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Beta-1,3 endo-glucanase with immune-enhancing activity andfrom Arca inflata Reeve and coding polynucleotide of beta-1,3 endo-glucanase

A technology of endoglucanase and immune enhancement, which is applied in the direction of enzymes, hydrolases, and microbial-based methods, and can solve problems that have not been reported in research

Active Publication Date: 2019-03-01
于荣敏
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

According to relevant literature, foreign scholars have characterized and expressed a polypeptide with antibacterial activity and immunomodulatory activity from the clam, but the research on the endoglucanase of the clam has not been reported.

Method used

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  • Beta-1,3 endo-glucanase with immune-enhancing activity andfrom Arca inflata Reeve and coding polynucleotide of beta-1,3 endo-glucanase
  • Beta-1,3 endo-glucanase with immune-enhancing activity andfrom Arca inflata Reeve and coding polynucleotide of beta-1,3 endo-glucanase
  • Beta-1,3 endo-glucanase with immune-enhancing activity andfrom Arca inflata Reeve and coding polynucleotide of beta-1,3 endo-glucanase

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Embodiment Construction

[0037] 1. Separation and purification of β-1,3 endoglucanase from cockles

[0038] Shell the cockles, take the contents, wash three times with low-temperature distilled water, weigh after drying, add extraction buffer (pH=8.0, 0.03mol / L, sodium phosphate buffer) according to the weight ratio of 1:3, Using a high-speed tissue masher, homogenate at 8000×g intervals for 3 minutes until fine and lump-free, place the homogenate in a low-frequency ultrasonic instrument for ultrasonic extraction for 40 minutes, and use a low-temperature high-speed centrifuge at 10000×g, 4°C, centrifuge for 30 minutes to remove After measuring the volume of the supernatant, place it in an ice bath environment and add ammonium sulfate with 70% saturation, continue stirring and salting out for 1 hour, remove the precipitate by high-speed centrifugation, take the supernatant, add 100 % saturation of ammonium sulfate, continue stirring and salting out for 1 hour, centrifuge at 10000×g, 4°C for 30 minutes,...

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Abstract

The invention relates to endo-glucanase with immune-enhancing activity and nucleotide sequences of genes coding the endo-glucanase, and belongs to the technical field of biology. The invention provides a beta-1,3 endo-glucanase protein sequence and nucleotide sequences of genes of beta-1,3 endo-glucanase. Beta-1,3 endo-glucanase is one of key enzymes in a glucan hydrolytic enzyme system. The beta-1,3 endo-glucanase is from Arca inflata Reeve, 333 amino acids are included, the total length of corresponding coding gene nucleotide is 999 bp, the content of (G+C) is 39.34%, and a novel glucan hydrolytic enzyme resource is achieved. Engineered strains constructed by using the enzyme gene nucleotide can high-efficiently express the beta-1,3 endo-glucanase, a Kd value of the beta-1,3 endo-glucanase is 13.09 muM when laminarin is used as a substrate, produced enzymic preparations can be used for food industry, medicine industry, fodder industry, washing industry and other industries, the problem of recycling of biomass waste being rich in glucan can be solved, and rich glucosyl-oligosaccharides can be obtained to generate considerable economic benefits.

Description

technical field [0001] The invention relates to a clam-derived endoglucan hydrolase beta-1,3 glucanase with immunoenhancing activity and its encoding gene nucleotide, which belongs to the field of biotechnology, and the produced enzyme preparation is used for food , medicine, feed, washing and other industries, not only can solve the problem of reuse of dextran-rich biomass waste, but also can obtain rich dextran oligosaccharides to generate considerable economic benefits. Background technique [0002] Glucan is the main component of plant and microbial cell walls, accounting for 30% to 50% of the total dry weight of plants. It is the most widely distributed and abundant renewable carbon source compound on the earth, accounting for 40% of the total biomass of the earth. According to reports, in my country, there are as many as 500 million tons of glucan-rich biomass such as crop straw, rice stalks, and kelp every year in China, and the plant biomass produced by photosynthesi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/24C12N15/56C12N15/70C12N1/21C12R1/19
CPCC12N9/2402C12N15/70
Inventor 于荣敏李春磊朱建华
Owner 于荣敏
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