JP6391020B2 - Method for producing natural pigment extract - Google Patents

Description

  The present invention relates to a method for producing a natural pigment extract, and more particularly to a method for producing a natural pigment extract using fruit peels.

Fruits such as apples, grapes, and strawberries are used for raw consumption and processed into juice, preservatives, dried fruits, etc. by squeezing, cooking and drying.
Fruit peels such as apples, grapes, and strawberries contain natural pigments (flavonoid compounds, carotenoid compounds, etc.), and this food is used to color processed foods.
As a method of extracting red pigment (anthocyanin pigment) contained in apple peel, there are a method of heating (boiling) apple peel and a method of using an extraction solvent such as methanol or ethanol. Moreover, the method (patent document 1) which extracts an anthocyanin type pigment | dye using an organic acid solution as an extraction solvent, the method (patent document 2) which refine | purifies a red pigment | flour fraction from the juice or extract obtained from the fruit of an apple, There is also a method using a food enzyme preparation (Non-patent Document 1).

JP 2013-166831 A JP-A-8-319433

Tsunetomo Matsuzawa: Creating a healthy and delicious “Nagano brand”-Regional contribution through training human resources for technological innovation-Journal of Biotechnology, Vol. 92, No. 12, pp.696-697 (2014)

When extracting and using pigments from fruits, it is of course possible to separate the pigment fraction and purify and use the pigment, but it is not necessary until the pigment is purified for use as a food material. It is sufficient if an extract from which the pigment is extracted is obtained.
In consideration of such applications, it is possible to easily obtain an extract of a dye without complicated steps such as purification of the dye, and the extract is used as it is by adding it to food. It is important to obtain a dye extract without using unsuitable acids that are not originally contained in fruits, acids that deviate from the original taste, or extraction solvents such as methanol. It is also possible to extract the pigment from the whole fruit, but it is more efficient to use the peel because most of it is present in the peel.

From the above viewpoint, a method for obtaining a pigment extract using an edible organic acid such as lactic acid or citric acid (Patent Document 1) is effective, but the pigment is further easily and efficiently extracted. Is required. In addition, the addition of an organic acid may require labeling on the final product depending on the amount added and use.
An object of this invention is to provide the natural pigment extract manufacturing method which can extract a pigment | dye efficiently and easily from fruit peels, such as an apple, a grape, and a persimmon.

The method for producing a natural pigment extract according to the present invention comprises heating a fruit peel to inactivate an endogenous enzyme in the fruit peel, and an enzyme reaction treatment for causing the enzyme to act on an object to be treated after the heating step. A step, a step of inactivating the enzyme of the object to be processed after the enzyme reaction treatment step, a step of solid-liquid separating the object to be processed after deactivating the enzyme and recovering the extract, and filtering the extract And a filtration step of recovering the natural pigment extract of the anthocyanin pigment contained in the fruit peel, and no extraction solvent other than water is used in any of the treatment steps .

Moreover, the method for producing a natural pigment extract according to the present invention is characterized in that, in the heating step, the heating temperature and the heating time of the object to be processed are 85 ° C. or more and 2 minutes to 10 minutes.
In addition, the method includes a concentration step of concentrating the natural pigment extract recovered by the filtration step.

Moreover, the method for producing a natural pigment extract according to the present invention uses apple peel as the fruit peel, and uses a complex enzyme mainly composed of protopectinase, cellulase, and hemicellulase as the enzyme. Thus, a natural pigment extract obtained by efficiently extracting pigment from apple peel can be obtained , and protopectinase IGA-C is suitable as the enzyme. In addition, grape skin was used as the fruit skin, and pigment was efficiently extracted from grape skin by using a complex enzyme mainly composed of cellulase, hemicellulase, and pectinase as the enzyme. A natural pigment extract can be obtained.

  According to the method for producing a natural pigment extract according to the present invention, a pigment can be efficiently and easily extracted from the fruit peels of fruits such as apples and grapes. The natural pigment extract obtained by the method of the present invention can be used as a food material as it is, and can be used for various purposes such as use for coloring jams and other foods such as yogurt. Can do.

It is a flowchart of the manufacturing process about the manufacturing method of a natural pigment extract. It is a photograph of the extract before filtration and centrifugation of enzyme-treated extraction of Fuji pericarp. It is the photograph of the extract after the centrifugation of the enzyme treatment extraction of Fuji peel.

(Natural pigment extract manufacturing process)
In FIG. 1, the manufacturing process about the manufacturing method of the natural pigment | dye extract which concerns on this invention is shown as a flowchart.
In the method for producing a natural pigment extract according to the present invention, a natural pigment extract is produced using fruit peels such as apple peel, grape peel, and persimmon peel. Below, the manufacturing method of the natural pigment | dye extract which concerns on this invention according to a manufacturing process is demonstrated.

(Preparation of fruit peel)
First, prepare the fruit peel, which is a material from which the pigment is extracted. Apples can be used in the manufacturing process of processed apple products, the peels obtained by peeling the apples with peels. As the persimmon skin, the pericarp obtained by peeling off the dried persimmon can be used. Grape can use the fruit juice residue after squeezing obtained in the process of producing grape juice or wine.
In the process of manufacturing processed apple products and the process of manufacturing dried persimmons, a large amount of peeled skin is discarded. In addition, fruit peel, which is a residue generated in the manufacturing process of juice and wine, which are processed grape products, is also discarded. According to the present invention, these discarded skins can be used effectively.

(Peel heating process)
The pericarp heating step is a step of heat-treating the prepared fruit pericarp for the purpose of reducing the influence of the endogenous enzyme of the pericarp on the natural pigment extract and the growth of microorganisms in the enzyme treatment reaction.
As a heating method, a method such as microwave, steam, or boiling can be used.
When adding water, it is preferable to add water 0.5 to 2 times, preferably 0.75 to 1.25 times the raw material ratio (weight).
The heating temperature and the heating time in the heating step may be 85 ° C. or higher and about 2 to 10 minutes in consideration of inactivation of the inherent enzyme and microbial contamination in the enzyme treatment.

(Cooling process)
A cooling process is a process of cooling the to-be-processed object after the said heat processing to the temperature suitable for an enzyme process. The temperature at which the enzyme reacts actively varies depending on the type of enzyme used in the enzyme reaction. Therefore, the object to be treated is cooled to a temperature suitable for the enzyme treatment according to the enzyme used.
In the examples, three types of enzymes were used: protopectinase IGA-C (IGA Bioresearch Co., Ltd.), Sumiteam SPC (Shin Nihon Chemical Industry Co., Ltd.), and acemocellulase KM (Kyowa Kasei Co., Ltd.). Of these three enzymes, the enzyme activity of protopectinase IGA-C is preferably 55 ° C or lower, more preferably 50 ° C or lower, and Sumiteam SPC is preferably 60 ° C or lower, more preferably 55 ° C. The acemocellulase KM is preferably 70 ° C. or lower, more preferably 65 ° C. or lower. Since the enzyme activation temperature varies depending on the enzyme used in this way, the treatment temperature of the object to be treated is adjusted according to the enzyme used.

(Enzyme reaction treatment)
The enzyme reaction treatment step is a step of cooling an object to be treated to an appropriate temperature for enzyme treatment, adding an enzyme to the object to be treated, and causing the enzyme to act on the object to be treated.
In the present invention, a complex enzyme containing an enzyme that degrades pectin contained in the plant wall, an enzyme that degrades cellulose, and an enzyme that degrades hemicellulose is used.
Enzymes that act on pectin are protopectinase that catalyzes the production of pectin from protopectin, pectinesterase that catalyzes the reaction of hydrolyzing pectin to produce pectinic acid, and acts on the hydrolysis of pectinic acid There is pectin polygalacturonase (pectinase).
Note that pectinase is a general term for a group of enzymes that degrade pectin contained in the cell walls of higher plants, and is mainly classified into the following three enzymes. Examples include polygalacturonase (hydrolyzing pectin acid), pectin lyase (decomposing pectin acid by elimination reaction), pectin esterase (decomposing pectin methyl ester bond) and the like.
Cellulase is an enzyme that degrades cellulose. This cellulase is not a single enzyme but a general term for several cellulases, and these enzyme groups cooperate to decompose cellulose. It is considered that the following three types of cellulases are involved in the mechanism by which natural cellulose is broken down to glucose. Cx-enzyme (CMCase) Randomly acts on non-crystalline cellulose to hydrolyze the polymer, producing cellodextrin, cellobiose and glucose), C1-enzyme (Avicerase) (acts on crystalline cellulose and non-crystalline And the like, β-glucosidase (decomposing cellobiose into glucose) and the like.
Hemicellulase is a general term for enzymes that hydrolyze hemicellulose. Hemicellulose is a generic name for polysaccharides extracted from plant tissues by alkali, and main polysaccharides include xylan, arabinoxylan, xyloglucan and glucomannan. Enzymes that hydrolyze these polysaccharides are generally referred to as hemicellulases. Typical enzyme names include xylanase and galactanase.

The amount of the enzyme added to the object to be processed may be 0.01% to 0.1% as a weight ratio with respect to the raw material (fruit peel).
The enzyme reaction may be a method in which the enzyme is added to the object to be treated and stirred, and then allowed to stand for a predetermined time. The time for performing the enzyme reaction is about 0.75 h to 18 h. By setting the temperature of the object to be treated below the temperature at which the enzyme is activated, the enzyme reaction proceeds only by leaving the object to be treated. By this enzyme reaction step, the pigment is extracted from the object to be treated.
It is known that anthocyanin pigments contained in the skin of apples, grapes and the like are generally easily decomposed at high temperatures. Therefore, it is possible to react the temperature of the object to be treated at the optimum temperature of each enzyme preparation. However, since the optimum temperature is relatively high, the method of performing the enzyme treatment for a long time with the object to be treated as low as possible. Is effective as a method for extracting anthocyanin pigments.

(Solid-liquid separation process)
The solid-liquid separation step is a step of recovering the extract by solid-liquid separation of the object to be processed that has undergone the enzyme reaction treatment. What is necessary is just to perform a solid-liquid separation process using a centrifuge, a decanter, and a press type squeezer. The solid-liquid separation method is not particularly limited.

(Enzyme deactivation process)
The enzyme deactivation process is a process of heating the extract obtained by solid-liquid separation and performing a process of deactivating the enzyme contained in the extract. The treatment for inactivating the enzyme by heating can be performed by heating the extract at 85 ° C. to 95 ° C. for about 3 minutes.

(Filtration process)
A filtration process is a process of filtering the extract after an enzyme deactivation process, and collect | recovering as a natural pigment extract. For the filtration step, an appropriate method such as a method using a mesh sieve, a centrifugal separation method, a ceramic filtration method, or a method using diatomaceous earth can be used.

The enzyme deactivation process described above is a process for preventing the enzyme from remaining in the extract. As a method for preventing the enzyme from remaining in the extract, apart from the method of heating the extract to inactivate the enzyme, when recovering the natural pigment extract in the filtration step, It is also possible to prevent the enzyme from remaining by filtering the enzyme and recovering the natural pigment extract. In order to separate the enzyme in the filtration step, it may be separated using a ultrafiltration membrane or a ceramic membrane.
When using the method of separating an enzyme in the filtration step, the enzyme may be separated in the filtration step without heating the extract obtained in the extraction step after the enzyme reaction treatment to deactivate the enzyme. Further, as described above, the filtration step can be used as a substitute for solid-liquid separation.

(Concentration process)
The natural pigment extract obtained in the filtration process can be used as it is, but the concentrated solution can be used for various purposes such as delay in fading of the pigment, storage, storage, and distribution. Is advantageous. Therefore, the natural pigment extract obtained in the filtration step is usually used after being concentrated.
As a method for concentrating the natural pigment extract, a method using a vacuum concentration kettle, UF membrane, RO membrane, or the like can be used.
In addition, when concentrating a natural pigment extract using a vacuum concentration kettle, the natural pigment extract is heated to 60 ° C. to 70 ° C. and concentrated in a vacuum kettle. Thus, when heating and concentrating in the concentration step, the concentration treatment can be performed also as the enzyme deactivation step described above. That is, in the concentration step, it is possible to concentrate the natural pigment extract while heating the filtrate obtained by filtering the extract to deactivate the enzyme.
When the concentration process also serves to deactivate the enzyme, it is not necessary to perform the enzyme deactivation process on the extract. In the filtration step, it is not necessary to separate the enzyme using a ultrafiltration membrane or the like, and a natural pigment extract containing the enzyme may be recovered.
The method of performing the treatment for inactivating the enzyme in the concentration step has the advantage that the heat treatment applied to the extract can be completed once.

(Filling / sterilization, storage process)
The concentrated solution after the concentration step is filled in a sealed container, heat-sterilized or hot-packed in a container after heat sterilization, cooled, and stored. Any container such as a plastic bag can be used as the sealed container.
Note that the concentrated concentrate of the natural pigment extract after the concentration step does not necessarily need to be sterilized. This filling and sterilization process is a process for reducing the microbial risk, and it is not necessary to add heat if the container is filled and frozen immediately after concentration. Heating at 65 ° C for about 10 minutes is necessary if the product is stored at room temperature or refrigerated without being frozen.

As an example of producing a natural pigment extract from the fruit peel, an experiment was carried out to produce a natural pigment extract using apple and grape peels as treatment objects. Hereinafter, experimental results will be described.
In addition, in order to analyze and compare the color tone and concentration of natural pigment extracts, L * a * b * values (Lab values) are measured using a colorimetric color difference meter, and the absorbance of the sample pigment substance is measured using a spectrophotometer. Was measured.
L * a * b * values are L * for lightness, a * and b * for chromaticity indicating hue and saturation, a * for red, -a * for green, b * for yellow, -b * indicates the blue direction. The color becomes brighter as the value increases, and the color becomes duller as the value becomes 0.

(Experiment with apple juice)
As an experiment prior to the production of a natural pigment extract from apple peel, an experiment was conducted to examine the difference in pigment content depending on the type of apple.
The apple juice used was varieties “Fuji”, “Akiei”, “Shinano Sweet” and “Kotama” that were squeezed together with the skin, and used the unadjusted apple turbid juice made at the fruit juice factory. .
In the color tone analysis, apple juice was filtered with a filter paper (ADVANTEC: No. 5A), and the filtrate was measured for Lab value with a colorimetric color difference meter (ZE-6000: manufactured by Nippon Denshoku Industries Co., Ltd.). In the analysis of absorbance, turbidity affects the measured value, so centrifuge at 12,000 rpm for 15 minutes, and the resulting supernatant is absorbed at a wavelength of about 520 nm using a spectrophotometer (V-630BIO: manufactured by JASCO). It was performed by the method of measuring.

Table 1 shows Lab values measured using a colorimetric colorimeter for the transmitted light of the filtrates of the four types of samples obtained by the above-described method.
The measurement results in Table 1 show that red balls are stronger in red than yellow, and the color brightness is lower than other three types of apples. Conversely, Fuji, Akiei and Shinano Sweet have a relatively strong yellow color and a bright color.

Table 2 shows the results of measuring the absorbance at a wavelength of 512 nm. Since anthocyanins in apples have strong absorption at 512 nm, the content of red pigment contained in apples can be estimated by examining the absorbance at 512 nm.
The measurement results in Table 2 indicate that among four types of Fuji, Akiei, Shinano Sweet, and Kodama, Kodama has the highest absorbance. From the absorbance measurement results, it is considered that among these apples, red balls contain the most red pigment.

(Apple: Example of natural pigment extract)
An example in which a natural pigment extract of apple is produced according to the method of the present invention using apple peels as a treatment target will be described.
Apple peel (Fuji) was finely cut, water (150 g) was added to the peel (150 g), sealed in a degassed state, and heated in boiling water for 3 minutes.

After the skin heat treatment, the container containing the object to be treated was cooled, cooled to below the optimum temperature in each enzyme reaction (about 40 ° C.), and then the enzyme was added.
The enzymes used in the experiment are three types: protopectinase IGA-C (manufactured by IGA Bioresearch Corporation), Sumiteam SPC (manufactured by Shin Nippon Chemical Industry Co., Ltd.), and acemocellulase KM (manufactured by Kyowa Kasei Co., Ltd.). In either case, 0.05% was added to the material to be treated in the raw material weight ratio.
After the enzyme is added and stirred, the container containing the object to be treated is allowed to stand in a room temperature environment to carry out the enzyme reaction treatment. The time for allowing the enzyme to act by allowing the treatment object to stand is 18 hours.

Next, the extract was sealed in a degassed state and heated in boiling water for 3 minutes to inactivate the enzyme.
Next, the processed material subjected to the enzyme reaction treatment was subjected to solid-liquid separation, and an extract was collected.
Table 3 shows the results of measuring Lab values using a colorimetric color difference meter for these two types of control and three types of enzyme-treated samples. Moreover, the extract obtained by the completely same process as the sample which performed the enzyme process except the process which adds an enzyme as an enzyme control section except what processed the skin heating process as a control section was used. That is, the control group is a product obtained by subjecting an object to be treated to an equal amount of water, followed by heating and cooling, and solid-liquid separation. In the enzyme control group, an equal amount of water was added to the object to be treated, and then heated and cooled. Then, the enzyme was not added, and the mixture was allowed to stand at the same temperature and for the same time as the enzyme treatment group.
The measurement results shown in Table 3 show that the red element shows a higher red element, especially protopectinase IGA, when the enzyme treatment is performed compared to the red element. It can be seen that the red color appears strongly in the sample using -C.

Table 4 shows the results of measuring the absorbance of the above five types of samples.
The measurement results in Table 4 indicate that the absorbance by the red pigment (anthocyanin) is increased by performing the enzyme treatment, that is, the red pigment is more effectively extracted by the enzyme treatment. In particular, the red absorbance of the sample using protopectinase IGA-C is high, and it can be said that the enzyme protopectinase IGA-C is effective in extracting the red pigment of apples.

  FIG. 2 shows the state of the extract before filtering and centrifuging the extract. The extract is slightly turbid and has an orangeish color, but the extract treated with the enzyme protopectinase IGA-C is slightly red.

  FIG. 3 shows a state after the extract is filtered with a filter paper. The extract that has been subjected to the enzyme treatment by removing the pulp has a deeper red color than the extract that has not been subjected to the enzyme treatment.

(Grape: natural pigment extract production example)
An example of producing a natural pigment extract of grapes according to the method of the present invention using grape skins as a treatment target will be described.
Grape peel (Concord) was finely cut, water (200 g) was added to the peel (200 g), placed in a heating pan, and heated at 85 ° C. for 5 minutes. Sealed in a degassed state and heated in boiling water for 3 minutes.

  After the heat treatment, the container containing the object to be treated is cooled, cooled to below the optimum temperature for each enzyme reaction (about 40 ° C), and then treated with apple protopectinase IGA-C (IGA Bio). Research Co., Ltd.), Sumiteam SPC (Shin Nihon Chemical Industry Co., Ltd.), Acremo Cellulase KM (Kyowa Kasei Co., Ltd.) are added at 0.05% by weight of the raw material, and left at room temperature for 18 hours. The enzyme reaction treatment was performed.

Next, the extract was sealed in a degassed state and heated in boiling water for 3 minutes to inactivate the enzyme.
Next, the processed material subjected to the enzyme reaction treatment was subjected to solid-liquid separation, and an extract was collected. In the analysis of the color tone, the Lab value was measured with a colorimetric color difference meter using a filter paper (ADVANTEC: No. 5A) filtered and diluted 10-fold. For the analysis of absorbance, a method of measuring absorbance at a wavelength of about 520 nm using a spectrophotometer (V-630BIO: manufactured by JASCO Corp.) after centrifuging at 12,000 rpm for 15 minutes and diluting the obtained supernatant 20 times I went there.

Table 5 shows the results of measuring Lab values using a colorimetric color difference meter for these two types of control and three types of enzyme-treated samples. Moreover, the extract obtained by the completely same process as the sample which performed the enzyme process except the process which adds an enzyme as an enzyme control section except what processed the skin heating process as a control section was used. That is, the control group is a product obtained by subjecting an object to be treated to an equal amount of water, followed by heating and cooling, and solid-liquid separation. In the enzyme control group, an equal amount of water was added to the object to be treated, and then heated and cooled. Then, the enzyme was not added, and the mixture was allowed to stand at the same temperature and for the same time as the enzyme treatment group.
The measurement results shown in Table 5 show that the red element shows that the a value is positive and the b value is negative in the case where the treatment is performed with the accremocellulase KM compared to the case where the enzyme treatment is not performed. From this, it can be seen that the purple color is extracted by the enzyme treatment.

Table 6 shows the results of measuring the absorbance at a wavelength of 530 nm using a spectrophotometer for the above five types of samples. Since anthocyanins have strong absorption around 520 nm, the content can be estimated by examining the absorbance at 530 nm.
The measurement results in Table 6 show that the absorbance by anthocyanins is increased by performing the acremcellulase KM treatment, that is, the acemocellulase KM treatment more effectively extracts the dye.

(Usage example of natural pigment extract)
Most apple jams in the market are yellow jams that are the same as the flesh color. The reason is that peeling is generally performed in the jam manufacturing process, and natural red pigment (anthocyanin) is not used. However, the image of apples is red, and consumer orientation towards red apple jam is high. So, we decided to develop a technically difficult red apple jam with no colorant added. By adding the “red bean” peel natural pigment extract obtained by the method for producing a natural pigment extract of the present invention to the jam, a red vivid apple jam was successfully developed.

The present invention provides a pigment extraction method that enables efficient and simple extraction of natural pigments contained in the peel. The natural pigment food material obtained by the method of the present invention has the advantage that the enzyme used for extraction is inactivated or removed, so that it can be handled as a processing aid, and there is no need to display additives. . Moreover, it is a material suitable for food without impairing the flavor of the original raw material. Therefore, it can be used as a useful material that can be used for various purposes other than jam.

Claims (6)

A heating step of heating the fruit peel and deactivating the endogenous enzymes of the fruit peel ;
An enzyme reaction treatment step in which an enzyme is allowed to act on the object to be treated after the heating step;
A step of deactivating the enzyme of the object to be treated after the enzyme reaction treatment step;
A step of solid-liquid separation of the object to be treated after inactivating the enzyme and recovering the extract;
Filtering the extract, and collecting a natural pigment extract of anthocyanin pigment contained in the fruit peel, and a filtration step ,
A method for producing a natural pigment extract, wherein no extraction solvent other than water is used in any of the treatment steps . 2. The method for producing a natural pigment extract according to claim 1 , wherein in the heating step, the heating temperature and the heating time of the object to be processed are 85 ° C. or more and 2 to 10 minutes . The method for producing a natural pigment extract according to claim 1, further comprising a concentration step of concentrating the natural pigment extract collected by the filtration step . Use apple peel as the fruit peel,
The method for producing a natural pigment extract according to any one of claims 1 to 3, wherein a complex enzyme mainly comprising protopectinase, cellulase, and hemicellulase is used as the enzyme . The method for producing a natural pigment extract according to claim 4 , wherein protopectinase IGA-C is used as the enzyme . Use grape skin as the fruit skin,
The method for producing a natural pigment extract according to any one of claims 1 to 3, wherein a complex enzyme mainly comprising cellulase, hemicellulase, and pectinase is used as the enzyme .