KR102466561B1 - Scaffold including bio-cellulose powders for manufacturing cultured meat and a preparation method thereof - Google Patents

Abstract

The present invention relates to a scaffold for manufacturing cultured meat including bio-cellulose powder, and a preparation method thereof. The present invention provides hydrogel scaffolds with new characteristics which are prepared by pulverizing microbial-derived bio-cellulose, and mixing the same with alginate, agar or carrageenan, which are raw materials conventionally used in producing hydrogel. The prepared hydrogel scaffolds can be usefully used as a scaffold for manufacturing cultured meat (cultured meat scaffold) by excellent cell adhesion and physical properties.

Description

Scaffold including bio-cellulose powders for manufacturing cultured meat and a preparation method thereof}

The present invention relates to a scaffold for producing cultured meat (cultured meat scaffold) containing bio-cellulose powder and a method for preparing the same.

For a long time, scientists have conducted various studies using a two-dimensional cell culture method using a Petri dish. However, there is a problem in that, when cells are cultured by a two-dimensional cell culture method, an environment different from an actual tissue is created, such as changing cell morphology or cell division progressing but gene expression may be different.

Methods for culturing cells similar to real tissues by 3D cell culture methods are being studied, and in particular, the importance of cell attachment scaffolds having a structure similar to the characteristics of extra cellular matrix (ECM) is emerging. .

As a three-dimensional cell culture scaffold, a hydrogel containing a large amount of water and having intermediate properties between liquid and solid is used. ) Depending on the structure, it has the property of being insoluble in water. Alginate, agar, carrageenan, and gelatin are the materials of hydrogel (natural polymeric gelling agent), and they have various forms and properties that differ depending on their ingredients, crosslinking, and manufacturing methods. A hydrogel scaffold is made with

Alginate, which is used as a material for hydrogels, is a natural polymer extracted from seaweed and has been used for quite a long time. When a hydrogel is prepared by adding a multivalent metal such as Ca ²⁺ and crosslinking alginate, it is attracting attention as a new culture technology because it can mimic the tissue of an actual organism.

As another ingredient, carrageenan is a natural polymer of the sulfated polysaccharide family extracted from red algae. It is tasteless and odorless, and has excellent water retention, so the viscosity does not change over time. It is widely used in the food industry, such as a gelling agent, thickener, stabilizer for beverages, emulsion stabilizer, fat replacer, and binder for meat products.

Agar (agar) is extracted from agar-agar, a seaweed, and is composed of 70% agarose and 30% agaropectin. As a low-calorie, high-fiber food material, it is recognized for its high potential as a substitute for fat and protein.

Alginate, carrageenan, and agar (agar), which are in the spotlight as raw materials for hydrogel (natural polymeric gelling agent), are simple to dissolve and shape, making it easy to create 2D or 3D structures. Since scaffolds are used as scaffolds for cell culture, cell adhesion is very low, making effective cell growth difficult.

On the other hand, since plant and seaweed-derived cellulose scaffolds use tissue itself, it is difficult to control the internal structure of the scaffold, and it is virtually impossible to control the components and contents. In addition, the decellularization process is very complicated, so there is a limit to its use as a scaffold for producing cultured meat.

Bio-Cellulose is a culture of microorganisms belonging to the genus Rhizobium sp., Agrobacter sp., Gluconacetobacter sp., and Acetobacter sp. Among them, strains of the genus Acetobacter are known to have the highest production efficiency.

Bio-cellulose produced through microbial culture is widely used in pharmaceuticals and cosmetics, and recently, its use fields, such as eco-friendly packaging materials, are gradually expanding.

The present inventors confirmed that bio-cellulose is harmless to the human body, can be ingested, and has excellent cell adhesion, so it is very suitable for realizing tissue. However, as a result of efforts to solve the problem that it is difficult to control the physical properties of bio-cellulose itself for use as a scaffold for producing cultured meat and that it takes a lot of time to produce bulky bio-cellulose, microbial-derived bio-cellulose is powdered and The present invention was completed by confirming that a scaffold for producing cultured meat could be produced by adding it to a hydrogel raw material.

An object of the present invention is to provide a composition for preparing a cultured meat scaffold comprising a microorganism-derived bio-cellulose powder, a natural polymer gelling agent, and a gelation accelerator.

Another object of the present invention is to provide a cultured meat scaffold prepared from the above composition.

Another object of the present invention is a method for producing a cultured meat scaffold comprising the following steps: (a) obtaining bio-cellulose by stationary culturing microorganisms in a medium containing peptone; (b) washing the bio-cellulose obtained in step (a) with distilled water and drying; (c) freezing the dried bio-cellulose and then pulverizing it at 10,000 to 15,000 rpm to obtain microorganism-derived bio-cellulose powder; (d) mixing the microorganism-derived bio-cellulose powder with a natural polymeric gelling agent; and (e) adding a gelation accelerator after lyophilizing the product of (d).

Another object of the present invention is to provide cultured meat prepared by culturing non-human animal-derived cells on the cultured meat scaffold prepared by the above method.

Advantages and features of the present invention, and methods of achieving them, will become clear with reference to the detailed description of the following embodiments taken in conjunction with the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below, but may be implemented in various different forms, only these embodiments are intended to complete the disclosure of the present invention, and are common in the art to which the present invention belongs. It is provided to fully inform the person skilled in the art of the scope of the invention, and the invention is only defined by the scope of the claims.

Terminology used herein is for describing the embodiments and is not intended to limit the present invention. In this specification, singular forms also include plural forms unless specifically stated otherwise in a phrase. As used herein, "comprises" and/or "comprising" does not exclude the presence or addition of one or more other elements other than the recited elements. Like reference numerals throughout the specification refer to like elements, and “and/or” includes each and every combination of one or more of the recited elements. Although "first", "second", etc. are used to describe various components, these components are not limited by these terms, of course. These terms are only used to distinguish one component from another. Accordingly, it goes without saying that the first element mentioned below may also be the second element within the technical spirit of the present invention.

Unless otherwise defined, all terms (including technical and scientific terms) used in this specification may be used with meanings commonly understood by those skilled in the art to which the present invention belongs. In addition, terms defined in commonly used dictionaries are not interpreted ideally or excessively unless explicitly specifically defined.

The present invention provides a composition for preparing a cultured meat scaffold comprising a microorganism-derived bio-cellulose powder, a natural polymer gelling agent, and a gelation accelerator.

The microorganism of the present invention is 1 selected from the group consisting of Rhizobium sp., Agrobacter sp., Gluconacetobacter sp., and Acetobacter sp. There may be more than one species. Preferably, it may be Acetobacter sp. or Gluconacetobacter sp., which has good production yield, but is not limited thereto.

The cultured meat of the present invention is also referred to as clean meat or cell-based meat or cultivated meat, and means suitable for human or non-human animals to eat or edible.

The scaffold of the present invention is a scaffold for producing cultured meat that has a structure similar to that of meat and has cell adhesion, made of a scaffold composition based on microbial-derived bio-cellulose powder, a natural polymeric gelling agent, and a gelation accelerator.

The bio-cellulose of the present invention may be produced in a medium containing animal peptone, bacterial peptone or vegetable peptone, preferably vegetable peptone, but is not limited thereto.

The animal peptone may be meat or casein.

The bacterial peptone may be bacto peptone.

The vegetable peptone is soy peptone, wheat peptone, broadbean peptone, potato peptone, pea peptone, papaic soy peptone and lupine It may be at least one selected from the group consisting of lupin peptone, preferably soybean peptone, but is not limited thereto.

The strains producing the microorganism-derived bio-cellulose of the present invention use glucose, sucrose, molasses, fructose, mannitol, and glycerol as carbon sources during growth. Persimmon juice and persimmon vinegar, apple juice, grape juice, beer waste liquid and coconut by-products are used as auxiliary carbon sources, and casein, peptone, yeast extract, etc. are used as nitrogen sources, but strains The optimal carbon source, supplementary carbon source, and nitrogen source are different depending on the

Establishing culture conditions with appropriate concentrations by selecting the optimum among various types of carbon sources, supplementary carbon sources, and nitrogen sources is not only important in terms of cost, but also prevents yield reduction due to lack of nutrients and prevents by-products caused by excessive consumption of carbon sources. This is important because it can produce yield improvement effects through accumulation inhibition.

In the present invention, when producing microbial-derived bio-cellulose using the Gluconacetobacter Xylinus subsp. Xylinus strain (accession number: KCCM 41431) strain, which has been confirmed to produce bacterial cellulose, for producing cultured meat The composition for preparing a cultured meat scaffold of the present invention was designed by studying culture conditions for a carbon source of glucose to have physical properties as a scaffold.

The natural polymer gelling agent (hydrogel raw material) of the present invention may include any one selected from the group consisting of alginate, agar, and carrageenan, but is not limited thereto.

The alginate of the present invention is a sticky mucilage component extracted from brown algae such as seaweed and kelp. It is insoluble in water, swells, and is a colloid, that is, a complex polysaccharide in which fine particles are aggregated or sprayed into a gas or liquid. It is used for various purposes such as a thickener, emulsion stabilizer, and gelling agent, and can be applied to various fields such as food, industry, and medicine. In a natural state, it exists in combination with sodium, calcium, potassium, etc., and when included in cosmetics, it has excellent adsorption properties, is effective in removing toxins, and has excellent moisturizing properties. When a hydrogel is prepared by adding a multivalent metal such as Ca ²⁺ and crosslinking alginate, the tissue of an actual organism can be simulated.

The agar (agar) of the present invention is a material extracted from agar-agar, which is a seaweed, and is freeze-dried material of agar-agar. It is dissolved in water and used for food or as an industrial material. It is a water-soluble, transparent, and soft substance. It is also used for the preparation of solid media in which microorganisms, plant tissues, and animal cells are fixed and cultured. As a low-calorie, high-fiber material, it has high potential as a substitute for fat and protein.

The carrageenan of the present invention is a substance extracted from red algae, and its main component is galactose. It has the property of completely dissolving in water of 70 to 80 ℃ or more. The carrageenan is a tasteless, odorless, light ivory-colored powder, and an aqueous solution in which carrageenan is dissolved becomes a viscous solution at room temperature or forms a gel. It has excellent water holding capacity and does not change its viscosity over time.

The gelation accelerator of the present invention may be a Group 1 or Group 2 metal salt. The gelation accelerator may be one or more metal salts selected from the group consisting of potassium chloride, calcium chloride, magnesium chloride and calcium lactate, but is not limited thereto.

The microorganism-derived bio-cellulose powder of the present invention may be obtained by freezing the microorganism-derived bio-cellulose, pulverizing at 10,000 to 15,000 rpm, and then separating the powder with a sieve having a diameter of 0.5 to 2 mm.

The composition for preparing a cultured meat scaffold of the present invention may include 1 to 20 parts by weight, preferably 1 to 10 parts by weight of the microorganism-derived bio-cellulose powder, based on 100 parts by weight of the composition for preparing cultured meat, and the natural polymer gel The agent may include 1 to 5 parts by weight, preferably 1 to 2 parts by weight, and the gelation accelerator may include 1 to 5 parts by weight, preferably 1 to 2 parts by weight, but is not limited thereto.

When the amount of the microorganism-derived bio-cellulose powder of the present invention is less than 1 part by weight based on 100 parts by weight of the composition for preparing cultured meat, the strength of the hydrogel itself is very weak and the amount of culturable cells is limited. In addition, if it exceeds 20 parts by weight, it is very difficult to maintain the shape of the hydrogel because the cellulose powder interferes with crosslinking of the hydrogel raw material.

The natural polymer gelling agent is less than 1 part by weight based on 100 parts by weight of the composition for preparing cultured meat, it is difficult to maintain the form of a hydrogel due to lack of components necessary for crosslinking, and when it is more than 5 parts by weight, a very hard hydrogel is formed As the size of the pores of the hydrogel decreases, the amount of water absorption decreases, making cell culture difficult.

In addition, the present invention provides a cultured meat scaffold prepared from the composition for preparing a cultured meat scaffold.

The cultured meat scaffold may include microbial-derived bio-cellulose powder and a natural polymeric gelling agent.

The cultured meat scaffold of the present invention can increase hardness, gumminess and chewiness of the cultured meat scaffold as the concentration of the bio-cellulose powder in the composition increases.

In addition, elasticity, cohesion, and elasticity of the cultured meat scaffold may decrease as the concentration of the bio-cellulose powder in the composition increases.

In addition, the present invention provides a method for producing a cultured meat scaffold.

The method for preparing the cultured meat scaffold may include the following steps:

(a) obtaining bio-cellulose by statically culturing microorganisms in a medium containing peptone;

(b) washing the bio-cellulose obtained in step (a) with distilled water and drying;

(c) freezing the dried bio-cellulose and then pulverizing it at 10,000 to 15,000 rpm to obtain microorganism-derived bio-cellulose powder;

(d) mixing the microorganism-derived bio-cellulose powder with a natural polymeric gelling agent; and

(e) freeze-drying the product of (d) and then adding a gelation accelerator.

The peptone is soy peptone, wheat peptone, broadbean peptone, potato peptone, pea peptone, papaic soy peptone and lupine peptone (lupin peptone) may be one or more selected from the group consisting of, but is not limited thereto.

The microorganism is one or more species selected from the group consisting of Rhizobium sp., Agrobacter sp., Gluconacetobacter sp., and Acetobacter sp. It may be, preferably Acetobacter sp. or Gluconacetobacter sp., but it may be, but is not limited thereto.

The natural polymer gelling agent may include any one selected from the group consisting of alginate, agar, and carrageenan, but is not limited thereto.

The gelation accelerator may be one or more metal salts selected from the group consisting of potassium chloride, calcium chloride, magnesium chloride and calcium lactate, but is not limited thereto.

In addition, the present invention provides cultured meat prepared by culturing non-human animal-derived cells on the cultured meat scaffold prepared by the above method.

The non-human animal-derived cells may be at least one selected from the group consisting of muscle stem cells, muscle cells, and progenitors thereof.

The myoblasts may be satellite cells or myoblasts, for example, myoblasts, but are not limited thereto.

The muscle cells may be myocytes or myotubes, for example, myocytes, but are not limited thereto.

The progenitor cells refer to cells capable of producing cells differentiated into multiple lineages, such as myoblasts, fibroblasts, adipocytes, stromal cells, pericytes, smooth muscle cells and endothelial cells. Progenitor cells differ from stem cells in that they do not typically have extensive self-renewal capacity.

The non-human animal-derived cells further include fat cells (fat cells or adipocytes) and/or progenitor cells thereof, stromal cells (connective tissue) and/or progenitor cells thereof, or endothelial cells (vascular cells) and/or progenitor cells thereof. It may include, but is not limited to.

Non-human animals of the present invention may be selected from the group consisting of cattle, sheep, pigs, poultry, shellfish and fish, but are not limited thereto.

The present invention provides a new hydrogel manufactured by pulverizing and pulverizing microbial-derived bio-cellulose and mixing it with raw materials used in conventional hydrogel production. It can be usefully used as a manufacturing scaffold (cultured meat scaffold).

1 is a schematic diagram showing a method of tertiary crushing of bio-cellulose.
2 is a diagram showing a scaffold for cultured meat prepared by mixing powdered bio-cellulose with alginate, agar or carrageenan.
Figure 3a is a diagram confirming the attachment and culture of muscle stem cells on a scaffold made of alginate and a scaffold to which bio-cellulose powder was added to alginate, and Figure 3b is a scaffold made of agar and bio-cellulose powder added to agar Figure 3c is a diagram confirming the attachment and culture of muscle stem cells on one scaffold, and Figure 3c is a diagram confirming the attachment and culture of muscle stem cells on a scaffold made of carrageenan and a scaffold to which bio-cellulose powder was added to carrageenan.
Figure 4 is a diagram showing the results of tracking the cell culturing ability through the ratio of fluorescence staining to the total area.
5 is a view showing the results of analyzing the cross-section of a scaffold containing alginate, agar or carrageenan and the cross-section of a scaffold in which alginate, agar or carrageenan and bio-cellulose powder are mixed.
6 is a view showing that a mixed scaffold was prepared by mixing 0, 1, 5, and 10 parts by weight of bio-cellulose powder and 2 parts by weight of alginate, agar, or carrageenan with respect to 100 parts by weight of the total composition.
7 shows the results of hardness, gumminess, chewiness, springiness, cohesiveness, and resilience of chicken, cow, and pork and bio-cellulose mixed hydrogel scaffold. It is a diagram showing changes in hardness, gumminess, chewiness, springiness, cohesiveness, and resilience depending on the concentration of bio-cellulose powder.

Hereinafter, the contents of the present invention will be described in more detail through the following examples and experimental examples. However, the scope of the present invention is not limited only to the following examples and experimental examples, and includes modifications of equivalent technical ideas.

Example 1. Production and Grinding of Bio Cellulose

For the production of bio-cellulose, a Gluconacetobacter Xylinus subsp. Xylinus strain (accession number: KCCM 41431) was distributed in the form of live cells from the Korea Microorganism Conservation Center and used.

The strain stored at -80 ° C was cultured in acetobacter solid medium (0.5% yeast extract, 0.5% soybean peptone, 2% glucose, 0.27% disodium phosphate, 0.15% citric acid and 2 % agar included) and cultured for 48 hours.

The strain cultured on the solid medium was inoculated with an acetobacter liquid medium (0.5% yeast extract, 0.5% soybean peptone, 2% glucose, 0.27% disodium phosphate, 0.15% citric acid) and Bio-cellulose was obtained by stationary culture for 72 hours or more. The obtained bio-cellulose was washed three times with distilled water to remove the culture medium contained in the bio-cellulose. Microorganisms were removed by boiling the bio-cellulose in a 0.1M NaOH solution, washed three times with distilled water to remove the 0.1M NaOH solution, and refrigerated in 30% ethanol.

To dry the stored bio-cellulose, it was washed three times with distilled water to remove 30% ethanol, and dried for 24 hours in a dryer (Dryoven) at 50 ° C. As moisture is removed from the dried bio-cellulose, it becomes very tough and hard to grind, making it very difficult to grind.

In order to overcome the characteristics of dried bio-cellulose and pulverize it, it was made hard using liquid nitrogen, and the primary pulverization was completed by filtering through a 1.5 mm sieve at 14,000 rpm in a Rotor mill (Pulverisette 14). In order to make the particle size of the firstly crushed cellulose smaller, the second grinding was carried out, and the bio-cellulose obtained in the first grinding was frozen again through liquid nitrogen and replaced with an impact bar ring to increase the grinding degree of the rotor mill. After grinding at 14,000 rpm, and sifting through a 0.12 mm sieve, the secondary grinding was completed. For additional pulverization, the secondary pulverized bio-cellulose powder was frozen again through liquid nitrogen, and the impact bar ring was replaced with an ACM ring, pulverized at 14,000 rpm, and then sieved through a 0.12 mm sieve to perform final tertiary pulverization (FIG. 1).

Example 2. Preparation of bio cellulose-hydrogel raw material composite scaffold

A new scaffold for cultured meat was prepared by mixing bio-cellulose powdered by the method of Example 1 and alginate, agar, or carrageenan previously used as a raw material for hydrogel (FIG. 2).

To prepare an alginate-bio-cellulose composite scaffold, 2 parts by weight of alginate based on 100 parts by weight of the total composition was dissolved in distilled water at room temperature for 30 minutes, and bio-cellulose powders of various concentrations were mixed.

The alginate-bio cellulose mixture was further ground and mixed for 3 minutes at 13,500 rpm using a homogenizer. The alginate-bio cellulose mixture was put in a mold and frozen at -80 ° C for 24 hours and then freeze-dried for 48 hours. For crosslinking of alginate, 1 part by weight of CaCl ₂ was treated with respect to 100 parts by weight of the total composition after primary freeze-drying, and washed with distilled water for 2 hours to remove CaCl ₂ , followed by freezing at -80 ° C for 24 hours and then 48 hours During freeze-drying, a scaffold was prepared.

In order to prepare agar-bio-cellulose and carrageenan-bio-cellulose composite scaffolds, first, 2 parts by weight of agar and carrageenan were dissolved in distilled water at 80 ° C. based on 100 parts by weight of the total composition. Dissolved agar and carrageenan were mixed with bio-cellulose powder of various concentrations, and then secondary pulverized and mixed for 3 minutes at 13,500 rpm using a homogenizer. Agar-Bio Cellulose and Carrageenan-Bio Cellulose mixture were put in a frame, frozen at -80 ° C. for 24 hours, and then lyophilized for 48 hours to prepare a scaffold.

Experimental Example 1. Muscle stem cell culture of bio-cellulose mixed scaffold

In order to confirm that the bio-cellulose powder mixture scaffold prepared by the method of Example 2 is more suitable for muscle cell culture than the scaffold made of only alginate, agar, or carrageenan, which is a hydrogel raw material, muscle stem was added to the bio-cellulose powder mixture scaffold. Cells were seeded. The muscle stem cells used in the experiment were non-human derived muscle stem cells, which were directly isolated from muscle tissues of animals such as cows, chickens or pigs through primary cell culture.

The muscle stem cells were seeded on each scaffold, and culture medium was additionally added after culturing for 3 hours so that they could adhere well. The muscle stem cells were cultured on the scaffold for the final 96 hours, and the survival of the muscle stem cells at 24 and 96 hours after culture was measured using LIVE&DEAD ASSAY (Thermofisher, L3223) reagent to confirm that the muscle stem cells were being cultured on the scaffold. confirmed using The LIVE&DEAD ASSAY reagent has the property of staining live cells green and dead cells red, so the culturing ability of muscle stem cells was confirmed through fluorescent staining photographs at 24 hours and 96 hours after culture.

As a result, scaffolds made of alginate, agar, or carrageenan are not cell-attached and cell-cultured, so green-stained cells are rarely observed, whereas in scaffolds with bio-cellulose powder added, all muscle stem cells Attachment and culture were confirmed (Figs. 3a to 3c).

In addition, as a result of tracking the cell culturing ability through the ratio of fluorescence staining to the total area, it was confirmed that the cell culturing ability significantly increased when the bio-cellulose powder was added (FIG. 4).

Through this, it shows that cell attachment and culture are possible by adding bio-cellulose powder to the disadvantages of scaffolds made of alginate, agar, or carrageenan, which are not cell attachment and culture.

Experimental Example 2. Cross-sectional analysis of bio-cellulose mixed scaffold

In order to confirm once again whether the result of culturing muscle stem cells through LIVE&DEAD ASSAY was the effect of adding bio-cellulose powder, a cross-section of the bio-cellulose mixed scaffold was analyzed.

In order to analyze the cross-section of the bio-cellulose mixed scaffold prepared by the method of Example 2, each scaffold was coated with platinum at 20 mA for 60 seconds, and the cross-section of the coated bio-cellulose mixed scaffold was scanned with SEM (Scanning Electron Microscopy) Observed through.

It was confirmed that the cross section of the scaffold containing 2 parts by weight of alginate, agar or carrageenan was regular and pores of a certain size were formed with respect to 100 parts by weight of the total composition.

As a result of analyzing the cross-section of a scaffold in which 2 parts by weight of alginate, agar or carrageenan and 2 parts by weight of bio-cellulose powder were mixed with respect to 100 parts by weight of the total composition, bio-cellulose was properly formed between the scaffolds made of alginate, agar or carrageenan. It was confirmed that the scaffold was positioned, and through cross-sectional analysis of the bio-cellulose mixed scaffold, it was confirmed that the attachment and culture of the muscle stem cells was the effect of the bio-cellulose powder located between the alginate, agar, or carrageenan scaffold (FIG. 5).

Experimental Example 3. Comparison of texture of bio-cellulose mixed scaffold according to addition of bio-cellulose powder

In order to confirm that the scaffold capable of attaching and culturing muscle stem cells due to the addition of bio-cellulose powder can control the texture according to the amount of bio-cellulose powder added, 0, 1, 5, and 10 parts by weight of bio Cellulose powder was mixed with 2 parts by weight of alginate, agar or carrageenan to prepare a mixed scaffold, and changes in physical properties were observed (FIG. 6).

As a result, it was confirmed that the strength of the fabricated scaffold increased as the hardness, gumminess, and chewiness increased in a concentration-dependent manner of the bio-cellulose powder. On the contrary, springiness, cohesiveness, and Resilience was confirmed to decrease (FIG. 7).

In addition, the results of hardness, gumminess, chewiness, springiness, cohesiveness, and resilience of chicken, cow, and pork were adjusted by adjusting the bio-cellulose powder content. It was confirmed that all values were included (FIG. 7).

Through this, the scaffold prepared by mixing 1 to 10 parts by weight of bio-cellulose powder with 2 parts by weight of alginate, agar or carrageenan with respect to 100 parts by weight of the total composition can reproduce the same texture of chicken, cow, or pork. indicates that there is

Based on the analysis results of the texture characteristics, it is shown that the physical properties of the bio-cellulose composite scaffold itself can be adjusted by the concentration of the bio-cellulose powder, and the actual texture of each meat and part can be reproduced by adding the bio-cellulose powder.

Although the embodiments of the present invention have been described with reference to the accompanying drawings, those skilled in the art to which the present invention pertains can be implemented in other specific forms without changing the technical spirit or essential features of the present invention. you will be able to understand Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.

Claims (19)

A composition for preparing a cultured meat scaffold comprising a bio-cellulose powder derived from Gluconacetobacter sp., a natural polymer gelling agent and a gelation accelerator, the bio-cellulose powder based on 100 parts by weight of the composition for preparing a cultured meat scaffold 1 to 20 parts by weight, 1 to 5 parts by weight of the natural polymer gelling agent and 1 to 5 parts by weight of the gelation accelerator, and the natural polymer gelling agent is composed of alginate, agar and carrageenan. A composition for preparing a cultured meat scaffold, characterized in that any one selected from the group.
delete delete According to claim 1,
The gelation promoter is a composition for preparing a cultured meat scaffold, characterized in that at least one metal salt selected from the group consisting of potassium chloride, calcium chloride, magnesium chloride and calcium lactate.
According to claim 1,
The bio-cellulose powder is a composition for preparing a cultured meat scaffold, characterized in that pulverized at 10,000 to 15,000 rpm after freezing the bio-cellulose.
According to claim 5,
The bio-cellulose is a composition for preparing a cultured meat scaffold, characterized in that produced in a medium containing vegetable peptone.
According to claim 6,
The vegetable peptone is soy peptone, wheat peptone, broadbean peptone, potato peptone, pea peptone, papaic soy peptone and lupine A composition for preparing a cultured meat scaffold, characterized in that at least one selected from the group consisting of peptone (lupin peptone).
According to claim 5,
The bio-cellulose is a composition for preparing a cultured meat scaffold, characterized in that produced in a medium containing glucose as a carbon source.
delete A cultured meat scaffold prepared from any one composition selected from claims 1 and 4 to 8.
According to claim 10,
A cultured meat scaffold, characterized in that the hardness, guminess and chewiness of the cultured meat scaffold increase as the concentration of the bio-cellulose powder in the composition increases.
According to claim 10,
Cultured meat scaffold, characterized in that elasticity, cohesiveness and elasticity of the cultured meat scaffold decrease as the concentration of the bio-cellulose powder in the composition increases.
(A) statically culturing Gluconacetobacter sp. in a medium containing peptone to obtain bio-cellulose;
(b) washing the bio-cellulose obtained in step (a) with distilled water and drying;
(c) freezing the dried bio-cellulose and then pulverizing it at 10,000 to 15,000 rpm to obtain bio-cellulose powder;
(d) mixing the bio-cellulose powder with a natural polymeric gelling agent; and
(e) lyophilizing the product of (d) and then adding a gelation accelerator, 1 to 20 parts by weight of the bio-cellulose powder with respect to 100 parts by weight of the product obtained by adding the gelation accelerator of (e); 1 to 5 parts by weight of the natural polymeric gelling agent and 1 to 5 parts by weight of the gelation accelerator, wherein the natural polymeric gelling agent is any one selected from the group consisting of alginate, agar, and carrageenan. Method for producing a cultured meat scaffold, characterized in that.
delete According to claim 13,
The peptone is soy peptone, wheat peptone, broadbean peptone, potato peptone, pea peptone, papaic soy peptone and lupine peptone Method for producing a cultured meat scaffold, characterized in that at least one selected from the group consisting of (lupin peptone).
delete Cultured meat prepared by culturing non-human animal-derived cells on the cultured meat scaffold prepared by the method of claim 13.
According to claim 17,
The non-human animal-derived cells are cultured meat, characterized in that at least one selected from the group consisting of muscle stem cells, muscle cells and progenitors thereof.
According to claim 17,
Cultured meat, characterized in that the non-human animal is selected from the group consisting of cattle, sheep, pigs, poultry, crustaceans and fish.

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