RT Journal Article SR Electronic T1 Insights into an alternative pathway for glycerol metabolism in a glycerol kinase deficient Pseudomonas putida KT2440 JF bioRxiv FD Cold Spring Harbor Laboratory SP 567230 DO 10.1101/567230 A1 Meg Walsh A1 William Casey A1 Shane T. Kenny A1 Tanja Narancic A1 Lars M. Blank A1 Nick Wierckx A1 Hendrik Ballerstedt A1 Kevin E O Connor YR 2019 UL http://biorxiv.org/content/early/2019/03/12/567230.abstract AB Pseudomonas putida KT2440 is known to metabolise glycerol via glycerol-3-phosphate using glycerol kinase an enzyme previously described as critical for glycerol metabolism (1). However, when glycerol kinase was knocked out in P. putida KT2440 it retained the ability to use glycerol as the sole carbon source, albeit with a much-extended lag period and 2 fold lower final biomass compared to the wild type strain. A metabolomic study identified glycerate as a major and the most abundant intermediate in glycerol metabolism in this mutated strain with levels 21-fold higher than wild type. Erythrose-4-phosphate was detected in the mutant strain, but not in the wild type strain. Glyceraldehyde and glycraldehyde-3-phosphate were detected at similar levels in the mutant strain and the wild type. Transcriptomic studies identified 191 genes that were more than 2-fold upregulated in the mutant compared to the wild type and 175 that were down regulated. The genes involved in short chain length fatty acid metabolism were highly upregulated in the mutant strain. The genes encoding 3-hydroxybutyrate dehydrogenase were 5.8-fold upregulated and thus the gene was cloned, expressed and purified to reveal it can act on glyceraldehyde but not glycerol as a substrate.