TWI600379B - Method of combining pressure and enzyme to extract hydrolysate of animal-derived composition - Google Patents

Description

Method for extracting animal component hydrolysate by combining pressure and enzyme

The invention relates to a method for extracting hydrolysate of animal components by combining pressure and enzyme, in particular to a method for extracting animal component hydrolysate by hydrolyzing enzymes under specific pressure.

Hydrolysates of animal ingredients are often used in a wide variety of different biotechnology products, such as foods with animal extracts, health products, makeup products, animal extract composts, and the like. Therefore, how to effectively prepare high-quality and high-concentration animal component hydrolysate has become the focus of research and development in this industry.

For example, collagen, which is often referred to today as a beauty product, is one of the hydrolysates of animal ingredients. Among them, collagen is a water-insoluble, fibrous glycoprotein, which accounts for 1/3 of the total amount of human protein, and is mainly distributed in connective tissue, and is a matrix that constitutes bones, cartilage, blood vessels, skin, and the like. Because collagen has quite a lot of effects, collagen raw materials are commonly used in oral nutrition or maintenance cosmetics. At first, most of them were extracted from animal tissues such as cattle, pigs, sheep, and birds. However, these animals may have diseases such as mad cow disease, foot-and-mouth disease and avian flu. Therefore, fish skin and fish scale collagen peptides are gradually being used in functional health foods.

When the conventional technique uses the fish scale to extract collagen, the fish scale material is first washed with water. Subsequent acid soaking, enzyme hydrolysis and other processes. Because the fish scales are not easily hydrolyzed, some conventional techniques also cook the fish scales at a high temperature of 100-121 ° C for 30 minutes before and after acid soaking, in order to facilitate hydrolysis of the fish scale materials. However, after high-temperature cooking of fish scale materials, not only will the nutritive value of the extract be lost, but also the taste may be poor.

In addition, traditional extraction methods often require a long period of time and a large amount of manpower, and the use of organic solutions can also cause environmental problems. Moreover, the conventional extraction method also has the disadvantages of low extraction efficiency and low nutritional value of the extract. Therefore, the development of extraction methods that can be applied to various animal tissues and have high extraction efficiency and extraction concentration must be beneficial to the development of related products.

In view of the above-mentioned problems of the prior art, it is an object of the present invention to provide a method for extracting an animal component hydrolysate in combination with pressure and an enzyme, comprising: pretreating an animal tissue to obtain a crude extract; The enzyme is added to the crude extract and hydrolyzed under a pressure of 0.2 to 400 MPa to obtain a hydrolyzed extract. The animal tissue may comprise animal epidermal tissue or meat tissue.

Preferably, in the extraction method disclosed in the present invention, the pre-treatment step may further comprise washing the animal tissue with a preset concentration of saline; removing the animal tissue by using a herbicide; and performing the animal tissue Pickling, and further obtaining a crude extract.

Most preferably, the hydrolysis step may comprise adding the crude extract to the solution at a ratio of 1:3 to 1:200 and hydrolyzing at a hydrolysis temperature of 10 to 70 ° C for 2 to 48 hours. Among them, the solution may be a vinegar solution or water.

Optimally, the enzyme may comprise a complex protease (protamex), an alkaline protease (alcalase), papain, flavourzyme, trypsin, bromelain, pepsin, synthetic protease, collagenase, saccharolytic enzyme, galactose, starch Enzyme, disaccharidase, glucose enzyme, glucanase, or chitinase. Among them, the enzyme concentration can be 0.001 to 10% w/v.

The extraction method disclosed in the present invention may further comprise a sterilization step of high pressure sterilization or high temperature sterilization. High-pressure sterilization can be carried out at a pressure of 200~800MPa for 1 minute to 30 minutes, while high temperature sterilization can be carried out at a temperature of 60~150 °C for 1 second to 2 hours.

Most preferably, in the extraction method disclosed in the present invention, the animal epidermal tissue may comprise from the carp, the mullet, the milkfish, the squid, and the squid.

Most preferably, the extraction method disclosed herein may further comprise filtering the hydrolyzed extract using a filter aid.

As described above, the method of extracting animal hydrolysate by combining pressure and enzyme according to the present invention may have one or more of the following advantages:

(1) The method of the present invention is a method in which the physical and chemical properties of animal tissues are hydrolyzed by using an enzyme under high pressure, whereby the concentration of the active material in the hydrolyzed extract can be increased.

(2) The method of the present invention can be hydrolyzed by different pressures for different animal tissues, and thus can be widely used for extracting different kinds of animal tissues, and is applied to research and development of various health foods or cosmetic products.

(3) The solvent used in the method of the present invention is water or vinegar solution, which is more environmentally friendly than the conventional method using an organic solvent.

(4) The method of the present invention does not require high temperature treatment, thereby avoiding the nutrition of the extract Problems such as loss of value or poor taste.

S1~S3, S11~S17, S31~S33‧‧‧ steps

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a flow diagram of a method for extracting hydrolysate of an animal component in combination with pressure and enzyme according to an embodiment of the present invention.

Figure 2 is a flow diagram of the prior processing steps in the method of the present invention.

Figure 3 is a flow diagram showing the steps of enzymatic hydrolysis of the crude extract by extraction of high pressure in the process of the present invention.

The invention will be further described in detail by the following preferred embodiments and the accompanying drawings. It should be noted that the experimental data disclosed in the following embodiments are for explaining the technical features of the present invention, and are not intended to limit the manner in which they can be implemented.

Fig. 1 is a flow chart showing a method of extracting an animal component hydrolysate in combination with pressure and an enzyme according to an embodiment of the present invention, and an embodiment of the present invention will be described in detail below with reference to Fig. 1.

Referring to Fig. 1, the extraction method of the present invention first performs a pretreatment step on animal tissues to obtain a crude extract (step S1). The animal tissue may comprise animal epidermal tissue, such as fish skin, chicken skin, pig skin or the like; or meat tissue, which may be derived from chicken, duck, pig, cow, sheep, turtle, fish, and the like. However, the invention is not limited thereto.

Next, the enzyme is added to the crude extract, and hydrolysis is carried out under a pressure of 0.2 to 400 MPa to obtain a hydrolyzed extract (step S3). Here, the type of the enzyme to be used can be adjusted according to the active substance to be extracted. For example, the enzyme may comprise a complex protease (protamex), an alkaline protease (alcalase), papain (papain), and a wind. Flavourzyme, trypsin, bromelain, pepsin, synthetic protease, collagenase, saccharolytic enzyme, galactose, amylase, disaccharidase, glucosidase, Portuguese A glycanase or a chitinase, and the enzyme concentration may be 0.001 to 10% w/v. However, the invention is not limited thereto.

Further, in the hydrolysis step of the step S3, the crude extract is further added to a solution in a ratio of 1:3 to 1:200, and is hydrolyzed at a hydrolysis temperature of 10 to 70 ° C for 2 to 48 hours. Here, the solution may comprise water or a vinegar solution.

In the extraction method of the present invention, the hydrolysis treatment may be sterilized by autoclaving or high temperature sterilization. Among them, high-pressure sterilization can be sterilized at a pressure of 200~800MPa for 1 minute to 30 minutes. The high temperature sterilization is carried out at a temperature of 60 to 150 ° C for 1 second to 2 hours.

Further, in the extraction method of the present invention, it may further comprise filtering the hydrolyzed extract with a filter aid (for example, diatomaceous earth).

Referring to Figure 2, there is shown a flow chart of the prior processing steps in the extraction process of the present invention. Here, the present embodiment will be described in detail by taking collagen in the extracted fish skin as an example. However, the invention is not limited thereto. Among them, the fish skin can be taken from various fish, such as: squid, squid, squid, squid, squid, and the like. In addition, fish skins that can be used to extract collagen generally refer to the epidermis, dermis, and derivatives thereof. Therefore, the commonly known fish skin or scales belong to the fish skin here.

In the pretreatment step, the extraction method of the present invention first washes the fish skin with fresh water (step S11), for example, washing the fish skin with tap water for about 15 to 30 minutes. Next, the fish skin is washed with water at a predetermined concentration of saline (step S13). Preset concentration of food The brine is a saline solution with a concentration ranging from 2% to 6%. When the fish skin is washed with saline, the ratio between animal tissue and saline is between about 1:2 and 1:20 (W/V). In addition, the number of flushes with saline can be a multiple number and may vary during the rinse. For example, the fish skin is rinsed twice with saline, and the period of each rinse is about 5 to 15 minutes.

Next, the fish skin is impregnated with a herbicide for about 1 to 20 hours (step S15), and the number of times of the immersion may be 1 to 3 times, and then washed with water and then pickled to obtain a crude extract (step S17).

According to a preferred embodiment of the present invention, when the fish skin is subjected to pickling treatment, an acidic substance such as vinegar, citric acid or malic acid having a concentration of 2% to 30% may be used to soak the fish skin for about 0.1 to 4 hours. After the acid pickling step is completed, the fish skin is rinsed again with water to remove the astringent-containing solution.

Thereafter, a solution (for example, vinegar or water) is further added to a container for removing the fish skin after the mash, and immersed at a low temperature (0 to 25 ° C) for about 0.1 to 24 hours. Among them, the ratio of fish skin to solution after mashing is about 1:3~1:200, and the pH value is adjusted to between 1~4 or 5~9.

Then, the impregnated fish skin is subjected to subsequent homogenization treatment, that is, the fish skin is muddy or emulsified into a crude extract. Among them, the time for homogenization treatment is about 1 to 30 minutes.

Please refer to Fig. 3, which is a process for the step of hydrolyzing the extract by subjecting the crude extract to enzymatic hydrolysis by extraction of high pressure. That is, further explanation will be given for step S3 shown in FIG. Herein, the collagen in the extracted fish skin is used as an example for detailed description. However, the invention is not limited thereto.

As shown in Fig. 3, the enzyme is added to the crude extract obtained through the pretreatment step to obtain a mixture (step S31). Here, the crude extract is a fish scale which has been soaked by vinegar. Since a part of collagen has been extracted by acid treatment, it is extracted by acid and crude; then the mixture is sealed in a vacuum bag and applied. The high pressure treatment is carried out to obtain a collagen hydrolyzed extract, that is, a collagen peptide (step S33).

In step S33, the pressure of the high pressure treatment is between 0.2 and 400 MPa, and is carried out at a temperature of 10 to 70 ° C for 2 to 48 hours.

Further, in the extraction method of the present invention, a herbicide and a filter aid (for example, diatomaceous earth) may be selectively used to remove the astringency of the fish skin. For example, the collagen hydrolyzed extract is filtered by a filter aid to remove the astringency of the collagen hydrolyzed extract.

Compared with the conventional technique of performing hydrolysis under normal pressure, when the extraction high pressure is used in the hydrolysis process, the produced collagen hydrolyzed extract does not have an offensive taste, and the mouthfeel is better.

In addition, by combining enzymes and high-pressure extraction methods, even the harder scales of the scales will be destroyed. According to experiments, only 1% of the soluble solids (Brix) content can be extracted when the collagen is extracted from the fish skin by conventional techniques. However, according to the hydrolyzed extract extracted by the combined enzyme and pressure extraction method disclosed in the present invention, 2.2% of the soluble solid matter can be extracted. That is to say, by using the enzyme of the present invention in combination with a high pressure extraction method, more than twice as much soluble solid matter can be extracted from the fish skin.

From this, it can be seen that according to the embodiment of the present invention, the fish skin can be hydrolyzed to a higher degree, and thus a relatively complete collagen component can be extracted. And the extract of the embodiment of the present invention The rate of taking is high, and the amount of small molecule peptide obtained is also large.

Since the molecular weight of the collagen peptide affects the rate of absorption through the intestines and skin, the smaller the molecular weight of the collagen peptide, the faster the intestinal absorption into the blood metabolism and across the skin barrier, and the more the dermis is activated. Fibroblasts, which in turn promote the biosynthesis of collagen. According to the concept of the present invention, the collagen extracted from the fish skin can be hydrolyzed by the enzyme to increase its water solubility and its molecular weight becomes smaller, which also increases the rate of intestinal absorption.

Furthermore, according to the embodiment of the present invention, the processing flow of the autoclaving can also be combined, so that the extraction process of the collagen does not need to use high temperature at all, so it can be applied to the production of high bioavailability undenatured collagen, and the extraction technique is further improved. It can enhance the overall nutritional and health value.

In another embodiment, the extraction of turtles as an animal tissue is performed. In this embodiment, the whole turtle is diced or crushed, and then added with appropriate water (ratio: 1:0 to 1:200), and then the enzyme is added, and the mixture is bagged and sealed to obtain a mixture and then placed at 0.2 to 600 MPa. The hydrolysis reaction is carried out in a water bath at a pressure of 1 to 80 °C.

Next, the enzyme is deactivated by heating, cooled by a cooling bath, and filtered. Here, filtration can be carried out using a filter aid such as diatomaceous earth or general filter paper. Secondly, the sterilization is carried out under high pressure or high temperature. The high pressure sterilization can be carried out at a temperature of 1 to 100 ° C for 1 second to 24 hours under a pressure of 1 to 1000 MPa, and the high temperature sterilization is performed at a temperature of 60 to 150 ° C for 1 second. ~2 hours. Thereby, a turtle hydrolyzed extract is obtained.

Comparing the soluble solids extracted from the turtle extracted by the above method with the soluble solids in the commercially available quercetin, the comparison results are shown in Table 1: Table 1

It can be seen from Table 1 that the soluble solids of the turtle-hydrolyzed extract extracted by the extraction method of the present invention is about twice as high as that of the commercially available quercetin, and the peptide content is more than 19 times higher.

From the above, it can be seen that the method of combining the enzyme and the pressure-extracting animal component hydrolyzate according to the present invention allows the animal tissue to be hydrolyzed to a higher degree, thereby obtaining a higher extraction rate. Furthermore, the method according to the present invention can be combined with a high-pressure sterilization process, so that the extraction process does not require the use of high temperature, so that it does not cause damage to heat sensitive proteins, and can improve the overall nutritional and health value, so it can be widely used. application.

The above is intended to be illustrative only and not limiting. Any equivalent modifications or alterations to the spirit and scope of the invention are intended to be included in the scope of the appended claims.

S1~S3‧‧‧ steps

Claims (11)

A method for extracting an animal component hydrolysate by combining pressure and an enzyme, comprising: performing a pretreatment step on an animal tissue to obtain a crude extract; and adding an enzyme to the crude extract at 5 to 400 MPa The hydrolysis is carried out under pressure to obtain a hydrolyzed extract, wherein the enzyme comprises a complex protease (protamex), an alkaline protease (alcalase), papain, trypsin, bromelain, pepsin. (pepsin), synthetic protease, collagenase, saccharide-degrading enzyme, galactosidase, amylase, disaccharidase, glucosease, glucanase, chitinase or a combination thereof, and the concentration of the enzyme is 0.001~ 10% w/v. The method of claim 1, wherein the animal tissue comprises an animal epidermal tissue or a meat tissue. The method of claim 1, wherein the pre-processing step comprises the step of homogenizing the animal tissue to obtain the crude extract. The method of claim 3, wherein the hydrolyzing step further comprises adding the crude extract to a solution at a ratio of 1:3 to 1:200, and hydrolyzing at a hydrolysis temperature of 10 to 70 ° C. ~48 hours. The method of claim 4, wherein the solution is a vinegar solution or water. The method of claim 1, further comprising a sterilization step. The method of claim 6, wherein the sterilization step comprises high pressure killing Bacteria or high temperature sterilization. The method of claim 7, wherein the autoclaving is performed at a pressure of 200 to 800 MPa for 1 minute to 30 minutes. The method of claim 7, wherein the high temperature sterilization is performed at a temperature of 60 to 150 ° C for 1 second to 2 hours. The method of claim 2, wherein the animal epidermal tissue comprises from the group of squid, squid, squid, squid, or squid. The method of claim 1, further comprising filtering the hydrolyzed extract with a filter aid.