KR101368731B1 - A method of manufacturing D-galactose for use of the production of D-tagatose from whey permeate or dried whey permeate - Google Patents

Abstract

According to the present invention,
Removing the solid precipitate from the solution of whey membrane permeate or whey membrane permeate powder;
Removing the protein from the whey membrane permeate or the solution of the whey membrane permeate powder from which the solid precipitate is removed; And
A method for preparing galactose from a whey membrane permeate or whey membrane permeate powder solution, comprising removing ash, salt or both from the protein-removed whey membrane permeate solution or whey membrane permeate powder solution. It is about.
The present invention also relates to a method for producing tagatose, comprising the step of isomerizing galactose produced by the method to obtain tagatose.

Description

A method of manufacturing D-galactose for use of the production of D-tagatose from whey permeate or dried whey permeate}

The present invention relates to a method for producing galactose, which is a raw material for producing tagatose, from whey permeate or whey permeate powder (hereinafter referred to as 'DWP'). Mass production of galactose and tagatose produced from inexpensive raw materials by removing impurities, proteins (including salt) and other solid precipitates from whey membrane permeate or whey membrane permeate powder To provide technology.

There is an increasing demand for low calorie functional sweeteners to prevent sugar excess calories, obesity and tooth decay.

Tagatose is an ideal sugar replacement sweetener that can satisfy consumers' desires, with a sweetness almost equivalent to sugar (about 90%) or a low-calorie sweetener that is about 30% sugar.

In addition, tagatose is a stereoisomer of galactose, which is known to be used as a low-calorie sugar substitute sweetener having low metabolic rate and low absorption rate in the body, without side effects. (See JECFA Sixty-third meeting, Geneva, 8-17 June 2004-Food Additives)

Tagatose absorbs only about 20% of the intake from the small intestine, and the remaining 80% is transported to the large intestine where the intestinal microbes live, selectively producing lactic acid bacteria, thereby producing short-chain fatty acids. butyrate) has a prebiotic property that produces large amounts (up to 50% of total short-chain fatty acids). Tagatose is a low-calorie natural sugar of 1.5 kcal / g, and is recognized as the General Recognized As Safe (GRAS) by the US FDA, and is used as a functional sweetener in foods, beverages, health foods, and dietary additives.

However, since tagatose is not commonly present in nature but is a rare sugar contained in dairy products and some plants, a mass production technique must be developed from inexpensive raw materials to be used as a low-calorie functional sweetener.

U.S. Patent No. 5002612 and U.S. Pat. Provided are methods for preparing D-tagatose.

Therefore, the key intermediate of production using tagatose enzymatic method is galactose, and until now, the supply of galactose through the decomposition of lactose is the only direction of technology development.

However, the price of lactose in the world market is not consistent with rising prices in the market due to various factors that cannot be quantified, such as crude oil production, milk powder demand, and changes in lactose consumption in third countries. It has a unique price pattern that repeats its decline. Raw material price fluctuations in these markets make it difficult to supply stable tagatose raw materials.

According to the present invention, when galactose is obtained from whey membrane permeate or whey membrane permeate powder generated during the manufacturing process of whey isolate protein which can be supplied at low cost, it is possible to supply more stable raw materials and stabilize price of tagatose. As a result, significant added value can be obtained.

U.S. Patent No. 5002612 U.S. Pat.No.5078796

none

Currently, the production of galactose, a raw material for the enzymatic production of tagatose, is dependent on lactose. However, lactose supply is unstable and the price of lactose is high, which directly affects the manufacturing cost of tagatose. Therefore, the provision of lactose and galactose of more stable low-cost raw materials is a prerequisite for mass production of tagatose. In the present invention, as a result of efforts to develop a low-cost alternative raw material as a raw material for the production of tagatose, a technology that can economically utilize whey membrane permeate or whey membrane permeation powder, which is a by-product of the production of whey isolate protein, which is not highly industrialized. We want to develop

Whey membrane permeate or whey membrane permeate contains protein, ash (including salt), and other impurities in addition to carbohydrate (lactose) components. To use whey membrane permeate or whey membrane permeate as a raw material for tagatose There is a need for a process for economically and effectively removing the above lactose non-components.

In the present invention, the precipitate is removed by inducing a solidification reaction without changing the sugar content and generating a reaction by-product from the dissolved solution of the whey membrane permeate or the whey membrane permeate through physical and chemical pretreatment such as pH adjustment and heat treatment. Economical protein removal was achieved by using activated charcoal without using a conventional relatively expensive adsorption method or ion chromatography method to remove proteins from the dissolved whey membrane permeate or the dissolved solution of whey membrane permeate powder.

In addition, the whey membrane permeate or whey membrane permeate powder solution in which the solid precipitate and the protein are removed as described above may be removed by electrodialysis to remove salts, ash or both, and the method according to the present invention may be Pretreatment of the precipitate and protein removal has the advantage of minimizing the degree of contamination of the electrodialysis membrane and increasing the desalination efficiency during electrodialysis.

The present invention develops a process for effectively and economically removing proteins, ash (including salts), and other solid precipitates, which are components other than lactose in whey membrane permeate or whey membrane permeate powder, which are by-products in the production of whey isolate proteins. This overcomes the supply and cost dependence of refined lactose and / or crystalline lactose, which is a raw material for tagatose production at present, and has an effect of directly or indirectly improving the production cost of tagatose.

The present invention also provides a technology that can economically utilize whey membrane permeate or whey membrane permeation powder, which are by-products of the production of whey isolate proteins, which have relatively high industrial utilization.

1 is a flow chart showing the overall process of the method for producing galactose according to an aspect of the present invention.
Figure 2 shows the result of the solidified precipitate dry weight removed as a result of 20% (w / v) DWP solution dissolved in pH 6.8, heat treatment, centrifugation at 70 ℃ for 30 minutes.
Figure 3 shows the deproteination rate results of protein removal after each powder activated carbon (A to F) treatment.
Figure 4a is a graph showing the result of protein removal rate according to the temperature and time condition changes after treatment with activated carbon A in the powdered activated carbon of Figure 3, Figure 4b, after the treatment with activated carbon B in the powdered activated carbon of Figure 3, changes in temperature and time conditions It is a graph showing the result of protein removal rate according to.
5 is a photograph showing deproteinization and decolorization results when activated carbon A is treated in the powdered activated carbon of FIG. 3.
6 is a graph showing the electrical conductivity and the rate of change of current according to the change in electrodialysis operation time.

According to the present invention,

Removing the solid precipitate from the solution of whey membrane permeate or whey membrane permeate powder;

Removing the protein from the whey membrane permeate or the solution of the whey membrane permeate powder from which the solid precipitate is removed; And

A method for preparing galactose from a whey membrane permeate or whey membrane permeate powder solution, comprising removing ash, salt or both from the protein-removed whey membrane permeate solution or whey membrane permeate powder solution. It is about.

The method for producing galactose according to the present invention is based on whey membrane permeate or whey membrane permeate powder generated as a by-product during the manufacturing process of whey isolate protein, and in the case of whey membrane permeate, solid precipitates are removed without further treatment. Can be carried out. When whey membrane permeable powder is used as a raw material, it can be dissolved in solvent water to prepare a whey membrane permeable powder solution for processing thereof, and solid precipitates can be removed therefrom.

As used herein, the term 'whey separated protein' refers to whey protein separated and recovered through a membrane permeation process using whey as a raw material.

The term 'whey membrane permeate' as used herein refers to a membrane permeate that is a by-product that occurs during the process of preparing the 'whey separated protein'.

As used herein, the term 'whey membrane permeation powder' refers to a powder obtained by concentrating and drying the whey membrane permeate.

The step of removing the solid precipitate produces a solid precipitate from a solution of whey membrane permeate or whey membrane permeate powder by a heat treatment process of 40 to 90 ° C., adjustment to pH 3 to 9, or both, and the resulting solid The precipitate can be carried out by separating and removing the precipitate by a conventional method such as centrifugation. The heat treatment at 40 to 90 ° C., the adjustment to pH 3 to 9, is performed without changing the sugar content or generating the reaction byproduct in the solution of the whey membrane permeate or the whey membrane permeate powder. Precipitating the solid component in the powder solution has the effect of removing the solid component without affecting the content or quality of the final product galactose. The temperature of the heat treatment process is preferably 50 to 80 ℃, the pH adjustment process may be preferably pH 4 to 8.

In the present invention, as the heat treatment temperature is increased, the precipitation degree of the solid insoluble material which is not a protein or a carbohydrate (lactose) component increases, and the precipitation degree of the solid insoluble material may increase as the pH is increased. Therefore, as the heat treatment temperature is raised and the pH is raised, precipitation of the solid insoluble substance may be accelerated, and the solid precipitate may increase significantly. In a preferred embodiment, the solid precipitate removal step may be carried out by warming the temperature of the heat treatment process to 60 to 80 ℃, adjust the pH to 5 to 8.

In the method for producing galactose according to the present invention, the protein can be removed from the solution of the whey membrane permeate or the whey membrane permeate powder from which the solid precipitate is removed. The protein removal step is advantageous in that economical protein removal is possible by using activated carbon without using a conventional relatively expensive adsorption method or ion chromatography method.

KB-B이고, B는 Norit사의 NORIT

CGSP, C는 Norit사의 NORIT

CASP, D는 Norit사의 NORIT

CA1, E는 Norit사의 A51, F는 Norit사의 Norit

SX plus가 사용되었다. 더욱 특히 바람직하게는 단백질 제거 효율 측면에서 A 또는 B를 사용할 수 있다. In the present invention, the step of removing the protein by treatment with activated charcoal is to treat the dissolved solution of whey membrane permeate or whey membrane permeate powder from which the solid precipitate is removed with activated charcoal and react at 20 to 90 ° C. for 10 minutes to 9 hours. It may include. The activated carbon may be preferably powdered activated carbon, for example, A to F, but is not limited thereto. Activated carbon A specified in the present invention is Norit DARCO

KB-B, B is NORIT of Norit Corporation

CGSP, C is Norit's NORIT

CASP, D is Norit's NORIT

CA1, E are Norit's A51, F is Norit's Norit

SX plus was used. More particularly preferably A or B can be used in terms of protein removal efficiency.

The activated carbon treatment conditions may be preferably reacted for about 3 hours to 8 hours at a temperature of 50 to 80 ℃, more particularly preferably about 4 hours to about 7 hours at a temperature of 55 to 75 ℃. .

In general, the longer the treatment time, the higher the reaction temperature, the higher the protein removal rate.

In the protein removal step, further decolorization may be performed together.

The method for producing galactose according to the present invention may further include removing ash, salts, or both from the whey membrane permeate from which the protein is removed or the solution of the whey membrane permeate powder. The removal of the ash, salt or both may preferably be carried out by electrodialysis. In the present invention, since the solid precipitate removal step and the protein removal step are performed before the electrodialysis, the degree of contamination of the electrodialysis membrane is minimized, thereby extending the life and improving the salt removal efficiency.

In the production method according to the present invention, lactose decomposition is performed in a solution of whey membrane permeate or whey membrane permeate powder before or after any one of the solid precipitate removal step, protein removal step, and ash and / or salt removal step. The addition of the enzyme may further comprise the step of producing galactose. Preferably, the step of adding the lactose degrading enzyme may be performed after the solid precipitate is removed and before the protein removal step.

LX 5000, 혹은 Maxilact

LG 5000등이 있다. The lactose degrading enzyme (Lactase) can use a known enzyme without limitation, for example, DSM's Maxilact

LX 5000, or Maxilact

LG 5000 and so on.

The concentration of the enzyme added in the step of adding lactose degrading enzyme is preferably 0.01 to 5 w / v%, it can be hydrolyzed through the enzyme using a fermenter of 35 to 39 ℃.

The process according to the invention may further comprise ion purification, concentration and chromatographic separation processes after the salt and / or ash removal step. Specifically, ion purification and concentration processes are included to separate, purify and recover galactose using SMB (Simulated Moving Bed) chromatography. Cation and anion purification chromatography, which typically uses ion removal, may be used, and may also be included before SMB chromatography separation and purification through a concentration process commonly used by those skilled in the art. Another aspect of the invention relates to a process for producing tagatose, comprising the step of isomerizing galactose produced by the method described above to obtain tagatose.

Hereinafter, the configuration and operation of the present invention through the preferred embodiment of the present invention will be described in more detail. It is to be understood, however, that the same is by way of illustration and example only and is not to be construed in a limiting sense.

Details that are not described herein will be omitted since those skilled in the art can sufficiently infer technically.

<Examples>

Example 1 Component Analysis of Whey Membrane Permeable Powder (DWP)

A component analysis in whey membrane permeable powder, a by-product generated during the manufacturing process of whey isolate protein, was performed to confirm the content and type of impurities in the raw materials. The results of the component analysis in the whey membrane permeable powder are shown in Table 1 below. It has a lactose content of over 80% in whey membrane permeable powder and contains proteins, fats, various inorganic salts and heavy metals in addition to sugars. The remaining components except the sugar component are to be removed using physical and chemical pretreatment.

Total ingredients Total carbohydrates 86.9% Total protein 3.6% Total fat 0.2% moisture - Ash 9.5% salt Calcium ions 742.99 mg / 100 g Magnesium ions 135.54 mg / 100g Sodium ions 562.99 mg / 100 g Potassium ions 2231.12 mg / 100g iron 0.22 mg / 100 g sign 113.1 mg / 100 g zinc 0.05 mg / 100 g Copper 0.03 mg / 100 g manganese 0.01 mg / 100 g Selenium 36.23 ug / 100g Party Lactose 85.1% Galactose 1.6%

[Based on% TS]

Example 2 Pretreatment Process for Removing Insoluble Matter in Whey Membrane Permeable Powder (DWP)

Whey membrane permeable powder (DWP) 4g of Hilmar Corporation was suspended and dissolved (~ 20%, w / v) in 20 ml of solvent water to prepare a whey membrane permeable powder solution. Physical and chemical pretreatment of the DWP solution was intended to remove only impurities by inducing solid insoluble precipitation without changing sugar content and generating reaction byproducts in DWP. In order to evaluate the removal of insoluble matter in the DWP solution according to the heat treatment and pH condition change, the prepared whey membrane permeate powder solution was heat-treated at 50-80 ° C. for 1 hour. In addition, in the case of sample preparation for each pH condition, the suspension was titrated to pH 4.0-8.0 with HCl and NaOH, and then treated at room temperature (˜25 ° C.) for 1 hour. Centrifugation was performed to recover the supernatant, pH (pH meter), conductivity (conductivity meter), Brix (Brix meter), lactose (HLC composition and quantitative analysis) and protein content (Bradford Protein Assay) Protein content quantitative analysis), the change in the degree of precipitation production was analyzed (Table 2).

As can be seen in Table 2, there was no change in lactose content and other protein content, Brix, pH, and conductivity according to the heat treatment (50-80 ° C.) temperature change. There was no significant change in lactose content, protein content, Brix, pH, and conductivity according to pH 4-8. The results show that there is no change in sugar content in the DWP solution upon pretreatment of the heat treatment temperature and pH condition changes, and also no protein denaturation or reaction byproducts.

However, as the heat treatment temperature increased, the precipitation of solid insoluble matter, not protein and carbohydrate (lactose) components, increased, especially the pH of 20% (w / v) DWP solution was increased. As a result, the precipitation of the insoluble material was accelerated, and the degree of precipitation of the insoluble material was significantly increased after centrifugation. In consideration of the enzymatic hydrolysis conditions of 20% (w / v) DWP solution, the temperature was adjusted to pH 6.8, followed by heat treatment and centrifugation at 70 ° C. for 30 minutes. (Dry precipitate weight / whey membrane permeate powder weight) could be removed (see FIG. 2).

Example 3 Enzymatic Hydrolysis Process

LG 5000을 사용하였다. 상기 실시예 2에서 전처리된 시료와 1%(w/v) 농도의 상기 효소를 넣고 38 ℃, 100rpm으로 조절된 발효조를 이용하여 효소가수분해를 실시하였다. Maxlact, a commercial enzyme for the enzymatic hydrolysis of lactose in DWP solution

LG 5000 was used. The enzyme was hydrolyzed using a fermenter adjusted to 38 ° C. and 100 rpm after putting the pretreated sample and the enzyme at a concentration of 1% (w / v) in Example 2.

To determine the degree of hydrolysis, the corresponding sample was taken at 0, 10, 24, 31, 48 and 58 hours after enzymatic hydrolysis was started to calculate the content of glucose and galactose produced. The content of glucose and galactose was quantitatively calculated using an HPLC equipped with a BioRad Aminex HPX-87C column.

Reaction condition Reaction time
(hr) Lactose content
(g / L) Glucose
(g / L) Galactose
(g / L) Total sugar content
(g / L) digestibility
(%) 20L DWP Solution
(200g DWP / L Water)
0 227.5 227.5 10 28.4 97.4 78.7 204.5 77.4 24 13.1 104.6 97.8 215.5 89.0 31 10.5 106.3 102.3 219.1 91.7 48 7.4 108.1 107.5 223.1 94.8 58 6.9 109.6 109.8 226.3     96.5

Example 4 Protein Removal and Decolorization in Whey Membrane Permeate Powder (DWP) Solution

We tried to remove protein and chromatic material through activated carbon treatment of DWP solution. In order to evaluate the possibility of protein removal after treatment with powdered activated carbon of each of A to F, DWP was suspended and dissolved in water to ˜20% (w / v), and the dissolved solution was titrated with NaOH at pH 6.8. Heat treatment was performed at about 70 ° C. for 30 minutes, and the supernatant was recovered after centrifugation and treated with 6 kinds of powder activated carbon (2 g powder activated carbon / 20% DWP solution L). After the reaction at room temperature and 50 ° C. for a certain time, the supernatant was recovered by centrifugation, and then the protein content change was quantitatively analyzed and the results are shown in FIG. 3.

KB-B 및 B로 명시한 Norit사의 NORIT

CGSP가 단백질 제거 효과 면에서 가장 우수한 결과를 나타내었다. 또한, 대부분의 활성탄에서 50℃에서 반응시간이 증가함에 따라 탈단백율이 증가하는 경향을 보였다. As can be seen in Figure 3, all six activated carbons of Company D A to E showed a protein removal effect, among which A DARCO Norit

Norit's NORIT as KB-B and B

CGSP showed the best results in terms of protein removal effect. Also, in most activated carbons, the deproteination rate increased with increasing reaction time at 50 ° C.

After treatment with powdered activated carbon, DWP was suspended and dissolved in water to ˜20% (w / v) in order to optimize protein removal rate and decolorization according to temperature and time conditions, and then the prepared solution was titrated with NaOH to pH 6.8, After heat treatment at 30 ° C. for 30 minutes, centrifugation, and supernatant recovery, the mixture was treated with two kinds of activated carbon A and B (2 g powder activated carbon / 20% DWP solution L) and reacted at 40-70 ° C. for 0-6 hours. . After centrifugation of the reaction solution, the supernatant was recovered, and quantitative analysis of protein content changes and qualitative evaluation of color change were performed (FIGS. 4 and 5).

As can be seen in Figure 4, after A activated carbon treatment (2g powder activated carbon / 20% DWP solution L) after about 6 hours at 60-70 ℃ reaction showed 43-44% deproteination rate, B activated carbon treatment (2g powder activated carbon) / 20% DWP solution L) showed a 40% deproteination rate when reacted at 60-70 ° C. for about 6 hours. In addition, it is shown in Figure 5 that the A-activated carbon treatment has a decolorizing effect.

Example 5 Desalting Electrodialysis

When the lactose hydrolyzate prepared by the deproteinization process of Example 4 was purified through a desalination electrodialysis apparatus, the economic efficiency of the desalination electrodialysis process such as the desalination rate of ions and the loss rate of organic sugars was examined. For this purpose, energy consumption and pollution level were investigated.

The electrodialysis machine uses a 3-compartment type device, and detailed electrodialysis machine specifications and operating conditions are shown in Table 4 below.

division Contents Device Micro acilyzer s3 Total effective film area 0.55 dm2 / cell cartridge AC-220-550 electrode Ti / Pt (Anode)
SUS316L (Cathode) Operating voltage 9V-12V Operating temperature 23 ℃ -32 ℃ Sample liquid 500 ml Electrode solution 5% Na ₂ SO ₄

As the ion exchange membrane, commercialized products (CMX, AMX, Astom, Japan) were used, and the characteristics of the ion exchange membrane used in the present invention are shown in Table 5 below.

Item Cation Exchange Membrane (CMX) Anion exchange membrane (AMX) Kinds Strongly acidic cation permeability Strong basic anion permeability Characteristic High mechanical strength (Na-form) High mechanical strength (Cl-form) Electrical resistance (Ω) 1.8 to 3.8 2.0-3.5 Burt strength ≥0.40 ≥0.30 Thickness (mm) 0.14-0.20 0.12-0.15

While measuring the conductivity value of the electrodialysis device in real time, when the conductivity value of the raw water falls to the desired value, the batch operation was performed continuously without chemical cleaning as a batch operation condition to stop the power supply to the stack of the electrodialysis device. (Water quality data of raw water and treated water), FIG. 6].

Item enemy Treated water Removal rate (%) pH 6.8 6.2  - Brix (%) 15.8 12.6 20.0 Electrical conductivity (㎲ / ㎝) 15600 300 98.10 Protein content (%) 0.23 0.18 21.74 Crude Fat Content (%) 0.20 0.15 25.00

Continuous operation
[batch count] Operating voltage
[V] Desalination Time
[min] Yield
[%] Ion removal rate
[%] Desalination Rate
[LMH] Energy consumption
[Wh] 1 time 9-12 56 77 98.1 9.9 6.9 Episode 2 9-12 50 80 97.6 11.3 4.6 3rd time 9-12 50 79 98.0 11.2 4.3 4 times 9-12 50 82 97.8 11.3 4.8 5 times 9-12 50 81 98.1 11.2 4.9

Table 7 shows the results of continuously operating the deproteinated lactose hydrolyzate without chemical cleaning.

Ion removal rate = 100-{(raw water electrical conductivity value / treated water electrical conductivity value) * 100}

As a result, the desalination rate of ions was 98% on average and the recovery rate of sugar was 80% on average.

It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present invention as defined by the following claims. As will be understood by those skilled in the art. It is therefore to be understood that the embodiments described above are in all respects illustrative and not restrictive.

Claims (12)

Removing a solid precipitate from a solution of whey membrane permeate or whey membrane permeate powder generated as a by-product during the production of whey isolate protein;
Removing the protein from the whey membrane permeate or the solution of the whey membrane permeate powder from which the solid precipitate is removed; And
A method for producing galactose from whey membrane permeate or whey membrane permeate powder solution, comprising removing ash, salt or both from the protein-removed whey membrane permeate solution or whey membrane permeate powder solution.
The solid precipitate of claim 1, wherein the step of removing the solid precipitate is carried out from a solution of whey membrane permeate or whey membrane permeate powder by a heat treatment process of 40 to 90 ° C., adjustment to pH 3 to 9, or both. Generating and removing it.
The method of claim 2, wherein the solid precipitate is an insoluble substance other than sugar in the group consisting of proteins, fats, salts, heavy metals, and sugars.
The method of claim 1, wherein removing the protein comprises treating activated carbon and reacting at 20 to 90 ° C. for 10 minutes to 8 hours.
The method of claim 4, wherein further decolorization is performed in the protein removal step.
The method of claim 4, wherein the activated carbon is powdered activated carbon.
The method of claim 1, wherein the removal of ash, salt, or both comprises performing by electrodialysis.
8. The method of any one of claims 1 to 7, further comprising adding lactose degrading enzyme to the solution of whey membrane permeate or whey membrane permeate powder to produce galactose.
The method of claim 8, wherein the reaction temperature and reaction time of the galactose production step are 35 to 39 ° C., and 50 to 60 hours, respectively.
A method for producing tagatose, comprising the step of isomerizing galactose produced by the method according to claim 8 to obtain tagatose.
8. The method of claim 1, further comprising ion purification, concentration, and chromatographic separation processes after removing the ash, salt, or both. 9.
Method for producing tagatose, comprising the step of isomerizing galactose produced by the method according to any one of claims 1 to 7 to obtain tagatose.

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