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Construction and characterization of nitrate and nitrite respiring Pseudomonas putida KT2440 strains for anoxic biotechnical applications
Ist Teil von
Journal of biotechnology, 2013-01, Vol.163 (2), p.155-165
Ort / Verlag
Netherlands: Elsevier B.V
Erscheinungsjahr
2013
Link zum Volltext
Quelle
MEDLINE
Beschreibungen/Notizen
► Two Pseudomonas aeruginosa cosmids containing gene clusters for nitrate or nitrite respiration were isolated. ► Transfer of these cosmids in the obligate aerobic Pseudomonas putida KT2440 generated two strains, which anaerobically reduced nitrate and nitrite, respectively. ► Microarray analysis confirmed expression of all structural, maturation and regulatory P. aeruginosa denitrification genes. ► Recombinantly produced P. aeruginosa enzyme systems allowed for anaerobic life of P. putida KT2440. ► Expression of P. putida genes encoding a putative copper stress response operon was observed.
Pseudomonas putida KT2440 is frequently used in biotechnical research and applications due to its metabolic versatility and organic solvent resistance. A major drawback for a broad application is the inability of the bacterium to survive and grow under anoxic conditions, which prohibits the production of oxygen-sensitive proteins and metabolites. To develop a P. putida strain, which is able to survive under anoxic conditions, the enzymatic systems of anaerobic nitrate and nitrite respiration were introduced into KT2440. For this purpose, two cosmids encoding all structural, maturation and regulatory genes for P. aeruginosa nitrate reductase (pNAR) and nitrite- and nitric oxide reductase (pNIR-NOR) were stably maintained in P. putida KT2440. Transcriptome analyses revealed expression of the encoded nar, nir and nor operons and accessory genes under anoxic conditions. The produced enzyme systems efficiently reduced nitrate or nitrite, respectively, sustaining anaerobic life of recombinant KT2440. Interestingly, anaerobic life of P. putida induced genes involved in arginine-fermentation and genes encoding a putative copper stress resistance operon.