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A strain of high-production rhamnogalacturonan hydrolase

A technology of galacturonic acid and rhamnose, applied in the field of microbial transformation, can solve the problems of unfavorable wide application, few strains, low yield and the like, and achieves the effect of promoting popularization and application and reducing production cost

Active Publication Date: 2019-11-08
QINGDAO VLAND BIOTECH GRP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, there are few bacterial strains producing rhamnogalacturonan hydrolase, and the yield is low, which is not conducive to the wide application of the enzyme. Therefore, it is important to screen microbial strains with high rhamnogalacturonan hydrolase production. significance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0029] (2.2) Preparation of sample solution:

[0030] Accurately draw 1ml of the enzyme solution, dilute it with 19ml of sodium acetate buffer, and use this solution as the mother solution, then dilute it with sodium acetate buffer to an appropriate multiple to measure the enzyme activity, so that the absorbance at 590nm is in the range of 0.2 to 1.4.

[0031] (2.3) Determination of enzyme activity:

[0032] Take 0.5ml of appropriately diluted enzyme solution in a test tube and place it at 40°C for 5 minutes;

[0033] Add AZCL-Rhamnogalacturonan tablet, let stand without stirring, after timing for 10 minutes, add 10ml of trisodium phosphate stop solution, vortex and immediately put it at room temperature;

[0034] Place at room temperature for 5 minutes, after cooling, vortex again, filter with Whatman No.1 (9cm) filter paper, and read at 590nm after filtration;

[0035] Add the stop solution to the enzyme solution in the blank tube first, and then add the AZCL-Rhamnogalactu...

Embodiment 2

[0042] Embodiment 2 mutagenesis screening

[0043] The applicant used Aspergiulls aculeatus (Aspergiulls aculeatus) DSM2344 as the starting strain, and further screened mutant strains with improved rhamnogalacturonate hydrolase production by ultraviolet mutagenesis.

[0044] 2.1 Determining the fatality rate

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PUM

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Abstract

The invention belongs to the technical field of microbial transformation, and specifically relates to a mutant strain of Aspergillus aculeatus with high production rate of rhamnogalacturonate hydrolase. The present invention uses Aspergiulls aculeatus DSM2344 as the starting strain, and obtains mutant strain Aspergiulls aculeatus T13 through screening by ultraviolet mutagenesis method, and its preservation number is CCTCC NO: M2017784. After 3.5 days of solid shake flask fermentation of the mutant strain, the enzyme activity of rhamnogalacturonate hydrolase reaches 63.7u / g, which is 41.6% higher than that of the original strain, and unexpected technical effects are achieved. The mutant strain of Aspergillus aculeatus provided by the invention can be widely used in the fermentative production of rhamnogalacturonan hydrolase, which is conducive to reducing the production cost of the enzyme and promoting the development of rhamnogalacturonan hydrolase in the food industry. promotion and application.

Description

technical field [0001] The invention belongs to the technical field of microbial transformation, and specifically relates to a mutant strain of Aspergillus aculeatus with high production rate of rhamnogalacturonate hydrolase. [0002] technical background [0003] The cell walls of higher plants are composed of cellulose, an intersupporting structure formed mainly of hemicellulose polymers, pectin matrix polymers, and spherical and non-spherical proteins. With the exception of globular proteins, all polymers play structural roles in the cell wall. As the cell wall matures, cross-links, interchain and intrachain connections are formed, and finally lignin is deposited on the cell wall, thereby making the cell wall very stable and making its components difficult to separate, digest, and handle. [0004] Pectin is a complex group of branched heteropolysaccharides that is abundant in the primary cell wall and in the interlayer between all plant cells. Pectin polymers are chemica...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14C12N9/24C12R1/66
CPCC12N9/2402C12Y302/01171C12N1/145C12R2001/66
Inventor 徐晓东周利伟张珍珍
Owner QINGDAO VLAND BIOTECH GRP