Protease composite enzyme and application thereof

A protease and compound enzyme technology, applied in the protease compound enzyme and its application fields, can solve the problems of decreased wool strength, not using chemical reagents, low market prospect, etc. Effects of skin damage and avoidance of environmental pollution

Inactive Publication Date: 2011-01-05
DONGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the development and application of wool fabric treatment process is mainly based on protease. Ordinary protease cannot treat the disulfide bonds rich in the outer scale layer of wool fiber, so redox pretreatment is necessary.
However, after chemical treatment, protease re-treatment of wool leads to a sharp decrease in wool strength. The main mechanism for this is that the hydrolysis of wool by protease is a relationship of "internal and external competition". Enzymes not only act on the surface of wool, but also penetrate into it. Into the interior of the wool fiber, the final result is that the strength of the wool fiber is greatly reduced
Due to the special stability of the disulfide bond in the scale layer of the outer layer of wool fiber, the effect of using conventional protease is not obvious, and sometimes it has no effect at all (Fan Zenglu et al., "Research on the Mechanism of Protease on Wool", 2002, "Acta Textile Science 》), so at present, the enzymatic treatment of woolen fabrics at home and abroad often adopts chemical pretreatment, which not only fails to achieve the purpose of not using chemical reagents, but also destroys the internal structure of woolen fabrics, and its actual application effect is still not ideal
In the fifth issue of "Jiangsu Silk" in 2006, the article "Application of protease in wool textile fiber modification" pointed out that pretreatment (oxidation or reduction) is a necessary step to modify wool textile with protease, and no pretreatment has been found so far. Protease Achieving Anti-felting Effect of Wool Fabric
People have also developed a resin finishing process to improve fiber strength, but this process causes serious environmental pollution and has not been widely used in practice.
Jason Patrick McDevitt disclosed in the patent US006051033A that the strength of wool is reduced by protease treatment, and then the peptide bond broken between the connecting proteins is treated with transglutaminase, thereby improving the protein fiber strength, but this process increases the processing flow and improves the Processing costs, the market prospect is not great, so there is no large-scale promotion and application at present

Method used

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  • Protease composite enzyme and application thereof
  • Protease composite enzyme and application thereof
  • Protease composite enzyme and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Stenotrophomonas maltophilia (Stenotrophomonas maltophilia) DHHJ (CGMCC No.2231) (a strain producing keratinase and its application, the application number is disclosed in 2007101719394) is used as the production strain, and the following medium is used for cultivation : Wool powder 2g, KCl 0.05g, NaCl 0.05g, disodium hydrogen phosphate 0.5g, potassium dihydrogen phosphate 0.05g, water 100ml, pH7.5, placed in a 500mL Erlenmeyer flask, at 40°C and 150rpm, Culture solution was obtained after constant temperature cultivation for 72 hours. Ultrasound (150W, 3 minutes) to break the wall, filter the bacteria, add 10%-40% ammonium sulfate, stir to dissolve, place at 4°C for 12 hours, centrifuge at 10000rpm for 10-20 minutes to obtain a precipitate, add 0.05mol / L pH7.8 Tris-HCl 100mL dissolved, lyophilized after dialysis to obtain protease compound enzyme enzyme powder;

[0046] After centrifugation, the protease activity was 14U / mL, and the disulfide bond reductase activity w...

Embodiment 2

[0049] Streptomyces olivaceus (Waksman) Waksman et Henrici) (CGMCCNo.4.943) (purchased from China Committee for the Collection of Microorganisms, Beijing) was used as the production strain, and the following medium was used for cultivation: soluble starch 2g, wool powder 1g, MgSO 4 ·7H 2 O 0.05g, KCl 0.05g, FeSO 4 ·7H 2 O 0.001g, K 2 HPO 4 0.05g, 100ml of water, pH 6.5, placed in a 500mL Erlenmeyer flask, under the conditions of 28°C and 150rpm, cultured at a constant temperature for 72 hours to obtain a culture solution. After centrifugation, the supernatant (extracellular fluid) was obtained, the protease activity was 12U / mL, and the disulfide bond reductase activity was 5U / mL. Utilize the extracellular enzyme lyophilized powder obtained from the fermentation broth, use it to treat wool fabric: lyophilized enzyme powder is dissolved in 0.05mol / L PBS (pH7.8) with 0.5%, bath ratio 1: 25, 50 ℃ for 70 minute.

[0050] Felt shrinkage (%)=(1-area of ​​wool fabric after trea...

Embodiment 3

[0056] Aspergillus carneus (Van.Teighem) Blochwitz (CGMCC No.3.3920) (purchased from China Committee for the Collection of Microorganisms, Beijing) was used as the production strain, and the following medium was used for cultivation: sucrose 2.0g, wool powder 1g, MgSO 4 ·7H 2 O 0.05g, KCl 0.05g, FeSO 4 ·7H 2 O 0.001g, K 2 HPO 4 0.1g, 100ml of water, pH 6.2, placed in a 500mL Erlenmeyer flask, under the conditions of 28°C and 150rpm, cultured at a constant temperature for 72 hours to obtain a culture solution. After centrifugation, the bacteria were obtained, and the wall was broken by ultrasonic wave (150W, 3 minutes) to obtain the intracellular enzyme, the protease activity was 7U / mL, the disulfide bond reductase activity was 8U / mL, and ammonium sulfate was added 10%-40% , stirred and melted, placed at 4°C for 24 hours, centrifuged at 10,000 rpm for 10 minutes to obtain a precipitate, added 0.05mol / L Tris-HCl 100mL with pH 7.8 to dissolve, dialyzed and freeze-dried to ob...

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Abstract

The invention relates to a protease composite enzyme and application thereof. A preparation method comprises the following steps of: inoculating microbes into a culture medium, placing the culture medium on a shaker at the temperature of between 25 and 40 DEG C and the rotating speed of 120 to 200 revolutions per minute to culture the microbes for 24 to 96 hours, performing enzyme or ultrasonic wall breakage or non wall breakage treatment, filtering bacteria, adding ammonium sulfate into the culture medium to settle protein, centrifuging the solution, adding Tris-HCl into the solution for dissolution, and freeze-drying the sediment after dialysis; or inoculating microbes into the culture medium, fermenting the culture medium for 23 to 72 hours at the temperature of 30 DEG C, culturing the microbes for 24 to 72 hours at the stirring speed of 120 to 600rpm, stopping fermentation, and filtering or centrifuging bacteria. The composite enzyme can be directly acted on various wool spinning fabrics and saves chemical reagent pretreatment and oxidation pretreatment in the conventional treatment process; the application method is simple and suitable for industrialized production; and the felting resistance and mechanical strength of the treated wool or wool fabrics are improved, and the felting rate of the treated wool fabric is less than 5 percent.

Description

technical field [0001] The invention belongs to the field of compound enzyme and its application, in particular to a protease compound enzyme and its application. Background technique [0002] Wool fabric is a high-grade product that is popular among people. It has advantages that other fiber fabrics cannot match, and its inherent properties make wool fabric a high-grade textile. Hand feel and dyeing performance are important characteristics of wool. However, the outer layer of wool has a dense and hydrophobic scale layer that is difficult to handle, which affects many indicators such as wool feel and dyeing and coloring. People have developed various processes to remove the scale layer for a long time. In the existing factory treatment process, chemical means are the main method, such as oxidizing the wool surface with chlorine water to improve the dyeing affinity or softening agent to improve the feel and characteristics. This treatment is a surface treatment because the ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/52C12N9/62D06M16/00C12S11/00C12R1/38C12R1/01C12R1/58C12R1/66D06M101/12
Inventor 曹张军蔡少博黄铮华张兴群周美华洪枫
Owner DONGHUA UNIV
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