Analysis of the molecular response of Pseudomonas putida KT2440 to the next-generation biofuel n-butanol

Publikations-Art

Zeitschriftenbeitrag (peer-reviewed)

Autoren

Simon, O., Klebensberger, J., Mükschel, B., Klaiber, I., Graf, N., Altenbuchner, J., Huber, A., Hauer, B., and Pfannstiel, J.

Erscheinungsjahr

2015

Veröffentlicht in

Journal of Proteomics

DOI

10.1016/j.jprot.2015.03.022

Seite (von - bis)

11-25

To increase the efficiency of biocatalysts a thorough understanding of the molecular response of the biocatalyst to precursors, products and environmental conditions applied in bioconversions is essential. Here we performed a comprehensive proteome and phospholipid analysis to characterize the molecular response of the potential biocatalyst Pseudomonas putida KT2440 to the next-generation biofuel n-butanol. Using complementary quantitative proteomics approaches we were able to identify and quantify 1467 proteins, corresponding to 28% of the total KT2440 proteome. 256 proteins were altered in abundance in response to n-butanol. The proteome response entailed an increased abundance of enzymes involved in n-butanol degradation including quinoprotein alcohol dehydrogenases, aldehyde dehydrogenases and enzymes of fatty acid beta oxidation. From these results we were able to construct a pathway for the metabolism of n-butanol in P. putida. The initial oxidation of n-butanol is catalyzed by at least two quinoprotein ethanol dehydrogenases (PedE and PedH). Growth of mutants lacking PedE and PedH on n-butanol was significantly impaired, but not completely inhibited, suggesting that additional alcohol dehydrogenases can at least partially complement their function in KT2440. Furthermore, phospholipid profiling revealed a significantly increased abundance of lyso-phospholipids in response to n-butanol, indicating a rearrangement of the lipid bilayer.