[Integration and expression of polyphosphate kinase gene in Pseudomonas putida]

Huan Jing Ke Xue. 2009 Oct 15;30(10):3011-5.
[Article in Chinese]

Abstract

In order to construct high accumulating-phosphate microorganism, the ppk gene from E. coli was inserted to the broad-host-range plasmid pBBR1MCS-2 to form plasmid pBBR1MCS-2-ppk. The complete ppk gene with promoter and terminator sequences from pBBR1MCS-2-ppk was then cloned and inserted to suicide plasmid pUTmini-Tn5 to form plasmid pUTmini-Tn5-ppk, which was transformed into Pseudomonas putida KT2440 by triparental conjugation. Finally, ppk gene was integrated into the chromosomal DNA of KT2440. The results of RT-PCR showed that the selected genetically engineered bacterium KT2440-PPK expressed ppk efficiently, while KT2440 as control expressed weakly. The results of artificial wastewater treatment showed after 1h inoculation, the concentration of poly-phosphate in KT2440-PPK came to the maximum approximately 3.05 mg/g, which was 15 times higher than that in KT2440 at the same experimental condition. And KT2440-PPK can remove more than 90% phosphate in artificial wastewater.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biodegradation, Environmental
  • DNA, Bacterial / genetics
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics*
  • Gene Expression Regulation, Bacterial
  • Mutagenesis, Insertional
  • Phosphorus / isolation & purification*
  • Phosphorus / metabolism
  • Phosphotransferases (Phosphate Group Acceptor) / biosynthesis
  • Phosphotransferases (Phosphate Group Acceptor) / genetics*
  • Plasmids / genetics
  • Pseudomonas putida / enzymology
  • Pseudomonas putida / genetics*
  • Waste Disposal, Fluid / methods

Substances

  • DNA, Bacterial
  • Phosphorus
  • Phosphotransferases (Phosphate Group Acceptor)
  • polyphosphate kinase