JP2852206B2 - Liquid food material manufacturing method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a liquid food material applicable to various foods such as beverages, juices and seasonings, and more specifically, it is produced directly from agricultural products such as vegetables and fruits by enzymatic treatment. The present invention relates to a liquid food material containing fructooligosaccharide or gluconic acid or both.

[0002]

BACKGROUND OF THE INVENTION Against the background of growing health consciousness, it is desired that foods based on agricultural products contain special ingredients which contribute to health.

[0003] For example, vegetable drinks have poor acidity,
Various measures have been taken to add acidity,
Traditionally, lemon juice and JP
This is a method in which an organic acid preparation such as citric acid described in 5-336936 is specially added from the outside. In addition, the addition of oligosaccharides having an action such as growth of bifidobacteria,
Β-carotene, which is considered to have anticancer activity, has also been added. However, these methods have disadvantages that they contain a large amount of additives and produce foods different from the original flavor of agricultural products.

[0004] Gluconic acid, which is one of the organic acids having a refreshing acidity, is widely used, for example, after production, purified and added to foods such as tofu and ice cream. However, there is no example of adding gluconic acid to a beverage such as vegetable juice or a seasoning for imparting acidity, and only calcium gluconate intended for a calcium source may be used. Recently, Asano et al. [No. 47
This gluconic acid has been reported to have a growth activity of bifidobacteria, and is attracting attention, according to the Annual Meeting of the Japanese Society of Nutrition and Food Science, p111, (1993)]. Therefore, if gluconic acid itself is present in beverages and seasonings, it is expected that nutritional value will be enhanced as well as taste, but such a food material has not been found.

It is well known that glucose (glucose) is oxidized by the action of glucose oxidase and catalase to produce gluconic acid, and several techniques related to the production of gluconic acid utilizing this enzyme reaction have been found. (JP-A-49-6193, JP-A-57-12)
277 etc.). However, there is no technology for producing a food material that imparts acidity by performing an enzymatic reaction using natural agricultural products as raw materials. For example, the above-mentioned JP-A-49-6-6
The technology described in 193 and JP-A-57-12277 is only for removing glucose from a synthetic sugar solution containing glucose.

Aspergillus niger (Aspergil
When an enzyme fructosyltransferase obtained from a microorganism such as lus niger) is allowed to act on a sucrose (sucrose) solution, fructose is added to the sucrose by 1-2%.
It is also known that a bound fructooligosaccharide can be obtained (JP-A-56-154967 and JP-B-63-6-6).
No. 2184). However, these technologies also use a synthetic sugar solution as a raw material, and convert sucrose as much as possible to produce sweeteners exclusively. However, it is not a technique for obtaining a food material with a balanced fructooligosaccharide and sucrose. Furthermore, this method is for the production of fructooligosaccharide alone, cannot produce gluconic acid, and is not a technique for producing gluconic acid as an acidity simultaneously with fructooligosaccharide.

[0007]

SUMMARY OF THE INVENTION Under the circumstances described above, the problem to be solved by the present invention, that is, the object of the present invention is to produce a new type of acidity from agricultural products such as vegetables and fruits. Gluconic acid or oligosaccharides,
Another object of the present invention is to develop a novel food material which does not add both of them from the outside, yet has a harmonized acidity and sweetness, and further has a growth activity of bifidobacteria different from fermented beverages.

[0008]

The present invention has achieved the above object by producing fructooligosaccharides and / or gluconic acid by causing an enzyme to act on a natural agricultural product itself.

Thus, according to one aspect of the present invention,
A sucrose (sucrose) -containing agricultural product is used as a raw material, and a fructosyltransferase-containing enzyme agent is allowed to act on the sucrose (sucrose) -containing enzyme product to produce fructooligosaccharide having one or two molecules of fructose bonded to sucrose. A method for producing a liquid food material is provided.

According to another aspect of the present invention, an agricultural product containing glucose (glucose) is used as a raw material, and an enzyme containing glucose and catalase is allowed to act on the raw material to produce gluconic acid. And a method for producing a liquid food material.

Further, in the most preferred embodiment of the present invention, a raw material agricultural product containing glucose and sucrose is treated with an enzyme agent containing glucose oxidase, catalase and fructosyltransferase to give gluconic acid and fructooligosaccharide. (1 to 2 molecules of fructose bonded to sucrose).

Agricultural products used as a raw material in producing the food material according to the present invention generally contain glucose or sucrose itself, but are not limited thereto, and a substance serving as a precursor thereof, that is, A raw material containing a substance capable of producing glucose or sucrose by the action of an appropriate enzyme can also be used.

For example, when using sugarcane as a raw material,
Sugar cane contains sucrose but not glucose. However, in the case of using an enzyme containing fructosyltransferase together with glucose oxidase / catalase, glucose is released from sucrose in the process of producing fructooligosaccharides by the action of fructosyltransferase even without glucose. From this, gluconic acid can be produced.

When cereals such as potatoes and rice are used as raw materials, starch is decomposed by the action of amylase to produce glucose, and gluconic acid can be produced therefrom.

Similarly, even if the raw material does not contain sucrose or contains only a trace amount, for example, when the raw material contains raffinose, sucrose will be produced by the action of α-galactosidase enzyme. It can also be produced to obtain fructooligosaccharides.

The agricultural products used as raw materials in the production of the liquid food material of the present invention are typically vegetables and fruits, but are limited to those generally included in these names. I can't. That is, the raw agricultural products in the present invention include cereals (rice, wheat, cereals, potatoes, etc.), beans, crafts (sugar cane, tea leaves, etc.), pastures, vegetables, fruit trees, root vegetables As well as those derived from plants that are sometimes referred to as leafy vegetables, flowers and vegetables, fruits and vegetables, flowers and the like, as well as those derived from animals such as honey, glucose, sucrose and / or Any one containing such precursor substances can be used. These are generally
It is made into an appropriate liquid (liquid state) such as a squeezed liquid, an extract, or a solution, and is subjected to treatment with a fructosyltransferase and / or a glucose oxidase / catalase-containing enzyme agent.

According to the present invention, a fructo-oligosaccharide having one or two molecules of fructose bound to sucrose, which is obtained by allowing a fructosyltransferase to act on an agricultural product containing sucrose or a substance which is a precursor of sucrose, -What is called kestose or nystose.

The present invention provides a liquid food material suitable for beverages having a high nutritional value and a good balance between sweetness and intestinal action by directly reacting fructosyltransferase on agricultural products containing sucrose. . Thus, according to a preferred embodiment of the present invention, when fructosyltransferase is allowed to act on a sucrose-containing agricultural product, the content of fructooligosaccharide (W / V%) formed and the content of sucrose (W / V%) are determined.
/ V%) is 1.0 or less. The food material thus obtained is preferably diluted so as to contain the fructooligosaccharide in an amount of 0.5% (W / V%) or more when the final beverage is obtained.

The fructosyltransferase for producing fructooligosaccharides used in the present invention includes Aspergillus niger IAM2020 and IAM209.
3, IFO31125, Rhizopus delema
r) Enzymes extracted from cells such as IFO4801 (these can be obtained from the Japan Society for Applied Microbiology and the Fermentation Research Institute) or commercially available enzymes (fructosyltransferase and β-D-fructo Furanosidase).

The present inventor has argued that, among these enzymes, Aspergillus niger IAM2020 was used in a medium containing no sucrose.
It has been found that the enzyme obtained by culturing is particularly excellent. That is, a medium to which sucrose, which is a substrate of this reaction, is added is usually used as a medium for culturing bacteria in producing a fructosyltransferase enzyme or the like. According to meat extract without added sucrose, peptone,
By using a medium containing NaCl, the fructose transfer activity (U _t ), total activity, and transfer activity (U _t ) /
An enzyme having a high hydrolysis activity (U _h ) is obtained, and a particularly active fructosyltransferase enzyme is obtained from IAM2020.

When a reaction is carried out using such a fructosyltransferase enzyme under appropriate reaction conditions, for example, at a temperature of 40 ° C., a vegetable beverage material containing fructooligosaccharide is obtained. The reaction can be performed in a batch system in which the enzyme is directly mixed with the raw agricultural product (eg, vegetable juice) and stirred. However, in consideration of the cost for mass production, a continuous reaction incorporating the immobilized enzyme using a bioreactor is considered. The method is preferred.

At this time, the production of fructo-oligosaccharides using the enzyme agent containing fructosyltransferase according to the present invention comprises immobilizing fructosyltransferase on cross-linked porous silica beads to carry out the enzymatic reaction. preferable. That is, agricultural products such as those used as raw materials in the present invention contain a large amount of solids, and their liquid properties are quite viscous. Therefore, when using a reactor, the immobilized material must have physical strength, but the physical strength of gel-like immobilized carriers such as alginate beads, chitosan beads, and carrageenan, which are commonly used in the past, are compared. Small and easy to channel. Thus, the present inventors have developed porous silica beads having a good physical strength (particle diameter of 1.05 to 1.05).
2.36 mm, average pore diameter 128 °, apparent porosity 74.
2 to 77.6%, pore volume 1.23 to 1.53 ml /
Using g), this was cross-linked with aminopropyltriethoxysilane and glutaraldehyde, and the enzyme was immobilized, incorporated into a bioreactor, and found to be able to react efficiently by passing a concentrated agricultural product-containing liquid through.

In producing a liquid food material exclusively containing fructooligosaccharides according to the present invention, the agricultural product (liquid) has a low acid content and therefore a pH of 4.5.
~ 7.0, preferably pH 5.5 ~
Particularly effective for 7.0 vegetable juice. An example of a most preferred agricultural product for use in such a method of the invention is vegetable juice, especially ginseng juice (carrot juice).
Preferred examples of other raw agricultural products include sugarcane juice.

In particular, when fructooligosaccharides are produced from agricultural products (liquid) in such a pH range, agricultural products (liquid) containing 10% or more sucrose are preferably concentrated agricultural products (liquid) containing 20% or more. ) Is desirable from the viewpoint of maintaining high enzyme reaction efficiency and activity.

Further, according to the present invention, by reacting glucose oxidase and catalase on a liquid food material containing glucose (or a precursor thereof), gluconic acid, which has not been used so far, is added as a sour taste. A new type of food material can be obtained.

When producing gluconic acid according to the present invention, any concentration of glucose can be used as long as it is present, but preferably a liquid agricultural product containing Brix 10 or more containing 0.5% or more of glucose, and A liquid agricultural product concentrated to the above concentration is desirable from the viewpoint of taste due to the balance between acid and sugar after the reaction.

Generally, such a glucose-containing raw material having a pH in the range of 3.0 to 7.0 is used. Since those having a pH of 4 or more have a low sourness, the generation of gluconic acid increases the sourness, and a liquid food material particularly suitable as a beverage can be obtained.

In general, it has been considered that the reaction by the glucose oxidase / catalase enzyme used in the present invention has a reduced activity in a low pH range and its stability is also deteriorated. However, the present inventor has found that a useful food material can be obtained even by using a low pH raw material such as fruit juice (for example, juice from kiwifruit, oyster, pear, etc.).

That is, by acting glucose oxidase / catalase on these agricultural products having a pH lower than 4 (but not less than 3), a food material having a stronger acidity and not suitable as a beverage but suitable as a seasoning can be obtained. . For example, a ponzu-like acidic seasoning can be produced by appropriately mixing such a liquid food material having an extremely strong acidity with soy sauce (for example, treated fruit juice: soy sauce = 8: 1). Conventional ponzu is citrus citric acid, and vinegar soy sauce is made of acetic acid, whereas the seasoning obtained using the liquid food material of the present invention is made from gluconic acid as a seasoning, Is a completely new acidic seasoning with different types and tastes.

Further, the food material of the present invention containing gluconic acid is used for producing wine by alcohol fermentation with yeast to produce wine, or vinegar or seasoning liquid having complex sourness obtained by acetic acid fermentation with acetic acid bacteria to give acetic acid. It can also be used for manufacturing.

The glucose oxidase for producing gluconic acid used in the present invention includes, for example, Aspergillus
Aspergillus niger (for example, IAM2094)
For example, a medium consisting of 10 g of glucose, 3 g of yeast extract, 3 g of malt extract, 5 g of peptone, and 1000 ml of distilled water can be used to prepare a filamentous fungus such as
Put into a 1-volume jar fermenter at a temperature of 30 ° C and a ventilation rate of 1
000 ml / min, stirring speed 300 rpm
There are bacterial cells obtained by culturing for 5 hours and treating with an organic solvent such as acetone, a crude enzyme solution extracted from the bacterial cells, and a commercially available glucose oxidase purified enzyme. When using glucose oxidase, it is better to mix a commercially available catalase purified enzyme in terms of removing hydrogen peroxide and improving the reaction efficiency. In the present invention, a commercial glucose oxidase / catalase mixed enzyme preparation for food addition in which glucose oxidase and catalase are mixed in advance can also be used. When such a glucose oxidase enzyme and a catalase enzyme are allowed to act on a liquid agricultural product or a concentrated solution thereof under aerobic conditions, for example, under conditions of rotating shaking treatment or aeration stirring treatment, a liquid food containing gluconic acid is obtained. Get the material in a short time.

The present inventors have also found that a food material containing both fructooligosaccharides and gluconic acid can be produced by first reacting fructosyltransferase and then glucose oxidase on the raw agricultural products. Thus, according to the present invention, as a raw material agricultural product containing glucose (or a precursor thereof), a liquid material also containing a fructooligosaccharide obtained as a result of acting fructosyltransferase as described above is used. The present invention provides a new type of liquid food material containing both gluconic acid and fructooligosaccharide, harmonizing acidity and sweetness, and also having a bifidobacterial growth effect.

Further, the present inventor has proposed that a mixed enzyme consisting of glucose oxidase, catalase and fructosyltransferase is allowed to act on a raw agricultural product containing both glucose and sucrose (or a precursor thereof) to thereby produce gluconic acid. And fructooligosaccharides can be obtained at the same time, and in particular, a method has been found in which such a reaction can be performed very effectively in a single step.

That is, although the production of fructooligosaccharides by fructosyltransferase does not require oxygen, the production of gluconic acid from glucose by glucose oxidase requires dissolved oxygen. The present inventor has found conditions under which this contradictory enzyme reaction can be performed extremely effectively. Thus, according to the most preferred embodiment of the present invention, the enzymatic reaction with glucose oxidase, catalase and fructosyltransferase proceeds with intermittent aeration with oxygen.

In the enzymatic reaction for ventilating the liquid agricultural raw material as in the present invention, the foaming phenomenon often becomes a problem, and in order to prevent this, it is essential to add an antifoaming agent.
However, when intermittent aeration with pure oxygen under pressure (for example, repeated aeration for 5 seconds and then a pause for 5 minutes and 55 seconds) is performed under agitation according to the present invention, foaming occurs even without adding an antifoaming agent. A sufficient oxygen supply can be ensured without occurring, gluconic acid can be efficiently produced, and fructooligosaccharide can be produced at the same time. In addition, even when gluconic acid is exclusively generated, intermittent aeration with pure oxygen may be performed.

The mixed enzyme used for simultaneously producing fructooligosaccharide and gluconic acid according to the present invention includes:
For example, Aspergillus nige
r) and other fungal cells obtained from appropriate culture media and culture conditions,
Alternatively, a mixture of a crude enzyme solution (containing fructosyltransferase and glucose oxidase) extracted from the cells and a commercially available catalase purified enzyme may be used. As another example, commercially available glucose oxidase containing fructosyltransferase for food addition
Catalase mixed crude enzyme preparations may be mentioned.

When such a mixed enzyme is allowed to act on a liquid agricultural product as a raw material under a condition in which dissolved oxygen is supplied, for example, under conditions of aeration and stirring, a liquid containing both fructooligosaccharide and gluconic acid is obtained. Obtain food ingredients. In the liquid food material obtained in this way, in the case of a liquid agricultural product in which the raw material is concentrated, the sweetness has been greatly reduced, so even if the dilution rate is reduced when reducing,
The balance between sweetness and sourness is achieved, and as a result, beverages and the like that are enriched with components and the like that contribute to health can be obtained. In the case of liquid agricultural products whose raw materials are still squeezed, the sweetness is reduced and the acidity is increased, so for example, Brix with water
By simply diluting it to about 12, it is possible to make a beverage that balances sweetness and sourness.

Moreover, since the production is carried out using a closed system of pure oxygen, no change or volatilization of aroma occurs, and a food material having the original aroma of the raw material can be obtained.

The enzyme is used by immobilizing it on a suitable insoluble carrier as described above, and after completion of the reaction, the carrier is removed from the reaction solution by an operation such as centrifugation or precipitation. Food materials can be manufactured.

In the present invention, agricultural products used for simultaneously producing fructooligosaccharides and gluconic acid have a low acid content, and thus have a pH in the range of 4.5 to 7.0, preferably pH 5.5. A liquid containing 0.5% or more of glucose (or a substance that is a precursor of glucose) or 10% or more of sucrose (or a substance that is a precursor of sucrose). Agricultural products are also desirable in terms of maintaining high enzyme reaction efficiency and activity. Examples of suitable raw agricultural products for use in the present invention include ginseng juice and sugarcane juice.

In order to carry out the method of the present invention described above, the reaction is usually stopped while checking the concentration of the reaction product, and the liquid material containing the gluconic acid or fructooligosaccharide at a concentration higher than the desired concentration. For example, a diluted product thereof is mixed with a raw liquid agricultural product or water to obtain a desired gluconic acid or fructooligosaccharide concentration. Thus, according to the present invention, gluconic acid directly produced from glucose (or a substance that is a precursor of glucose) in agricultural products and / or sucrose (or a substance that is a precursor of sucrose) in agricultural products And a beverage containing fructooligosaccharides directly produced from. The beverage thus obtained has a harmonized acidity and sweetness, and also has a growth effect of bifidobacteria,
It is of high quality different from fermented beverages.

As described above, the liquid food material obtained according to the present invention has a unique flavor by being mixed with another material (eg, soy sauce) or subjected to another treatment (eg, acetic acid fermentation). It can also be used to prepare seasonings and the like.

In any case, the food material of the present invention is produced directly from the raw agricultural product by the enzyme, and retains the original flavor of the raw material as much as possible.

[0044]

EXAMPLES The present invention will be described in more detail with reference to the following Examples, which should not be construed as limiting the present invention.

Example 1 Preparation of Food Material Containing Gluconic Acid

Aspergillus niger
iger) IAM2094 (obtained from the Japan Society for the Promotion of Applied Microbiology) was prepared using a 2000 ml jar fermenter in a medium comprising 10 g of glucose, 3 g of yeast extract, 3 g of malt extract, 5 g of peptone, and 1000 ml of distilled water. Culturing was performed for 15 hours at a temperature of 30 ° C., an aeration rate of 1000 ml / min, and a stirring speed of 300 rpm. After the culture, the cells were collected by filtration, ground in a mortar while adding MacCluben buffer solution at pH 6.0, and centrifuged at 10,000 rpm for 10 minutes to obtain 20 ml of a supernatant.

2 ml of this supernatant and 3 mg of a commercially available catalase purified enzyme (Wako Pure Chemical Industries, origin: Bovineliver)
100 ml of McCluben buffer solution containing 10% glucose and pH 6.0, and 25 ml of concentrated ginseng juice of Brix42 and 75 ml of water as a raw material produce reduced dilute ginseng juice 100
Each of the flasks was added to a 500 ml Erlenmeyer flask with a folded cap and the reaction was started with a rotary shaker at 150 rpm. Table 1 shows the results of measuring the content of gluconic acid produced over time by high performance liquid chromatography. As is clear from this, it was recognized that gluconic acid was produced in both the buffer solution and the ginseng juice as time passed. The liquid food material thus produced had a refreshing acidity of gluconic acid and was suitable as a delicious beverage.

[0048]

[Table 1]

Example 2 Preparation of Food Material Containing Gluconic Acid

Commercially available glucose oxidase for food additives
Catalase mixed crude enzyme preparation (Amano Pharmaceutical, origin: Aspergillu
s niger, trade name: Hyderase 15) 0.1 g to 5%
100 ml of McCluben's buffer solution containing pH 6.0 glucose and 25 ml of concentrated ginseng juice of Brix42 as a raw agricultural product and 100 ml of reduced diluted ginseng juice of 75 ml of water were added to a 500-ml ml-folded conical Erlenmeyer flask, respectively. The reaction was started similarly.

Table 2 shows the measurement results. As can be seen, it was observed that gluconic acid was produced in the buffer and ginseng juice over time.
The liquid food material derived from ginseng thus produced had a refreshing acidity of gluconic acid and was suitable as a delicious beverage.

[0052]

[Table 2]

[Example 3] Preparation of food material containing gluconic acid

0.1 g of a crude glucose oxidase / catalase mixed enzyme preparation for food addition containing fructosyltransferase used in Example 2 was added to 100 ml of squeezed liquid of oyster, pear, kiwifruit and sugarcane, respectively. The reaction was carried out in the same manner as in Example 2 using a conical flask equipped with a rotary shaker at 150 rpm.

Table 3 shows the change in concentration in each liquid.
As can be understood from the table, gluconic acid was produced from fruits such as kiwifruit, and a liquid food material having a strong acidity with a pH of 4 or less and suitable for production of acidic seasonings and the like was obtained.

It should be noted that when sugarcane squeezed liquid is used as a raw material, gluconic acid is produced even without glucose, and gluconic acid is produced by glucose released from sucrose in the process of fructooligosaccharide production. Understood. Thus, the liquid food material obtained from this sugarcane contains both gluconic acid and fructooligosaccharide, and for example, becomes a delicious beverage by being diluted with water.

[0057]

[Table 3]

Example 4 Acetic Acid Fermentation of Enzyme-treated Solution

100 ml of the liquid food material (enzyme-treated liquid) derived from the oyster, pear and kiwi fruit obtained in Example 3 was added to 50
In a 0 ml Erlenmeyer flask, add 2 m of ethyl alcohol
After the addition, 5 ml of a precultured acetic acid bacterium IFO3283 solution was inoculated and cultured with shaking at 30 ° C. The change in the acidity of the fermentation
As shown in Table 4, 1 ml of the fermentation broth was titrated with 0.1 N NaOH using phenolphthalein as an indicator. The fermentation was terminated on the ninth day, and the organic acid composition was determined by high performance liquid chromatography. The results shown in Table 5 were obtained. It has been found that this food material after the acetic acid fermentation treatment contains acetic acid in addition to gluconic acid, and is suitable as a vinegar and a seasoning liquid.

[0060]

[Table 4]

[0061]

[Table 5]

Example 5 Preparation of Highly Active Enzyme for Producing Fructooligosaccharides Using three types of media having the following compositions,
Aspergillus niger IAM202
0, 30 (obtained from the Japan Society for the Promotion of Applied Microbiology)
After culturing the mixture at 9 ° C for 9 days, the cells were collected, ground with a homogenizer, and allowed to stand for 2 days. The digested solution was filtered through a 0.45 μm filter to obtain a crude enzyme solution. The effect of the medium was examined by comparing the transfer activity and the total activity per gram of wet cells. Medium: sucrose 5.0W / V%, peptone 1W
/ V%, meat extract 0.7W / V%, NaCl 0.3W /
V% medium: peptone 1W / V%, meat extract 1W / V%, N
aCl 0.5 W / V% medium: sucrose 5.0%, yeast extract 3.6
%: CMC 0.5%

As a result, as shown in Table 6, the transfer activity (Units / g) and total activity (Units) per cell obtained were 2.18 and 23.3 in the medium, 5.90 and 23.3 in the medium. 55.1, the medium was 0.17 and 4.33, and the use of a meat extract medium without sucrose added resulted in a 2.7-fold increase in the transfer activity per cell as compared with a conventional medium containing sucrose. , 2.4 times in total activity,
An enzyme having a 35-fold and 13-fold transfer activity can be obtained in comparison with the above.

[0064]

[Table 6]

Example 6 Preparation of Food Material Containing Fructooligosaccharide

Aspergillus niger IAM2020 strain as prepared in Example 5 was cultured in a medium containing 1% meat extract, 1% peptone, and 0.5% NaCl to obtain cells in a stationary phase, and a buffer solution was prepared. And triturate with an ultra homogenizer to obtain a digestive juice. The crude enzyme solution is obtained by a conventional method such as filtration with a filter and salting out with ammonium sulfate. The optimum pH of the crude enzyme solution is 5.5, the reaction temperature is 40 to 50 ° C., and the sucrose concentration of the substrate and the production amount show a proportional relationship.

As a raw agricultural product, 100 g of concentrated carrot juice (carrot juice) which had been heated and sterilized until it reached 80 ° C. (ultimately sterilized) was placed in an Erlenmeyer flask, and the extracted crude enzyme 50 U _t (15.6 ml, 3.2 U _t / ml), and the reaction was carried out at 40 ° C with gentle stirring using a stirrer. Samples were taken over time and the sugar composition was analyzed by liquid chromatography. 1, 2, and 3 show that sucrose in ginseng juice is converted to fructooligosaccharides such as kestose and nystose over time, as shown in the time-dependent changes in the sugar analysis liquid chromatogram and the sugar composition. Was done.

The sugar composition of the first concentrated carrot juice was fructose 7.0%, glucose 9.8%, sucrose 15.1%, and kestose 2.7%. After 6 hours of reaction, the sugar composition was fructose 6. 9%, glucose
5%, sucrose 8.7%, kestose 7.3%,
Nistose was 0.12%. At this time, the ratio of the produced fructooligosaccharide (kestose, nystose) to sucrose in the reaction product was 0.85. The concentrated juice was reduced and reduced by a factor of 5 to 7 to obtain a juice containing 1.5 to 1.0% of oligosaccharides of kestose and nystose.

Example 7 Preparation of Gluconic Acid-Containing Food Material Using Fructooligosaccharide-Containing Vegetable Juice as a Raw Material

6.5% fructose, 10.4% glucose, 3.8% sucrose, 3.0% kestose, 0% gluconic acid, 2.8% malic acid, obtained by the method described in Example 6, 500 g of 100 ml of concentrated ginseng juice having a composition of citric acid 0.3% as a raw agricultural product
A commercially available mixed glucose oxidase / catalase crude enzyme preparation for food addition (Amano Pharmaceutical, origin:
Aspergillus niger, trade name: Hyderase 15) 0.1
g and 10 mg of a commercially available glucose oxidase purified enzyme (Wako Pure Chemical Industries, origin: Aspergillus niger) and a commercially available catalase purified enzyme (Wako Pure Chemical Industries, origin: Bovine liver)
A mixture of 3 mg was added, and the mixture was reacted for 7 hours in the same manner as in Example 1.

As a result, in the case of a commercial glucose oxidase / catalase mixed crude enzyme preparation for food addition, 7.2% of fructose, 10.2% of glucose, and 3.
3%, kestose 3.9%, gluconic acid 1.7%, malic acid 2.7%, citric acid 0.3%, commercial glucose oxidase purification enzyme and commercial catalase purification enzyme 7.0% fructose, glucose 6.4%, sucrose 3.2%, kestose 3.3%, gluconic acid 4.6%,
A concentrated ginseng juice having a composition of malic acid 2.8% and citric acid 0.3% was obtained. A two- to three-fold dilution of these ginseng juices is rich in β-carotene and has a refreshing acidity in which gluconic acid and ginseng original malic acid harmonize with each other, and is suitable as a delicious beverage. I was

Example 8 Preparation of Food Material Containing Gluconic Acid and Fructooligosaccharide

Aspergillus niger
ger IAM 2094).
ml (5.7 × 10 ⁷ spores) of glucose 10 g, yeast extract 3 g, malt extract 3 g, peptone 5 g, distilled water 1
8 50 containing 160 ml of medium consisting of 000 ml
A 0 ml Erlenmeyer flask was inoculated and cultured at 30 ° C. with shaking. The bacterial cells after 15 hours of culture were collected by filtration, washed 3 times with Makkuruben buffer pH 6.0, sucrose _{150g / l, MgSO 4 · 7H} 2
O 0.15 g, Na ₂ HPO ₄ 5.4 g, KH ₂ PO ₄
A culture medium obtained by dissolving 3.8 g in 1000 ml with distilled water and the obtained washed cells were put into a 2000 ml jar fermenter, and cultured at a temperature of 30 ° C., an aeration rate of 1000 ml / min, and a stirring speed of 300 rpm. After 15 hours, the cells were similarly filtered and washed, crushed in a mortar, dried with acetone, and crushed with a crusher to obtain 2.2 g of the cells.

0.5 g of the dried and ground cells and a commercial catalase purified enzyme (Wako Pure Chemical Industries, origin: Bovine liver) 1
0 mg of concentrated ginseng juice of Brix42 as a raw agricultural product 100
The mixture was added to a 500-ml Erlenmeyer flask with a folded fold and the reaction was started in the same manner as in Example 1. After 19 hours, the compositions of the organic acid and the sugar were measured by high-performance liquid chromatography. As a result, at the start of the reaction, concentrated carrot juice having a composition of 0.1% fructooligosaccharide (kestose) and 0% gluconic acid was converted to 3.4% fructooligosaccharide (kestose),
The composition changed to 2.3% gluconic acid. It was found that the use of a cell, which is one of the crude enzymes, produces not only gluconic acid but also kestose having the growth activity of bifidobacteria, that is, oligosaccharide. The ginseng juice diluted 4 to 5 times had a refreshing acidity in which gluconic acid and ginseng original malic acid were harmonized, and was suitable as a delicious beverage.

Example 9 Simultaneous Production of Gluconic Acid and Fructooligosaccharides with Intermittent Aeration

Commercially available glucose oxidase / catalase mixed crude enzyme preparation containing foodstuff containing fructosyltransferase (Amano Pharmaceutical, origin: Aspergillus niger,
Product name: Hyderase 15) 1 g as Br
ix40 was added to a 2000 ml jar fermenter containing 1000 ml of concentrated ginseng juice, sealed, pressurized to 1.2 atm with pure oxygen, and pure oxygen in the head space of the jar fermenter was pumped at 500 ml / min with an air pump. After aeration for 5 seconds using a G4 glass filter at a flow rate,
While repeating the operation of stopping for 5 minutes and 55 seconds, a temperature of 30 to
The reaction was started at 60 ° C. with a stirring speed of 100 rpm. Further, in order to compare with the conventional oligosaccharide production conditions, a test was conducted at a temperature of 40 ° C. under conditions where dissolved oxygen was not supplied efficiently, that is, under conditions where aeration was not performed.
Table 7 shows the results of measuring gluconic acid and fructooligosaccharide after the reaction for 7 hours. Table 8 shows the sugar and organic acid compositions after 7 hours of the reaction when pure oxygen was intermittently aerated at 40 ° C.

As is evident from the above, without aeration, oligosaccharides were produced but no gluconic acid was produced. If aeration is performed during the reaction,
Even in the case of using a jar fermenter form used for industrially performing a large amount of reaction, it was recognized that foaming did not occur and gluconic acid was generated in a short time by intermittently aeration of pure oxygen. . Furthermore, it has been found that this method using a crude enzyme agent produces oligosaccharides in addition to gluconic acid.

When gluconic acid and oligosaccharides are produced simultaneously, the temperature is suitably from 40 to 50 ° C., preferably 40 ° C. The reaction solution obtained under the aeration conditions at 40 ° C. has an acid content about twice that of the stock solution compared to the stock solution, has a total sugar content of at least 6% lower, and has a low sweetness of 2% kestose. Because of the above content, the sweetness was further reduced. As a result, even when the reduction rate was reduced, a balance between sweetness and sourness was achieved, and a beverage enriched in β-carotene could be obtained. In fact, this concentrated ginseng juice diluted 3 to 4 times is rich in β-carotene, rich in red-orange color, and the refreshing acidity and sweetness of gluconic acid and ginseng's original malic acid are harmonized, delicious, Suitable as a beverage.

[0079]

[Table 7]

[0080]

[Table 8]

Example 10 Preparation of Fructooligosaccharide-Containing Food Material Using Immobilized Enzyme

The commercially available silica beads were treated with aminopropylethoxysilane and glutaraldehyde, and the crude enzyme of Aspergillus niger IAM2020 was immobilized by a cross-linking method. Next, in a single 200 ml Erlenmeyer flask, 50 g each of concentrated carrot juice (Brix42) sterilized at 80 ° C. and raw temperature as a raw agricultural product and 5.04 g of immobilized enzyme.
(Total activity 20.5 Ut, sterilized with 70% ethanol) was added, and the mixture was stirred gently with a stirrer to carry out a batch reaction in a thermostat at 20 ° C. and 40 ° C. Samples were taken over time to analyze the sugar composition. As shown in Table 6, it was observed that sucrose in ginseng juice was converted to fructooligosaccharides such as kestose and nystose over time.

The ratio of the produced fructooligosaccharides of kestose and nystose to sucrose after 3 hours was 0.39 at 40 ° C., 0.29 at 20 ° C. and 40 ° C.
The reaction having higher fructose transferability than the reaction at 20 ° C. was obtained. The concentrated juice at this time was diluted 5-fold to obtain a juice containing 0.7% of oligosaccharide.
The reaction was further continued, and after the reaction was completed for 27 hours, the enzyme was recovered, washed 2-3 times with a 10 mM buffer solution, and the residual activity was measured. As a result, it was 60%.

[0084]

[Table 9]

Example 11 Production of Fructooligosaccharide-Containing Food Material Using Packed Reactor The crude enzyme of Aspergillus niger IAM2020 was immobilized on silica beads by the same method as in Example 10, and this was incorporated into the filled reactor. 80 as raw material
Continuous processing was carried out using concentrated carrot juice (Brix42) sterilized at ℃ and heated to a raw material. Fig. 4 shows the structure of the reactor
The operating conditions are shown below.

Immobilized enzyme weight: 455 g, total activity: 3
62 units (U _t ), bead capacity: 405 ml, reactor capacity: 430 ml (inner diameter: 45 mm, height: 27
0mm), Model: Packed bed type (upward flow type)

FIG. 5 shows the course of operation. As a result of operating while changing the liquid sending speed, about 35 ml / h (SV = 0.086,
Note: Space velocity SV = f / v (hr ^-1 ), reactor volume is v (here, bead volume), and inflow of supplied substrate solution is f. ), About half of the sucrose in the raw material could be converted to fructooligosaccharide. Table 10 shows the raw materials and the sugar composition after conversion. After operation, remove beads and remove 10 mM
After washing with a buffer, the residual activity of the beads was measured.
%Met.

[0088]

[Table 10]

Example 12 Production of Fructooligosaccharide-Containing Food Material by Rotating Column Reactor

An immobilized enzyme (enzyme activity 1.
82U _t / g) incorporated into a rotary column reactor, 80 ° C. as a raw material agricultural, concentrated carrot juice was ItaruAtsushi sterilized (Brix42) as a raw material was subjected to continuous processing. The configuration of the reactor is such that the packed reactor is replaced with a rotating column reactor. The conditions are as follows.

Immobilized enzyme weight: 97.2 g, total activity:
177 units (U _t ), bead capacity: 87 ml, reactor capacity: 510 ml, model: rotary column type, column capacity 170 ml

FIG. 6 shows the course of operation. First, the tank is filled with concentrated juice, and after 13 hours, the liquid sending speed is 10
It was operated continuously for 20 days at ml / h (SV = 0.115). As shown in Table 11, the raw material and the sugar composition after conversion (sum of the total amount of the processing solution), fructooligosaccharides (kestose, nystose) were produced.

After the operation, the beads were removed and washed with a 10 mM buffer, and the residual activity of the beads was measured. As a result, 67% (1.
22 U _t / g).

[0094]

[Table 11]

Example 13 Simultaneous Production of Gluconic Acid and Fructooligosaccharides with Intermittent Aeration

As in Example 9, the enzyme treatment was performed while intermittent aeration was performed. However, sugarcane squeezed liquid was used as a raw material agricultural product. Table 12 shows the concentration changes of sugar and organic acid in the liquid.
Shown in Gluconic acid is produced even though the raw sugar cane does not contain glucose. Thus, a food material containing both gluconic acid and fructooligosaccharides and suitable as a delicious beverage was obtained from sugarcane.

[0097]

[Table 12]

[0098]

According to the present invention, a novel liquid food material containing gluconic acid and / or fructooligosaccharide using only agricultural products as raw materials and retaining the original flavor of the raw materials can be obtained. The liquid food material of the present invention is, for example, a liquid food material that has enhanced sourness and reduced sweetness, and is suitable for use as a beverage having a refreshing acidity and sweetness harmonizing with the activity of proliferating Bifidobacteria. Furthermore, the liquid food material of the present invention can also be used for preparing a new flavor seasoning.

[Brief description of the drawings]

FIG. 1 is a liquid chromatograph of the sugar composition of carrot juice concentrated as a raw material for producing a liquid food material suitable as a beverage according to the present invention as shown in Example 6.

FIG. 2 is a liquid chromatograph of the sugar composition of the concentrated carrot juice after the enzymatic reaction of Example 6.

FIG. 3 is a graph showing a change in the sugar composition of a reaction solution of the crude enzyme and the concentrated carrot juice of Example 6.

FIG. 4 is a view showing the configuration of a filling-type reactor used for producing the liquid food material of the present invention as shown in Example 11.

FIG. 5 is a view showing the progress of operation by the packed reactor of Example 11;

FIG. 6 is a diagram showing an operation course when producing a liquid food material according to the present invention using a rotary column reactor as shown in Example 12.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Packing type reactor (upflow type) 2 Silica immobilized enzyme 3 Raw material tank 4 Product tank 5 Constant temperature bath (-1 degreeC) 6 Temperature controller 7 Pump 8 Heat exchanger 9 Hot water circulation device 10 Sampling hole 11 Air filter

──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Masanori Furuta 4-6-304 Hakomatsu, Higashi-ku, Fukuoka City, Fukuoka Prefecture (72) Inventor Osamu Ota 4-67 Okamoto, Kasuga-shi, Fukuoka Prefecture (72) Inventor Mitsuru Suenaga Saga 2317 Eguchi, Kita-moyasu-cho, Sanyo-gun, Prefecture (72) Inventor Tsuyoshi Yamaguchi 296-87, Oji-Tohara, Chikushino-shi, Fukuoka (58) Field surveyed (Int.Cl. ⁶ , DB name) A23L 2/00-2/84

Claims (13)

(57) [Claims] 1. A sucrose or an agricultural product containing a substance which is a precursor of sucrose is used as a raw material, and an enzymatic agent containing a fructosyltransferase is acted on the raw material, so that one or two molecules of fructose are bound to the sucrose. A method for producing a fructooligosaccharide-containing liquid food material, which comprises producing fructooligosaccharide. 2. A generated method for manufacturing a liquid food material according to claim 1, wherein the ratio of fructooligosaccharide pair sucrose, characterized in that the set to be 1.0 or less. 3. Fructosyltransferase is expressed in Aspergillus niger (Aspegillus niger) on a medium without sucrose.
rgillus niger) IAM2020 claim 1 or 2 wherein <br/> method liquid food material in the manufacturing, characterized in that is obtained by culturing. 4. The process for producing liquid food material according to claim 3, characterized in that to fix the fructosyltransferase to porous silica beads crosslinking treatment performed enzymatic reactions. 5. A liquid food material manufacturing method according to any one of claims 1 to 4, wherein the raw material agricultural is carrot juice. 6. A method of manufacturing a liquid food material according <br/> to any one of claims 1 to 4, wherein the raw material agricultural is sugarcane juice. 7. Gluconic acid characterized by producing gluconic acid by using glucose or an agricultural product containing a substance that is a precursor of glucose as a raw material, and causing an enzyme agent containing glucose oxidase and catalase to act on the raw material. A method for producing a liquid food material containing: 8. A material agricultural products containing glucose or glucose precursor material, according to claim 7, characterized in that the liquid food material containing fructooligosaccharide obtained according to any of claims 1 to 6 Production method of liquid food material. 9. material agricultural products containing glucose, a manufacturing method of a liquid food material according to claim 7, characterized in that the pressed juice of the fruit. 10. A raw material comprising glucose or a substance that is a precursor of glucose and sucrose or an agricultural product containing a substance that is a precursor of sucrose, which contains glucose oxidase, catalase, and fructosyltransferase. A method for producing a liquid food material containing gluconic acid and fructooligosaccharide, which comprises reacting an enzyme agent to produce gluconic acid and fructooligosaccharide having one or two molecules of fructose bonded to sucrose. While performing intermittent aeration with 11. Oxygen, glucose oxidase, a manufacturing method of a liquid food material according to claim 10, wherein the advancing the enzyme reaction of catalase and fructosyltransferase. 12. The method of claim 10 or 11 liquid food material manufacturing method according to, characterized in that the raw agricultural is carrot juice. 13. The method of claim 10 or 11 liquid food material manufacturing method according to, characterized in that the raw agricultural is sugarcane juice.