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The E ntner– D oudoroff pathway empowers P seudomonas putida KT 2440 with a high tolerance to oxidative stress
Author(s) -
Chavarría Max,
Nikel Pablo I.,
PérezPantoja Danilo,
Lorenzo Víctor
Publication year - 2013
Publication title -
environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.954
H-Index - 188
eISSN - 1462-2920
pISSN - 1462-2912
DOI - 10.1111/1462-2920.12069
Subject(s) - pseudomonas putida , biology , biochemistry , escherichia coli , nad+ kinase , catabolism , oxidative stress , oxidative phosphorylation , pentose phosphate pathway , microbiology and biotechnology , metabolism , enzyme , glycolysis , gene
Summary Glucose catabolism of P seudomonas putida is carried out exclusively through the E ntner– D oudoroff ( ED ) pathway due to the absence of 6‐phosphofructokinase. In order to activate the E mbden– M eyerhof– P arnas ( EMP ) route we transferred the pfkA gene from E scherichia coli to a P . putida wild‐type strain as well as to an eda mutant, i.e. lacking 2‐keto‐3‐deoxy‐6‐phosphogluconate aldolase. PfkA E . coli failed to redirect the carbon flow from the ED route towards the EMP pathway, suggesting that ED was essential for sugar catabolism. The presence of PfkA E . coli was detrimental for growth, which could be traced to the reduction of ATP and NAD ( P ) H pools along with alteration of the NAD ( P ) H / NADP + ratio. P seudomonas putida cells carrying PfkA E . coli became highly sensitive to diamide and hydrogen peroxide, the response to which is very demanding of NADPH . The inhibitory effect of PfkA E . coli could in part be relieved by methionine, the synthesis of which relies much on NADPH . These results expose the role of the ED pathway for generating the redox currency ( NADPH ) that is required for counteracting oxidative stress. It is thus likely that environmental bacteria that favour the ED pathway over the EMP pathway do so in order to gear their aerobic metabolism to endure oxidative‐related insults.