JP6140587B2 - Method for producing polysaccharide-containing composition derived from cassis fruit - Google Patents

Description

  The present invention relates to a method for producing a polysaccharide-containing composition derived from a cassis fruit, and more specifically, a cassis fruit or a processed product thereof using at least one polysaccharide-degrading enzyme selected from a glucan-degrading enzyme and a pectin-degrading enzyme. The present invention relates to a method for producing a polysaccharide-containing composition derived from a cassis fruit, comprising a hydrolysis step.

  In recent years, research on the physiological activity of plant-derived polysaccharides has become active in the field of food, drink, cosmetics, and pharmaceutical industry, and various methods for efficiently obtaining polysaccharides from plants have been reported.

  For example, a method for producing a polysaccharide having an effect of improving allergic constitution by causing cellulase to act on an extraction residue derived from coffee beans has been reported (Patent Document 1).

  In addition, a method for producing a saccharified plant material in which the concentration of oligosaccharide or polyphenol is increased by treating a heat-treated product of apple or other fruit juice residue with a mixed enzyme of cellulase and pectinase has been reported. (Patent Document 2).

As for cassis, various methods for obtaining the polysaccharide component have been studied. Cassis is known as black currant with its Japanese name, black currant, and is widely cultivated mainly in North America, Europe, New Zealand, etc., and is a very well-known material in the food industry such as alcohol and confectionery. It is. Polysaccharides derived from black currant is ca ssis p oly s accharide (CAPS ) and also called, (see Patent Documents 3 and 4) that have been reported to have excellent physiological activities such as anti-tumor effect or anti-allergic effect.

  As a specific method for obtaining a cassis-derived polysaccharide, for example, there is a method of purifying cassis juice with a cation exchange resin and an anion exchange resin and obtaining a flow-through fraction obtained by passing through a C-18 column. It has been reported (Patent Document 3).

  In addition, some of the inventors of the present invention have obtained an average molecular weight of about 10,000 to 10,000 having excellent physiological activity such as immunoregulatory activity by partially digesting a polysaccharide-containing fraction isolated from cassis juice with β-galactosidase. It has been reported that 40,000 polysaccharide-containing compositions were obtained (Patent Document 4).

  However, in order to produce a cassis-derived polysaccharide-containing composition having excellent physiological activity on an industrial scale, a more efficient and stable production method is still required.

JP 2007-217466 A JP 2011-148724 A JP 2004-107660 A International Publication No. 2007/125823

  The present inventors have now industrially produced the amount of β-galactosidase described in Patent Document 4 for the purpose of scaling up the production of a cassis-derived polysaccharide-containing composition having excellent physiological activity to an industrial scale. The experiment of Test Example 1 to be described later was conducted by adjusting to a low level suitable for the above. As a result, in order to obtain a polysaccharide-containing composition having an average molecular weight of about 10,000 to 40,000 as described in Patent Document 4, a step of preheating the cassis raw material and inactivating the endogenous enzyme before the addition of the enzyme is required. It became clear that In order to suppress industrial equipment investment, manufacturing labor, and material costs, it is preferable to produce a polysaccharide-containing composition using a small amount of enzyme without performing pre-heating treatment before enzyme addition. Therefore, as a result of further intensive studies, the present inventors have applied a specific polysaccharide-degrading enzyme other than β-galactosidase, without performing a preheating treatment before adding the enzyme and using a small amount of enzyme, It has been found that a desired cassis-derived polysaccharide-containing composition can be produced efficiently and stably. The present invention is based on such knowledge.

  Therefore, the present invention is intended to efficiently and stably produce a cassis-derived polysaccharide-containing composition having excellent physiological activity without performing a pre-heating treatment prior to enzyme addition and using a small amount of enzyme. That is the purpose.

According to the present invention, the following (1) to (10) are provided.
(1) A method for producing a polysaccharide-containing composition comprising a step of hydrolyzing a cassis fruit or a processed product thereof using at least one polysaccharide-degrading enzyme selected from a glucan-degrading enzyme and a pectin-degrading enzyme.
(2) The manufacturing method as described in (1) whose usage-amount of polysaccharide degrading enzyme is 0.001-0.5% with respect to the total mass of a cassis fruit or its processed material.
(3) The production method according to (1) or (2), which does not include a step of heating the cassis fruit or a processed product thereof to inactivate an endogenous enzyme of the cassis fruit before the hydrolysis step.
(4) The production method according to any one of (1) to (3), wherein the polysaccharide degrading enzyme is β-glucanase or pectinase.
(5) The production method according to any one of (1) to (4), wherein β-galactosidase is further used in the hydrolysis step.
(6) The production method according to any one of (1) to (5), wherein the average molecular weight of the cassis polysaccharide is 10,000 to 40,000.
(7) The production method according to any one of (1) to (6), wherein the processed product is a cassis fruit extract, a cassis juice, a cassis crushed product, or a cassis puree.
(8) A polysaccharide-containing composition derived from cassis fruit, obtained by the production method according to any one of (1) to (7).
(9) A food or drink comprising the cassis polysaccharide-containing composition according to (8).
(10) The food or drink according to (9), which is a beverage.

  According to the present invention, it is possible to efficiently and stably produce a cassis-derived polysaccharide without using a small amount of enzyme without preheating the cassis fruit or its processed product before addition of the enzyme. INDUSTRIAL APPLICABILITY The present invention is advantageous in reducing industrial equipment investment, manufacturing labor, or material costs in the industrial scale production of cassis polysaccharide.

Detailed description of the invention

Method for producing a cassis-derived polysaccharide-containing composition The method for producing a cassis-derived polysaccharide-containing composition of the present invention comprises hydrolyzing a cassis fruit or a processed product thereof using at least one polysaccharide-degrading enzyme capable of degrading glucan or pectin. It is characterized by comprising the step of decomposing.

  In the present invention, the varieties of cassis are not particularly limited, and generally available varieties such as Ben Ard, Ben Rua, Scope Giant and the like can be used. The cassis fruit of the present invention may be a fruit from which the skin, leaves, tree, etc. have been removed or a fruit containing the above. In addition, the collected cassis fruit may be used as it is, may be a dried product, may be a refrigerated product or a frozen product, but is preferably a frozen product.

  In the present invention, the cassis fruit may be used as a processed product. The processed cassis fruit may be a cassis fruit extract using an aqueous medium (water or the like), a cassis pulverized product containing all components in the cassis fruit, or a processed product containing fiber such as cassis puree.

  Further, in the present invention, the amount of cassis fruit used is not particularly limited and can be appropriately determined in consideration of the type of raw material, the type of enzyme, reaction conditions, etc., but the method of the present invention is on an industrial scale. It is preferable when using a scaled-up amount. Specifically, the amount of cassis fruit used is, for example, about 100 kg to 200 t, preferably about 1 t to 100 t, and more preferably about 10 t to 50 t.

  In the present invention, the glucan-degrading enzyme or pectin-degrading enzyme used for the hydrolysis may be any polysaccharide-degrading enzyme capable of degrading glucan or pectin, such as α-glucanase, β-glucanase, pectinase, and polygalacturonase. , Pectinesterase, and the like, preferably β-glucanase or pectinase. These polysaccharide degrading enzymes may be used alone or in combination of two or more.

  In the method of the present invention, another polysaccharide-degrading enzyme can be used together with the polysaccharide-degrading enzyme. Preferable examples of such polysaccharide degrading enzymes include β-galactosidase. When β-galactosidase is used in combination with the polysaccharide-degrading enzyme of the present invention, the amount of β-galactosidase used can be reduced and the hydrolysis time can be shortened as compared with the case of β-galactosidase alone. This is advantageous.

  The amount of polysaccharide-degrading enzyme used is not particularly limited and can be determined appropriately in consideration of the raw material and enzyme type, reaction conditions, etc., but it should be a small amount considering the cost of materials on an industrial scale. Is preferred. The amount of such polysaccharide-degrading enzyme used is preferably 0.001 to 0.5% (w / w), more preferably 0.01 to 0.00%, based on the total mass of the cassis fruit or processed product thereof. It is 05% (w / w), More preferably, it is 0.02-0.03% (w / w). In addition, the amount of polysaccharide-degrading enzyme used is preferably less than 0.06% with respect to the total mass of the cassis fruit or processed product thereof.

  In the present invention, when hydrolysis is performed using a plurality of glucan-degrading enzymes or pectin-degrading enzymes, the amount of polysaccharide degrading enzyme used represents the total amount of enzymes that decompose a plurality of glucans or pectin.

  According to a preferred embodiment of the present invention, the hydrolysis step can be performed with a small amount of the enzyme described above without heating the cassis fruit or processed product thereof to deactivate the endogenous enzyme before the hydrolysis step. . Therefore, this invention is preferable when suppressing the capital investment for performing the preheating process of a cassis fruit or its processed material.

  In the present invention, the specific conditions of the hydrolysis step are not particularly limited as long as the polysaccharide-containing composition of the present invention can be obtained, and depending on the type of polysaccharide-degrading enzyme and the content of the desired cassis-derived polysaccharide, etc. It can set suitably as follows.

  The reaction temperature in the hydrolysis step is preferably performed at a low temperature because the polysaccharide-degrading enzyme can be deactivated by a high-temperature treatment (eg, 70 ° C. or higher) in cassis. The hydrolysis temperature is, for example, about 40 to 60 ° C., preferably about 45 to 55 ° C.

  In the present invention, the hydrolysis reaction time is not particularly limited, but is, for example, within 10 hours, preferably 0.1 to 8 hours, and more preferably 1 to 6 hours. .

  In addition, the hydrolysis reaction is stopped, for example, by boiling for 5 to 10 minutes and heating for 10 to 40 minutes (for example, 70 to 100 ° C., preferably 80 to 90 ° C.) in consideration of the thermal stability of the produced polysaccharide. Etc. Sterilization may be performed by this heating operation.

  In the hydrolysis step, for example, the peak on the saccharide (monosaccharide to oligosaccharide) side of a low molecular weight having a molecular weight of less than about 1,000 is not considered, and the peak on the polysaccharide side (MW> 1,000) is a desired size. At that time, the enzyme reaction is stopped, and the desired polysaccharide-containing composition can be obtained. At this time, after starting the hydrolysis reaction, a part of the reaction solution is sampled over time and analyzed by gel filtration analysis by high performance liquid chromatography (HPLC), or a simple method that can reflect gel filtration analysis The time of reaction stop can be monitored while analyzing by the method. In the molecular weight measurement and monitoring as described above, a differential refraction detector is applied to a high-performance liquid chromatography (HPLC) detector according to the description in Test Examples 1 and 2 described later, and sugars (monosaccharides, disaccharides, Oligosaccharides and polysaccharides) can be detected.

  Moreover, in the manufacturing method of this invention, the process of refine | purifying the polysaccharide containing composition obtained by a hydrolysis process may be included as needed. In the present invention, the purification method is not particularly limited, and for example, a small amount of insoluble precipitate generated in the polysaccharide-containing composition by boiling or the like may be removed by centrifugation. Further, after removing the insoluble precipitate, for example, a polysaccharide-containing composition is purified using an ultrafiltration membrane having a fractional molecular weight of about 1,000, and a low molecular weight (monosaccharide to oligosaccharide) having a molecular weight of less than 1,000 is purified. ) May be removed from the composition.

  According to the present invention, a cassis-derived polysaccharide-containing composition can be obtained by the above-described production method. In the polysaccharide-containing composition of the present invention, the average molecular weight of the polysaccharide is preferably 10,000 to 40,000, more preferably 15,000 to 25, in gel filtration analysis by high performance liquid chromatography (HPLC). 000.

The polysaccharide-containing composition of the present invention may be in any form of liquid, solid, or semi-solid. For example, polysaccharide-containing composition of the present invention, cassis polysaccharide: can be provided in the form of (CAPS ca ssis p oly s accharide ) juice containing (CAPS juice).

  In addition, the polysaccharide-containing composition of the present invention may be used as it is in the state obtained by the above production method, but may be used as a dried product after lyophilization treatment or the like.

  Moreover, the polysaccharide-containing composition of the present invention may contain a cassis-derived protein, a polyphenol compound, or the like in addition to the polysaccharide.

  Moreover, the polysaccharide-containing composition of the present invention can be provided as an oral composition as it is or by appropriately containing a pharmaceutically or food hygienically acceptable additive.

  The content of the cassis-derived polysaccharide in the polysaccharide-containing composition of the present invention is not particularly limited, but is usually 0.001 to 50% by weight, preferably about 0.01 to 10% by weight, based on the dry weight. It is. In addition, the polysaccharide-containing composition of the present invention has a unit intake of 0.1 to 100 g, preferably 0.5 to 50 g, more preferably 1 to 15 g as a fruit juice in terms of Brix 15 per day per adult. Or in an oral intake form.

  The form of the polysaccharide-containing composition of the present invention is not particularly limited, and may be liquid, semi-liquid, solid or paste.

  According to the preferable aspect of this invention, the said polysaccharide containing composition can be provided as food-drinks. The food or drink of the present invention is not particularly limited, but a cassis-derived polysaccharide composition prepared as it is as a food or drink; food hygiene-acceptable additions such as various proteins, sugars, fats, trace elements, and vitamins And those obtained by further adding a product to a polysaccharide composition; those obtained by adding a cassis-derived polysaccharide composition to a general food or drink. Moreover, according to the preferable aspect of this invention, food-drinks are drinks.

  The foods and drinks of the present invention are classified as beauty foods / health foods, functional foods, foods for specified health use, dietary supplements, foods with a disease risk reduction label, or foods for the sick. Are also included.

  Examples of the present invention are shown below, but the present invention is not limited to these Examples.

Test example 1
About 3 kg of frozen cassis fruit (New Zealand Ben Rua) was transferred under refrigerated conditions at 1 ° C. and stored overnight. Next, in a half-thawed state, the cassis fruit with ice attached was pulverized by a roll mill.

  Next, 400 g of the cassis pulverized material obtained by the pulverization treatment was placed in a stainless steel vessel at 81 ° C. and stirred for about 12 minutes, and was heated to 75-80 ° C. (hereinafter also referred to as “preheating”). The preheated cassis pulverized product was allowed to stand after 3 minutes, transferred to ice and further cooled.

Next, 360 g of the preheated cassis pulverized product was stirred and heated to a reaction temperature of 42 ° C. Next, β-galactosidase (Sumilac L, manufactured by Shin Nippon Chemical Co., Ltd.) was added to 0.6% (w / w) with respect to the total amount of the crushed cassis product obtained, and stirred for 1 minute. Next, the obtained reaction product was allowed to stand, sampled over time, and the molecular weight analysis of the polysaccharide was performed. After the sampling, β-galactosidase was inactivated by setting the hot water temperature at 80 ° C. for 35 minutes.
Next, a centrifugal supernatant of the obtained sample is obtained, and molecular weight analysis of the polysaccharide is performed by gel filtration analysis using high performance liquid chromatography (HPLC) (column: OHpak SB-804 HQ (manufactured by Showa Denko KK)). Went.

  Moreover, also about the cassis ground material which is not pre-heated as mentioned above, except the addition amount of (beta) -galactosidase being 0.6 or 0.03% (w / w), it follows the same procedure as the pre-heated cassis ground material. The experiment was conducted.

The results were as shown in Table 1.

  As shown in Table 1, when carrying out enzymatic degradation of cassis fruit using 0.6 or 0.03% (w / w) of β-galactosidase, a molecular weight of 10,000 to 40, No fruit juice (CAPS fruit juice) containing 000 polysaccharides was obtained. On the other hand, when the preheating step was performed, fruit juice (CAPS fruit juice) containing a polysaccharide having a molecular weight of 10,000 to 40,000 was obtained after 4 hours of reaction time.

Test example 2
About 3 kg of frozen cassis fruit (New Zealand Ben Rua) was transferred under refrigerated conditions at 1 ° C. and stored overnight. Next, in a half-thawed state, the cassis fruit with ice attached was pulverized by a hammer mill.

Next, without preheating the cassis pulverized material obtained by the pulverization treatment, β-galactosidase (Sumilac L, manufactured by Shin Nippon Chemical Co., Ltd.), β-glucanase (Viscozyme L, manufactured by Novozymes), pectinase ( One or two kinds of sucrase (registered trademark) S, manufactured by Mitsubishi Chemical Foods Co., Ltd.) were added, heated to a reaction temperature of 48 ° C., and stirred for 1 minute. The obtained reaction product was allowed to stand and sampled over time, and then each enzyme was inactivated by treatment at a hot water temperature of 80 ° C. for 35 minutes.
Next, a centrifugal supernatant of the obtained sample is obtained, and molecular weight analysis of the polysaccharide is performed by gel filtration analysis using high performance liquid chromatography (HPLC) (column: OHpak SB-804 HQ (manufactured by Showa Denko KK)). Went.

The molecular weight analysis results of the polysaccharide are shown in Table 2 below.

As shown in Table 2, when β-galactosidase (Sumilacto L) was used without pre-heating treatment, CAPS juice was obtained at an enzyme concentration of 1% (w / w) (sample number 2), while an enzyme concentration of 0 In 2% (w / w) (sample number 1), no CAPS juice was obtained.
On the other hand, when β-glucanase (Viscozyme L) or pectinase (Sucrase S) is used without performing preheating treatment (sample numbers 3, 4, and 5), each is 0.05% (w / w) or less. CAPS juice was obtained at a low enzyme concentration.
When β-galactosidase (Sumilacto L) and pectinase (Sucrase S) were used in combination (Sample No. 6), CAPS juice was obtained at a lower enzyme concentration than β-galactosidase alone (Sample No. 2).

Claims (3)

A method for producing a polysaccharide-containing composition derived from cassis fruit, comprising a step of hydrolyzing cassis fruit or a processed product thereof using at least one polysaccharide-degrading enzyme selected from glucan-degrading enzyme and pectin-degrading enzyme. And
The manufacturing method whose average molecular weight of the polysaccharide derived from the said cassis fruit is 10,000-40,000.   The manufacturing method of Claim 1 whose usage-amount of a polysaccharide decomposing enzyme is 0.001-0.5% with respect to the total mass of a cassis fruit or its processed material.   The manufacturing method of Claim 1 or 2 which does not include the process which heats a cassis fruit or its processed material and inactivates the endogenous enzyme of a cassis fruit before the said hydrolysis process.