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Phosphatase FM2382 and coding gene and application thereof

A technology of phosphatase and phosphohydrolase, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as non-degradation, human and animal health threats, etc.

Active Publication Date: 2016-05-11
深圳中科欣扬生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These phosphorus-containing compounds are generally environmentally toxic, relatively stable in chemical properties, not easy to be degraded, and easy to accumulate in humans and animals through the food chain, posing a great threat to the health of humans and animals

Method used

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  • Phosphatase FM2382 and coding gene and application thereof
  • Phosphatase FM2382 and coding gene and application thereof
  • Phosphatase FM2382 and coding gene and application thereof

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0085] Embodiment 1, the acquisition and functional verification of phosphatase

[0086] 1. Preparation of phosphatase

[0087] 1. Acquisition of FM2382 protein and its coding gene

[0088] The DNA molecule shown in the 61-930th position of sequence 1 from Fulvimarinamanganoxydanssp.nov.8047 (CGMCC1.10972) is named gene FM2382, and the protein encoded by this gene is named FM2382, and the amino acid sequence of FM2382 is as sequence 2 in the sequence listing Shown in bits 21-309 of .

[0089] 2. Obtaining the recombinant vector

[0090] Replace the DNA fragment between the NdeI and HindIII recognition sequences of the pET-28a(+) vector with the gene FM2382 shown in the 61-930th position of sequence 1, keep the other sequences of the pET-28a(+) vector unchanged, and obtain the recombinant Vector, the recombinant vector is named pET-FM2382. In pET-FM2382, the expression of gene FM2382 is driven by T7 promoter, and pET-FM2382 can express the protein shown in Sequence 2.

[00...

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Abstract

The invention discloses a phosphatase FM2382 and a coding gene and the application of the phosphatase FM2382. The phosphatase FM2382 is (A1), (A2), (A3) or (A4) as follows: an amino acid sequence of the (A1) is protein of a sequence 2; the amino acid sequence of the (A2) is protein of the sequence 2 from the 21th site to the 309th site; the (A3) is protein which is obtained by replacing and / or missing and / or adding one or several amino acid residues in the sequence 2 or the amino acid sequences of the sequence 2 from the 21th site to the 309th site, has the same functions and is derived by the (A1) or the (A2); the (A4) is fused protein which is obtained by connecting a label to the N end or / and the C end of the (A1), (A2) or (A3). An experiment shows that the phosphatase FM2382 disclosed by the invention has the activity of phosphohydrolase; the phosphatase FM2382 is easy to prepare and good in stability, middle and high temperature can be tolerated, the appropriate temperature is 45 DEG C, not only can phosphor compounds be degraded, but also halogen compounds can be degraded.

Description

technical field [0001] The invention relates to a phosphatase FM2382 and its coding gene and application in the field of biotechnology. Background technique [0002] Organophosphorus compounds are widely used as pesticides and pharmaceutical intermediates. These phosphorus-containing compounds are generally environmentally toxic, relatively stable in chemical properties, not easy to be degraded, and easy to accumulate in humans and animals through the food chain, posing a great threat to the health of humans and animals. [0003] Phosphatase (Phosphatase, EC3.1.3) can catalyze the hydrolysis of phosphate esters or phosphoric anhydrides. Its mechanism of action is to catalyze the dephosphorylation of organophosphorus compounds. By hydrolyzing phosphate monoesters, the phosphate groups are released from the compounds, and at the same time, phosphate ions and free hydroxyl groups are generated. Phosphatase can relieve or reduce the toxicity of organophosphorus pesticides by r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/16C12N15/55
CPCC12N9/16C12Y301/03
Inventor 董志扬陈林材董亮董欣欣
Owner 深圳中科欣扬生物科技有限公司