KR20240009128A - Immobilized enzyme complex, manufacturing method thereof and method for preparing non-roasting coffee using the same - Google Patents

Abstract

The present invention relates to a polysaccharide complex in which enzymes are fixed to improve taste, aroma, color, etc. by adding it to green coffee bean extract to produce coffee without a roasting process, a method of manufacturing the same, and a method of manufacturing non-roasted coffee using the same. will be.
The present invention proposes a new coffee processing method that completely replaces the roasting process, ensuring the sensory characteristics such as taste, aroma, and color formed by roasting, while fundamentally eliminating the possibility of forming acrylamide, a suspected carcinogen, and reducing the cost required for roasting. An enzyme-immobilization complex that saves energy and thereby minimizes carbon emissions from coffee production, a manufacturing method thereof, and a manufacturing method of non-roasted coffee using the same are provided.

Description

Enzyme-immobilized complex, manufacturing method thereof, and manufacturing method of non-roasting coffee using the same {IMMOBILIZED ENZYME COMPLEX, MANUFACTURING METHOD THEREOF AND METHOD FOR PREPARING NON-ROASTING COFFEE USING THE SAME}

The present invention relates to a polysaccharide complex in which enzymes are fixed to improve taste, aroma, color, etc. by adding it to green coffee bean extract to produce coffee without a roasting process, a method of manufacturing the same, and a method of manufacturing non-roasted coffee using the same. will be.

According to the International Coffee Organization (ICO), global coffee consumption is 9,980,760 tons as of 2021, steadily increasing by more than 1% per year on average. In this way, coffee has established itself as a common favorite food for people around the world.

Coffee beans, the raw material of coffee, contain chlorogenic acid, a polyphenol component known for its antioxidant/anticancer and cholesterol biosynthesis inhibition effects. It is known that 10g of Robusta coffee green beans contains up to 600 mg of chlorogenic acid, and Arabica coffee beans contain up to 300 mg.

Coffee is widely consumed in the form of powder or beverage through a 'roasting' process in which coffee beans (green coffee beans) are processed at high temperatures. However, chlorogenic acid, which is contained in large amounts in green coffee beans, loses its activity or is destroyed due to heat denaturation during the process of roasting coffee beans and making coffee beans. This worsens depending on the degree of roasting, and in the case of dark roasting, which is the roasting level preferred in many countries, including Korea, to produce a high body, it is known that more than 90% of the chlorogenic acid contained in green coffee beans is lost.

The flavor enhancement phenomenon that occurs during the roasting process, such as the unique sweetness, rich aroma, vivid color, and heavy texture, occurs when light green green beans are roasted and turn into dark brown beans. A reaction occurs and melanoidin, a browning substance, is formed.

However, during the roasting process, not only melanoidin, a beneficial substance that gives aroma and taste to coffee and has antioxidant effects, but also 'acrylamide', which is presumed to be a carcinogen, is formed. Accordingly, the European Union (EU) and the Korean Ministry of Food and Drug Safety have notified relevant regulations regarding acrylamide, which is generated when raw materials containing carbohydrates are processed at high temperatures, and are legally regulating recommended standards in foods.

In reality, it is very difficult to remove acrylamide, which is produced along with melanoidin, a biologically active and flavor-enhancing ingredient, during the roasting process. Therefore, there is a need for a new coffee processing method that can reduce the production of suspected carcinogens while maintaining taste, aroma, and various physiological activity effects.

Additionally, as coffee consumption increases, a huge amount of carbon dioxide is emitted during the roasting process of heating coffee beans. According to researchers at University College London in the UK, carbon emissions per 1 kg of coffee are 15.3 kg, which is more than half of the carbon emissions of beef.

Countries around the world, including Korea, have recently declared carbon neutrality, and companies are making efforts to reduce greenhouse gas emissions. Accordingly, the need for new coffee processing methods that can fundamentally eliminate or reduce carbon emissions generated during the coffee roasting process is increasing.

According to this technical need, various technical attempts have been made to reduce the acrylamide content generated by roasting through the addition of additives or post-processing processes, but there is a limitation in that it is not a method that fundamentally/originally prevents acrylamide from being generated. there was.

Japanese Patent Publication No. 2006-0503592

In order to solve the above-mentioned problems, the present invention proposes a new coffee processing method that completely replaces the roasting process, thereby securing the sensory characteristics such as taste, aroma, and color formed by roasting, while reducing the possibility of generating acrylamide, a presumed carcinogen. The purpose is to provide an enzyme-immobilization complex that fundamentally eliminates the energy required for roasting and thereby minimizes carbon emissions from coffee production, a manufacturing method thereof, and a manufacturing method of non-roasted coffee using the same.

In order to achieve the above technical problem, the present invention according to a preferred embodiment includes the steps of preparing an enzyme-immobilization complex in which polyphenol oxidase is immobilized on viscopolysaccharide (S10); Preparing green bean extract by adding hot water to green coffee bean powder and filtering it (S20); reacting the green bean extract with an enzyme-immobilized complex (S30); and recovering the enzyme-immobilized complex from the reactant (S40). It provides a method for producing non-roasted coffee using an enzyme-immobilized complex including a.

At this time, the polyphenol oxidase may be any one or a mixture of two or more selected from the group consisting of laccase, tyrosinase, catechol oxidase, and ferroxidase. Among these, laccase is more preferable.

The viscopolysaccharide is a group consisting of alginate, chitosan, hyaluronic acid, cellulose, ulvan, gelatin, starch, and fluran. It may be any one or a mixture of two or more selected from among them, and alginate is more preferable.

The enzyme-immobilized complex preferably has the shape of a bead.

In the step (S10) of preparing the enzyme-immobilized complex, the aqueous solution in which the polyphenol oxidase and viscopolysaccharide are dissolved is dropped into a solution containing divalent transition metal ions to form a bead-shaped stabilized agglomerate. This may be a step in preparing an enzyme-immobilization complex.

Specifically, the step of preparing the enzyme-immobilization complex (S10) involves dissolving sodium alginate as the viscopolysaccharide and laccase as the polyphenol oxidase in an aqueous solution containing divalent transition metal ions. It is preferable that this step is to prepare an alginate-transition metal complex in which bead-shaped laccases stabilized in aggregated form are fixed by dropping them into the solution.

At this time, the weight ratio between viscopolysaccharide and polyphenol oxidase in the aqueous solution may be 5:1 to 15:1.

The divalent transition metal may be copper (Cu(II)), and the solution containing the divalent transition metal ion is preferably a copper(II) sulfate (CuSO ₄ ) solution.

The present invention provides, according to another preferred embodiment, non-roasted coffee using an enzyme-immobilized complex prepared by the above-described production method.

Meanwhile, according to another preferred embodiment, the present invention is an additive added to green bean extract extracted from unroasted coffee beans to enhance the taste, aroma, and color of the green bean extract, and is a chelator between viscopolysaccharide and divalent transition metal. An enzyme-immobilization complex is provided, characterized in that polyphenol oxidase is immobilized within a network three-dimensional structure having a bead shape stabilized in an agglomerated form through a chemical reaction.

At this time, the polyphenol oxidase may be any one or a mixture of two or more selected from the group consisting of laccase, tyrosinase, catechol oxidase, and ferroxidase. Among these, laccase is more preferable.

The viscopolysaccharide is a group consisting of alginate, chitosan, hyaluronic acid, cellulose, ulvan, gelatin, starch, and fluran. It may be any one or a mixture of two or more selected from among them, and alginate is more preferable.

Additionally, the divalent transition metal may be copper (Cu(II)).

Meanwhile, according to another preferred embodiment, the present invention includes the steps of mixing viscopolysaccharide and polyphenol oxidase and dissolving them in water (S1); Dropping the prepared aqueous solution into a solution containing divalent transition metal ions to form a bead-shaped enzyme-immobilized complex stabilized in an agglomerated form (S2); and washing and drying the enzyme-immobilized complex (S3).

At this time, the polyphenol oxidase may be any one or a mixture of two or more selected from the group consisting of laccase, tyrosinase, catechol oxidase, and ferroxidase. Among these, laccase is more preferable.

The viscopolysaccharide is a group consisting of alginate, chitosan, hyaluronic acid, cellulose, ulvan, gelatin, starch, and fluran. It may be any one or a mixture of two or more selected from among them, and alginate is more preferable.

Additionally, the solution containing the divalent transition metal ion may be a copper(II) sulfate (CuSO ₄ ) solution.

In addition, in the step of dissolving in water (S1), the weight ratio between the mixed viscopolysaccharide and polyphenol oxidase is preferably 5:1 to 15:1.

According to the present invention as described above, a new method that excludes roasting or heating is adopted in the coffee processing process to fundamentally suppress the generation of acrylamide, a presumed anti-cancer substance.

In addition, through the oxidation polymerization reaction between the chlorogenic acid component in the green bean extract and polyphenol oxidase, a similar or equivalent level of taste, aroma, and flavor can be formed without roasting.

The enzyme-immobilized composite beads used in coffee processing have high industrial utility because they can be easily recovered and reused by a relatively simple physical method after being used in green coffee extract processing.

In addition, because the roasting process through heating is omitted, the resulting use of energy and related resources can be reduced, and carbon emissions can be dramatically reduced, making it possible to produce coffee, for which demand is steadily increasing, in an eco-friendly manner.

The effects of the present invention are not limited to those mentioned above, and may be clearly understood by those skilled in the art from the description throughout the specification, but also include other effects that are not explicitly mentioned.

Figure 1 shows a general bead (Control, left) made using alginate and copper solution, and an enzyme-immobilized complex bead (Laccase, right) according to the present invention in which laccase enzyme was immobilized during the manufacturing process. This is a photo of the exterior.
Figure 2 is a photograph of the appearance of green bean powder (left) and roasted bean powder (right).
Figure 3 is a photograph of coffee being extracted using green coffee bean powder and coffee bean powder, respectively (left) and the resulting extract liquid (right).
Figure 4 is a photograph of green coffee extract (left) and coffee bean extract (right) to which enzyme-immobilized complex beads according to the present invention were added in a falcon tube.
Figure 5 is a photograph of green bean extract and green bean extract to which enzyme-immobilized beads according to the present invention were added (left), and a photograph of their reaction on a roller (right).
Figure 6 is a photograph of comparative examples and examples after 12 hours of reaction.
Figure 7 is a photograph of the enzyme-immobilized complex according to the present invention before (left) and after 12 hours of reaction (right).

Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the attached drawings. The advantages and features of the present invention and methods for achieving them will become clear by referring to the embodiments described in detail below along with the accompanying drawings. However, the present invention is not limited to the embodiments disclosed below and may be implemented in various different forms, and the present embodiments are merely intended to ensure that the disclosure of the present invention is complete and to provide common knowledge in the technical field to which the present invention pertains. It is provided to fully inform those who have the scope of the invention, and the present invention is only defined by the scope of the claims. Like reference numerals refer to like elements throughout the specification.

Unless otherwise defined, all terms (including technical and scientific terms) used in this specification may be used with meanings that can be commonly understood by those skilled in the art to which the present invention pertains. Additionally, terms defined in commonly used dictionaries are not interpreted ideally or excessively unless clearly specifically defined. The terms used in this specification are for describing embodiments and are not intended to limit the invention. As used herein, singular forms also include plural forms, unless specifically stated otherwise in the context.

As used in the specification, “comprises” and/or “comprising” refers to the presence of one or more other components, steps, operations and/or elements. or does not rule out addition.

First, the present invention provides a method for producing non-roasted coffee, that is, excluding the roasting process.

The method for producing coffee according to a preferred embodiment includes an enzyme-immobilization complex preparation step (S10), a green bean extraction step (S20), a reaction step (S30), and a recovery step (S40).

The complex preparation step (S10) is a step of preparing a complex in which polyphenol oxidase is immobilized using polyphenol oxidase and viscopolysaccharide. Specifically, polyphenol oxidase and viscopolysaccharide are first mixed with water (distilled water) and completely dissolved while stirring to create an enzyme-polysaccharide aqueous solution (S1).

The resulting enzyme-polysaccharide aqueous solution is slowly added dropwise (drop by drop) into a solution containing divalent transition metal ions using a syringe, pipette, dropper, or dropper, forming round beads in the transition metal solution. Molding (S2).

Forming into a bead shape is the result of self-structuring or self-assembly resulting from a chelating reaction between a polysaccharide material and a divalent transition metal. Making it into the shape of a bead and using it in coffee processing is advantageous in ensuring high reactivity, leaving no reactants in the final product (coffee), and easily recovering it for reuse.

After going through the above process, a chelating reaction between the viscopolysaccharide and the divalent transition metal forms a three-dimensional network structure in the shape of a bead that is stabilized in an agglomerated form, and polyphenol oxidase is inside the bead. By immobilization, the enzyme-immobilization complex according to the present invention is prepared. Specifically, the polyphenol oxidase is trapped between the three-dimensional network-shaped viscopolysaccharide (alginate)-divalent transition metal (copper) structure, that is, the enzyme stays through a pocket-like three-dimensional structure. It can be said that a space for unique functions is provided.

The molded enzyme-immobilized complex is washed to remove unreacted substances (copper, etc.) adhering to the surface of the composite bead and dried (S3) to be used in the next step.

Polyphenol oxidase oxidizes chlorogenic acid, a polyphenol component contained in coffee beans (green beans) or green bean extract (a reaction that polymerizes monomers by removing their hydrogen with an oxidizing agent), making them similar to roasted coffee beans even though they are not roasted. It is a core material that makes it possible to achieve the same level of taste, aroma, and color.

Polyphenol oxidase enzymes that can be used include laccase, tyrosinase, catechol oxidase, and ferroxidase, which can be used alone or in combination of two or more. You can use it.

At this time, laccase can be adopted as the polyphenol oxidase. Lacase is a polypinol oxidase containing four copper ions and is generally known as an enzyme that decomposes lignin. Lacase is an enzyme that can oxidize almost all compounds with similar structures, including p-polyphenol, can remain stable in the air for a long time, and has environmentally friendly characteristics as the only product after the reaction is water. Lacase is the most preferred example of the polyphenol oxidase enzymes available in terms of effectiveness according to the present invention.

Mucilpolysaccharide is a material that self-assembles into a three-dimensional structure with transition metal ions and constitutes the skeleton of the enzyme-immobilization complex according to the present invention, and is a substance of proteoglycan, an acidic mucopolysaccharide complexed with protein. It is a substance that exists in the form of a substance that exists in the epidermis or connective tissue of most living organisms and is involved in the support or flexibility of tissues.

The viscopolysaccharides and biocompatible natural polymer materials that can be used include alginate, chitosan, hyaluronic acid, cellulose, ulvan, gelatin, starch, and There are fluran, etc., and they can be used alone or in combination of two or more.

At this time, alginate, more specifically alkaline or alkaline earth metal alginate, can be used as the viscous polysaccharide. Alginate is a polysaccharide acid contained in the cell walls of brown algae plants such as seaweed and kelp, and is a type of viscous polysaccharide with high viscosity.

Alginic acid or alginate changes from sol to gel by chemical change, but has irreversible properties in which the reverse reaction does not occur, so it is widely used for encapsulation/formulation and enzyme immobilization/stabilization. The cation attached to the carboxyl group of the alginate chain is very stable against heat and solvents, and is inexpensive, eco-friendly, and non-toxic, making it very useful as a food additive.

As mentioned earlier, the divalent transition metal is a key element that combines with viscopolysaccharide to form a three-dimensional structure. When the viscopolysaccharide solution is dropped into an aqueous transition metal solution, the transition metal and viscopolysaccharide become entangled like a net, forming a round bead. It will be achieved.

In addition, cupric copper (Cu(II)) can be used as a divalent transition metal, and more preferably, cupric copper sulfate (CuSO ₄ ) solution can be used as a solution containing divalent transition metal ions. . Divalent copper ions effectively induce a chelating reaction with alginate, a viscopolysaccharide, allowing the enzyme-immobilization complex to come together stably to form round beads. After bead formation, components that did not participate in the binding are removed by washing. It has characteristics that make it easy to remove.

Meanwhile, the amount of viscopolysaccharide and polyphenol oxidase mixed in the complex preparation step (S10) is increased by adding more viscopolysaccharide based on weight. Specifically, it is desirable to control the weight ratio between viscopolysaccharide and polyphenol oxidase at a level of 5:1 to 15:1, that is, 5 to 15 times the weight of viscopolysaccharide compared to polyphenol oxidase.

If it is controlled to less than 5 times and the polyphenol oxidase is excessive or the viscopolysaccharide is insufficient, the economic efficiency in terms of enzyme use is low and the specific gravity of the viscopolysaccharide is low, making it difficult to form a stable bead shape smoothly. On the other hand, if polyphenol oxidase is insufficient or viscopolysaccharide is excessive by more than 15 times, the amount of polyphenol oxidase, which plays a key role in processing non-roasted coffee, is insufficient, resulting in improvement of taste, aroma, and color. The effect will not be sufficient.

In addition, in the complex preparation step (S10), the concentration of the divalent transition metal in the solution containing the divalent transition metal (preferably cupric copper) into which the viscopolysaccharide and polyphenol oxidase aqueous solution is dropped is at the level of 0.05M to 0.5M. It is desirable to be controlled.

If the concentration of the transition metal is less than 0.05M, the chelating reactivity with viscopolysaccharide and beading or self-assembly are weak, so the strength and holding power of the bead are not sufficient. If the concentration is more than 0.5M, it is not possible to obtain a fairly hard bead. However, during the reaction with green coffee extract, the structure of the enzyme-immobilized complex bead is destroyed or disintegrated due to the high concentration of transition metals, causing excess copper to be eluted, causing the color of the final reactant to turn blue, resulting in poor food compatibility. .

The enzyme-immobilized complex prepared through the complex preparation step (S10) is prepared by preparing a green bean extract, specifically a green bean extract obtained by hot-water extraction of unroasted green bean powder through filter paper (S20), and then adding the enzyme-immobilization complex thereto. React (S30).

In this process, the color of the green bean extract changes to that of roasted coffee bean extract due to an enzymatic reaction (specifically, oxidation polymerization reaction) between the polyphenol oxidase (preferably laccase) fixed in the complex and the chlorogenic acid contained in the green bean extract. It is browned to a similar or darker color, and sensory characteristics such as taste and aroma are improved.

The amount of polyphenol oxidase in the enzyme-immobilized complex added to the green bean extract is preferably controlled to 0.25 to 1.25 parts by weight based on the total weight (100 parts by weight) of the green bean extract that is the input object. Considering this input ratio, the enzyme-immobilized complex The immobilized complex can be prepared or the dosage of the prepared complex can be determined.

If the amount of polyphenol oxidase added is less than 0.25 parts by weight, the reaction time of the green bean extract is very slow, which reduces process efficiency. If it exceeds 1.25 parts by weight, the reaction rate converges to a certain level, resulting in a large reaction rate. There is a problem of low economic feasibility due to excessive use of enzymes because the synergistic effect cannot be enjoyed.

Finally, in order to commercialize coffee, the enzyme-immobilized complex that has completed the reaction is recovered through a physical separation process such as filtration (S40). The enzyme-immobilized complex formed into beads can be easily separated and recovered, and the recovered complex beads do not lose their chemical activity and can be reused after a simple process such as washing, making them highly industrially usable and economical.

The present invention provides non-roasted coffee produced by the above-described production method according to another preferred embodiment. Coffee made in this way does not contain acrylamide, a suspected carcinogen, but has sensory and visual properties similar to or superior to roasted coffee beans. In addition, since there is no roasting (heating) process in the coffee processing process, energy can be saved and greenhouse gas emissions are minimized, making it eco-friendly.

Hereinafter, the present invention will be described in more detail using examples and comparative examples. However, the following examples and comparative examples are for illustrative purposes only, and the scope of the present invention is not limited thereto.

Preparation of regular beads and enzyme-immobilized complex beads

0.45 g of sodium alginate was added to 100 ml distilled water and completely dissolved with a stirrer (solution 1). In addition, 0.45 g of sodium alginate and 50 mg of Trametes versicolor laccase derived from the fungus Trametes versicolor were added to 100 ml of distilled water and completely dissolved with a stirrer (solution 2).

The solutions 1 and 2 were slowly added dropwise to 3 ml of 0.05 M cupric sulfate (CuSO ₄ ) solution using a syringe to form beads. The obtained beads were thoroughly washed with distilled water at least five times to completely remove copper, etc. that did not participate in the binding. Finally, the normal beads (Control) obtained from the first solution and the enzyme-immobilized complex beads (Laccase) obtained from the second solution are shown in Figure 1.

It can be seen that alginate and copper, which are polysaccharide substances, are stably aggregated through a chelation reaction to form round beads. No special differences in appearance were observed between regular beads and enzyme-immobilized composite beads. The obtained beads were dried for 2 hours at room temperature to remove moisture and used in the experiment.

By manufacturing in the form of such beads, the enzyme can be fixed inside to induce a smooth reaction with chlorogenic acid in the green coffee extract in the future, while also facilitating recovery of the beads after use.

Manufacturing of green coffee beans and coffee bean extract

Green coffee beans (Ethiopian Yirgacheffe Koke Honey) were ground in a commercial blender and filtered through a 2 mm sieve to obtain powder form. The coffee beans used were commercial coffee beans (completely roasted) of the Brazilian Cerrado variety, ground to espresso size. The green coffee bean powder and coffee bean powder used in the experiment are shown in Figure 2.

You can see that green coffee beans have a light ocher color, and coffee beans that have gone through a roasting process in which green coffee beans are heated and roasted have a dark brown color. This is known to be the result of the Maillard reaction, which is well known for adding taste and aroma to food, and the proteins and polysaccharides contained in green coffee undergo an oxidative polymerization process to produce melanoidin.

Coffee filter paper was placed on a glass funnel, 5 g of green beans and coffee bean powder were placed in each filter paper, and coffee was extracted by slowly pouring 180 ml of distilled water heated to 95 degrees in 5 portions. Figure 3 shows hot water extraction of green coffee bean powder and coffee bean powder, respectively, and the result (extract liquid).

The green coffee bean extract was light green and had a high acidity and had a sour scent, while the coffee bean extract was dark brown and had a fragrant and deep-bodied scent. This difference is also because a chemical reaction occurs in the green coffee beans through the Maillard reaction, causing changes in color and aroma.

Comparative experiment of reaction between extract and beads

The extracted green bean extract and coffee bean extract were cooled to room temperature and then used in the experiment. 30 ml of each was placed in a 50 ml falcon tube, and 1 g of the regular beads and enzyme-immobilized complex beads prepared above were added and reacted on a roller. The experiment was repeated twice. Figure 4 shows green coffee extract and coffee bean extract with added beads.

Each extract to which regular beads and enzyme-immobilized complex beads were added was continuously reacted by mixing with a roller at room temperature. At this time, it was confirmed that the color of the specimen to which the enzyme-immobilized complex beads were added became noticeably darker after 30 minutes of reaction. This is the result of an enzymatic chemical reaction occurring between specific components of green coffee beans due to enzyme-fixed beads. The reaction of adding beads to green coffee extract is shown in Figure 5.

After reacting for 12 hours, the beads added to each test subject were recovered using filter paper, and the filtered extract was obtained for each test subject. Referring to Figure 6, in the case of green coffee extract, the color of the specimen reacted with the enzyme-immobilized complex beads according to the present invention was noticeably darker compared to the specimen reacted with regular beads. Likewise, in the case of coffee bean extract, it can be seen that the color of the extract reacted with the enzyme-immobilized complex beads is darker than that of regular beads.

In addition, as a result of a sensory test conducted between non-roasted coffee prepared according to a preferred embodiment of the present invention, regular coffee beans, and regular Dutch coffee (conducted on a total of 8 people, see Table 1), the appearance (color) etc.) were very similar to regular coffee, many said the flavor was similar to regular Dutch coffee, and the taste was also quite similar to regular Dutch coffee.

In contrast to green bean extract, which is lacking in sour taste, body, and sweet and savory taste, green bean extract reacted with enzyme-immobilized complex beads has a body similar or equivalent to coffee bean extract (hot water extracted coffee or Dutch coffee) and a savory and sweet flavor. I could taste it.

very similar Similarity commonly dissimilar very similar vs.
common
coffee beans
coffee appearance 7 One - - - incense One 3 4 - - taste One 2 4 One - vs.
common
Dutch
coffee appearance 4 2 2 - - incense 4 4 - - - taste 5 3 - - -

Recovery of beads after reaction

When the beads used in the reaction for 12 hours were recovered, they were confirmed to have changed to black compared to their appearance before the reaction (see Figure 7). The recovered beads can be reused after being washed in distilled water or alkaline solution (approximately pH 9.0). There are several methods of fixing enzymes to a support, such as bonding them to filters or porous materials in addition to hydrogel beads such as the present invention, but the method of the present invention is a very useful method for reusing enzymes.

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

Claims (24)

Preparing an enzyme-immobilized complex in which polyphenol oxidase is immobilized on viscopolysaccharide (S10);
Preparing green bean extract by adding hot water to green coffee bean powder and filtering it (S20);
reacting the green bean extract with an enzyme-immobilized complex (S30); and
Recovering the enzyme-immobilized complex from the reaction product (S40);
Method for producing non-roasted coffee using an enzyme-immobilized complex comprising. According to paragraph 1,
The polyphenol oxidase is characterized in that it is one or a mixture of two or more selected from the group consisting of laccase, tyrosinase, catechol oxidase, and ferroxidase. Method for producing non-roasted coffee using an enzyme-immobilized complex. According to paragraph 2,
A method of producing non-roasted coffee using an enzyme-immobilized complex, wherein the polyphenol oxidase is laccase. According to paragraph 1,
The viscopolysaccharide is a group consisting of alginate, chitosan, hyaluronic acid, cellulose, ulvan, gelatin, starch, and fluran. A method of producing non-roasted coffee using an enzyme-immobilized complex, characterized in that it is one or a mixture of two or more selected from among. According to paragraph 4,
A method of producing non-roasted coffee using an enzyme-immobilized complex, wherein the viscopolysaccharide is alginate. According to paragraph 1,
A method for producing non-roasted coffee using an enzyme-immobilized complex, wherein the polyphenol oxidase is laccase and the viscopolysaccharide is alginate. According to paragraph 1,
A method of producing non-roasted coffee using an enzyme-immobilized complex, wherein the enzyme-immobilized complex takes the shape of a bead. According to paragraph 1,
The step of preparing the enzyme-immobilized complex (S10) is,
An enzyme characterized in that it is a step of preparing a bead-shaped enzyme-immobilized complex stabilized in an agglomerated form by dropping the aqueous solution in which the polyphenol oxidase and viscopolysaccharide are dissolved into a solution containing divalent transition metal ions. -Method for manufacturing non-roasted coffee using an immobilized complex. According to paragraph 1,
The step of preparing the enzyme-immobilized complex (S10) is,
An aqueous solution containing sodium alginate as the viscopolysaccharide and laccase as the polyphenol oxidase was dropped into a solution containing divalent transition metal ions to form a stabilized bead-shaped bead. A method for producing non-roasted coffee using an enzyme-immobilized complex, characterized in that it is a step of preparing an alginate-transition metal complex to which laccase is immobilized. According to clause 8 or 9,
A method for producing non-roasted coffee using an enzyme-immobilized complex, characterized in that the weight ratio between viscopolysaccharide and polyphenol oxidase in the aqueous solution is 5:1 to 15:1. According to clause 8 or 9,
A method for producing non-roasted coffee using an enzyme-immobilized complex, wherein the divalent transition metal is copper (Cu(II)). According to clause 8 or 9,
A method for producing non-roasted coffee using an enzyme-immobilized complex, wherein the solution containing the divalent transition metal ion is a copper (II) sulfate (CuSO ₄ ) solution. Non-roasted coffee using an enzyme-immobilized complex prepared by the production method of any one of claims 1 to 12. An additive added to green bean extract extracted from unroasted coffee beans to enhance the taste, aroma, and color of the green bean extract,
An enzyme-immobilized complex characterized in that polyphenol oxidase is immobilized within a network three-dimensional structure in the form of a bead that is stabilized in an aggregate form through a chelation reaction between viscopolysaccharide and a divalent transition metal. According to clause 14,
The viscopolysaccharide is a group consisting of alginate, chitosan, hyaluronic acid, cellulose, ulvan, gelatin, starch, and fluran. An enzyme-immobilization complex, characterized in that it is one or a mixture of two or more selected from among. According to clause 14,
The polyphenol oxidase is characterized in that it is one or a mixture of two or more selected from the group consisting of laccase, tyrosinase, catechol oxidase, and ferroxidase. Enzyme-immobilization complex. According to clause 14,
An enzyme-immobilized complex characterized in that the viscopolysaccharide is alginate and the polyphenol oxidase is laccase. According to clause 14,
An enzyme-immobilization complex, characterized in that the divalent transition metal is copper (Cu(II)). Mixing viscopolysaccharide and polyphenol oxidase and dissolving them in water (S1);
Dropping the prepared aqueous solution into a solution containing divalent transition metal ions to form a bead-shaped enzyme-immobilized complex stabilized in an agglomerated form (S2); and
Washing and drying the enzyme-immobilized complex (S3);
Method for producing an enzyme-immobilized complex comprising. According to clause 19,
The viscopolysaccharide is a group consisting of alginate, chitosan, hyaluronic acid, cellulose, ulvan, gelatin, starch, and fluran. A method for producing an enzyme-immobilized complex, characterized in that it is one or a mixture of two or more selected from among. According to clause 19,
The polyphenol oxidase is characterized in that it is one or a mixture of two or more selected from the group consisting of laccase, tyrosinase, catechol oxidase, and ferroxidase. Method for producing an enzyme-immobilized complex. According to clause 19,
A method for producing an enzyme-immobilized complex, characterized in that the viscopolysaccharide is alginate and the polyphenol oxidase is laccase. According to clause 19,
A method for producing an enzyme-immobilized complex, characterized in that the solution containing the divalent transition metal ion is a copper (II) sulfate (CuSO ₄ ) solution. According to clause 19,
In the step of dissolving in water (S1),
A method for producing an enzyme-immobilized complex, characterized in that the weight ratio between the mixed viscopolysaccharide and polyphenol oxidase is 5:1 to 15:1.