JP2002051706A - Method for producing high purity vegetable protein material - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing a purified vegetable protein, and more particularly, to provide a method for producing high purity vegetable protein isolates and concentrates. SOLUTION: This method for producing a purified vegetable protein material with a low concentration of ribonucleic acids is characterized by comprising a step of forming an aqueous slurry of a vegetable protein material, a 2nd step wherein the slurry is treated with an acid phosphatase at a temperature and a pH for a time effective to decompose the ribonucleic acids in the vegetable protein material, and a 3rd step of washing the thus treated slurry.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to a method for producing a purified plant protein substance, and more particularly to a method for producing a high-purity plant protein isolate and a concentrate.

TECHNICAL BACKGROUND Many food and beverage products use plant materials such as soybeans, beans, and peas.
s), and other legumes, and protein supplements from oilseed, such as rapeseed. Vegetable protein materials, especially soy, are used in infant formulas. The purpose of this vegetable protein supplement in infant formulas is to increase the nutritional value of the formula and to provide a protein content close to that of human milk. However, commercially available protein concentrates and isolates contain some impurities that are undesirable in products such as infant formulas. Undesirable specific impurities in plant protein isolates and concentrates are not available for human anabolism, which is associated with phytic acid, phytate, ribonucleic acid, ash, and phytic acid, phytate or ribonucleic acid Inorganic substances such as phosphorus, calcium, chlorine, iron, zinc,
And copper. It would be desirable to provide a method of reducing the concentration of these impurities in plant protein isolates and concentrates, particularly for use in products such as infant formulas.

It is also of interest to reduce the concentration of phytic acid, also known as inositol hexaphosphate, and phytate, a salt of phytic acid, in plant protein material. Because phytic acid and phytate
This is because there is a tendency to form a complex with the protein and the polyvalent metal cation to reduce the nutritional value of the plant protein material. Extensive efforts have been made to reduce the levels of phytic acid and phytate in plant protein materials. For example, U.S. Pat.
No. 8,765 discloses an aqueous slurry of a phytate-containing material with a low p.
A method is described for separating phytate and manganese from protein and dietary fiber by treatment with alumina at H. The alumina is then separated from the protein and fiber material along with the phytate associated with the alumina. U.S. Pat.No. 2,732,395 granted to Bolley et al., U.S. Pat.No. 4,072,670 granted to Goodnight et al., U.S. Pat.No. 4,008,795 granted to Goodnight et al. U.S. Pat.No. 4,091,120 and deRha
British Patent No. 1,574,110 to m) teaches a method for removing phytic acid and phytate from protein materials, all by various precipitation and differential solubility separation techniques.

[0003] Other methods for reducing the concentration of phytic acid and phytate in plant protein materials utilize enzymes to degrade phytic acid or phytate. European Patent Application No. 0380343A2 discloses a method for producing phytate-free or low-phytate soy protein isolates and concentrates, in which phytic acid and phytate-degrading enzymes (hereinafter `` phytase '') are disclosed.
Is added to the soy protein isolate or concentrate at a temperature of 20-60 ° C and pH 2-6 to degrade phytic acid and phytate in the protein material. U.S. Pat.No. 4,642,236 granted to Friend et al., U.S. Pat.No. 3,733,207 granted to McCabe, and Japanese Patent Application Laid-Open No. 8 (1996) -214787, all use phytase, soybeans. A method for degrading phytic acid and phytate in proteins is disclosed. Phytase enzyme formulations are used to produce plant protein materials because they are inexpensive and readily available commercially.
Particularly useful.

[0004] The phytase enzyme is a phosphate monoester hydrolase (IUB3.1.3) and can usually be derived from microbial or fungal sources, such as Aspergillus and Rhizopus species. Commonly used phytase enzyme compositions typically comprise, as the main phytase enzyme, the enzyme 3-phytase (myo-inositol-
Hexakisphosphate 3-phosphohydrolase (IUB
3.1.3.8)). Some, but not all, phytase enzyme compositions contain a sufficient amount of the enzyme acid phosphatase (orthophosphate monoester phosphohydrolase (IUB3.1.3.2)) to effect the degradation of phytic acid and phytate. The phytase enzyme composition is considered not to reduce the concentration of ribonucleic acid and bound inorganic substances in plant protein substances because the most common phytases, especially 3-phytases, do not degrade ribonucleic acid structures. Have been. Ribonuclease enzyme compositions are known to open and degrade ribonucleic acids and can be used to reduce the concentration of ribonucleic acids in plant proteins, but such enzyme compositions are quite expensive. Yes, and it is not possible to use purified plant protein materials on the scale required for industrial production. It is desirable to reduce the concentration of ribonucleic acid material and bound minerals in plant proteins at a cost that allows the method to be used on an industrial scale.

[0005]

SUMMARY OF THE INVENTION In one aspect, the present invention provides a method for reducing the concentration of ribonucleic acid and minerals associated with ribonucleic acid in a plant protein material. In this method, a vegetable protein material is prepared and dispersed in an aqueous solution to form a slurry. The slurry is treated with an enzyme formulation containing acid phosphatase at a pH, temperature and time effective to substantially reduce the concentration of the ribonucleic acid in the plant protein material. Next, the slurry treated with the enzyme is washed to obtain a plant protein substance containing ribonucleic acid at a low concentration. In one preferred aspect of the invention, the mineral content of the plant protein material is reduced by treating the slurry of the plant protein material with an enzyme formulation containing acid phosphatase.

In another preferred embodiment of the invention, the vegetable protein material is soy protein, the pH at which the slurry is treated with the enzyme formulation is from about 3 to about 6, and the slurry is treated with the enzyme. The processing temperature of the formulation is about 20 to about 70 ° C
And the time for treating the slurry with the enzyme formulation is from about 30 minutes to about 4 hours. After treating the enzyme-treated slurry with the enzyme formulation, the slurry is washed. In yet another preferred embodiment, the slurry is enzymatically treated and washed, then heat treated, and the heat treated slurry is dried. In another aspect, the present invention relates to a method for producing phytic acid, phytate,
Methods for reducing the concentration of ribonucleic acids and phytic acid, phytate, and minerals associated with ribonucleic acids are provided. In this method, a vegetable protein material is prepared and dispersed in an aqueous solution to form a slurry. This slurry is then treated with an enzyme formulation comprising acid phosphatase and phytase, which is effective to substantially reduce the concentration of phytic acid, phytate and the ribonucleic acid in the plant protein material.
Process, pH, temperature and time.

[0007]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention relates to the discovery that the acid phosphatase enzyme unexpectedly breaks down ribonucleic acids and thus uses them to degrade ribonucleic acid substances in plant protein substances on an industrial scale. And that the concentration can be reduced and the inorganic and ash bound to the ribonucleic acid material can be removed. Some commercially available phytase enzyme formulations include acid phosphatase, but to date, acid phosphatase has been useful in the degradation of ribonucleic acid and the concentration of ribonucleic acid in plant protein material has been reduced by acid phosphatase treatment. Was not recognized. Acid phosphatase is able to degrade phytic acid, phytate and ribonucleic acid, and thus acid phosphatase
It can be used to degrade and reduce the concentration of phytic acid, phytate and ribonucleic acid, or it can be used in combination with other phytases.

The starting material used in the process according to the invention is a vegetable protein concentrate or vegetable protein isolate. As used herein, and according to conventional definitions, a plant protein material (material) is a plant protein material containing 65% to 90% protein on a dry weight basis, and a plant protein isolate. The body is a vegetable protein material that contains at least 90% protein on a dry weight basis. Vegetable protein concentrates and isolates are readily available commercially. For example, a soy protein isolate that can be used in the methods of the present invention is a protein soy protein isolate from Protein Technologies International, St. Louis, Mo.
(Technologies International, Inc.) and trade name SU
Commercially available under PRO ^R 500E and SUPRO ^R 620.
Vegetable protein concentrates and vegetable protein isolates can be prepared by conventional methods. The vegetable protein concentrate is usually (i) leached vegetable protein material with an aqueous solution having a pH near the isoelectric point of the protein, (ii)
It is prepared by extracting the vegetable protein material with aqueous alcohol, or (iii) denaturing the vegetable protein material by wet heating and then extracting the modified vegetable protein material with water.

In one preferred embodiment, a soy protein concentrate is prepared for use in the method of the present invention. Commercially available defatted soy flakes (hulled and defatted soy) have a pH near the isoelectric point of the soy protein, preferably from about 4 to about 5, most preferably from about 4.4 to about 4.6.
Wash with an aqueous solution having a different pH. This aqueous acid solution is made from the soy protein, water-soluble carbohydrates, inorganic substances,
Leach phenolic and other non-protein substances. Vegetable protein isolates are produced by extracting vegetable protein material with an aqueous alkaline solution to solubilize the protein material. This extract of solubilized proteinaceous material is then separated from insoluble plant material, such as cellulose and other plant fibers. next,
The pH of the protein extract is adjusted to a value near the isoelectric point of the protein to precipitate the protein. The precipitated protein is separated from the solution by filtration or centrifugation, and the protein material is separated from the water-soluble carbohydrates, minerals, phenolic substances and other non-protein substances remaining in the solution. Next, the separated protein is washed with water to obtain the isolated protein.

In a most preferred embodiment, a soy protein isolate is prepared for use in the method of the present invention. Commercially available defatted soy flakes are used as starting material. Preferably, the soy flakes are treated with a sulfite, such as sodium sulfite, to provide flow properties and anti-
Improve microbial properties. The soy flakes are extracted with an aqueous alkaline solution, preferably an aqueous sodium hydroxide solution having a pH of about 8 to about 11. Preferably, the weight ratio of the extract to the soy flake material is from about 5: 1 to about 16: 1. This extract is separated from insoluble materials such as soybean fiber and cellulose by filtration or centrifugation and decantation of the supernatant extract from the insoluble materials. The pH of the separated extract is a value near the isoelectric point of the soy protein, preferably about 4 to about 5, most preferably about 4.
4 to about 4.6, a suitable acid, preferably hydrochloric acid, sulfuric acid, nitric acid,
Alternatively, adjust with acetic acid to precipitate the soy protein material.
The precipitated protein material is separated from the extract, preferably by centrifugation or filtration. The separated protein material is washed with water, preferably at a water to protein material weight ratio of about 5: 1 to about 12: 1 to obtain the soy protein isolate.

An aqueous slurry of the vegetable protein concentrate or vegetable protein isolate (hereinafter generally referred to as "protein material") is formed by mixing the protein material with water. Preferably, the slurry is about
It should contain from 2% to about 30% of the protein material, and more preferably from about 5% to about 20% by weight, and most preferably from about 10% by weight.
It should contain from about% to about 18% of the protein material. Then
This slurry is then treated with an enzyme formulation containing acid phosphatase (orthophosphate monoester phosphohydrolase (IUB3.1.3.2)), an acid phosphatase concentration effective to substantially reduce the concentration of ribonucleic acid in the protein material, temperature,
Treat for pH and time. This enzyme formulation containing acid phosphatase is from a microbial or fungal source, such as Aspergillus and Rhizopus species. A preferred source of this acid phosphatase useful in the method of the present invention is the fungus Aspergillus niger. Phytase enzyme formulations derived from Aspergillus niger and containing acid phosphatase are commercially available.

The enzyme formulation is added to the slurry in a sufficient amount to achieve an acid phosphatase concentration effective to degrade and substantially reduce the ribonucleic acid present in the protein material. I do. Preferably, at least a majority of the ribonucleic acids present in the initial plant protein material are degraded by the acid phosphatase enzyme. Here, the term “most” is defined as being 50% or more. More preferably, the acid phosphatase is at least 60% ribonucleic acid in the plant protein material, even more preferably at least 70% ribonucleic acid in the protein material, even more preferably at least 70% of the ribonucleic acid in the protein material.
Degrades 80% of the ribonucleic acid and most preferably substantially all of the ribonucleic acid in the protein material. To effectively degrade and reduce the concentration of ribonucleic acid in the protein material, the enzyme formulation requires a sufficient amount of acid phosphatase or acid phosphatase and another phytase, e.g. -Phytase (myo-inositol-
Hexakisphosphate 3-phosphohydrolase (IUB
3.1.3.8)), thus degrading the ribonucleic acid and substantially reducing its concentration. Preferably, the enzyme formulation is characterized in that the acid phosphatase is present in the slurry at about 0.1% to about 10% by dry weight of the protein material, more preferably at about 0.3% of the protein material by dry weight. % To about 5%, and most preferably about 0.5% to about 3% of the protein material on a dry weight basis.
Present in a certain amount.

In a most preferred embodiment of the present invention, the enzyme formulation degrades and reduces the concentration of phytic acid and phytate and ribonucleic acid. Preferably, the enzyme formulation degrades at least a majority of the phytic acid and phytate, where the majority is defined as at least 50%, more preferably at least 75% of the phytic acid and phytate, More preferably, at least 85% of the phytic acid and phytate are degraded, and most preferably substantially all of the phytic acid and phytate are degraded by the enzyme formulation. In order to effectively degrade ribonucleic acid, phytic acid and phytate in the protein substance and effectively reduce the concentration thereof,
The enzyme formulation comprises a sufficient amount of acid phosphatase, or acid phosphatase and another phytase, such as
It must include a combination with 3-phytase (myo-inositol-hexakisphosphate 3-phosphohydrolase (IUB3.1.3.8)), which must degrade the ribonucleic acid, phytic acid and phytate. In a most preferred embodiment, the enzyme formulation is characterized in that the acid phosphatase and 3-phytase are present in the slurry in an amount of about 0.1% to about 10% by dry weight of the protein material, more preferably on a dry weight basis. About 0.3% to about 5% of the protein material, and most preferably about 0.3% of the protein material on a dry weight basis.
0.5% to about 3% is added to be present.

The activity of the enzyme formulation should be effective in degrading ribonucleic acid, phytic acid and phytate, and substantially reducing its concentration. The enzyme formulation is preferably present in an amount of about 400 to 1 kg protein solids.
It has an activity in the range of about 1400 kilophytase units (KPU / kg protein solids), more preferably about 600 to about 1200 KPU /
It has an activity in the range of kg protein solids, and most preferably has an activity of about 1000 KPU / kg protein solids. One kilophytase unit is equal to 1000 phytase units, where one phytase unit is used under standard conditions (40
C., pH 5.5, and a 15 minute incubation), equal to the amount of enzyme that releases 1 nM of inorganic phosphate from sodium phytate per minute. The activity of the enzyme formulation includes the acid phosphatase activity and any other phytase enzyme activity contained in the enzyme formulation.

[0015] The pH of the slurry treated with the enzyme formulation
Is the pH at which the enzyme formulation effectively degrades ribonucleic acids, and preferably the pH at which the enzyme formulation also degrades phytic acid and phytate. The acid phosphatase enzyme has been found to degrade ribonucleic acids very effectively at a pH of about 4.5, and the phytase enzyme degrades phytic acid and phytate very effectively at a pH of about 5.3. This is known in the art. In one preferred embodiment, the pH of the slurry treated with the enzyme formulation is from about 3 to about 6, more preferably about 3.
5 to about 5.5, and even more preferably about 4 to about 5, and most preferably about 4.4 to about 4.6. PH of this slurry
Is adjusted with a suitable acidic reagent such as hydrochloric acid, sulfuric acid, nitric acid or acetic acid, or a suitable basic reagent such as sodium hydroxide, calcium hydroxide or ammonium hydroxide as necessary to achieve a predetermined pH. it can.

The temperature of the slurry treated with the enzyme formulation should be such that the enzyme in the enzyme formulation effectively degrades ribonucleic acid and preferably also degrades phytic acid and phytate. is there. Preferably, the temperature of the slurry should be high enough to maximize the enzymatic degradation of the ribonucleic acid, phytic acid and phytate,
It should not be high enough to inactivate the enzyme or degrade the protein material in the slurry. In one preferred embodiment, the temperature at which the slurry is treated with the acid phosphatase-containing enzyme formulation is from about 20 to about 70 ° C, more preferably from about 30 to about 60 ° C, and most preferably about 40 to about 60 ° C.
~ 55 ° C.

The time of treating the slurry with the enzyme formulation is such that the enzyme effectively degrades ribonucleic acid and effectively reduces its concentration, and preferably also reduces the phytic acid content in the plant protein material. And phytate should be sufficient to be able to break down and reduce its concentration. Preferably, the slurry is treated at an effective pH and temperature for about 30 minutes to about 4 hours, more preferably for about 45 minutes to about
Treat with the enzyme formulation for about 3 hours and most preferably for about 1 hour to about 2 hours. Subsequent to treatment of the plant protein material with the enzyme formulation, the plant protein material is washed to remove the degraded material, ash, and minerals.
Preferably, the vegetable protein material is washed with water by diluting the material with water and centrifuging the diluted slurry. More preferably, the plant protein material, e.g., diluting the plant protein material slurry with water, centrifuging the diluted slurry with a disk centrifuge, and then centrifuging the slurry with a ball centrifuge Wash twice.

Most preferably, the pH of the slurry during the washing step, after the decomposition of the ribonucleic acid, phytic acid and phytate, is set to a value near the isoelectric point of the plant protein material, The loss of Degradation of the ribonucleic acid, phytic acid and phytate can result in a shift in the isoelectric point of the protein substance. For example, soy protein containing ribonucleic acid, phytic acid and phytate has an isoelectric point of about pH 4.5,
After the enzymatic decomposition of these substances, they have an isoelectric point of about pH 5.1. The pH of the slurry can be adjusted, if necessary, to about the isoelectric point of the protein material with a suitable acidic or basic material prior to washing the protein material.

The washing is carried out with a sufficient amount of washing water, preferably having a pH adjusted to about the isoelectric point of the protein, to obtain the degraded ribonucleic acid, and preferably the degraded ribonucleic acid. Phytic acid and phytate are removed from the plant protein material. In one preferred embodiment, at least a majority of the degraded ribonucleic acid, phytic acid and phytate present in the initial plant protein material are removed by the method of the present invention. Here, the term “most” is defined as 50% or more. More preferably,
The method of the present invention is effective to remove at least 60% of the degraded ribonucleic acid, phytic acid and phytate present in the plant protein material, and more preferably present in the plant protein material. At least 70% of the degraded ribonucleic acid, phytic acid and phytate,
And even more preferably, the degraded ribonucleic acid, at least 80% of the phytic acid and phytate present in the plant protein material, and most preferably the degraded ribonucleic acid present in the plant protein material, Substantially all of the phytic acid and phytate are removed.

After washing, the purified plant protein material is
It can be recovered by drying. In one preferred embodiment, the purified plant protein material is recovered by spray drying according to known spray drying techniques.
The plant protein material, including low concentrations of ribonucleic acid and preferably low concentrations of phytic acid and phytate, may be further processed, if necessary, to obtain a purified protein material having improved functional properties. It is possible. The purified protein material slurry can be heat treated to denature the protein and to sterilize the protein material. Preferably, the slurry is heat treated by jet cooking and flash cooled by discharge from the jet cooker into a vacuum chamber according to known jet cooking techniques. Most preferably, this slurry of purified protein material is heated under pressure to about 140 to about 160 ° C.
At a temperature of about 1 to 15 seconds. In a most preferred embodiment, the pH of the protein slurry is
Is neutralized to a pH of about 6 to about 8 with a suitable basic reagent, preferably an aqueous sodium hydroxide / potassium hydroxide solution,
It assists in the treatment of the heat-treated protein material.

Heat treated or unheated,
Subjecting the purified protein material to enzymatic hydrolysis,
The viscosity of the protein material can also be reduced. Enzymatic hydrolysis is particularly preferably performed after the heat treatment of the protein substance. This is because the denatured protein material is more viscous than a similar protein material that has not been subjected to heat treatment. Slurry of the purified protein material,
Known commercially available enzymes can provide effective pH, temperature, enzyme concentration and activity, and time treatment to hydrolyze the protein material. The pH at which the enzymatic hydrolysis is performed depends on the particular protease enzyme used. The protease enzyme should be selected to carry out the hydrolysis, having a known pH range effective for the enzyme to hydrolyze the protein, and the hydrolysis of the purified protein material should be It should be performed within the known effective pH range of the enzyme. In one preferred embodiment, the protease enzyme Bromelain is used at a pH of about 4 to about 9.

[0022] The concentration and activity of the protease should be sufficient to achieve the desired degree of hydrolysis of the protein. Preferably, the protease comprises from about 0.1% to about 10% of the protein material by dry weight, and more preferably from about 0.5% to about 5% of the protein material by dry weight.
The protease is added to the purified proteinaceous material slurry so as to be present in an amount. Further, preferably, the protease has about 1000 to 4000 tyrosine units per gram (TU / g), and more preferably about 2000 to about 3000
It should have an activity of TU / g, where 1 TU / g is incubated at the optimal pH of the protease for 15 minutes at 30 ° C.
Equivalent to enzyme activity releasing 1 μM tyrosine per minute.

The temperature of the slurry treated with the protease should be such that the protease effectively hydrolyzes the purified proteinaceous material. Preferably, the temperature of the slurry is high enough to maximize enzymatic hydrolysis of the protein material, but not high enough to deactivate the enzyme. In one preferred embodiment, the temperature at which the slurry is treated with the protease is about
15 to about 75C, more preferably about 30 to about 65C, and most preferably about 40 to about 55C. The time for treating the slurry with the protease is such that the enzyme removes the protein material from the slurry.
The value should be sufficient to allow hydrolysis to a given degree of hydrolysis. Preferably, the slurry is
Treat with the protease at effective pH and temperature for about 15 minutes to about 2 hours, more preferably about 30 minutes to about 1.5 hours, and most preferably about 45 minutes to about 1 hour. After the enzymatic hydrolysis is complete, the slurry is heated to the temperature at which the protease is inactivated, for example, by heating the slurry to 75 ° C.
The reaction is stopped by heating at a temperature above.

The hydrolyzed and purified plant protein material may be heat treated, if necessary, to sterilize the protein material and, if the protein material has not been previously heat treated, the hydrolyzed protein material The substance can be modified. Preferably, the slurry is heat treated by jet cooking according to known jet cooking techniques and flash cooled by discharge from a jet cooker to a vacuum chamber. Most preferably, the hydrolyzed and purified proteinaceous material is compressed under pressure from about 140 to
Heat treat at about 160 ° C. for about 1 to 15 seconds. After enzymatic hydrolysis and optional heat treatment, the hydrolyzed and purified protein material is recovered from the slurry by drying the protein material. In a preferred embodiment, the hydrolyzed and purified plant protein material is recovered by spray drying the protein material according to known spray drying techniques.

[0025]

The following examples are given to illustrate the methods of the present invention, but it is not to be understood that the invention is limited to these illustrative methods. Example 1 Purified plant protein isolate is produced according to the method of the present invention. 243 lbs of soy protein isolate is converted to 2959 lbs.
To form a soy protein isolate slurry containing 7.6% solids. The pH of the slurry is adjusted to 4.5 with hydrochloric acid and the temperature of the slurry is raised to 50 ° C. An enzyme formulation containing acid phosphatase and one phytase and having an activity of 1000 KPU / kg curd solids is added to the slurry. The slurry is treated with the enzyme formulation for 2 hours, after which the pH of the slurry is
Adjust to 5.1 with an alkaline mixture containing potassium hydroxide and sodium hydroxide. Next, this slurry is
Dilute to 4.2% solids and wash in a ball centrifuge. 275 lbs of this washing slurry is neutralized with an alkaline mixture containing potassium hydroxide and sodium hydroxide. The neutralized material is heat treated by jet cooking at 150 ° C and cooled to 53 ° C by discharging into a vacuum chamber having a pressure of about 26 Torr. The heat treated slurry is then spray dried to recover 15.5 lb of purified soy protein isolate. Example 2 A hydrolyzed and purified plant protein isolate is formed according to the method of the present invention. 1015 lb, 15.5% solids
The purified soy protein isolate containing (about 157 lb of purified soy protein material) is adjusted to pH 7.4 with 1400 ml of a sodium hydroxide / calcium hydroxide mixture. This slurry is
Jet cook at 150 ° C. for 9 seconds and flash cool by discharging into a vacuum chamber. 725 lb of the slurry is treated with the protease enzyme bromelain. The enzyme has an activity of 2500 TU / g, which is added to the slurry at a concentration of 0.29% of the protein material in the slurry on a dry weight basis. The temperature of the enzyme treated slurry is maintained at about 50 ° C. during the enzyme treatment. The processing period is 40 minutes. After the enzymatic treatment, the slurry is cooled to 16 ° C. and a further 190 ml of a sodium hydroxide / calcium hydroxide mixture are added to the slurry. The slurry is then jet cooked at 150 ° C. for 9 seconds and flash cooled by discharging into a vacuum chamber. The slurry is then dried to recover 75 lbs of hydrolyzed and purified soy protein isolate. Example 3 The effect of enzyme activity on ribonucleic acid concentration and phytic acid concentration in a soybean protein isolate is examined. here,
The soy protein isolate is purified by a method performed according to the present invention. A soy protein isolate slurry is formed by mixing the soy protein isolate with water adjusted to pH 4.5 with hydrochloric acid. Here, the total solids in the slurry is present in an amount of about 8.5% of the slurry by weight. The slurry is heated at a temperature of 50 ° C. Two samples of this slurry are prepared from the protein isolate slurry for enzymatic degradation of ribonucleic acid and phytic acid. The first sample was 1530 lb and contained 8.66% total solids by weight, and the second sample was 151
0 lb and contains 8.66% total solids by weight. An enzyme formulation containing acid phosphatase and one phytase enzyme is added to each slurry sample. 80
An enzyme formulation with an activity of 0 KPU / kg curd solids is added to the first sample. An enzyme formulation having an activity of 1400 KPU / kg curd solids is added to the second sample. The samples are reacted with the enzyme formulation for one hour. Following enzymatic treatment of the samples, the samples are thoroughly washed and the enzymes are thermally inactivated by jet cooking at a temperature of 150 ° C. By discharging these samples into the vacuum chamber,
Flash cool to 50 ° C. The cooled samples are then spray dried.

A control sample having a total solids content of 7.6% is prepared from the initial protein slurry for comparison purposes. The control sample is heated at 50 ° C. for 1 hour and then washed thoroughly. The washed control sample is jet cooked at 150 ° C. and then flash cooled to 52 ° C. in a vacuum chamber. Next, the control sample is dried. The samples are analyzed to determine the ribonucleic acid content and the phytic acid content of the sample. The results of this analysis are shown in Table 1 below.

[0027]

[Table 1]

[0028] These results clearly show that acid phosphatase and phytase are involved and that the activity is 800 and 1400 KP
The enzyme formulation with U / kg curd solids has been shown to be effective in substantially reducing ribonucleic acid content in soy protein isolates. These results also indicate that the enzyme formulation is quite effective in reducing the phytic acid content of the protein isolate. Example 4 The effect of pH on the enzymatic degradation of ribonucleic acid and phytic acid in soy protein isolates by an enzyme formulation containing acid phosphatase and one phytase enzyme is investigated. Here, the soy protein isolate is purified according to the present invention. By mixing sufficient soy protein isolate and water adjusted to pH 4.5 with hydrochloric acid to form a slurry containing about 8% soy protein isolate by weight, the soy protein isolate is To form a slurry. The slurry is heated to a temperature of 50 ° C.

[0029] Two samples of the slurry, having a total solids content of 8% by weight, are prepared from the protein isolate slurry for enzymatic degradation of ribonucleic acid and phytic acid. The pH of the first sample is adjusted to 5.1 with a potassium hydroxide / sodium hydroxide mixture. The pH of the second sample remains at 4.5. The samples are then treated for two hours with an enzyme formulation containing an acid phosphatase enzyme and one phytase enzyme and having an activity of 1400 KPU / kg curd solids. Following the enzymatic treatment of these samples, the samples are thoroughly washed and the enzymes are thermally inactivated by jet cooking at a temperature of 150 ° C. The samples are flash cooled to 50 ° C. by draining into a vacuum chamber. The cooled sample is then spray dried.

A control sample having a total solids content of 7.6% is prepared from the initial protein slurry for comparison. Run this control sample at 50 ° C
Heat for 1 hour, then wash thoroughly. The washed control sample is jet cooked at 150 ° C. and then flash cooled to 52 ° C. in a vacuum chamber.
Next, the control sample is spray-dried. The samples are analyzed to determine the ribonucleic acid content and the phytic acid content of the sample. The results of these analyzes are shown in Table 2 below.

[0031]

[Table 2]

[0032] These results indicate that the enzyme formulations containing acid phosphatase and phytase were pH 4.5 and pH 5.1.
, Both phytic acid and ribonucleic acid content in soy protein isolates are shown to be effective in substantially reducing. The reduction in ribonucleic acid content in the protein isolate is particularly effective at pH 4.5. Example 5 The effect of enzyme treatment time on the enzymatic degradation of ribonucleic acid and phytic acid in soy protein isolates by an enzyme formulation containing acid phosphatase and phytase enzymes is investigated. Here, the soy protein isolate is purified according to the present invention. With enough soy protein isolate and hydrochloric acid
A slurry of the soy protein isolate is formed by mixing with water adjusted to pH 4.6 to form a slurry containing about 8% of the soy protein isolate by weight. The slurry is heated at a temperature of 50 ° C.

Two samples of the slurry are prepared for the enzymatic degradation of the ribonucleic acid and phytic acid. The first slurry sample is 3202 lb and on a weight basis
Contains 7.6% total solids. The weight of the second sample is 1530 l
b and contains 8.6% total solids by weight. 1400 containing acid phosphatase and phytase enzymes
An enzyme formulation having an activity of KPU / kg curd solids is added to the first sample and the second sample. The first sample is treated with the enzyme formulation for one hour, and the second sample is treated with the enzyme formulation for two hours.
Following the enzymatic treatment of the samples, the samples are thoroughly washed and the enzyme is
Thermally deactivated by jet cooking at 150 ° C. The samples are flash cooled to 50 ° C. by draining into a vacuum chamber. The cooled sample is then spray dried.

A control sample having a total solids content of 7.6% is prepared from the initial protein slurry for comparison purposes. The control sample is heated at 50 ° C. for 1 hour and then thoroughly washed. The washed control sample is jet cooked at 150 ° C. and then flash cooled to 52 ° C. in a vacuum chamber. Then
The control sample is spray dried. The samples are analyzed to determine the ribonucleic acid content and the phytic acid content of the sample. The results of these analyzes are shown in Table 3 below.

[0035]

[Table 3]

Treatment of soy protein isolate with an enzyme formulation containing acid phosphatase and phytase enzymes for one hour or two hours reduces the ribonucleic acid content and phytic acid content in the protein isolate substantially. It is effective in reducing the total. Two hours of processing, compared to one hour of processing,
Increases the rate of decrease in phytic acid content in the protein isolate, but does not show a significant increase in the rate of decrease in ribonucleic acid content in the protein. Example 6 The effect of temperature on the enzymatic degradation of ribonucleic acid and phytic acid in soy protein isolates by an enzyme formulation containing acid phosphatase and phytase enzymes is investigated. Here, the purification of the soy protein isolate is performed according to the present invention. By mixing a sufficient soy protein isolate with water adjusted to pH 4.5 with hydrochloric acid to form a slurry containing about 8% by weight of the soy protein isolate, a soy protein isolate is formed. Prepare a slurry of the debris.

A first sample of the slurry containing 8% total solids by weight is prepared from the protein isolate slurry for enzymatic degradation of ribonucleic acid and phytic acid. This first sample is adjusted to a temperature of 50 ° C. A second sample of the slurry containing 4% total solids by weight is prepared from the protein isolate slurry. This second sample is adjusted to a temperature of 38 ° C. The samples are then treated for two hours with an enzyme formulation containing an acid phosphatase enzyme and a phytase enzyme and having an activity of 1400 KPU / kg curd solids. Following enzymatic treatment of these samples, the samples are thoroughly washed and the enzymes are thermally inactivated by jet cooking at a temperature of 150 ° C. The samples are flash cooled to 53 ° C. by draining into a vacuum chamber.
These cooled samples are then spray dried.

A control sample having a total solids content of 7.6% is prepared from the initial protein slurry for comparison. The control sample is heated at 50 ° C. for 1 hour and then thoroughly washed. The washed control sample is jet cooked at 150 ° C. and then flash cooled to 52 ° C. in a vacuum chamber. Then
The control sample is spray dried. The samples are analyzed to determine the ribonucleic acid content and the phytic acid content of the sample. The results of these analyzes are shown in Table 4 below.

[0039]

[Table 4]

The treatment of the soy protein isolate at 38 ° C. and 50 ° C. with an enzyme formulation comprising acid phosphatase and phytase substantially reduces the ribonucleic acid content and phytic acid content in the protein isolate. It is effective to reduce to. Treatment at 50 ° C. increases the rate of decrease in ribonucleic acid content and phytic acid content as compared to treatment at 38 ° C. Example 7 The effect of enzymatic degradation of phytic acid and ribonucleic acid in a soy protein isolate by an enzyme formulation containing an acid phosphatase enzyme and a phytase enzyme on the mineral content in the protein is examined. In particular, it affects the concentration of calcium, iron, magnesium, sodium, zinc, copper, potassium, manganese, and phosphorus in a soy protein isolate.
The effect of the enzymatic degradation will be examined.

Sufficient soy protein isolate and pH with hydrochloric acid
Mix with water adjusted to 4.6 to form a slurry containing about 8% of the soy protein isolate by weight. From this slurry, a sample is prepared for enzymatic degradation of phytic acid and ribonucleic acid. Where the weight of the sample is
3202 lb and has a total solids concentration of 7.6% by weight. The sample is heated to a temperature of 50 ° C. An enzyme formulation comprising an acid phosphatase enzyme and a phytase enzyme and having an activity of 1400 KPU / kg curd solids is added to the sample and the sample is treated with the enzyme formulation for 1 hour. Following enzymatic treatment of the sample, the sample is thoroughly washed and the enzyme is thermally deactivated by jet cooking the sample at 150 ° C. The sample is flash cooled to 53 ° C. by draining into a vacuum chamber. Next, the cooled sample is spray-dried.

A control sample having a total solids content of 7.6% is prepared from the initial protein slurry for comparison. The control sample is heated at 50 ° C. for 1 hour and then thoroughly washed. The control sample is jet cooked at 150 ° C and then flash cooled to 52 ° C in a vacuum chamber. The control sample is then spray dried. The samples are analyzed to determine the calcium, iron, magnesium, sodium, zinc, copper, potassium, manganese, and phosphorus content of the samples. The results of these analyzes are shown in Table 5 below.

[0043]

[Table 5] Table 5: Unit: ppm

The treatment of a soy protein isolate with an enzyme formulation containing acid phosphatase and phytase can be accomplished by treating the calcium, iron, magnesium, sodium, zinc, copper, potassium, manganese, and phosphorus contents in the protein isolate. Is effective in reducing the This enzymatic treatment is particularly effective in reducing the sodium, potassium, and phosphorus content in the protein material. Those skilled in the art will appreciate that various modifications can be made to the disclosed invention without departing from the spirit of the invention. The present invention is not limited to the described embodiments, which are given for illustrative purposes only.
Rather, the invention is limited only by the following claims and equivalents thereof.

────────────────────────────────────────────────── ───

[Procedure amendment]

[Submission date] December 27, 2000 (200.12.
27)

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] Claims

[Correction method] Change

[Correction contents]

[Claims]

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Theodore M. Won, United States 63201 Missouri Weatherwood 802 (72) Inventor David A. Singer United States Missouri 63112 St. Louis Lindel Boulevard 6127 (72) Inventor Santa H. Linn United States Missouri 63017 Town and Country Millfield Court 820 F-term (reference) 4B064 AG01 CB05 CC06 CC07 CC09 CD20 CE16 DA10

Claims (80)

[Claims] 1. A method for producing a purified plant protein material having a low ribonucleic acid concentration, comprising the steps of forming an aqueous slurry of the plant protein material, and subjecting the slurry to an acid phosphatase enzyme. Treating at a temperature, pH and time effective to degrade ribonucleic acid therein, and washing the treated slurry to produce a plant protein material containing low levels of ribonucleic acid. The above method, characterized in that: 2. The method according to claim 1, wherein the plant protein material is a plant protein concentrate or a plant protein isolate. 3. The method according to claim 2, wherein said plant protein material is a soy protein concentrate or a soy protein isolate. 4. The method according to claim 1, wherein the slurry comprises about 2% to about 30% by weight.
2. The method of claim 1, comprising a range of the protein material. 5. The method of claim 5, wherein the slurry comprises about 5% to about 20% by weight.
5. The method of claim 4, comprising a range of the protein material. 6. The method of claim 1, wherein the slurry comprises about 10% to about 18% by weight.
5. The method according to claim 4, comprising in the range of%. 7. The method of claim 1, wherein treating the slurry with the enzyme is effective to degrade most of the ribonucleic acids in the plant protein material. 8. The washing of the enzyme-treated slurry is effective to remove the degraded ribonucleic acid and to obtain a plant protein substance from which most of the ribonucleic acid has been removed.
The described method. 9. The method of claim 1, wherein treating the slurry with the enzyme is effective to degrade at least 60% of the ribonucleic acids in the plant protein material. 10. The washing of the enzyme-treated slurry is effective for removing the degraded ribonucleic acid and obtaining a plant protein substance from which at least 60% of the ribonucleic acid has been removed.
The method according to claim 9. 11. The method of claim 1, wherein treating the slurry with the enzyme is effective to degrade at least 70% of the ribonucleic acids in the plant protein material. 12. The washing of the enzyme-treated slurry is effective for removing the degraded ribonucleic acid and obtaining a plant protein substance from which at least 70% of the ribonucleic acid has been removed.
The method of claim 11. 13. The method of claim 1, wherein treating the slurry with the enzyme is effective to degrade at least 80% of the ribonucleic acids in the plant protein material. 14. The method of claim 13, wherein washing the enzyme-treated slurry is effective to remove the degraded ribonucleic acid and to obtain a plant protein material from which at least 80% of the ribonucleic acid has been removed. the method of. 15. The method of claim 1, wherein treating the slurry with the enzyme is effective to degrade substantially all ribonucleic acids in the plant protein material. 16. The method according to claim 15, wherein washing the enzyme-treated slurry is effective for removing the degraded ribonucleic acid and obtaining a plant protein substance from which substantially all of the ribonucleic acid has been removed. the method of. 17. The method of claim 1, wherein treating the slurry with the enzyme is effective to degrade phytic acid and phytate in the plant protein material. 18. The washing of the enzyme-treated slurry,
18. The method of claim 17, wherein said degraded phytic acid and phytate are removed and said phytic acid and phytate are removed and said method is effective in obtaining a plant protein material. 19. The method of claim 17, wherein the slurry has a pH in the range of about 3 to about 6.
2. The method according to claim 1, wherein the method is treated with acid phosphatase. 20. The method of claim 19, wherein said slurry is treated with acid phosphatase at a pH in the range of about 3.5 to about 5.5. 21. The method of claim 21, wherein the slurry has a pH in the range of about 4 to about 5.
20. The method according to claim 19, wherein the method is treated with acid phosphatase. 22. The method of claim 19, wherein said slurry is treated with acid phosphatase at a pH in the range of about 4.4 to about 4.6. 23. The method of claim 1, wherein said slurry is treated with acid phosphatase at a temperature ranging from about 20 to about 70 ° C. 24. The method of claim 23, wherein said slurry is treated with acid phosphatase at a temperature in the range of about 40 to about 55 ° C. 25. The method of claim 1, wherein the slurry is treated with acid phosphatase, wherein the acid phosphatase has an activity in the range of about 600 KPU / g curd solids to about 1400 KPU / g curd solids. . 26. The method of claim 1, wherein said slurry is treated with acid phosphatase, wherein said acid phosphatase is present in said slurry in an amount of about 0.1% to about 10%, based on the dry weight of said proteinaceous material. the method of. 27. The method of claim 26, wherein the slurry is treated with an acid phosphatase, wherein the acid phosphatase is present in the slurry in an amount of about 0.3% to about 5%, based on the dry weight of the protein material. the method of. 28. The method of claim 1, wherein said slurry is treated with acid phosphatase for about 30 minutes to about 4 hours. 29. The method of claim 28, wherein said slurry is treated with acid phosphatase for about 45 minutes to about 3 hours. 30. The method of claim 1, further comprising drying the treated and washed slurry to obtain a purified plant protein material. 31. The method of claim 1, further comprising the step of heat treating the treated slurry. 32. The method further comprises the step of treating the washed and acid phosphatase treated vegetable protein slurry with a protease enzyme at a temperature, pH, and time sufficient to hydrolyze the proteins in the slurry. Claim
Method according to 1. 33. The method of claim 32, wherein said protease enzyme is present in said slurry at a concentration of about 0.1% to about 10%, based on the dry weight of the protein in said slurry. 34. The method according to claim 32, further comprising a step of heat-treating the hydrolysis slurry. 35. The method of claim 32, further comprising the step of drying the hydrolyzed protein material after hydrolysis with the protease enzyme. 36. The method of claim 1, wherein said treating said vegetable protein material slurry with acid phosphatase and washing said treated slurry is effective in reducing the mineral content in said vegetable protein material. 37. A method for producing a purified plant protein material having low ribonucleic acid, phytic acid and phytate concentrations, comprising: forming an aqueous slurry of the plant protein material; Treating the ribonucleic acid, phytic acid and phytate in the plant protein material with a temperature, pH and time effective to degrade the ribonucleic acid, phytic acid and phytate in the plant protein material; The above method, comprising the step of producing a vegetable protein material comprising an acid and phytate. 38. The method of claim 37, wherein said plant protein material is a plant protein concentrate or a plant protein isolate. 39. The method of claim 38, wherein said plant protein material is a soy protein concentrate or a soy protein isolate. 40. The slurry comprises from about 2% to about 3% by weight.
38. The method of claim 37, wherein said method comprises 0% of said proteinaceous material. 41. The slurry comprises from about 5% to about 2% by weight.
42. The method of claim 40, wherein said method comprises 0% of said proteinaceous material. 42. The slurry comprises from about 10% to about 10% by weight.
42. The method of claim 40, wherein the method comprises 18% of the protein material. 43. The method of claim 37, wherein treating the slurry with the enzyme is effective to degrade most of the ribonucleic acids in the plant protein material. 44. The method of claim 43, wherein washing the enzyme-treated slurry is effective to remove the degraded ribonucleic acid to obtain a plant protein material from which most of the ribonucleic acid has been removed. Method. 45. The method of claim 37, wherein treating the slurry with the enzyme is effective to degrade at least 60% of the ribonucleic acids in the plant protein material. 46. The washing of the enzyme-treated slurry removes the degraded ribonucleic acid and removes at least 60% of the ribonucleic acid.
46. The method of claim 45, wherein said method is effective to obtain a plant protein material from which is removed. 47. The method of claim 37, wherein treating the slurry with the enzyme is effective to degrade at least 70% of the ribonucleic acids in the plant protein material. 48. The washing of the enzyme-treated slurry removes the degraded ribonucleic acid and removes at least 70% of the ribonucleic acid.
48. The method of claim 47, wherein said method is effective in obtaining a plant protein material from which is removed. 49. The method of claim 37, wherein treating the slurry with the enzyme is effective to degrade at least 80% of the ribonucleic acids in the plant protein material. 50. The washing of the enzyme-treated slurry is effective to remove the degraded ribonucleic acid and obtain a plant protein material from which at least 80% of the ribonucleic acid has been removed. The described method. 51. The method of claim 37, wherein treating the slurry with the enzyme is effective to degrade substantially all ribonucleic acids in the plant protein material. 52. The washing of the enzyme-treated slurry is effective for removing the degraded ribonucleic acid to obtain a plant protein substance from which substantially all of the ribonucleic acid has been removed.
52. The method of claim 51. 53. The method of claim 37, wherein treating the slurry with the enzyme is effective to degrade phytic acid and phytate in the plant protein material. 54. The washing of the enzyme-treated slurry,
54. The method of claim 53, wherein said degraded phytic acid and phytate are removed and effective to obtain a phytic acid and phytate-free plant protein material. 55. The method of claim 53, wherein treating the slurry with the enzyme is effective to degrade at least a majority of the plant protein material. 56. The washing of the enzyme-treated slurry is effective to remove the degraded phytic acid and phytate to obtain a vegetable protein material from which at least a majority of phytic acid and phytate have been removed. Claim 55
The method described in. 57. The method of claim 53, wherein treating the slurry with the enzyme is effective to degrade at least 75% of phytic acid and phytate in the plant protein material. 58. The washing of the enzyme-treated slurry removes the degraded phytic acid and phytate to remove at least 75% of phytic acid and phytate.
58. The method of claim 57, which is effective for obtaining a plant protein material. 59. The method of claim 53, wherein treating the slurry with the enzyme is effective to degrade at least 85% of phytic acid and phytate in the plant protein material. 60. The washing of the enzyme treated slurry removed the degraded phytic acid and phytate to remove at least 85% of phytic acid and phytate.
60. The method of claim 59, which is effective for obtaining a plant protein material. 61. The method of claim 53, wherein treating the slurry with the enzyme is effective to degrade substantially all phytic acid and phytate in the plant protein material. 62. The washing of the enzyme treated slurry is effective to remove the degraded phytic acid and phytate to obtain a plant protein material from which substantially all of the phytic acid and phytate have been removed. 62. The method of claim 61. 63. A slurry having a pH in the range of about 3 to about 6.
38. The method according to claim 37, wherein the method is treated with acid phosphatase and phytase. 64. The method of claim 63, wherein said slurry is treated with acid phosphatase and phytase at a pH in the range of about 3.5 to about 5.5. 65. The slurry, wherein the pH is in the range of about 4 to about 5.
64. The method according to claim 63, wherein the treatment is performed with an acid phosphatase and a phytase. 66. The method of claim 63, wherein said slurry is treated with acid phosphatase and phytase at a pH in the range of about 4.4 to about 4.6. 67. The method of claim 37, wherein the slurry is treated with acid phosphatase and phytase at a temperature in a range from about 20 to about 70 ° C. 68. The method of claim 67, wherein said slurry is treated with acid phosphatase and phytase at a temperature in the range of about 40 to about 55 ° C. 69. The slurry is treated with an enzyme formulation comprising acid phosphatase and phytase, wherein the enzyme formulation has an activity ranging from about 600 KPU / g curd solids to about 1400 KPU / g curd solids. 38. The method of claim 37, comprising: 70. The slurry is treated with an enzyme formulation comprising acid phosphatase and phytase, wherein the enzyme formulation has an amount of about 0.1% to about 10%, based on the dry weight of the proteinaceous material, in the slurry. 38. The method of claim 37, wherein 71. The method of claim 70, wherein said enzyme formulation is present in said slurry in an amount of about 0.3% to about 5%, based on the dry weight of said proteinaceous material. 72. The method of claim 37, wherein the slurry is treated with an enzyme formulation comprising acid phosphatase and phytase for about 30 minutes to about 4 hours. 73. The method of claim 72, wherein said slurry is treated with said enzyme formulation for about 45 minutes to about 3 hours. 74. The method of claim 37, further comprising drying the treated and washed slurry to obtain a purified plant protein material. 75. The method according to claim 37, further comprising a step of heating the enzyme-treated slurry. 76. The method according to claim 76, further comprising the step of treating the washed and enzyme-treated vegetable protein slurry with a protease enzyme at a temperature, pH, and time sufficient to hydrolyze the proteins in the slurry. Item 36. The method according to Item 37. 77. The method of claim 76, wherein said protease enzyme is present in said slurry at a concentration of about 0.1% to about 10%, based on the dry weight of the protein in said slurry. 78. The method of claim 76, further comprising the step of heat treating said hydrolyzed slurry. 79. The method of claim 76, further comprising the step of drying said hydrolyzed protein material after hydrolysis with said protease enzyme. 80. The method of claim 37, wherein the treating the vegetable protein material slurry with acid phosphatase and phytase and washing the treated slurry is effective in reducing the mineral content in the vegetable protein material. the method of.