JP3201811B2 - Foods and beverages containing low molecular weight pectic acid - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a food or drink containing a low-molecular-weight pectic acid which has been reduced in molecular weight while maintaining its physiological activity as a dietary fiber.

[0002]

BACKGROUND OF THE INVENTION Dietary fiber is defined as an indigestible component in food that cannot be digested by human digestive enzymes. Not only plant cell wall components such as cellulose, lignin and pectin, but also non-digestible components such as chitin and chitosan are widely used. Contains digested organic matter. In recent years, they have been shown to have various effects such as a blood cholesterol lowering effect, including an effect of improving bowel movement, and also play an important role in preventing adult diseases.

[0003] Among these dietary fibers, pectic substances such as pectin and pectic acid have a strong activity as dietary fiber, and have various effects such as improving bowel movement, suppressing the increase in blood cholesterol level, suppressing the formation of gallstones, and suppressing hypertension. The effect has been reported. Conventionally, pectin has been used as a stabilizer in the food industry for jams, fruit jellies, drink yogurt, lactic acid bacteria drinks, etc.
Because of the above-mentioned effects, it is expected to be added to food and drink as food fiber.

[0004] Pectic substances are present in the form of a complex called protopectin by binding to cellulose in immature fruits or plants, and are particularly contained in large amounts in citrus fruits, apples, and karin. This protopectin is insoluble, but is partially hydrolyzed to soluble pectin or pectic acid when the fruit matures.

[0005] Pectin is mainly composed of polygalacturonan, which is a polymer of galacturonic acid, and generally has a portion of the carboxyl group of galacturonic acid that is methylesterified. Pectic acid is obtained by demethylating this pectin. Demethyl esterification is
It can also be carried out by a method using an alkali (JP-A-63-89501) or an enzymatic method (JP-A-52-121398).

However, the conventional pectic acid thus obtained generally has the properties of low solubility, high viscosity, and strong gelling ability. Therefore, pectic acid has various effects as described above,
Because of its properties, it can be added only to foods and drinks in small amounts, and it has been difficult to incorporate into foods and drinks an amount that is expected to have an activity as a dietary fiber.

[0007]

SUMMARY OF THE INVENTION Accordingly, an object of the present invention is to provide a food or drink containing low-molecular-weight pectic acid having low viscosity and high solubility and maintaining physiological activity as a dietary fiber. It is in.

[0008]

Means for Solving the Problems To solve the above-mentioned problems, the present invention comprises 0.01 to 50% by weight of a low-molecular pectic acid obtained by allowing endo-type polygalacturonase to act on pectic acid. Providing food and drink.

The endo-type polygalacturonase can be produced from the genus Kluyveromyces or the genus Geotricum.

Hereinafter, the present invention will be described in detail.

The present inventor has studied the mode of degradation of pectic acid by many pectinolytic enzymes. As a result, when the endo-type polygalacturonase is acted on pectic acid, the degradation of pectic acid having two molecular weight peaks occurs. I found that I could get something. This decomposition product has a small molecular weight of 13
It consists of a mixture of polysaccharides of the order of ten thousand and polysaccharides with a large molecular weight of about 8,000. This degradation product is a polysaccharide having the same physiological activity as pectic acid except that it is reduced in molecular weight, that is, a low molecular weight pectic acid.

In obtaining this low-molecular-weight pectic acid, first, an endo-type polygalacturonase is allowed to act on pectic acid.

The pectic acid may be of natural origin or may be obtained by subjecting pectin to demethylesterification by a conventional method. As pectin, any pectin can be used as a raw material, and its origin is not limited. Accordingly, many fruits such as lemon pectin and apple pectin which are generally known can be used.

In causing the endo-type polygalacturonase to act on pectic acid, any of a purified product, a culture supernatant (crude enzyme solution) or a processed product thereof may be used.

In general, endo-type polygalacturonase is
It is distributed in bacteria, microorganisms, higher plants, etc., and any of these can be purified and used. That is, the culture supernatant from which the cells have been removed from the culture solution of the microorganisms and the like is subjected to ammonium sulfate precipitation to salt out only the protein, which is separated by charge using an ion exchanger, and further subjected to gel filtration to determine the molecular weight. Purification is performed by a general enzyme purification step of separation.

A commercially available pectinase may be used, but also in this case, purification is required to remove pectin esterase and hemicellulase present in the pectinase.

By the way, it has been found that when yeast belonging to the genus Kluyveromyces or the genus Geotricum is used, the culture supernatant can be directly used for the enzyme reaction without performing purification treatment. A yeast belonging to the genus Kluyveromyces or the genus Geotricum is a yeast having an ability to produce only an endopolygalacturonase, and an endopolygalacturonase is an extracellular enzyme secreted outside the body. Therefore, when these yeasts are used, the culture supernatant can be used as it is as a crude enzyme solution. Normal,
These yeasts are cultured on an agar medium, further subjected to main culture and cultured in large scale, and the resulting culture is centrifuged to remove the cells, thereby obtaining a culture supernatant. One such yeast, Kluyveromyces fragilis (Kluy
veromyces fragilis) JTF-1 is
On October 11, 1991, Geotricum candidum JTF
-2 was deposited with the National Institute of Microbial Industry and Technology on December 19, 1991 under the name of Microbial Laboratory Bacteria No. 12670.

It is more preferable to use a dialysis-treated culture supernatant obtained by simply subjecting the culture supernatant to a simple treatment such as dialysis, ultrafiltration, ion exchange, or gel filtration.
This is because these treatments can remove the yeast odor and make the liquid color transparent.

Either the endo-type polygalacturonase thus purified or the culture supernatant obtained using the specific yeast of the present invention can be used. The obtained culture supernatant can be directly used for the enzymatic reaction, thereby simplifying the enzymatic purification step, which is more preferable.

The low molecular pectic acid is obtained by reacting the purified product, the culture supernatant obtained as described above, or the processed product thereof with a suspension of pectic acid suspended in a buffer such as acetic acid. Can be

Here, the reaction time and the quantitative ratio of the endo-type polygalacturonase to pectic acid are not particularly limited since the decomposition of pectic acid stops at a molecular weight of about 130,000 or 8,000 even when it is allowed to act to the decomposition limit. The reaction time is preferably 12 to 48 hours, and the quantitative ratio of the yeast culture supernatant to 1 part by weight of pectic acid is preferably 5 to 20 parts by weight. Also,
The reaction temperature and pH are preferably such that the reaction proceeds sufficiently and the endo-type polygalacturonase is not inactivated, that is, 30 to 60 ° C and pH 3.0 to 5.0, respectively.

The decomposed product of pectic acid thus obtained may be used after drying as it is, or may be further treated.

In the case of further treatment, a purification step such as dialysis or ultrafiltration is performed to remove galacturonic acid and its oligosaccharides in the decomposition product, and acetic acid used as a buffer during the enzyme reaction. It is made into a powder by a precipitation step using an organic solvent such as ethanol, acetone or the like, or a drying step such as freeze-drying or spray-drying, and then used.

The two low molecular pectic acid having a molecular weight of the order and 8,000 1 30,000 by the above method is obtained, the production rate is 13% and 87% by weight. This low molecular weight pectic acid is one of the physiological activities of dietary fiber, despite being smaller than the existing pectic acid as a food additive and having a considerably lower viscosity than the original pectic acid and having a higher solubility. The effect of improving bowel movement is maintained.

Further, since the low-molecular-weight pectic acid has the above-mentioned properties, the low-molecular-weight pectic acid has such an amount that it can maintain the physiological activity as a dietary fiber, which was impossible in the past, ie, 0.01 to 0.01%.
50% by weight, preferably 0.1 to 5% by weight can be contained in various foods and drinks such as juice, candy, bread, jam and the like.

The food or beverage containing the low molecular weight pectic acid exhibits improved physical properties and texture different from the case where the existing pectic acid is added, even with the above content.

[0027]

The present invention will be described in more detail with reference to the following examples.

All parts and percentages in the examples are by weight.

Example 1 Preparation and Analysis of Lemon Pectic Acid a) Preparation of Crude Enzyme Solution Kluyveromyces fragilis JTF-1 was sucrose 2%
In potato decoction agar slant medium (pH 5.0) containing
After culturing at 7 ° C. for 24 hours, one loop of bacterial cells was put on 5% glucose, 0.2% ammonium phosphate, 0.1% potassium phosphate, 0.1% magnesium sulfate and 0.4% yeast extract. Inoculate 50 ml of medium containing
For 3 days. This culture was inoculated into 1 liter of a medium having the same composition, and further cultured at 27 ° C. for 3 days. The culture was centrifuged at 13,000 rpm for 10 minutes, and the cells were removed to obtain a culture supernatant.

B) Preparation of Lemon Pectic Acid Lemon pectin (100 g) was dissolved in 0.05N NaOH and reacted at 4 ° C. for 90 minutes for demethyl esterification. After the reaction, the pH was immediately adjusted to 4.8 with acetic acid to prepare a 4 liter lemon pectic acid solution.

C) Preparation of Lemon Low-Molecular Pectic Acid (i) One liter of the culture supernatant prepared in a) was added to the 2.5% lemon pectic acid solution prepared by the method of b), and
The reaction was performed at 24 ° C. for 24 hours. After concentrating the obtained reaction solution with a rotary evaporator, it was dialyzed overnight against 100 times the amount of deionized water of the sample solution, and further freeze-dried to obtain 35.21 g of lemon low molecular weight pectic acid.

(Ii) Low molecular weight pectic acid was prepared in exactly the same manner as (i) except that ultrafiltration (excluded molecular weight 10,000) was used instead of dialysis. 46.91 g of lemon low molecular weight pectic acid was obtained.

D) Analysis of Lemon Low Molecular Weight Pectic Acid The lemon low molecular weight pectic acid obtained in (i) and (ii) was subjected to the following measurements (1) to (4).

(1) Measurement of molecular weight TSK-G was obtained from the obtained lemon low molecular weight pectic acid.
The main peak was measured by gel filtration with 4000 PW, and the molecular weight was calculated using pullulan (STANDARD P-82, Showa Denko) as a standard sample.

(2) Measurement of the ratio of galacturonic acid: neutral sugar Lemon low-molecular-weight pectic acid was completely decomposed by dolyserase and then measured by HPLC analysis using a Shodex SH-1821 column (Matsuhashi, Inoue, Hatanaka, Journal of the Japanese Society of Agricultural Chemistry, 65, p399 (199
1)).

(3) Measurement of Viscosity 1 First, a 5% solution of lemon low molecular weight pectic acid was prepared, and its viscosity was measured with an E-type viscometer.

Table 1 shows the results of the above (1) to (3).

[0038]

[Table 1] * The low-molecular-weight pectic acid obtained in Example 1c) (i) had a molecular weight of 13.6 × 10 ⁴ of 13% and a molecular weight of 0.8 × 10 ⁴ of 87%.

(4) Measurement of Viscosity 2 The viscosities of lemon low molecular weight pectic acid and, for comparison, lemon pectic acid were measured with an E-type viscometer (50 rpm). The result is shown in FIG. From this, the viscosity of pectic acid was considerably reduced.

(5) Effect of improving bowel movement 4 week-old male SD rats were bred on solid feed (Oriental Yeast Solid Feed MF) for 4 days, and divided into 4 groups of 5 rats each. Thereafter, each group was fed a feed having the composition shown in Table 2 below and a solid feed, and bred for 9 days.
Day stool was collected. The results obtained are shown in Table 3 below. On the basis of the softness of the stool of the solid feed, the cured food was defined as-and the softened one was evaluated as +.

[0041]

[Table 2]

[0042]

[Table 3] From the above results, it was found that the low-molecular-weight pectic acid of the present invention has a stool softening effect similarly to pectic acid, and is effective in improving bowel movement.

Example 2 Use of Low-Molecular Pectic Acid a) 30% Apple Juice Juice 1 part of the low-molecular pectic acid obtained in c) (ii) of Example 1 was mixed with 6 parts of 5-fold concentrated apple juice and granulated sugar. 1
0 parts, 0.2 parts of DL-malic acid, 0.02 parts of trisodium citrate and 83 parts of distilled water to finally give a 30% apple juice containing 1% by weight of low molecular weight pectic acid. Produced.

The juice containing low molecular weight pectic acid is
It exhibited smooth physical properties.

B) Hard candy Apple-type hard candy was prepared in the following manner using the composition raw materials shown in Table 4 below containing 1 part of the low molecular weight pectic acid obtained in c) (ii) of Example 1.

[0046] Sugar, starch syrup and water were mixed and heated to 110 ° C. A low molecular weight pectic acid dissolved in a small amount of water was added and the mixture was boiled down to 147 ° C. Citric acid, fragrance and dye were added, mixed, cooled and molded. As a control, a sample to which 1 part of pectin was added was prepared and compared. Result is,
The results are shown in Table 5 below.

[0047]

[Table 4]

[0048]

[Table 5] When 1% by weight of pectin was added to the candy raw material, the pectin remained as a stalk and did not disperse well, whereas when the same amount of low molecular weight pectic acid was added, it dispersed well.
It could be easily processed. In addition, the candy added with pectin had a strong sour taste and had an unpleasant taste, whereas the candy added with low molecular weight pectic acid also had a good taste.

C) Loaf of bread Loaf of bread was prepared using the ingredients shown in Table 6 below.

2.5 parts of the low molecular weight pectic acid obtained in c) (ii) of Example 1 were dissolved in water in advance and mixed with the raw materials shown in Table 6 below except for dry yeast. This mixture is mixed with a Sanyo bread machine (SPM-B
It was put into the bread case of 1), dry yeast was added, kneaded, fermented and baked into bread. Also, as a control,
Bread from which the low molecular weight pectic acid was removed from the raw material composition shown in Table 6 below was baked.

[0051]

[Table 6] The results of the sensory test are shown in Table 7 below. There was almost no difference in baking between the bread containing 0.5% by weight of low-molecular-weight pectic acid and the control, but softness was added to bread in the group added with low-molecular-weight pectic acid.

[0052]

[Table 7] Example 3 Preparation of apple low molecular weight pectic acid In exactly the same manner as in Example 1 c) (ii) except that apple pectic acid prepared from apple pectin by the method of Example 1 b) was used instead of lemon pectic acid. , An apple low molecular weight pectic acid was prepared. Molecular weight 13.2 × 10
^4. A low molecular pectic acid of 0.8 × 10 ⁴ was prepared.

Example 4 Preparation of low molecular weight pectic acid using dialysis culture supernatant In place of the culture supernatant prepared by the method of Example 1, 1 liter of this culture supernatant was further added to a 0.025 M acetate buffer (pH
4.8) A low-molecular-weight pectic acid was prepared in exactly the same manner as in c) (ii) of Example 1, except that the culture supernatant obtained by dialyzing 300 liters overnight was used. Molecular weight 1
3.6 × 10 ⁴ and 0.8 × 10 ⁴ low molecular weight pectic acids could be prepared.

Example 5 Geotricum Candy Dam J
Preparation of low molecular weight pectic acid using TF-2 A low molecular weight pectic acid was prepared in exactly the same manner as in Example 4 except that Geotricum candy dam JTF-2 was used instead of Kluyveromyces fragilis JTF-1.
Low-molecular-weight pectic acids having a molecular weight of 13.3 × 10 ⁴ and 0.8 × 10 ⁴ could be prepared.

[0055]

The low molecular weight pectic acid obtained according to the present invention has low viscosity, high solubility, and retains the physiological activity of dietary fiber such as improvement of bowel movement. Can be easily added to the extent that the physiological activity can be added. Further, beverages containing the low-molecular-weight pectic acid of the present invention, foods and drinks such as bread, improved physical properties,
Has texture and the like.

[Brief description of the drawings]

FIG. 1 is a view showing a viscosity curve of a low molecular weight pectic acid of the present invention.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61K 31/732 A61P 1/00 A61P 1/00 1/14 1/14 C08B 37/06 C08B 37/06 C12P 19/14 Z C12P 19/14 A23L 2/00 G (72) Inventor Susumu Maeda 6-2 Umegaoka, Midori-ku, Yokohama-shi, Kanagawa Prefecture, Japan Eating Habits Research Institute (72) Inventor Chitose Hatanaka 3120-4 Nakasho, Kurashiki-shi, Okayama Prefecture Address (56) References JP-A-2-72842 (JP, A) JP-A-3-151854 (JP, A) JP-A-2-286058 (JP, A) JP-A-61-231966 (JP, A) JP-A-2-295441 (JP, A) JP-A-1-108983 (JP, A) JP-A-56-26901 (JP, A) Fermentation Engineering Vol. 62, No. 1 (1984) p. 1-7 (58) Field surveyed (Int. Cl. ⁷ , DB name) A23L 1/308 C08B 37/06 JICST file (JOIS) JICST file (JAFIC)

Claims (1)

(57) [Claims] 1. A pectic acid containing Kluyveromyces spp.
Is characterized by containing 0.1 to 5 % by weight of low-molecular-weight pectic acid obtained by reacting endo-type polygalacturonase produced from Geotricum yeast.
A method for producing a food or drink containing low molecular weight pectic acid .