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Streptomycete 2-nitropropane dioxyenase gene and its application

A technology of dioxygenase and nitropropane, applied in application, genetic engineering, oxidoreductase and other directions, can solve the problem of not many structural genes, and achieve the effect of improving the environment

Inactive Publication Date: 2003-03-19
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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Problems solved by technology

It has been found in microorganisms such as Mrakhansenula, Bacillus subtilis and Streptomyces, and their optimal substrates are 2-nitropropane, nitroethane, etc. These enzymes were isolated and purified and deduced Amino acid sequence, but there are not many reports about the structural genes of these enzymes, especially in Streptomyces and even Actinomyces.

Method used

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  • Streptomycete 2-nitropropane dioxyenase gene and its application
  • Streptomycete 2-nitropropane dioxyenase gene and its application
  • Streptomycete 2-nitropropane dioxyenase gene and its application

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Embodiment 2

[0043] The cells were suspended in 0.2M pH7.0 phosphate buffer solution, the cells were ultrasonically disrupted (100W, 15min), centrifuged at 10000rpm for 15min, and the supernatant was taken to obtain the enzyme stock solution. After measuring its activity, it was found that the cell disruption solution containing the recombinant plasmid produced a positive reaction, that is, a clear purple red color appeared rapidly, while the control strain containing only the empty vector pET23b and the blank control without bacteria did not have this color reaction. It shows that the expression product of npD gene can oxidize the substrate. Using different nitroalkane compounds as substrates, it was found that it has different degrees of biological activity on 1-nitropropane, 2-nitropropane, and nitroethane, and its activity on nitromethane is relatively low (such as Figure 5 ). Embodiment 2: Separation, purification and biological activity of 2-nitropropane dioxygenase

[0044] The o...

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Abstract

The present invention relates to the application of environmental microbe. Especially, by means of molecular technology, one new gene is cloned from Streptomyces ansochromogenes 7100 and the gene encodes one kind of 2-streptomycete dioxyenase. The encoded product has great value in biologically degrading industrial pollutant nitroalkane and in improving environment.

Description

technical field [0001] This invention belongs to the field of applied environmental microorganisms, specifically, a new gene is cloned from Streptomyces chromogenes by molecular biology techniques, and the product encoded by it is a kind of 2-nitropropane dioxygenase, It has great application value in the biodegradation of industrial pollutants nitroalkanes and can be used to improve the environment. technical background [0002] Nitroalkane oxidase is a large class of enzymes whose substrates are mainly nitroalkanes, denitrates these compounds and generates corresponding aldehydes and ketones, accompanied by the generation of nitrous acid. Among them, 2-nitropropane dioxygenase (2-Nitropropane Dioxygenase) is an important nitroalkane oxidase. It has been found in microorganisms such as Mrakhansenula, Bacillus subtilis and Streptomyces, and their optimal substrates are 2-nitropropane, nitroethane, etc. These enzymes were isolated and purified and deduced However, there are...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N15/63C12N15/70
Inventor 谭华荣张集慧马文勃
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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