JPH06240297A - Washing assistant containing immobilized enzyme - Google Patents

Abstract

PURPOSE:To stabilize the enzymatic activity by immobilizing a proteolytic enzyme and at least another digestive enzyme on a water-soluble organic high- molecular polymer. CONSTITUTION:A specified amount of a water-soluble organic high-molecular polymer such as a methyl vinyl ether/maleic anhydride copolymer is dissolved in a certain amount of a phosphate buffer solution to form a solution (A). A specified amount of a proteolytic enzyme such as alkali protease is dissolved in a certain amount of a phosphate buffer solution to form a solution (B). A specified amount of at least one digestive enzyme selected from among a fat or oil decomposition enzyme such as lipase, a starch decomposition enzyme such as amylase and like enzymes is dissolved in a certain amount of a phosphate buffer solution to form a solution (C). Equal volumes of solutions A, B and C are mixed with each other, and the resulting mixture is subjected to an enzyme immobilization reaction at 20 deg.C for about 48 hours to immobilize the enzymes simultaneously. Thus, the objective assistant having high enzymatic activity and high deterging power can be obtained.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to an enzyme-immobilized cleaning agent which is effective in maintaining the activity of the enzyme alone in the enzyme-containing cleaning agent, while at the same time preventing the cannibalization inactivation of the enzymes by proteolytic enzymes. It is related to agents.

[0002]

2. Description of the Related Art It is a well known fact that enzymes that decompose proteins and lipids have been excellent as cleaning aids, and enzyme-containing detergents containing these enzymes are used in various fields. Widely used in.

However, among these enzyme-containing detergents, especially those containing a proteolytic enzyme and another enzyme, the enzyme-containing liquid detergent loses the solution stability during long-term storage, and thus the modified enzyme is denatured. However, deactivation occurs, and the deactivated enzyme is more easily affected by the proteolytic enzyme, so that the deactivation proceeds and the detergency decreases. In a detergent solution containing proteolytic enzymes,
During soaking and washing, or during washing, the combined enzymes are deactivated by the proteolytic enzyme, and the enzyme activity is remarkably reduced, so that the detergency is lowered. There were drawbacks such as.

[0004]

DISCLOSURE OF THE INVENTION The present invention has the above-mentioned problems, namely, loss of an enzyme due to a proteolytic enzyme during storage when a proteolytic enzyme and another enzyme are mixed as a cleaning aid. An object of the present invention is to provide an enzyme-immobilized cleaning aid that solves the problems of activity, problems of deactivating enzymes due to proteolytic enzymes during immersion washing and washing, and problems of reduced detergency.

[0005]

To achieve the above object, the present invention has the following constitution.

That is, it is an enzyme-immobilized cleaning aid characterized in that a water-soluble organic polymer is immobilized with a protease and at least one kind of digestive enzyme other than the protease. .

The present invention provides the stability and detergency of an enzyme which cannot be achieved only by using a combination of a proteolytic enzyme and another enzyme.
This is made possible by immobilizing a proteolytic enzyme and a digestive enzyme on a water-soluble organic polymer.

When an enzyme is immobilized on a water-soluble organic polymer, high enzyme activity and stability are expressed, and at the same time, since the immobilized enzyme itself is water-soluble, a high affinity with a substance to be decomposed is to be maintained. You can

Further, the water-soluble organic polymer as the immobilization carrier is in a form in which the enzyme is released alone when the immobilization carrier itself is decomposed by proteolytic enzyme or other immobilizing digestive enzyme, Since other enzymes may be directly inactivated by the effect of proteolytic enzymes, the water-soluble organic polymer according to the present invention is a polymer that is not decomposed by proteolytic enzymes or other immobilized digestive enzymes. Is preferred.

The water-soluble organic polymer used in the present invention may be any water-soluble organic polymer, but among them, a methyl vinyl ether / maleic anhydride copolymer immobilizes an enzyme reaction. Is not decomposed by, and the binding between the enzyme and the immobilized carrier is easy,
It is preferably used because it is excellent from the viewpoint of high maintainability of enzyme activity after immobilization.

When used in combination with each of the proteolytic enzyme and at least one or more digestive enzymes other than the proteolytic enzyme immobilized alone, the proteolytic enzyme is immobilized but is released. In this state, the cannibalism and inactivation of the enzymes are likely to occur, but the proteolytic enzyme and at least one or more digestive enzymes other than the proteolytic enzyme are simultaneously immobilized on the water-soluble organic polymer to release the enzyme. The cannibalism and inactivation due to the above are less likely to occur, and the activity retention is greatly improved.

In the present invention, the use of a proteolytic enzyme as an enzyme to be immobilized decomposes a protein which acts as a binder of a stain substance in daily life stains and facilitates effective removal of stains on a fiber cloth. It is important because you can.

As a proteolytic enzyme, when washing,
Included are any proteolytic enzymes known to readily degrade dirt and stains. Suitable proteolytic enzymes among the proteolytic enzymes include alkaline proteases that exhibit high activity in the neutral to alkaline region.

The use of at least one type of digestive enzyme other than the proteolytic enzyme in the present invention is because most of the stain components related to daily life stains are stains of the biological system and food system, and these stains are removed. It is important because it is efficiently removed by a synergistic effect with a proteolytic enzyme.

The digestive enzyme is not particularly limited, and various lipases that hydrolyze fats and oils, amylases that hydrolyze starch and the like can be mentioned.

Since the proteolytic enzyme and other digestive enzymes are immobilized on the water-soluble organic polymer, the cleaning aid of the present invention having the above-mentioned constitution is modified or lost by conventional methods. By suppressing the activity, the inactivation of the compounded enzyme by the proteolytic enzyme, and the reduction of the detergency, it is possible to maintain high enzyme activity and detergency.

[0017]

EXAMPLES Next, the present invention will be described more specifically by way of examples.

The measuring method in the examples was carried out by the following method, and the measurement results are shown in Table 1. In the examples,% means% by weight.

(1) Enzyme Stability in Solution After standing for a certain period of time, it was calculated based on the residual activity of the digestive enzyme not decomposed by the proteolytic enzyme in the solution.

As the digestive enzyme, lipase B-2 (manufactured by Nagase Seikagaku Co., Ltd.) which is an oil and fat degrading enzyme (lipase), and spitase LH (manufactured by Nagase Seikagaku Corporation) which is a starch degrading enzyme (amylase) were used. , The activity per unit weight of these digestive enzymes alone is 100,
It was calculated as the relative activity of the enzyme in the washing solution per unit weight.

As one measure of the liquid stability, the turbidity of the washing liquid due to the enzyme (protein) which has been denatured and insolubilized after being left for a certain period is classified into four levels (⊚: not cloudy, ○: slightly). Turbid, △: Turbid, ×:
It was judged to be very cloudy).

(2) Detergency with an enzyme-immobilized cleaning aid A cloth was immersed in a contaminated liquid and air-dried to give a contaminated cloth.

An enzyme-immobilized cleaning aid was added to a commercial detergent (no added enzyme) cleaning solution, and after the contaminated cloth was washed for a certain period of time, the degree of stain remaining on the cloth was evaluated using a staining gray scale.

Condition Cloth: Polyester taffeta Warp, weft; 75 denier x 36 filament Weave density; Warp 98 x Width 84 / inch Unit weight; 64g / m ² Contamination liquid: Raw egg, potato starch against fresh egg 1 %,
Carbon black with 1% added to raw eggs Detergent concentration: Zab (manufactured by Kao Corporation) subjected to enzyme deactivation treatment at 100 ° C for 3 minutes 2g / l Enzyme-immobilized washing aid: proteolysis Based on the amount of enzyme,
Add proteolytic enzyme to 0.5g / l Washing condition: Wash at 40 ° C for 25 minutes, rinse for 10 minutes

Example 1 Proteolytic enzyme "Bioprase APL-30" (manufactured by Nagase Seikagaku Co., Ltd.) 20 g / l phosphate buffer solution Other digestive enzyme "lipiase B-2" (oil and fat degrading enzyme; Nagase Biochemical Industry) 20g / l phosphate buffer "Spitase LH" (starch-degrading enzyme; manufactured by Nagase Seikagaku Corporation) 20g / l phosphate buffer Water-soluble organic polymer "GANTREZ AN" (methyl vinyl ether / Maleic anhydride copolymer; manufactured by GAF, MW: 20,000) 80 g / l phosphate buffer is mixed in equal volumes, and the enzyme immobilization reaction is performed at 20 ° C. for 48 hours to obtain an enzyme immobilization washing aid. Obtained.

As shown in Table 1, the residual activity of the immobilized digestive enzyme was as follows:
It was as high as 85% and amylase: 80%, and no turbidity was observed in the washing solution.

Further, when the gray scale of residual stains after washing was judged to be grade 5, an enzyme-immobilized washing aid having a high washing effect was obtained.

Example 2 Proteolytic enzyme "Bioplase APL-30" (manufactured by Nagase Seikagaku Co., Ltd.) 15 g / l phosphate buffer solution Other digestive enzyme "lipiase B-2" (oil and fat degrading enzyme; Nagase Biochemical Industry) 15g / l phosphate buffer solution Water-soluble organic polymer "GANTREZ AN" (methyl vinyl ether / maleic anhydride copolymer; GAF, MW: 20,000) 60g / l phosphate buffer solution Were mixed in equal volumes and the enzyme immobilization reaction was carried out at 20 ° C. for 48 hours to obtain an enzyme-immobilized cleaning aid according to the present invention.

As shown in Table 1, the residual activity of the immobilized digestive enzyme was as follows:
It was as high as 85%, and no turbidity was observed in the washing solution.

As a result of the absence of amylase in the digestive enzyme, an enzyme-immobilized cleaning aid having a slightly low cleaning effect of 4 to 5 was obtained by the gray scale classification of residual stains after washing.

Example 3 Three types of enzyme solutions were prepared based on Example 1, and
As a water-soluble organic polymer, glutaraldehyde (25% aqueous solution): 320 g / l phosphate buffer was mixed in equal volumes, and the enzyme immobilization reaction was carried out at 20 ° C. for 48 hours. I got an agent.

As shown in Table 1, the residual activity of the immobilized digestive enzyme was as follows:
It was as high as 75% and amylase: 75%, and a little turbidity was observed in the washing solution.

Due to the difference in the water-soluble organic polymer, the activity of the immobilized digestive enzyme was reduced, so that the level of the residual stain after washing was graded as a grade 4 in the gray scale classification, and the enzyme immobilization was a little less effective. A cleaning aid was obtained.

Example 4 Two kinds of enzyme solutions prepared in accordance with Example 2 were mixed with glutaraldehyde (25% aqueous solution): 240 g / l phosphate buffer as a water-soluble organic high-molecular polymer in equal volumes. The enzyme immobilization reaction was carried out at 20 ° C. for 48 hours to obtain an enzyme-immobilized cleaning aid according to the present invention.

As shown in Table 1, the residual activity of the immobilized digestive enzyme was determined by lipase even after standing for a certain period (30 days):
It was as high as 75%, and the turbidity was a little observed in the cleaning solution.

Due to the difference in the water-soluble organic polymer, the activity of the immobilized digestive enzyme decreased and the amount of amylase not present in the digestive enzyme. As a result, an enzyme-immobilized cleaning aid having a fourth grade and a low cleaning effect was obtained.

Comparative Example 1 Proteolytic enzyme "Bioprase APL-30" (manufactured by Nagase Seikagaku Co., Ltd.) 10 g / l phosphate buffer solution Water-soluble organic polymer "GANTREZ AN" (methyl vinyl ether / maleic anhydride copolymer) Polymer; manufactured by GAF, MW: 20,000) 40 g / l phosphate buffer was mixed in equal volumes, and the enzyme immobilization reaction was performed at 20 ° C. for 48 hours to obtain an enzyme immobilization substance.

As shown in Table 1, no turbidity was observed in the washing solution even after standing for a certain period of time (30 days), but since there was no immobilized digestive enzyme, the gray scale classification of residual stain after washing was judged. In the case of 3, an enzyme-immobilized substance having a third grade and no cleaning effect was obtained.

Comparative Example 2 Proteolytic enzyme "Bioplase APL-30" (manufactured by Nagase Seikagaku Corporation) 30 g / l phosphate buffer water-soluble organic polymer "GANTREZ AN" (methyl vinyl ether / maleic anhydride) Acid copolymer; manufactured by GAF, MW: 20,000) 120 g / l phosphate buffer was mixed in equal volumes, and enzyme immobilization reaction was carried out at 20 ° C. for 48 hours (solution A).

On the other hand, other digestive enzymes “lipiase B-2” (oil and fat degrading enzyme; manufactured by Nagase Seikagaku Co., Ltd.) 30 g / l phosphate buffer water-soluble organic polymer “GANTREZ AN” (methyl vinyl ether / Maleic anhydride copolymer; manufactured by GAF, MW: 20,000) 120 g / l phosphate buffer was mixed in equal volumes, and enzyme immobilization reaction was carried out at 20 ° C. for 48 hours (solution B).

Further, starch degrading enzyme "Spitase LH" (manufactured by Nagase Seikagaku Corporation) 30 g / l phosphate buffer water-soluble organic polymer "GANTREZ AN" (methyl vinyl ether / maleic anhydride copolymer; GAF, MW: 20,000) 120 g / l phosphate buffer was mixed in equal volumes, and the enzyme immobilization reaction was carried out at 20 ° C. for 48 hours (solution C).

Mix the above three preparation liquids A, B and C,
An enzyme-immobilized substance was obtained.

As shown in Table 1, the residual activity of the immobilized digestive enzyme after standing for a certain period (30 days) was 6 in each case.
It decreased to 0%, and turbidity was observed in the cleaning liquid.

Although each enzyme is immobilized, the immobilized enzymes are in a state of being released from each other, so that the activity decreases due to cannibalism. An enzyme-immobilized substance having no grade and no washing effect was obtained.

Comparative Example 3 Proteolytic enzyme "Bioplase APL-30" (manufactured by Nagase Seikagaku Corporation) 15 g / l phosphate buffer Other digestive enzymes "lipiase B-2" (oil and fat degrading enzyme; Nagase Seibutsu) Chemical Industry Co., Ltd.) 15 g / l phosphate buffer “Spitase LH” (starch-degrading enzyme; Nagase Seikagaku Co., Ltd.) 15 g / l phosphate buffer mixed in equal volumes, totaling 30 ml, mixed enzyme solution Got

As shown in Table 1, the residual activity of the digestive enzyme was as low as 40% for lipase and 40% for amylase after standing for a certain period (30 days), and the washing solution was very cloudy.

Since the enzyme was not immobilized, the enzyme was largely inactivated by the proteolytic enzyme, and when the gray scale of residual stains after washing was judged to be the second grade, an enzyme-immobilized substance having no washing effect was obtained.

[0048]

[Table 1]

[0049]

EFFECTS OF THE INVENTION As is apparent from the above description, the present invention can prevent the inactivation of an enzyme by a proteolytic enzyme during storage when a proteolytic enzyme and another enzyme are mixed as a conventional washing aid. It has a remarkable effect on problems, such as problems of deactivating enzymes due to proteolytic enzymes during washing by dipping and washing, and problems of deterioration of detergency, and high enzyme activity and detergency can be maintained.

Claims (2)

[Claims] 1. An enzyme-immobilized cleaning aid characterized in that a water-soluble organic polymer is immobilized with a proteolytic enzyme and at least one or more digestive enzymes other than the proteolytic enzyme. 2. The enzyme-immobilized cleaning aid according to claim 1, wherein the water-soluble organic polymer is a copolymer of methyl vinyl ether and maleic anhydride.