KR20200084267A - Functional composition comprsing low molecular collagen peptide extract from skate skin and Functional food comprising the same - Google Patents

Abstract

The functional ingredient composition for body fat reduction according to the present invention contains low-molecular collagen peptide resulting from skate skin hydrolysis as an active ingredient. The low-molecular collagen peptide has a protein content of 90 wt% or more and contains 5.8 to 8.9 mg/g of a first compound represented by the following chemical formula 1 and 190 to 286 mg/g of a second compound represented by the following chemical formula 2.

Description

Development and industrialization of functional raw materials for reducing body fat through preclinical and human application experiments of small molecule-derived collagen peptides derived from skatefish {Functional composition comprsing low molecular collagen peptide extract from skate skin and Functional food comprising the same}

The present invention relates to a functional raw material composition for reducing body fat, which contains a low molecular weight collagen peptide obtained by hydrolyzing a cuttlefish skin as an active ingredient, and to develop and industrialize a functional raw material for reducing body fat through preclinical and human application experiments of a small molecule collagen peptide derived from a cutlet skin. It is about.

Skate (Skate) is a favorite food in Korea and has various pharmacological effects such as digestive function and appetite enhancement, immunity improvement and anti-aging, skin elasticity maintenance, and arthritis prevention. , Simmered redfish, simmered redfish (liver), etc. are ingested without special processing.

Due to the functional characteristics and peculiar palatability of redfish, it has escaped the form of small consumption in the Jeonnam region in the past, and its demand has soared nationwide, and it is currently being consumed in large-scale marts and home shopping.

Fish shells and visceral tissues contain large amounts of collagen components, and unlike conventional collagen derived from skins and bones of cattle and pigs, they are reported to be safe because they do not have the risk of human metastasis such as mad cow disease or foot and mouth disease.

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

Korean Registered Patent No. 10-1774701

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

In addition, the present invention is an experiment for extracting collagen peptides by molecular weight by using shells, which are by-products generated during processing of redfish, to produce low-molecular-weight collagen peptides. 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 the above object, the functional raw material composition for reducing body fat according to an embodiment of the present invention includes a low-molecular collagen peptide obtained by hydrolyzing skatefish shell as an active ingredient, and has a protein content of 90% by weight or more,

A low-molecular-weight collagen peptide containing the first compound 5.8 to 8.9 mg/g represented by the following Chemical Formula 1 and the second compound 190 to 286 mg/g represented by the following Chemical Formula 2 is included as an active ingredient.

[Formula 1]

[Formula 2]

In the body fat reducing functional raw material composition according to an embodiment of the present invention, the free amino acids of the low-molecular-weight collagen peptides are 6 to 8% by weight of the first compound and the second compound 19 based on 100% by weight of the total of 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 reducing functional raw material composition according to an embodiment of the present invention, the constituent amino acids of the low molecular weight collagen peptide are Aaspartic acid 5.36 to 5.90% by weight, Threonine 3.38 to 3.72% by weight, Serine 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% Can.

In the body fat reducing functional raw material composition according to an embodiment of the present invention, the free amino acid of the low molecular weight collagen peptide is based on 100% by weight of the total free amino acid, Phosphoserine 0.03 to 0.05% by weight, Taurine 0.01 to 0.02% by weight, Aspartic 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 containing the above-described body fat reducing 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 power in the body.

In addition, when the functional food according to the present invention is taken mid/long-term, it may have an effect on reducing body fat, and it is safe to consume 2,000mg/day for adults for a long period of time.

In addition, despite the mass production of the body fat reducing functional raw material composition according to the present invention, the yield of the product appeared to be about 17%, showing a considerably high yield when considering the water content of the raw material of the skate.

In addition, the present invention is extremely low in use, so the collagen peptide is extracted by using a by-product of redfish that has been disposed of, so it has the effect of preventing the 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.
Figure 2 is a view showing the content change, yield shown in accordance with the 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 examples and drawings are provided as examples to sufficiently convey the spirit of the present invention to those skilled in the art. In addition, unless otherwise defined in the technical terms and scientific terms used in the present invention, those skilled in the art to which this invention belongs have the meanings commonly understood, and the present invention in the following description and accompanying drawings Descriptions of well-known functions and configurations that may unnecessarily obscure the subject matter are omitted.

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

In addition, unless otherwise indicated in this specification,% means weight%.

1. Manufacturing and industrialization of low molecular collagen peptides

For the production of low-molecular-weight collagen peptides, collagen peptides were extracted for each molecular weight using shells, a by-product generated during the processing of redfish. As a test to confirm the possibility of industrial application using the same, through experiments such as general ingredients and molecular weight, amino acid composition, deodorization, etc. Industrial applicability was reviewed.

end. Materials and methods

(1) Hydrolysis of raw material of redfish skin

As shown in Figures 1 and 2, for the hydrolysis of the raw material of the shellfish, the raw material of the shellfish generated during the shell stripping process during the raw fish processing process of Yeongsan Hongeo Co., Ltd. ) Was collected (S10).

Subsequently, after washing three times with running water, it was stored frozen at -25°C or lower (S20).

In order to prepare a low molecular collagen peptide, the shell was placed in a high-pressure reactor (autoclave), and high-pressure hot water was extracted at 121°C for 90 minutes (S30), followed by commercial proteolytic enzymes Alcalase, Flavorzyme and Protamex (Novozyme, Denmark). Hydrolysis was carried out for each condition (S40).

(2) Preparation of collagen peptides by molecular weight

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

I. Measurement of physicochemical properties

(1) Measurement of general components

The general ingredients were measured according to the general test method of the Food Code (Food Code, 2013). Moisture content was measured by atmospheric pressure drying method, crude protein content by micro kjeldahl method, crude fat content by Soxhlet method, and ash content by direct ashing 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, hydrolyzed, and then nitrogen gas was replaced. After standing for 22 hours in an oven at 110° C., the mixture was evaporated under reduced pressure and dried. After diluting with 0.02N hydrochloric acid, it was filtered with 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, the column was TSKgel SuperMultipore®PW-H (6.0 mm ID Х 15 cm, Distilled Water, Japan), and injected with 2 µl of a sample to pH 7.0 buffer (0.1M KH2PO4 + 10%) at a rate of 1 mL/min. NaOH) was measured at 25°C while flowing.

la. Ultrafiltration

1K Da. of the extract extracted from the redfish skin using Flow 20 using an ultrafilter (EMP Series, EMP Tech, Korea). The filtration membrane (Pall corporation, USA) was passed. Filtration was performed until no more filtrate came out, and after the filtration was completed, the filtered liquid and the unfiltered liquid were separated to prepare a sample.

hemp. Human body application test

The human application test was conducted after the clinical trial monitoring plan was established and approved by the IRB of the Clinical Review Committee, followed by the Helsinki Declaration, and was conducted in compliance with the KGCP.

Before screening to select the candidates, a description of the test was distributed to all applicants, and after sufficient explanation, the consent was signed by the applicant's voluntary intention and a copy was provided to each candidate.

bar. Capsule manufacturing for human body application testing

Preparation of capsules for the human body application test was prepared in the formulation ratio of Table 1 by a manufacturer specialized in health functional food. In order to facilitate filling of the capsule with 99% of redfish collagen peptide, silicon dioxide, a safe food additive that does not cause metabolism when absorbed by the body, was used. In addition, placebo was made from 100% starch.

[Table 1] Capsule raw material mixing ratio

[Table 2] Information of human application test food

2. Results and Discussion

end. Physicochemical properties

(1) General ingredients

Table 3 shows the general content of the skate skin collagen peptide. Collagen peptides had a protein content of about 95%, fat content of about 0.1%, ash content of about 1.2%, and carbohydrates of 0%. These results were judged to be due to the fact that the shell and the redfish meat were completely separated during the peeling process during the manufacturing process of the aged redfish sashimi, and most of the ingredients in the redfish shell 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 analysis results of the constituent amino acids of the low-molecular-weight collagen peptide prepared from the skate. In the S40 process, as a hydrolysis enzyme, 0.1 to 0.5% by weight of Alcalae and 0.1 to 0.5% by weight of Flavorzyme are added at the same time, or hydrolysis by adding 0.1 to 0.5% by weight of Alcalae and 0.1 to 0.5% by weight of Protamex at the same time It is a result of producing a collagen peptide. As shown in Table 4, as a result of measurement of the constituent amino acids, the glycine content was highest regardless of the molecular weight, and then the content was highest in the order of glutamic acid, alanine, and arginine. 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.

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

(3) free amino acids

Table 5 shows the results of the analysis of the free amino acid content of the low-molecular-weight collagen peptide prepared from the skate. As a result of free amino acids, the content of glycine and hydroxy proline was about 22.0% and 7.0%, which showed typical collagen protein characteristics, and the amino acid composition according to 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 specifications and test methods for indicator components (or functional components)

-Specification and rationale for indicator component (or functional component)

○ The index components of the skate skin collagen peptide were set to three types: protein content of 90% or higher, Hydroxy proline 7% or higher, and Glycin 20% or higher.

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

○ It is desirable to set the specification of the content of the indicator component in the range of 80 to 120% of the results analyzed by the accredited institution.

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

-Indicator component (or functional component) standard product information

○ Hydroxy proline

○ Glycine

-Basis for setting index components

○ Glycine: Summary table of accredited test institutes

○ Hydroxy proline: Summary table of official test report

■ Test report summary table

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

In order to investigate the effect of oral intake of collagen peptides extracted from skatefish skin on body fat reduction, we tested whether there was a difference between the groups in the basal characteristics prior to administration of the test/control foods to the human subjects. As shown in Table 8, as a result, there was no statistically significant difference in gender, age, weight, height, etc. in both the ITT and PP groups, so it can be seen that the prior homogeneity of the two groups is recognized, and randomization is statistically appropriate. Is judged.

[Table 8] Study patients baseline characteristics

As shown in Table 9 and Table 10, the change in body fat percentage (%) through the primary efficacy evaluation variable DEXA and the secondary efficacy evaluation variable before and after administration of test/control food (Visit2, week0) and Visit3 after administration (week6) and Visit4 (week12). For each measured value and the degree of change compared to baseline (Visit2), the values between groups and within groups (by visit time) are 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, as a result of the primary efficacy endpoint analysis, both the ITT and PP analysis groups showed a significant decrease in the test group in the in-group comparison of changes in body fat percentage (%) measured by DEXA. The group showed the amount of change. Therefore, it is thought that the daily intake of test foods can help improve body fat loss.

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

In addition, the pre/post comparison within each group showed a tendency to decrease in the test group, and the amount of change in the test group showed clinical significance.

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. It is thought to help decrease.

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

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

As shown in Table 11, there were 1 case (indigestion) of adverse reactions during the test period, but the reported adverse reactions were resolved, and it was determined that there was no connection with the food for testing. As a result of safety evaluation through clinical laboratory tests and physical examinations, there was no basis for judging that they were clinically dangerous. Therefore, it is judged that it is safe to ingest 2,000mg/day of adult-based collagen peptides derived from the skin of the skates for mid- to long-term.

[Table 11] Atypical response report

All. Mass production of collagen peptides and related applications

(1) Mass production of collagen peptides

Mass production was performed at the factory site using mass-produced equipment using optimized conditions in the laboratory. The raw material of the scallop shell generated during the processing of the aged scallop ash is pretreated by washing in running water, and subjected to a pressure heat treatment at 120° C. for 2 hours to gelatinize the raw material, and then molecular weight 1,000 Da. In the above case, the enzyme was hydrolyzed for 2 hours by adding 0.1% (w/w) of Alcalase and Flavorzyme to the input raw material at 50°C at a temperature, 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 Flavorzyme, respectively, to the feedstock. After completion of the 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 completed, it was filtered through a filter press to remove activated carbon, concentrated to 30 brix in a vacuum concentrator to increase the concentration of solid content in the liquid, and then spray dried to commercialize it. The yield of the product appeared to be about 17%, which showed a fairly high yield when considering the water content of the raw material of the skate. Based on this, a product standard was prepared.

(2) Application product production

end. Low molecular collagen peptide powder products

Mass production of collagen peptides in the field, and mass-produced products launched and sold products packaged in units of 100 g.

I. Redfish collagen peptide capsule products for human testing

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

All. Redfish collagen peptide (health functional food) capsule product

A health functional food was prepared using the redfish collagen peptide as the main ingredient. It was manufactured as a capsule-type health functional food after being approved by the Food and Drug Administration at a mixing 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 spirit included in the present invention. Therefore, the embodiments disclosed in the present specification are not intended to limit the technical spirit of the present invention, but to explain the present invention, it is obvious that the scope of the technical spirit of the present invention is not limited by these embodiments. Within the scope of the technical spirit included in the specification and drawings of the present invention, modifications and specific embodiments that can be easily inferred by those skilled in the art are all included in the scope of the present invention. Should be interpreted.

Claims (5)

As a functional raw material composition for reducing body fat, comprising a low-molecular-weight collagen peptide obtained by hydrolyzing the redfish skin as an active ingredient,
Low-molecular-weight collagen peptides having a protein content of 90% by weight or more and containing the first compounds 5.8 to 8.9 mg/g represented by the following Chemical Formula 1 and the second compounds 190 to 286 mg/g represented by the following Chemical Formula 2 as active ingredients A functional raw material composition for reducing body fat.
[Formula 1]

[Formula 2]

According to claim 1,
The free amino acid of the low molecular weight collagen peptide, based on 100% by weight of the total free amino acid, 6 to 8% by weight of the first compound and 19 to 23% by weight of the second compound, body fat reducing functional raw material composition.
According to claim 1,
Constituent amino acids of the low-molecular-weight collagen peptide, based on 100% by weight of the total amount of the constituent amino acids, Aaspartic acid 5.36 to 5.90% by weight, Threonine 3.38 to 3.72% by weight, Serine 9.77 to 10.71% by weight, Glutamic acid 9.77 to 10.71% by weight, Glycine 21.27~23.64%, Alanine 7.65~8.51%, Valine 2.88~3.17%, Methionine 1.72~2.16%, Isoleucine 2.34~2.58%, Leucine 3.50~3.87%, Tyrosine 0.73~0.86%, Phenylalanine 2.07 ~2.28% by weight, Lysine 3.54~3.88% by weight, Ammonia 0.84~0.92% by weight, Histidine 1.49~1.62% by weight and Arginine 7.11~7.93% by weight Body fat reducing functional raw material composition.
According to claim 1,
The free amino acid of the low molecular collagen peptide is, based on 100% by weight of the total free amino acid, 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 7.74% by weight, body fat reduction functional raw material composition.
Functional food comprising a functional raw material composition for reducing body fat according to any one of claims 1 to 4.

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