Overexpression of isocitrate lyase is an important strategy in the survival of Pseudomonas fluorescens exposed to aluminum.
Isocitrate lyase, ICL (EC 4.1.3.1), an enzyme that cleaves isocitrate into succinate, and glyoxylate appears to play a pivotal role in the detoxification of aluminum (Al) in Pseudomonas fluorescens. Here, we present evidence that the 4-fold increase in ICL activity observed in Al-stressed cells is due to the overexpression of this enzyme. Blue-Native-PAGE, Western blotting, and spectrophotometric experiments revealed that ICL is optimally expressed at 35 h of growth in Al-stressed cells. However, following the immobilization of Al, at 60 h of growth, the level of the enzyme decreases markedly. This enzyme that exists as a homotetramer with a molecular mass of approximately 133 kDa appears to be transcriptionally regulated. The overexpression of ICL may be a specific response to Al-stress as P. fluorescens grown in the presence of such metals as Ga3+, Pb2+, and Ca2+ does not undergo any significant increase in ICL activity. Thus, these findings support the notion that the overexpression of ICL plays a pivotal role in the survival and in the increased oxalogenesis observed in Al-stressed P. fluorescens.
Hamel
Robert
R
Department of Chemistry and Biochemistry, Laurentian University, Sudbury, Ont., Canada.
Appanna
Vasu D
VD
Viswanatha
Thammaiah
T
Puiseux-Dao
Simone
S
D000535:Aluminum / Q000378:metabolism / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression / Q000187:drug effects; D006038:Glyoxylates / Q000378:metabolism; D007522:Isocitrate Lyase / Q000096:biosynthesis* / Q000378:metabolism; D008670:Metals / Q000494:pharmacology; D019906:Nuclear Magnetic Resonance, Biomolecular; D011551:Pseudomonas fluorescens / Q000166:cytology / Q000187:drug effects / Q000201:enzymology / Q000502:physiology*; D019802:Succinic Acid / Q000378:metabolism
D006038:Glyoxylates; D008670:Metals; D019802:Succinic Acid; D000535:Aluminum; D007522:Isocitrate Lyase; C031150:glyoxylic acid