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Strain for producing ratgalacturonorhamnan hydrolase at high yield

A technology of galacturonic acid and rhamnose half, which is applied in the field of mutant strains of Aspergillus aculeatus, can solve the problems of few strains, unfavorable wide application, low yield and the like, and achieves the effects of reducing production costs and promoting popularization and application.

Active Publication Date: 2018-05-04
QINGDAO VLAND BIOTECH GRP +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] 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
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Effect test

preparation example Construction

[0028] (2.2) Preparation of sample solution:

[0029] 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.

[0030] (2.3) Determination of enzyme activity:

[0031] Take 0.5ml of appropriately diluted enzyme solution in a test tube and place it at 40°C for 5 minutes;

[0032] 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;

[0033] 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;

[0034] Add the stop solution to the enzyme solution in the blank tube first, and then add the AZCL-Rhamnogalactu...

Embodiment 2

[0041] Embodiment 2 mutagenesis screening

[0042] 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.

[0043] 2.1 Determining the fatality rate

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Abstract

The invention belongs to the technical field of microbial transformation and particularly relates to an Aspergiulls aculeatus mutant strain for producing ratgalacturonorhamnan hydrolase at high yield.According to the Aspergills aculeatus mutant strain disclosed by the invention, Aspergiulls aculeatus DSM2344 is used as an original strain and an ultraviolet mutagenesis method is adopted to selectfor obtaining a mutant strain Aspergiullus aculeatus T13 with a preservation number of CCTCC NO: M2017784. After a mutant strain solid is subjected to solid shake flask fermentation for 3.5 days, theenzyme activity of the ratgalacturonorhamnan hydrolase reaches 63.7u / g, which is increased by 41.6 percent compared with that of an original strain; unexpected technical effects are achieved. The Aspergiulls aculeatus mutant strain provided by the invention can be widely applied to fermentation production of the ratgalacturonorhamnan hydrolase; the production cost of the enzyme is favorably reduced and promotion and application of the ratgalacturonorhamnan hydrolase in the food industry are promoted.

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. technical background [0002] 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. [0003] 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 chemically dist...

Claims

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Application Information

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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
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