66 wt%, Valine 2.93~3.34 wt%, Methionine 1.67~2.15 wt%, Isoleucine 2.20~2.45 wt%, Leucine 3.32~3.67 wt%, Tyrosine 0.81~0.91 wt%, Phenylalanine 2.02~2.23 wt%, Ammonia 0.71 to 0.74 wt%, Hyroxylysine 0.09 to 1.05 wt%, Ornithine 0.03 to 0.04 wt%, Lysine 3.22 to 3.49 wt%, Histidine 1.26 to 1.38 wt%, and Arginine 7.05 to 7.84 wt%.

In addition, the present invention provides a functional food comprising the above-described body fat reduction functional raw material composition.

The functional raw material composition for reducing body fat according to the present invention can improve the effect of reducing body fat by maximizing absorption in the body.

In addition, when the functional food according to the present invention is taken for a medium/long term, it may have an effect on reducing body fat, and it is safe even if 2,000 mg/day for adults is consumed for a medium/long term.

In addition, despite the mass production of the functional raw material composition for reducing body fat according to the present invention, the yield of the product was about 17%, which shows a considerably high yield when considering the moisture content of the raw material of skate skin.

In addition, since the present invention extracts collagen peptides using by-products of skateboards that have been disposed of due to their extremely low availability, there is an effect of preventing waste of resources and reducing the cause of environmental pollution.

1 is a process flow chart showing a manufacturing process of a functional raw material composition for reducing body fat according to an embodiment of the present invention.
2 is a view showing a change in content and yield according to a manufacturing process of a functional raw material composition for reducing body fat according to an embodiment of the present invention.

Hereinafter, the present invention will be described in detail. The following embodiments and drawings are provided as examples in order to sufficiently convey the spirit of the present invention to those skilled in the art. In addition, unless there are other definitions in the technical and scientific terms used in the present invention, the present invention has the meanings commonly understood by those of ordinary skill in the art to which this invention belongs, and the present invention in the following description and accompanying drawings Descriptions of known functions and configurations that may unnecessarily obscure the gist of are omitted.

In describing the present invention, the low molecular weight collagen peptide is about 500 to 2,000 Da. It means a peptide having a molecular weight.

In addition, unless otherwise specified in the specification,% means% by weight.

1. Manufacturing and industrialization of small molecule collagen peptides

For the production of low molecular weight collagen peptides, collagen peptides were extracted by molecular weight by using the skin, a by-product generated during processing of skates, and this is an experiment to confirm the industrial applicability. Through experiments such as general ingredients, molecular weight, amino acid composition, and deodorization. The industrial applicability was reviewed.

end. Materials and methods

(1) Hydrolysis of raw materials of skate skin

As shown in Figs. 1 and 2, for hydrolysis of the raw materials of skate skin, the raw material of the skate skin generated during the peeling process during the process of processing the skates of our company (Youngsan Skate Co., Ltd.) (consists of 100% skate skin) ) Was collected (S10).

Then, after washing with running water three times, it was stored frozen at -25 ℃ or less (S20).

To prepare the low molecular weight collagen peptide, the skin was put in a high-pressure reactor (autoclave) and extracted with high-pressure hot water at 121°C for 90 minutes (S30), and then commercial proteolytic enzymes Alcalase, Flavourzyme, and Protamex (Novozyme, Denmark) were used. Hydrolyzed for each condition (S40).

(2) Preparation of collagen peptides by molecular weight

After the hydrolysis was completed, after going through a deodorizing process for removing fish odor using activated carbon and an ion exchange resin (S50), impurities were removed through vacuum filtration (S60). The filtered liquid was cut-off based on a molecular weight of 1,000 Da. by fractionating by molecular weight using an ultrafiltration device or by fractionating by molecular weight according to the enzyme type and hydrolysis time. The liquid that passed the molecular weight cut-off was concentrated (S70), lyophilized (S70) (SFDSM06, Samwon Co,. Korea), and used in the experiment, and industrial production was commercialized using a spray dryer.

I. Measurement of physicochemical properties

(1) Measurement of general ingredients

General ingredients were measured according to the general test method of the Food Code (Food Code, 2013). Moisture content was measured by normal pressure drying method, crude protein content was measured by micro kjeldahl method, crude fat content was measured by Soxhlet method, and ash content was measured by direct painting method. Each measurement was repeated three times to obtain the average.

(2) amino acid analysis

Hydrochloric acid was added to a certain amount of the dried sample, followed by hydrolysis, nitrogen gas was substituted, and left to stand in an oven at 110° C. for 22 hours, followed by evaporation under reduced pressure and dried. It was diluted with 0.02N hydrochloric acid, filtered through a 0.45 μm filter, and a certain amount was taken and analyzed using an automatic amino acid analyzer (L-8800 Hitachi Co,. Japan).

All. Molecular weight measurement

The molecular weight of the collagen peptide was measured using GPC (Gel Permeation Chromatography, Japan), and the standard product was a PEG calibration kit (agilent technologies, United Kingdom). At this time, TSKgel SuperMultipore®PW-H (6.0 mm ID Х 15 cm, Distilled Water, Japan) was used as the column, and 2 µl of the sample was injected into a buffer of pH 7.0 at a rate of 1 mL/min (0.1M KH2PO4 + 10%). NaOH) was measured at 25°C while flowing.

la. Ultrafiltration

Using an ultrafilter (EMP Series, EMP Tech, Korea), the extract extracted from the skate shell was used as Flow20 to 1K Da. It was passed through a filtration membrane (Pall Corporation, USA). Filtration was performed until no more filtrate came out, and a sample was prepared by separating the filtered liquid and the unfiltered liquid after the filtration was finished.

hemp. Human body application test

Human application trials were conducted after establishing a clinical trial monitoring plan, approved by the Institutional Review Board, and approved by the Institutional Review Board, followed the Declaration of Helsinki, and followed the Pharmaceutical Clinical Trial Management Standard (KGCP).

Before screening to select subjects, explanations for this test were distributed to all applicants, and after sufficient explanations, consent was signed by the applicant's own voluntary intention, and a copy was provided to each subject.

bar. Capsule manufacturing for human application testing

The preparation of capsules for human application test was prepared in the blending ratio of Table 1 by a health functional food manufacturer. In order to smoothly fill the capsule with 99% of skate collagen peptide, silicon dioxide, a safe food additive that does not cause metabolism during absorption in the body, was used. In addition, placebo was prepared from 100% starch as a raw material.

[Table 1] Blending ratio of raw materials for capsules

[Table 2] Information on food for human application test

2. Results and Discussion

end. Physicochemical properties

(1) General ingredients

The content of general components of the skate skin collagen peptide is shown in Table 3. Collagen peptides were found to have about 95% protein, about 0.1% fat, about 1.2% ash, and 0% carbohydrates. This result was judged to be because the skin and skate meat were completely separated during the peeling process during the manufacturing process of aged skate sashimi, and most of the ingredients in the skate skin were composed of protein.

[Table 3] The result of proximate analysis in collagen peptide of skate skin

(2) constituent amino acids

Table 4 shows the results of analysis of the amino acid content of the low-molecular collagen peptide prepared from skate skin. In the S40 process, as a hydrolytic enzyme, 0.1-0.5% by weight of Alcalae and 0.1-0.5% by weight of Flavourzyme are added at the same time to hydrolyze, or 0.1-0.5% by weight of Alcalae and 0.1-0.5% by weight of Protamex are simultaneously added and hydrolyzed. This is the result of preparing a collagen peptide. As shown in Table 4, as a result of measuring the constituent amino acids, the glycine content was the highest regardless of the molecular weight, followed by glutamic acid, alanine, and arginine in order. The molecular weight and amino acid composition did not show a significant difference, but the lower the molecular weight, the higher the amino acid content was measured.

[Table 4] Analysis of constituent amino acid with molecular weights of collagen peptide

(3) free amino acids

Table 5 shows the results of analysis of free amino acid content of low molecular weight collagen peptides prepared from skate skin. As a result of free amino acids, the contents of glycine and hydroxy proline were about 22.0% and 7.0%, indicating the characteristics of a typical collagen protein, and the amino acid composition according to the molecular weight did not show a big difference. The content was high.

[Table 5] Analysis of free amino acids in skate skin collagen peptide

(4) Data on the standards and test methods for indicator components (or functional components)

-Standards and grounds for indicator components (or functional components)

○ The index components of skate skin collagen peptide were set to be 90% or more of protein content, 7% or more of hydroxy proline, and 20% or more of Glycin.

[Table 6] Analysis results of index components for collagen peptide extract from skate skin

○ It is desirable to set the standard for the content of indicator components in the range of 80 to 120% of the results analyzed by an accredited institution.

[Table 7] Analysis results of index components for the collagen peptide extract of skate skin

-Indicator component (or functional component) standard product information

○ Hydroxy proline

○ Glycine

-Grounds for setting indicator components

○ Glycine: Summary table of the report of an accredited testing institution

○ Hydroxy proline: Summary table of official test institute report

■ Summary table of test report

I. The Effect of Oral Intake of Skate Skin-derived Low Molecular Collagen Peptide on Body Fat Reduction

In order to investigate the effect of oral intake of collagen peptides extracted from skate shells on body fat reduction, we tested whether there was a difference between groups in the basal characteristics before administration of the test/control food for this human test subject. As shown in Table 8, as a result, there was no statistically significant difference in sex, age, weight, height, etc. in both the ITT group and the PP group, so it can be seen that prior homogeneity of the two groups is recognized, and randomization is statistically appropriate. I think it is.

[Table 8] Study patients baseline characteristics

As listed in Tables 9 and 10, for the change in body fat percentage (%) through DEXA, the primary efficacy endpoint, and Visit3 before and after administration of the test/control food (Visit2, week0) for the secondary efficacy endpoint. It was measured at (week6) and Visit4 (week12). For each measurement value and the degree of change compared to the baseline (Visit2), the values between groups and within groups (by visit time) were also compared and shown in a table.

[Table 9] Comparison between and within each groups (ITT population)

[Table 10] Comparison between and within each groups (PP population)

As shown in Table 9 and Table 10, the results of the primary efficacy evaluation variable analysis showed a significant decrease in the test group in the comparison of the change in body fat percentage (%) measured by DEXA in both the ITT and PP analysis groups. It showed the amount of change in the group. Therefore, it is believed that the daily use of test foods helps improve body fat reduction.

In the results of the secondary efficacy endpoint analysis, the test group significantly decreased compared to the control group in the comparison of the change in body fat mass (g) measured by DEXA.

In addition, in the before/after comparison within each group, there was a tendency to decrease in the test group and the amount of change in the test group was considered to be clinically significant.

As a result of analysis of body fat mass (g) through DEXA, which was set as an evaluation index for body fat reduction, the test group significantly decreased compared to the control group, and the extent of the reduction was also judged to be clinically significant, so the test food (collagen peptide derived from skate skin) was It is thought that it helps to reduce.

Therefore, it is judged that this test food can have an effect on reducing body fat when taken in the medium and long term.

On the other hand, as a safety evaluation, 45 subjects in the test group and 45 subjects in the control group who took the test food at least once were evaluated for safety by evaluating vital signs and clinical laboratory tests during the test period.

As shown in Table 11, there was one case (dyspepsia) in which an adverse reaction occurred during the test period, but the reported adverse reaction was resolved, and it was judged that there was no association with the test food. As a result of safety evaluation through clinical laboratory test values and physical examination, there was no basis for judging that it was clinically hazardous. Therefore, it is judged that it is safe to ingest 2,000mg/day of the collagen peptide derived from the skate skin for an adult standard for a medium or long period.

[Table 11] Atypical response report

All. Mass production of collagen peptides and production of related applications

(1) Mass production of collagen peptides

Mass production facilities were mass-produced at the factory site using conditions optimized in the laboratory. The raw materials of skate skins generated during the processing of aged skates are pretreated by washing them with running water, and subjected to pressure heat treatment at 120°C for 2 hours to gelatinize the raw materials, and then have a molecular weight of 1,000 Da. In the above case, 0.1% (w/w) of Alcalase and Flavourzyme were added to each of the raw materials at 50° C., and the enzyme was hydrolyzed for 2 hours, and the molecular weight was 1.000 Da. The following products were enzymatically hydrolyzed for 6 hours by adding 0.2% (w/w) of Alcalase and Flavourzyme respectively to the input raw materials. After completion of hydrolysis, the residue was removed through primary filtration, and activated carbon was added for deodorization, followed by stirring at 75° C. for 30 minutes. After the deodorization was finished, it was concentrated to 30 brix in a vacuum concentrator through microfiltration using a filter press to remove activated carbon, and then increased the concentration of the solid content in the liquid, and spray dried to commercialize it. The yield of the product was about 17%, which was considerably high when considering the moisture content of the raw materials of the skate skin. Based on this, a product standard was prepared.

(2) Application product production

end. Low molecular weight collagen peptide powder product

Collagen peptides were mass-produced on-site, and mass-produced products were released and sold in units of 100g.

I. Skate Collagen Peptide Capsule Product for Human Test

For the human test, a capsule product filled with 100% of Hongo collagen peptide was produced, and the same amount of placebo product for the control test was produced.

All. Skate Collagen Peptide (Health Functional Food) Capsule Product

A health functional food was produced using skate collagen peptide as the main ingredient. It was manufactured as a capsule-type health functional food after obtaining approval from the Food and Drug Administration at a blending ratio of 1.7% vitamin and 70% collagen peptide.

The embodiments described in the present specification and the accompanying drawings are merely illustrative of some of the technical ideas included in the present invention. Accordingly, it is obvious that the embodiments disclosed in the present specification are not intended to limit the technical idea of the present disclosure, but to explain the technical idea, and thus the scope of the technical idea of the present disclosure is not limited by these embodiments. Modifications and specific embodiments that can be easily inferred by those of ordinary skill in the art within the scope of the technical idea included in the specification and drawings of the present invention are included in the scope of the present invention. It will have to be interpreted.

Claims (5)

As a functional raw material composition for reducing body fat comprising a low molecular weight collagen peptide obtained by hydrolyzing skate skin as an active ingredient,
As an active ingredient, a low-molecular collagen peptide containing a protein content of 90% by weight or more and containing 58-89 mg/g of the first compound represented by the following Formula 1 and 190-286 mg/g of the second compound represented by the following Formula 2. Including,
The low molecular weight collagen peptide is
Collecting raw materials of skate skin (consisting of 100% skate skin) generated during the skate processing process and storing them in a frozen manner;
Putting the skin in a high-pressure reactor and extracting hot water at 121° C. for 90 minutes;
Hydrolyzing the proteolytic enzymes Alcalase and Protamax by adding 0.5% of each of the raw materials for 2 hours;
Deodorizing using activated carbon and ion exchange resin;
Removing impurities through vacuum filtration; And
Concentrating and freeze-drying after cut-off based on molecular weight of 1,000 Da.; It is manufactured through
The free amino acids of the low molecular weight collagen peptide include 6 to 8% by weight of the first compound and 19 to 23% by weight of the second compound, based on 100% by weight of the total free amino acids,
The constituent amino acids of the low molecular weight collagen peptide are Aaspartic acid 5.36 to 5.90 wt%, Threonine 3.38 to 3.72 wt%, Serine 9.77 to 10.71 wt%, Glutamic acid 9.77 to 10.71 wt%, Glycine 21.27 based on 100 wt% of the total amino acids. ~23.64 wt%, Alanine 7.65~8.51 wt%, Valine 2.88~3.17 wt%, Methionine 1.72~2.16 wt%, Isoleucine 2.34~2.58 wt%, Leucine 3.50~3.87 wt%, Tyrosine 0.73~0.86 wt%, Phenylalanine 2.07~ 2.28 wt%, Lysine 3.54 to 3.88 wt%, Ammonia 0.84 to 0.92 wt%, Histidine 1.49 to 1.62 wt%, and Arginine 7.11 to 7.93 wt% A functional raw material composition for reducing body fat, characterized in that it comprises.

[Formula 1]

[Formula 2]

delete delete The method of claim 1,
The free amino acid of the low molecular weight collagen peptide is 0.03 to 0.05% by weight of Phosphoserine, 0.01 to 0.02% by weight of Taurine, 6.46 to 7.11% by weight of Aspartic acid, 6.10 to 6.85% by weight of Hydroxy proline, Threonine 3.29~3.42 wt%, Serine 5.38~5.71 wt%, Glutamic acid 9.93~10.78 wt%, Proline 8.28~9.02 wt%, Glycine 19.97~22.09 wt%, Alanine 6.92~7.66 wt%, Valine 2.93~3.34 wt%, Methionine 1.67~2.15 wt%, Isoleucine 2.20~2.45 wt%, Leucine 3.32~3.67 wt%, Tyrosine 0.81~0.91 wt%, Phenylalanine 2.02~2.23 wt%, Ammonia 0.71~0.74 wt%, Hyroxylysine 0.09~1.05 wt%, Ornithine 0.03 -0.04% by weight, Lysine 3.22 to 3.49% by weight, Histidine 1.26 to 1.38% by weight, and Arginine 7.05 to 7.84% by weight, comprising a body fat reduction functional raw material composition.
Functional food comprising the functional raw material composition for reducing body fat according to any one of claims 1 and 4.

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