JP2016067290A - Production method of composition containing mannose and/or mannooligosaccharide - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for reducing a release of by-product which becomes a load in the subsequent complicated purification process, by more selectively releasing mannose and/or mannooligosaccharide from a natural product.SOLUTION: The invention provides a production method for a composition containing mannose and/or mannooligosaccharide by the action of hemicellulase to mannan, glucomannan, galactomannan, or a natural product containing one or more kinds of them as a constituent sugar. The method is characterized by performing the heat treatment of the hemicellulase before or at the same time of the action of the hemicellulase.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a method for producing a mannose and / or mannooligosaccharide-containing composition, and more particularly useful as a functional food and feed for preventing infection of harmful bacteria, or a synthetic raw material for mannitol and pharmaceuticals. The present invention relates to a method for producing a mannose and / or mannooligosaccharide-containing composition.

  As a method for producing mannose, a method of producing mannose from glucose using a molybdate as a catalyst has been proposed (see, for example, Patent Document 1). There has also been proposed a method for obtaining mannose by obtaining mannose-containing liquid by allowing mannose isomerase to act from fructose as a raw material (see, for example, Patent Documents 2 to 4). These methods are difficult to use in foods because they use chemical substances and microorganisms that have little or no track record in foods. Alternatively, since the yield of mannose is low, it is not a sufficient production method from the viewpoint of inexpensive production.

  On the other hand, a method for producing mannose and / or mannooligosaccharide by acid degradation or enzymatic degradation of mannan, glucomannan, galactomannan, or a natural product containing them has been proposed (see, for example, Patent Documents 5 and 6). .) The former method of acid decomposition requires a large amount of energy because heating is required, or a large amount of salt is generated by neutralization treatment, so it was not a preferable method from the viewpoint of environmental load and cost. . The latter enzymatic degradation method is a highly safe production method with a high yield of mannose and / or mannooligosaccharides, but the enzyme used is a cell itself that produces hemicellulase or a commercially available industrial hemicellulase. Yes, because it contains not only mannan-degrading enzyme activity that produces mannose and / or manno-oligosaccharides, but also various enzyme activities, components other than mannose and / or manno-oligosaccharides are liberated, compared to other chemical production methods. The thing of a miscellaneous composition will be obtained. Therefore, there has been a problem that a complicated purification work for increasing the purity of mannose and / or mannooligosaccharide is essential.

  As described above, many methods for producing a mannose and / or mannooligosaccharide-containing composition have been proposed. However, in order to obtain a mannose and / or mannooligosaccharide having a high purity, any production method increases the production cost. This is a factor that hinders the practical use of high-purity mannose and / or mannooligosaccharide-containing compositions.

JP-A-4-368347 JP-A-4-218370 JP-A-6-292578 JP-A-8-9986 JP 2000-70000 A Japanese Unexamined Patent Publication No. 11-46787

  An object of the present invention is a method for producing mannose and / or mannooligosaccharide by allowing hemicellulase to act on mannan, glucomannan, galactomannan, or a natural product containing one or more of them as a constituent sugar. By heating the hemicellulase before or simultaneously with the cellulase, mannose and / or mannooligosaccharides are more selectively released from the natural product, and a by-product that becomes a burden on the complicated purification process that follows. An object of the present invention is to provide a method for producing a mannose and / or mannooligosaccharide-containing composition with reduced release of substances.

  As a result of diligent study to solve the above problems, the present inventors have found that the activity of hemicellulase that releases mannose and / or mannooligosaccharide is more than the activity of releasing constituent sugars other than mannose and / or mannooligosaccharide. It was found that the thermal stability is high. Then, mannose and / or mannooligosaccharides can be more selectively released from the natural product by heating the hemicellulase before or simultaneously with the action of hemicellulase on the natural product, followed by It has been found that the liberation of by-products, which is a burden on a complicated purification process, is suppressed, and further studies have been made to complete the present invention.

That is, the gist of the present invention is the following (1) to (5).
(1) A method for producing a mannose and / or mannooligosaccharide-containing composition by allowing hemicellulase to act on mannan, glucomannan, galactomannan, or a natural product containing one or more of them as a constituent sugar. A method for producing a mannose and / or mannooligosaccharide-containing composition, wherein the hemicellulase is heat-treated before or simultaneously with the action of hemicellulase.
(2) The method for producing a mannose and / or mannooligosaccharide-containing composition according to (1), wherein the temperature at which hemicellulase is allowed to act on the natural product is not more than the optimum temperature of the hemicellulase.
(3) The method for producing a mannose and / or mannooligosaccharide-containing composition according to (1) or (2), wherein the hemicellulase is mannanase or mannosidase.
(4) The method for producing a mannose and / or mannooligosaccharide-containing composition according to any one of (1) to (3), wherein the hemicellulase is derived from Aspergillus or Trichoderma.
(5) The mannan, glucomannan, galactomannan, or a natural product containing one or more of them as a constituent sugar is a palm kernel or coconut, (1) to (4) A method for producing a mannose and / or mannooligosaccharide-containing composition.

  According to the present invention, by using heat-treated hemicellulase, mannose and / or mannooligosaccharide can be more selectively released from the natural product. Specifically, the galactose ratio can be 1.0% or less, preferably 0.5% or less, more preferably 0.3% or less, as the sugar purity in the obtained composition. Therefore, it is possible to easily and efficiently produce a composition containing high-purity mannose and / or mannooligosaccharide that can be used safely in food. Moreover, the manufacturing method of this invention can be applied also to industrial manufacture.

FIG. 1 shows that in hemicellulase, the activity of releasing mannose and / or mannooligosaccharide has higher heat resistance than the activity of releasing byproducts such as glucose other than mannose and / or mannooligosaccharide. FIG.

Hereinafter, the present invention will be described in detail.
The mannan, glucomannan, galactomannan of the present invention or a natural product containing one or more thereof as a constituent sugar is not particularly limited as long as it contains mannan, glucomannan, galactomannan as a constituent sugar. , For example, palm kernel, coconut palm (copra) or oiled residue (meal) thereof, tsukunei, yam, konjac potato, locust bean, guar bean, coffee bean and the like. The natural product may be a natural product itself or a purified product. A mixture thereof may be used.

  The hemicellulase used in the present invention is not particularly limited as long as it is derived from a microorganism that produces mannan-degrading enzymes such as mannanase (mannase), mannosidase, and the microorganisms include Bacillus subtilis. Subtilis), filamentous fungi (Aspergillus acureitans, Aspergillus awamori, Aspergillus niger, Aspergillus usami, Humicola insolens, Trichoderma harzianum, Trichoderma Koningi, Trichoderma longibrachiatum, Trichoderma bili (Corticium, Pycnopoulos coccineum) and the like. Among these microorganisms, bacteria derived from the genus Aspergillus are preferable because of their high ability to produce mannan degrading enzymes, and in particular, mannan degrading enzymes derived from Aspergillus niger (IFO6661, IFO4417, IFO31012, IFO8541, IFO31125) are preferable. As commercially available hemicellulases derived from Aspergillus niger used in the present invention, cellulosin GM5 (manufactured by HBI Corporation, titer 10,000 units / g), Sumiteam ACH (manufactured by Shin Nippon Chemical Industry Co., Ltd., titer 50,000) Unit / g), Sumiteam AC (manufactured by Shin Nippon Chemical Industry Co., Ltd., titer 1,500 units / g), and hemicellulase derived from Trichoderma viride are cellulosin TP25 (manufactured by HI Corporation, titer 25,000 units). / G). Among these, from the viewpoint of high mannan degrading enzyme activity, cellulosin GM5 and Sumiteam ACH are more preferable.

  In the present invention, it is necessary to heat-treat the hemicellulase before or simultaneously with the action of the hemicellulase on the natural product. From the viewpoint of further suppressing the release of by-products to be described later, It is preferable to heat-treat in the absence of the natural product before allowing the hemicellulase to act on the product.

  The by-product referred to in the present invention is a saccharide such as galactose or glucose other than hemicellulose mannose, peptides or amino acids that are degradation products of proteins contained in natural products, glycerol, natural pigments or saccharides. A colored substance such as a Maillard reaction product produced by a complex reaction of amino acids and the like. Since these by-products become a burden in purification using ion exchange resin, activated carbon, column chromatography, etc., as a result, it can be a factor of lowering the purity of the target substance. In the present invention, the action of hemicellulase that has been heat-treated on the natural product suppresses the liberation of the by-product, resulting in improved purification efficiency and mannose and / or in the resulting composition. Or since the purity of manno-oligosaccharide becomes high, quality improves. In particular, among the sugar mixture components obtained by the conventional methods of acid degradation and enzymatic degradation of natural products, especially galactose is very difficult to separate and purify because it can only be separated by column chromatography with low processing efficiency. Met. The composition of the present invention is extremely useful because it can reduce the burden of the complicated purification process that follows since the ratio of galactose is extremely suppressed.

  Examples of the heat treatment method of the hemicellulase include a method of directly heating the enzyme powder itself, a method of performing a heat treatment after forming an aqueous suspension, or a method of performing a heat treatment by introducing an enzyme into the reaction composition. Any of these methods can be used for the heat treatment, but it is preferable to apply heat uniformly to the enzyme using an incubator or a warm bath, or to heat the enzyme as a water suspension. The heat treatment method is preferably heat treatment in the absence of the natural product.

  When the hemicellulase enzyme powder itself is directly heat-treated, the heat treatment temperature is preferably 30 to 100 ° C, more preferably 50 to 90 ° C, and still more preferably 60 to 90 ° C. The heat treatment time is preferably 0.1 to 5 hours, more preferably 0.5 to 4 hours, and even more preferably 1 to 3.5 hours.

  When heat-treating hemicellulase in a suspended state in water, the heat treatment temperature is preferably in the range of the lower limit of the optimum temperature range of mannan degrading enzyme activity to the deactivation start temperature of mannan degrading enzyme activity + 5 ° C. Preferably, it is more preferably carried out in the range of the lower limit of the optimum temperature range of mannan degrading enzyme activity + 5 ° C or higher to the deactivation start temperature of mannan degrading enzyme activity. The heat treatment time can be appropriately set depending on the mannan degrading enzyme species, etc., for example, preferably 1 minute or more, preferably 5 minutes to 40 hours, more preferably 10 minutes to 2 hours, most preferably 15 minutes to 1 hour. .

  Further, when the hemicellulase is allowed to act on the natural product and at the same time the hemicellulase is heat-treated, from the viewpoint of further reducing the activity of liberating byproducts other than the mannose and / or mannooligosaccharide, mannan degrading enzyme Lower limit of optimum temperature range of activity + 5 ° C or higher-Heat treatment is preferably performed in the range of deactivation start temperature of mannan degrading enzyme activity + 5 ° C, lower limit value of optimum temperature range of mannan degrading enzyme activity + 10 ° C or higher It is more preferable to heat-treat in the range of the deactivation start temperature of mannan degrading enzyme activity.

  The optimum temperature range of mannan degrading enzyme activity in the present invention is that the ability to liberate mannose when a locust bean gum (pH 5.0) is used as a substrate and is allowed to act at a temperature of 40 ° C. for 1 minute is 90% or more. A temperature range.

  As the release reaction conditions for the mannose and / or mannooligosaccharides that allow the above-mentioned heat-treated hemicellulase to act on the natural product, optimum conditions can be selected according to each enzyme. The reaction temperature is desirably carried out under conditions that do not deactivate the enzyme. For example, the reaction temperature is 30 ° C to 75 ° C, preferably 45 ° C to 70 ° C, and more preferably 50 ° C to 65 ° C. Usually, the optimum temperature range of the enzyme is evaluated by the free activity when it is allowed to act at a specific temperature for 1 minute as described above. However, in the method of the present invention, hemicellulase is heat-treated and is a by-product. In view of the balance between enzyme deactivation due to high temperature and release of mannose and / or manno-oligosaccharide, the lower limit of the optimum temperature range is preferable. A temperature that is 1 ° C. or more lower than the lower limit of the optimum temperature range is more preferred, and a temperature that is 5 ° C. or more lower than the lower limit of the optimum temperature range is even more preferred.

  The pH at the time of the free reaction can be selected under optimum pH conditions according to the enzyme. Depending on the type of enzyme used, for example, the pH is preferably 2-9, more preferably pH 2.5-8, and even more preferably pH 3.0-6.0. In addition, although the reaction time depends on conditions such as the amount of enzyme to be used, for example, it is usually preferable to set the reaction time between 3 hours and 48 hours.

  By causing the above-described heat-treated hemicellulase to act on the mannan, glucomannan, galactomannan, or a natural product containing one or more thereof as a constituent sugar, a by-product other than mannose and / or mannooligosaccharide As a result, mannose and / or mannooligosaccharide can be more selectively released from the natural product.

  The amount of hemicellulase that acts on mannan, glucomannan, galactomannan, or a natural product containing one or more of them as a constituent sugar can be arbitrarily selected from the viewpoint of reaction conditions. For example, the natural product (substrate) ) 1 to 500 units per gram are preferable, and 10 to 100 units are more preferable.

  A sugar solution containing mannose and / or mannooligosaccharide obtained by allowing the above hemicellulose to act on a natural product can be further purified using activated carbon, an ion exchange resin, and various chromatographies using them. It can also be solidified by conventional methods such as spray drying and crystallization.

  Hereinafter, the present invention will be described specifically by way of examples. The present invention is not limited to these examples. In addition, the analysis of the various components of a comparative example and an Example was implemented by the method shown below.

Analysis of saccharides (1) Standard substance:
Mannose (trade name “D-(+) Mannose Special” manufactured by Junsei Co., Ltd.)
Glucose (trade name “D-(+) glucose special grade” manufactured by Junsei Co., Ltd.)
Galactose (trade name “D-(+) Galactose special grade” manufactured by Ishizu Pharmaceutical Co., Ltd.)
β1-4 Mannobiose (trade name “β1-4 mannobios” manufactured by Funakoshi Co., Ltd.)
β1-4 Mannotriose (Funakoshi Co., Ltd., trade name “β1-4 mannotriose”)
(2) Analysis method High performance liquid column chromatography method 1 (analysis of mannose, glucose, β1-4 mannobiose, β1-4 mannotriose)
Analysis column: Aminex HPX-87H (manufactured by Bio-Rad)
Column temperature: 60 ° C., mobile phase: 0.005 M sulfuric acid, flow rate: 0.6 mL / min, detection: differential refractometer High-performance liquid column chromatography method 2 (analysis of galactose)
Analysis column; Aminex HPX-87P (Bio-Rad)
Column temperature: 60 ° C., mobile phase: water, flow rate: 0.6 mL / min, detection: differential refractometer

(3) Analysis of amino acids Fully automatic amino acid analyzer (JLC500 / V, manufactured by JEOL Ltd.)
Standard substance: Amino acid (trade name "Amino acid mixed standard solution AN-II" manufactured by Shimadzu Corporation)
Amino acids (trade name “Amino Acid Mixed Standard Solution Type B” manufactured by Shimadzu Corporation)

(4) Color analysis Spectrophotometric method.
The value obtained by subtracting Abs 720 nm from the Abs 420 nm value in an optical path length 1 cm cell was defined as the coloring degree.

Reference example 1 (cleaning of palm kernel meal)
Water and sulfuric acid were added to 450 g of palm kernel meal (produced in Indonesia) to adjust the pH to 1.0, which was adjusted to 4 times the volume of palm kernel meal. After heat treatment at 90 ° C. for 5 hours, a sodium hydroxide solution was added to adjust the pH to 4.0 ± 0.1. The solid content was collected by suction filtration, added water and sucked to remove the salt. The mass of the collected solid (raw material after washing) was 756 g, and the moisture content was 5%.

Reference Example 2 (Relationship between various enzyme activities of hemicellulase and temperature)
To 16.8 g of the raw material after washing obtained in Reference Example 1, 0.05 g of Cellulosin GM5 manufactured by HI Corporation, titer 10,000 units / g, optimum temperature range 60-78 ° C.) and 16.8 g of water were added and stirred. The reaction was carried out for 20 hours at each reaction temperature (58, 62, 65, 68 ° C.). The amount of mannose, the amount of β1-4 mannobiose, the amount of glucose and the amount of galactose contained in this reaction supernatant were measured by the above methods.

  The obtained results are shown in FIG.

  Hemicellulase includes not only mannan degrading enzyme activity for releasing mannose and / or mannooligosaccharide in the natural product, but also various enzyme activities for releasing sugars such as glucose and galactose. As shown in Table 1, it was found for the first time that the activity of releasing mannose and / or mannooligosaccharide has higher heat resistance than the activity of releasing byproducts such as glucose other than mannose and / or mannooligosaccharide. According to this result, high-purity mannose and / or mannooligosaccharide, which can be used safely in food, is obtained by heat-treating the hemicellulase before or simultaneously with the action of hemicellulase on the natural product. It was suggested that the composition can be produced easily and efficiently.

Comparative Example 1
To 16.8 g of the washed raw material obtained in Reference Example 1, 0.05 g of cellulosin GM5 (manufactured by HI Corporation, titer 10,000 units / g, optimum temperature range 60-78 ° C.) and 16.8 g of water were added, The reaction was carried out at 58 ° C. for 20 hours under stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose contained in this reaction supernatant, the amount of glucose, the amount of galactose, the total amino acid, and the coloration were measured by the above methods.

Comparative Example 2
To 16.8 g of the raw material after washing obtained in Reference Example 1, 16.8 g of water and Sumiteam AC (manufactured by Nippon Chemical Co., Ltd., titer 2000 knit / g, optimum temperature range 52 to 65 ° C.) 0.05 g and water 16.8 g was added and reacted for 20 hours at 58 ° C. with stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose contained in this reaction supernatant, the amount of glucose, the amount of galactose, and coloring were measured.

Comparative Example 3
16.8 g of water and Cellulosin TP25 (manufactured by HI Corporation, titer 25000 units / g, optimum temperature range 58-62 ° C.) 0.05 g and water 16. 8 g was added and reacted for 20 hours at 58 ° C. with stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose contained in this reaction supernatant, the amount of glucose, the amount of galactose, and coloring were measured.

Comparative Example 4
To 5.0 g of copra meal, 0.05 g of cellulosin GM5 (manufactured by HI Corporation, titer 10000 units / g, optimum temperature range 60-78 ° C.) and 16.8 g of water are added and reacted at 58 ° C. for 20 hours with stirring. I let you. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose contained in this reaction supernatant, the amount of glucose, the amount of galactose, and the total amount of amino acids were measured.

Comparative Example 5
Cellulosin cellulosin GM5 (manufactured by HI Corporation, titer 10,000 units / g, optimum temperature range 60-78 ° C.) 0.025 g was added to 5 g of locust bean gum, and the total weight was adjusted to 100 g with water. The reaction was carried out at 58 ° C. with stirring for 20 hours. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose and galactose contained in the reaction supernatant were measured.

Comparative Example 6
To 16.8 g of the washed raw material obtained in Reference Example 1, 0.05 g of Sumiteam ACH (Hemicellulase manufactured by Shin Nippon Chemical Industry Co., Ltd., titer 50000 units / g, optimum temperature range 50 to 80 ° C.) and 16.8 g of water And allowed to react at 58 ° C. for 20 hours under stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose and galactose contained in the reaction supernatant were measured.

Example 1
Cellulosin GM5 (mannanase manufactured by HBI Co., Ltd., titer 10,000 units / g, optimum temperature range 60-78 ° C.) was dissolved in water at a concentration of 1% by mass and heated in a 62 ° C. hot bath for 16 hours. 11.8 g of water and 5 g of the aforementioned heat-treated enzyme solution were added to 16.8 g of the washed raw material obtained in Reference Example 1, and the mixture was reacted at 58 ° C. for 20 hours with stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, and galactose in the supernatant of this reaction was measured.

Example 2
Cellulosin GM5 (mannanase manufactured by HBI Co., Ltd., titer 10,000 units / g, optimum temperature range 60-78 ° C.) was dissolved in water at a concentration of 1% by mass, and placed in a 70 ° C. warm bath for 30 minutes and heated. 11.8 g of water and 5 g of the aforementioned heat-treated enzyme were added to 16.8 g of the washed raw material obtained in Reference Example 1, and the mixture was reacted at 58 ° C. for 20 hours with stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, galactose, total amino acid, and coloring of this reaction supernatant were measured.

Example 3
Cellulosin GM5 (mannanase manufactured by HBI Co., Ltd., titer 10,000 units / g, optimum temperature range 60-78 ° C.) was heated in a 90 ° C. incubator for 3 hours in the form of powder. 16.8 g of water and 0.05 g of the aforementioned heat-treated enzyme were added to 16.8 g of the washed raw material obtained in Reference Example 1, and reacted at 58 ° C. for 20 hours with stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, and galactose in the supernatant of this reaction was measured.

Example 4
Cellulosin GM5 (mannanase manufactured by HBI Co., Ltd., titer 10,000 units / g, optimum temperature range 60-78 ° C.) was dissolved in water at a concentration of 1% by mass and heated in a warm bath at 80 ° C. for 1 minute. To 16.8 g of the washed raw material obtained in Reference Example 1, 11.8 g of water and 5 g of the aforementioned heat-treated enzyme solution were added, and the mixture was reacted at 58 ° C. for 20 hours with stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, and galactose in the supernatant of this reaction was measured.

Example 5
To 16.8 g of the raw material after washing obtained in Reference Example 1, 0.05 g of cellulosin GM5 (mannanase manufactured by HBI Co., Ltd., titer 10,000 units / g, optimum temperature range 60 to 78 ° C.) and 16.8 g of water were added. The mixture was placed in a hot bath at 70 ° C. for 30 minutes and subjected to heat treatment. The reaction was carried out at 58 ° C. for 20 hours under stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, and galactose in the supernatant of this reaction was measured.

Example 6
To 16.8 g of the washed raw material obtained in Reference Example 1, 11.8 g of water and 50 mg of cellulosin GM5 (mannanase manufactured by HBI Co., Ltd., titer 10,000 units / g, optimum temperature range 60 to 78 ° C.) were added and stirred. And reacted at 65 ° C. for 20 hours. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, and galactose in the supernatant of this reaction was measured.

Example 7
Sumiteam AC (manufactured by Shin Nippon Chemical Co., Ltd., titer 2000 knit / g, optimum temperature range 52-65 ° C.) was dissolved in water at a concentration of 1% by mass and heated in a 65 ° C. bath for 30 minutes. 11.8 g of water and 5 g of the aforementioned heat-treated enzyme solution were added to 16.8 g of the washed raw material obtained in Reference Example 1, and reacted at 58 ° C. for 20 hours with stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, galactose, and coloring of the reaction supernatant were measured.

Example 8
Cellulosin TP25 (HBI xylanase, titer 25000 units / g, optimum temperature range 58-62 ° C.) was dissolved in water at a concentration of 1% by mass and heated in a 65 ° C. hot bath for 30 minutes. The above-mentioned heat-treated enzyme solution 5 g and 11.8 g of water were added to 16.8 g of the washed raw material obtained in Reference Example, and the mixture was reacted at 58 ° C. for 20 hours with stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, galactose, and coloring of the reaction supernatant were measured.

Example 9
Cellulosin GM5 (manufactured by HBI Co., Ltd., titer 10,000 units / g, optimum temperature range 60-78 ° C.) was dissolved in water at a concentration of 1% by mass and heated in a 65 ° C. bath for 30 minutes. To 5 g of copra meal, 2.5 g of the aforementioned heat-treated enzyme and 22.5 g of water were added, and the mixture was reacted at 58 ° C. for 20 hours with stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, galactose, and coloring of the reaction supernatant were measured.

Example 10
Cellulosin GM5 (manufactured by HBI Co., Ltd., titer 10,000 units / g, optimum temperature range 60 to 78 ° C.) was dissolved in water at a concentration of 1% by mass, and heated in a 65 ° C. hot bath for 30 minutes. 2.5 g of the aforementioned heat-treated enzyme and 92.5 g of water were added to 5 g of locust bean gum, and reacted at 58 ° C. for 20 hours under stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, and galactose in the supernatant of this reaction was measured.

Example 11
Sumiteam ACH (manufactured by Nippon Chemical Industry Co., Ltd., hemicellulase, titer 50000 units / g, optimum temperature range 50-80 ° C.) is dissolved in water at a concentration of 1% by mass, placed in a 65 ° C. warm bath for 10 minutes and heated. did. 5 g of the aforementioned heat-treated enzyme solution and 11.8 g of water were added to 16.8 g of the washed raw material obtained in Reference Example, and the mixture was reacted at 60 ° C. for 20 hours with stirring. The amount of mannose, β1-4 mannobiose and β1-4 mannotriose, glucose, and galactose in the supernatant of this reaction was measured.

  The obtained results are shown in Table 1.

As shown in Table 1, in Examples 1 to 11 in which hemicellulase was heat-treated before or simultaneously with the action of hemicellulase on a natural product containing mannan or the like, Comparative Examples 1 to 6 without heat treatment In comparison with the above, the release of by-products such as glucose, galactose and amino acids was largely suppressed, and a composition with improved purity of mannose and / or mannooligosaccharide could be obtained. In particular, in Examples 1, 2, 4, and 5 in which a specific natural product was used as a raw material, and heat treatment was performed under specific conditions before the Aspergillus niger-derived hemicellulase was allowed to act on the natural product, Generation was extremely suppressed. Then, it was confirmed that the release of glucose and galactose was greatly reduced and the purity of mannoses was improved as compared with the case where hemicellulase was not heat-treated. Moreover, total amino acids and coloring were also reduced, and it was confirmed that the load during purification was reduced.

Claims (5)

Mannan, glucomannan, galactomannan, or a method for producing mannose and / or manno-oligosaccharide by reacting hemicellulase with a natural product containing one or more of them as a constituent sugar, before the hemicellulase is allowed to act Or the manufacturing method of the mannose and / or mannooligosaccharide containing composition characterized by heat-processing the said hemicellulase simultaneously with making it act. The method for producing a mannose and / or mannooligosaccharide-containing composition according to claim 1, wherein the temperature at which hemicellulase is allowed to act on the natural product is not more than the lower limit of the optimum temperature range of the hemicellulase. . The method for producing a mannose and / or mannooligosaccharide-containing composition according to claim 1 or 2, wherein the hemicellulase is mannanase or mannosidase. The method for producing a mannose and / or mannooligosaccharide-containing composition according to any one of claims 1 to 3, wherein the hemicellulase is derived from Aspergillus or Trichoderma. The mannose and / or the mannose according to any one of claims 1 to 4, wherein the mannan, glucomannan, galactomannan, or a natural product containing one or more of them as a constituent sugar is a palm kernel or coconut palm. Or the manufacturing method of a manno-oligosaccharide containing composition.

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