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Multi-enzyme complex capable of efficiently degrading gluten protein and application of multi-enzyme complex

A technology of gluten protein and complex, which is applied in the field of multi-enzyme complexes that efficiently degrade gluten protein, can solve the problem that gluten protein is difficult to be completely hydrolyzed, achieve effective degradation, efficient degradation, and reduce safety risks

Pending Publication Date: 2022-05-06
NANCHANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the problem that gluten protein is difficult to be completely hydrolyzed in the background technology, the object of the present invention is to provide a multi-enzyme complex for efficiently degrading gluten protein and its application

Method used

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  • Multi-enzyme complex capable of efficiently degrading gluten protein and application of multi-enzyme complex
  • Multi-enzyme complex capable of efficiently degrading gluten protein and application of multi-enzyme complex
  • Multi-enzyme complex capable of efficiently degrading gluten protein and application of multi-enzyme complex

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: the preparation of bacillus cereus multienzyme complex

[0028] (1) In SCB solid medium (starch 10g·L -1 , Casein 3.0g·L -1 、KNO 3 2.0g·L -1 , NaCl2.0g·L -1 、K 2 HPO 4 2.0g·L -1 , MgSO 4 0.05g·L -1 , CaCl 2 0.02g·L -1 , FeSO 4 0.01g·L -1 , pH value 7.2) was inoculated with Bacillus cereus AFA01, cultured at 37°C for 2 days, the cells were collected, and the cells were resuspended with 50 mM Tris-HCl (pH 8.5).

[0029] (2) Suspend the cells and sonicate on ice (300W, 2s, interval 3s, total time 15min), add CHAPs to make the final content reach 1.2% (w / v), place at 4°C for 1h, mix every 10min. Then centrifuge at 12000g for 10min, take the supernatant, and pass through a 0.22μm filter membrane.

[0030] (3) The DEDA FF anion exchange column was equilibrated with 20 mM Tris-HCl buffer (pH 8.5, containing 0.5% w / v CHAPs). After loading the sample, carry out gradient elution with 0-0.6M NaCl, and collect the active components.

[0031] (4) T...

Embodiment 2

[0035] The gliadin decomposition experiment was carried out with the multi-enzyme complex obtained in Example 1.

[0036] The concentration of gliadin is 0.5 mg / mL, the concentration of multi-enzyme complex is 0.01 mg / mL (0.01‰), the enzymatic hydrolysis temperature is 37°C, and the pH of the buffer is adjusted to 7.0. Sampling was used to determine the change of gliadin, and the detection method was referred to "Research on the Detection Method of Zein" published by Liu Zhenyu et al. see results figure 1 , it can be found from the figure that the use of multi-enzyme complex can reduce the gliadin content to 5% within 30 minutes, and reduce the gliadin content to below 2% in 120 minutes.

Embodiment 3

[0038] The celiac disease peptide decomposition experiment was carried out with the multi-enzyme complex obtained in Example 1.

[0039] The celiac peptide (33-mer) concentration was 0.5 mg / mL, the multi-enzyme complex concentration was 0.005 mg / mL, the enzymatic hydrolysis temperature was 37°C, and the buffer was adjusted to pH 7.0. Samples were taken to determine the degradation of 33mer, and the detection method was referred to "Detection of serum peptides in patients with lung squamous cell carcinoma by MALDI-TOF-MS and analysis of its correlation with chemotherapy efficacy" published by Zhao Guanhua et al. ("Chinese Journal of Lung Cancer", Issue 5, 2017). see results figure 2 , it can be seen from the figure that the multi-enzyme complex can degrade all the complete 33-mer within 5 minutes, and the remaining polypeptides are basically less than 9 peptides at 30 minutes, and the celiac T cell epitope in the 33-mer is destroyed.

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Abstract

The invention provides a multi-enzyme complex capable of efficiently degrading gluten protein. The multi-enzyme complex is a multi-enzyme complex derived from bacillus cereus AFA01. The multi-enzyme complex not only can efficiently degrade gluten protein, but also can efficiently degrade celiac disease causing peptides and destroy T cell epitopes in the celiac disease causing peptides. The enzyme preparation prepared from the multi-enzyme complex can be used as an oral enzyme preparation for assisting human body in digesting gluten protein and is used for assisting in treating celiac disease; the strain can also be used in food processing, can more effectively degrade gluten protein, and is expected to provide safe and reliable hypoallergenic food for celiac disease patients.

Description

technical field [0001] The invention relates to the technical fields of enzyme preparation and food, in particular to a multi-enzyme complex for efficiently degrading gluten protein and its application. Background technique [0002] Wheat is a food crop widely grown in the world, and it is also one of the eight common food allergens reported by FAO (1995). It can cause a variety of gluten-related diseases, among which celiac disease is a chronic small intestinal inflammatory disease caused by gluten, with a global incidence of about 1%. Celiac disease has seriously affected the quality of life of some people, even endangering their lives, and has become one of the unavoidable global food safety problems. [0003] There is no specific treatment for celiac disease, and the only reliable treatment is a lifelong strict gluten-free diet. Therefore, the processing and preparation of low-allergenic wheat products has received much attention. At present, commonly used desensitiza...

Claims

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Application Information

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IPC IPC(8): C12N9/00C12N1/20A61K38/54A61P1/00A23L5/20A23L33/195C12R1/085
CPCC12N9/00C12N1/20A61K38/54A61P1/00A23L5/25A23L33/195A23V2002/00A23V2200/30A23V2250/546
Inventor 陈红兵袁娟丽芦军武涌高金燕李欣佟平孟轩夷谢彦海
Owner NANCHANG UNIV
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