WO2000062623A1

WO2000062623A1 - Process for producing soybean protein

Info

Publication number: WO2000062623A1
Application number: PCT/JP2000/002403
Authority: WO
Inventors: Kazunobu Tsumura; Tsutomu Saitoh; Wataru Kugimiya
Original assignee: Fuji Oil Company, Limited
Priority date: 1999-04-16
Filing date: 2000-04-13
Publication date: 2000-10-26
Also published as: AU3677500A; JP2000300185A

Abstract

A process whereby soybean protein containing a reduced amount of phytic acid and having a high solubility and a good taste can be produced on an industrial scale. This process comprises treating undenatured soybean protein or soybean protein with a low extent of denaturation with a microbial-origin phytic acid decomposing enzyme having an activity within a pH range of from 6 to 9 to thereby decompose phytic acid in a pH range of from 6 to 9.

Description

Description Protein ¾it method Technical field

The present invention is based on phytic acid: ^: protein! Background art on the lit method

Contains many high-quality proteins and has long been used as an excellent protein source. In particular, the separated protein has a high protein content and various functions such as emulsifiability, gel stiffness, and ifc'l ^ l. Therefore, as a food or food modifying material, meat products, water fibers, etc. It is widely used in foods, prepared foods, bread, and ingredients for kin. Moreover, its prosperity is high, making it an ideal protein source. It contains about 1% of phosphorus, of which 70 to 80% is said to exist as phytin. Phytic acid (myo-inositol hexarate) is a nutrient ≤: it forms chelate bonds with essential mineral components (calcium, magnesium, iron, iron, etc.) to form degradable ^! It is preferable to remove or remove these occupations because they may reduce the absorption of these occupations or have an effect on consumption.

Therefore, many attempts have been made to convert the phytic acid in it. To reduce phytic acid:: ^: washing with water or salt coat near the isoelectric point of protein (near pH 5) (^ 6-693445, JP (Japanese Patent Application Laid-Open No. Hei 7-222715) has been proposed. However, these methods are not only production but also require large amounts of waste liquid treatment, and are not suitable for commercial production. Is inappropriate.

—On the other hand, there is also a method of performing angle division with an enzyme that decomposes phytic acid (Fiyuichize). A method of treating with soybean protein-derived phytase near or below the isoelectric point (pH 2 to 6) (Japanese Patent Application Laid-Open No. 4-5003002 No. 艮), alcohol conversion '^! : Protein (NS I 30 or less) near isoelectric point (pH 5

.5) method (Japanese Patent Laid-Open No. 6-86640, a method in which the soybean milk is treated with the phyase of a plant ^^ (^ F1-277706 ^), Treatment of pasteurized commercial bean paste L using neutral and active fibrose (Japanese Patent Application Laid-Open No. Hei 6-38 745). However, in the above method, Shima Almost ^: The protein is in the haze, in the vicinity or at the lower pH range (pH 2 to 6), and according to the knowledge of the present inventors, the solubility In addition, it is difficult to increase the protein to act on, and the efficiency is low, or the action tends to take a long time to cause the effect.

It is an object of the present invention to produce a soybean protein having good phytic acid solubility and good solubility and flavor. Invention 鹏

The present inventors have studied to improve the above, and as a result, when the pH is 6 or more, the phytic acid is treated by treating the protein with an enzyme having phytate tolerance. The inventors have found that it is possible to obtain a significantly improved solubility and a good protein of 1% and a protein, and have made the present invention true. That is, the present invention is directed to reacting an enzyme having phytic acid content and properties with iS ^! Fe ^ protein in a pH range of 6 to 9 to the iS ^! Fe ^ protein. It is a method of protein. And the phytic acid boiling enzyme used here is an enzyme having an activity whose pH is in the range of 6 to 9, particularly a ferment-derived enzyme ¾ ^ 'preferred U ヽ. Description

The protein may be soymilk, soymilk, m, or protein containing protein, but the protein is used for the sake. If the ^ protein that is excessively denatured by the addition is treated with phytic acid,

In particular, if the protein s is high, such as in the case of isolated ^ protein or soy milk, ^ processing and processing will be difficult. For example, to remove phytic acid from the soybean protein, extract it with water at pH 7 to 8, and separate it into a water-insoluble fraction (okara) and a water-soluble fraction (soybean milk). The soluble fraction is made equal at pH around 4.5 and separated into an insoluble fraction (force) and a soluble fraction (whey) to obtain an acid curd. In the step of neutralizing the suspension and performing sterilization, it can be efficiently performed in the step of extracting soymilk from the soybean or the step of neutralizing the acid card. At this time, t ^ S to be used is preferably not accompanied by protein denaturation, or so-called 1 «^: ^: which has been subjected to a processing treatment in which protein denaturation is the nucleus.

Specifically, low-temperature extraction was performed using n-hexane as an extraction solvent! ^ ^ Beans are suitable as a raw material, and particularly, NSI (nitrogen solubility coefficient) is 60 or more, preferably 80 or more. I prefer ^^. ^ The force of denaturation of a protein in a solution containing the protein by heating or the like can be determined by differential scanning calorimetry (DSC) analysis of the protein (Nagano et al. J. Agric. Food Chem., 40. 941-944 (1992)). According to this analysis ^ method, for example, in the case of native isolated ^ protein, While the endothermic peak powers of 7S globulin and 11S globulin, which are the main constituents of β¾, are recognized, the endothermic peaks of ^ ^ of the soybean protein are affected by the denaturation of fiber. Since it cannot be moved, the presence or absence of denaturation can be easily determined.

In the present invention, the pH power of phytic acid degradation is particularly important, and is difficult at pH 6 to 9, preferably above pH 6.0, more preferably at pH 6.2 to 8.5, and even more preferably at pH 6.5 to 7.5. Good to do. The solubility of soy protein cut at a pH of less than 6 is reduced, resulting in poor strength. Further, if the pH exceeds 9.0, J »becomes poor, which is not preferable.

Therefore, it is necessary that the enzyme used for $ M in the present invention is capable of degrading a medium of pH 7 or more, ie, T ′ phytin and phytin. Phytin and phytin ^^ can be used as enzymes which can be used in animals, plants, and yeasts. For example, if it is derived from a product, it may be of the genus Aspergillus (Aspergillus), the genus Penicillium (Penici 11 i urn), the genus Bacillus (Baci 11 us), the genus Pseudomonas (Pseudomonas), or the genus Saccharomyces ( Saccharomyces), Kluyveromyces II, and Neurospora produced from Neurospora, and enzymes expressing these enzyme genes in other microorganisms. Phosphatase capable of decomposing phytic acid and phytin ^ is also included.

The enzyme can be used regardless of the powder or liquid form. 0.01% to 10Sfi%, preferably 0.05%, of crude protein S3 in soybean protein

22% by weight, more preferably 0.1 to 1 ^, but the enzyme titer is 0.1 to: I 0

Preferably - Five -

0.5-2

It is preferred to add soy protein filterase. In addition, the fermentation contains 4 mM sodium phytate, 0.2 MT Tris-HC 1

(pH6. 5) 0. 5ml, distilled water 0. 4 ml及Hi solution 0. 1 ml or Ranaru Si ^ liquid 37 ^e was C for 30 minutes SJ ^ are the stops adding 10% TCA 1. 0ml. The amount of phosphorus in the reaction solution was measured by a Fiske-Sububowow method. As stated above, 1 unit (U) of enzyme that produces 1 micromol of «phosphoric acid» per minute in cows was used.

Enzyme SJ ^ of ^ ^ and time, can be the enzyme used, usually 2 0 ° C~80 ^e C, preferably 30; C~ ~ 70 ° C, more preferably, 40 ° 60. React with C for 103 hours, preferably 30 minutes to 1 hour. In addition, it is possible to achieve a target by passing ^^ through a column on which immobilization is performed. The fraction obtained in this way is neutralized, sterilized and purified by the method of ^. In addition, if there is ^, the fats and oils and Z or milk <are added to the protein before or after the sterilization step, and a certain amount is added after the ^ !: step, or during the i ^ step. It is optional to perform ^ ¾ with a protein component. Furthermore, the decomposed product may be removed by precipitation separation at UF and isoelectric point. In addition, the amount of phytin can be measured by a known method, for example, Cereal Chem., 63, 475 (1980).

The soy protein obtained by the present invention has a phytin ^ * content of 0.2% ^ J¾J¾ under fiber, 0.1%, whereas the normal protein content of the protein contains about 2% phytic acid. ^ ¾ It is greatly reduced and the original machine food (solubility etc.) and the flavor are impaired, so it can be used for various food ingredients ^ and it is also preferred in terms of: ^ Protein power can get.

Hereinafter, the mode of the present invention will be specifically described using difficult examples. However, the technical scope of the present invention is not limited to these examples.

^ I ^ cs (Nitrogen soluble; NS 192) Add 12 dragons of water to the ima part, extract at room temperature, pH 7 for 1 hour, centrifuge to obtain soybean milk, pH with fiber However, the fiber was spliced to 4.5, and a distant '«fraction (hereinafter referred to as M¾» card) was obtained. Add water to the acid curd, thoroughly disperse (crude protein content 10%), test ρ to 6.5 with caustic soda, and apply phytase (trade name “Sumiteam ス”, Shin Nihon k 日本 t) to the crude protein. Then, 0.4% by weight (4 U g crude ^^ protein) was added, and the mixture was stirred at 40 for 1 hour to make the enzyme difficult. SJ ^ ¾, reconditioning the _P H to 7.0 caustic soda ^, sterilized 140 ° C, 15 seconds, which was injection ^燥to obtain a protein.

麵 Example 2

Add water to the acidified card of HJ Example 1 and disperse it sufficiently (crude protein content: 10%). Separate to pH = 7.0 with caustic soda, and add phytase (trade name “Sumiteam PHYJ, Shin Nihon I ^”) to crude protein. 0.4% by weight

Add 40. The mixture was stirred for 1 hour at C to remove the enzyme. The SCT was sterilized at 140 ° C for 15 seconds, and then subjected to: ieS protein was obtained.

Table 1 summarizes the obtained: ^ !: protein phytin ^ ft and solubility, 1).

Examples 1, 2

Water was added to the acid salt obtained in Example 1 and thoroughly dispersed (crude protein content: 1 o £ ft%), and the pH was adjusted to 5.5 (Comparative Example 1) and 5.0 (Comparative Example) with sodium bicarbonate. After 2), 0.4% by weight (4UZg crude soy protein) of phytase (trade name "Sumiichi PHYJ, Shin Nihon I ^; fc ^") was added to the crude protein weight, and the mixture was added at 40 C for 1 hour. Stir and carry out each enzymatic reaction -7-

Was. Anti] ^, 140 after readjusting the pH to 7.0 with caustic soda. C, sterilized for 15 seconds and dried to obtain protein

Each got: ^! : Table 1 summarizes the phytin age and solubility of protein.

(table 1)

®¾ DH Phytin ^ Amount Solubility (NSI) m

6.5 <m \ l) o.o 5% 85

7.o cm m 2) 0.06% 87 m

5.5 (赚 Example 1) 0.04% 65 Somewhat bad

5. 0¾ Example 2) 0.04% 55 ae There is excellent solubility and flavor, and phytic acid is added to the protein at 50% at 6Rh as described above.

Difficult case 3

Ryujin 1 soy milk (pH 7.0), phytase (trade name "Sumichi-M PHY")

(4 U

Add 40. The mixture was stirred at C for 1 hour, and the enzyme reaction was interwoven. Anti! ^, Immediately set the pH to 4.5 with the fiber, centrifuged to obtain the acid カード ^ card, disperse this in water, 雄 to pH 7.0 with a soda, 140. C, sterilized for 15 seconds, and sprayed: ^ to obtain protein. The obtained soybean protein had a phytic acid content of 0.04% .m. (NS I) 89 and a good flavor.

Disgusting example 4

Bacillus subtilis isolated from Ichisaka bean was purified to 0.1 ltt% CaC 12 One eight one

, 1M:% D-mannose, 1% glucose, 1. Yeast extract, 2.5 £ 4% Heart 'inhibition broth (manufactured by Difco) 液 ^ ground (pH 7.4) The mixture was cultured at 37 ° C with shaking for 5 days to obtain a culture__L clear solution. The #JL clarified solution was filtered with a filter fiber (fraction ^ 10,000) in m ^, and a partially purified fraction of phytase was obtained on DEAE-Toyopearl column chromatography (40 U / m 1).

Add water to the acid salt of m ι and disperse it thoroughly (crude protein ι o

Then, the pH was adjusted to 7.5 with sodium hydroxide, and jfaBacillus genus P was purified by adding 10 UZg & ^: bean protein to the crude protein. The mixture was stirred for 1 hour at C, and the enzyme was difficult. It was sterilized with Sl¾, 140 for 15 seconds, and sprayed to obtain protein. The obtained: ^: protein had a phytin ^ content of 0.03%, solubility (NSI) of 90 and good flavor.

Difficult case 5

: Was immersed in water overnight, and ^ *: ^ (water working 50%). This ^: is placed in a basket, separated into soy milk and okara, and the resulting soy milk (solid content 9S »%, crude protein 4.5 mm%) is concentrated in a vacuum to produce fiber soy milk (solid content 13%, 6.5 g of crude protein was prepared, and ^ g soymilk (pH 6.2) was mixed with Fissease (trade name: Sumiteam PHY) and Shin Nippon Toridakune 0.4 wt. %

Add 40. Stirred at C for 1 hour. Immediately, the pH was adjusted to 7.0 with caustic soda and sterilized at 140 ° C for 15 seconds. The obtained ugly soybean milk had a phytin age of 0.07%, solubility (NS I) of 86, and female gender.

Comparative Example 3

^: 95 prepared in the same manner as in Difficult Example 5. C, heat for 5 minutes, immediately 2 o. After cooling to c, the mixture was separated into soymilk and okara. The resulting soymilk (solid content 9%, crude protein 4.5Sfi%) was vacuumed. This concentrated soymilk (solid content 13%, crude protein 6.5 Sft%, pH 6.2) was supplemented with phytase (trade name “Sumiteam PHY”, Shin Nihon <b¾t¾) at 0.4% by weight (4UZg crude) based on the crude protein weight. ^: Protein) 40. Stirred at C and started the enzyme. Immediately after the start, the viscosity increased and gradually became a gel. Therefore, implementation with heated concentrated soymilk was difficult.

Claims

The scope of the claims

1. The method of making a protein is to allow the protein to have an enzyme having a phytic acid content of j ^ iS in the range of 6 to 9 with a high pH.

2. The method according to claim 1, wherein the enzyme having a phytic acid content is an enzyme having an activity in a pH range of 6 to 9.

3. The method according to claim 1 or 2, wherein the enzyme is derived from an enzyme having phytic acid production.