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An ascorbate peroxidase mutant maapx1 m36k and its application

A technology of ascorbic acid peroxide and mutants, applied in the direction of oxidoreductase, application, DNA/RNA fragments, etc., can solve problems such as no corresponding reports

Active Publication Date: 2019-11-15
SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The activity of APX in plants is regulated by various post-translation, but there is no corresponding report on how to increase the activity of APX

Method used

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  • An ascorbate peroxidase mutant maapx1 <sup>m36k</sup> and its application
  • An ascorbate peroxidase mutant maapx1 <sup>m36k</sup> and its application
  • An ascorbate peroxidase mutant maapx1 <sup>m36k</sup> and its application

Examples

Experimental program
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Embodiment 1

[0039] 1. Sampling: The selected banana (Musa genus) species is Brazilian banana, collected from Nansha City, Guangdong Province.

[0040] 2. Cloning of MaAPX1 gene

[0041] The total RNA in the banana peel was extracted by the conventional hot boric acid method, and the extracted total RNA was extracted according to the kit PrimeScript TM The method provided by RT Master Mix (Perfect Real Time, RR036A, TaKaRa) was used to reverse transcribe RNA into cDNA. Reaction system: X μL RNA (<500 ng), 2 μL 5×Mix, Y μL RNase Free water and mixed to make the total volume 10 μL, Then place it in a constant temperature water bath at 37°C for 15 minutes, heat shock in a metal bath at 85°C for 5 seconds, and cool at 4°C.

[0042] 3. Construction of MaAPX1 expression strain

[0043] According to the MaAPX1 gene sequence design in the banana genome (Banana Genome Hub), the primers containing the nde I restriction site and the primers of the mutation site, the primer sequences are as shown i...

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Abstract

The invention provides an ascorbate peroxidase mutant MaAPX1 M36K The invention and its application relate to the technical field of biotechnology. Mutant MaAPX1 of the present invention M36K The amino acid sequence of is shown in SEQ ID NO.3. The present invention expresses mutant MaAPX1 through prokaryotic M36K Reconstituted protein, mutant MaAPX1 discovered by enzyme activity assay M36K The catalytic efficiency has been increased by nearly 5 times, which can provide technical reference for further research and application of APX1.

Description

technical field [0001] The invention belongs to the technical field of biotechnology, in particular to an ascorbate peroxidase mutant MaAPX1 M36K and its application. Background technique [0002] Reactive oxygen species (ROS) are by-products of cellular metabolism, mainly including H 2 o 2 , superoxide anion, and singlet oxygen play an important role in signal transduction and homeostasis. However, during environmental stress or aging, excessive accumulation of ROS can cause oxidative damage to macromolecules such as proteins, DNA, and lipids, which in turn leads to loss of structure and function, and may even lead to cell death. To combat oxidative stress, organisms have evolved complex protection systems, such as reactive oxygen species scavenging enzymes and macromolecular repair systems. [0003] There are antioxidant enzyme systems such as superoxide dismutase, ascorbic acid peroxidase (APX), catalase, glutathione peroxidase and peroxidase in plants. In addition, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/08C12N15/53C12N15/70C12N1/21C12R1/19
CPCC12N9/0065C12N15/70C12Y111/01011C12N9/0004C12N15/52
Inventor 段学武肖璐蒋国祥蒋跃明严慧玲李志伟曾睛丁晓春
Owner SOUTH CHINA BOTANICAL GARDEN CHINESE ACADEMY OF SCI
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