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Bacterial strain for generating rhamnolipid biosurfactant and generated microbial inoculum thereof

A biological surface, rhamnolipid technology, applied in the field of biological high, can solve the problems of instability of plasmids, introduction of antibiotic markers, etc., to achieve the effect of protecting the ecological environment, promoting degradation and stable quality

Active Publication Date: 2011-01-19
JIANGSU SENTAY ENVIRONMENTAL SCI & TECH CO LTD
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  • Application Information

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Problems solved by technology

So far, plasmids are still used as vectors to construct genetically engineered bacteria. This method has the disadvantages of plasmid instability and the introduction of antibiotic markers.

Method used

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  • Bacterial strain for generating rhamnolipid biosurfactant and generated microbial inoculum thereof
  • Bacterial strain for generating rhamnolipid biosurfactant and generated microbial inoculum thereof
  • Bacterial strain for generating rhamnolipid biosurfactant and generated microbial inoculum thereof

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Embodiment Construction

[0014] Primers were designed according to the sequence of the gene cluster related to rhamnolipid synthesis (rhlABRI), and the forward primer was 5′-AGAT GCGGCCGC TTCGACACCGGAAACC-3′; reverse primer is 5′-AGAT GCGGCCGC For GGCATGGCGACTCCTC-3', a NotI restriction site was introduced at both ends of the primer (the restriction site is underlined). Using the genomic DNA of Pseudomonas aeruginosa BSFD5 as a template, the complete sequence of the gene cluster related to rhamnolipid synthesis was amplified. It was sequenced and registered in GenBank (accession number: HM190303). and connect it to the vector pUTTns to construct pUTTns-rhlABRI, and transform it into E.coli DH5α λpir E.coli DH5α λpir -pUTTns-rhlABRI. The rhlABRI gene cluster was integrated into the chromosome of Pseudomonas putida KT2440 by triparental conjugation, and the resistance gene was lost by screening the zygote in LB medium supplemented with 10% (w / v) sucrose. A genetically engineered bacterium KT2440-...

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Abstract

The invention provides a bacterial strain KT2440-rhlABRI capable of generating a rhamnolipid biosurfactant and the generated microbial inoculum thereof, which belong to the field of biological high technology. The microbial inoculum is obtained by integrating a gene cluster rhlABRI related to the synthesis of the rhamnolipid biosurfactant in Pseudomonas aeruginosa BSFD5 into chromosomes of a type strain KT2440 of Pseudomonas putida by an unmarked homologous recombination method. The microbial inoculum product can be directly used for promoting the degradation of polycyclic aromatic hydrocarbons pyrene in soil and can be applied to the bioremediation of soil polluted by polycyclic aromatic hydrocarbons.

Description

1. Technical field [0001] The invention provides a bacterial strain capable of producing rhamnolipid biosurfactant and the microbial agent produced therefor, belonging to the field of biological high technology. It uses microbiological methods to promote the degradation of polycyclic aromatic hydrocarbons in soil, and can be applied to the bioremediation of polycyclic aromatic hydrocarbon-contaminated soils. 2. Background technology [0002] Polycyclic aromatic hydrocarbons (Polycyclic Aromatic Hydrocarbons, PAHs) are a class of refractory organic pollutants that widely exist in the environment. In recent years, with the continuous acceleration of the development and utilization of petroleum, coal, natural gas and other resources, the polycyclic aromatic hydrocarbons in the environment have a tendency to increase. Due to the potential toxicity, carcinogenicity and mutagenicity of polycyclic aromatic hydrocarbons, they have obvious ecological risks to human health and the ec...

Claims

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Application Information

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IPC IPC(8): C12N1/21B09C1/10A62D3/02C12R1/40A62D101/20
Inventor 洪青王倩蒋建东李荣李顺鹏
Owner JIANGSU SENTAY ENVIRONMENTAL SCI & TECH CO LTD
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