9). Pyruvate formate lyase produces acetyl-CoA and formate from
pyruvate. Only in 23K, the pflAB genes encoding formate C-acetyltransferase and its activating enzyme involved in formate formation were strongly up-regulated (4.0 and 1.7, respectively). This strain was the only one to strongly induce L-lactate oxidase encoding genes which are responsible for conversion of lactate to acetate when oxygen is present (Table 1). In 23K and LS 25, the ppdK gene coding for the pyruvate phosphate dikinase involved in regenerating PEP, was induced, as was also lsa0444 encoding a putative malate dehydrogenase that catalyzes the conversion
of malate into oxaloacetate using NAD+ and vice versa (Table 1). During growth on ribose, this website L. sakei was shown to require thiamine (vitamine CP673451 research buy B1) [15]. The E1 component subunit α of the PDC, as well as Pox and Xpk, require thiamine pyrophosphate, the active form of thiamine, as a coenzyme [54]. This could explain the induction of the thiMDE operon and lsa0055 in LS 25, as well as lsa0980 in 23K, encoding enzymes involved in thiamine uptake and biosynthesis (Table 1). The up-regulation of lsa1664 (1.1-1.6) encoding a putative dihydrofolate reductase involved in biosynthesis of riboflavin (vitamin B2) in all the strains could indicate a requirement for flavin nucleotides as enzyme cofactors. Riboflavin is the precursor for flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) redox cofactors in flavoproteins, and the E3 component of PDC as well as glycerol-3-phosphate dehydrogenase encoded from
the up-regulated glpD, are among Parvulin enzymes requiring FAD. Another cofactor which seems to be important during growth on ribose is lipoate, click here essential of the E2 component of the PDC. An up-regulation of lplA (1.0 – 1.6) encoding lipoate-protein ligase, which facilitates attachment of the lipoyl moiety to metabolic enzyme complexes, was seen in all the strains, allowing the bacterium to scavenge extracellular lipoate [55, 56]. Nucleoside catabolism The L. sakei genome contains a multiplicity of catabolic genes involved in exogenous nucleoside salvage pathways, and the bacterium has been shown to catabolize inosine and adenosine for energy [7]. Three iunH genes are present in the 23K genome, which encode inosine-uridine preferring nucleoside hydrolases responsible for conversion of inosine to ribose and purine base. The iunH1 gene was up-regulated in all the strains when grown on ribose (1.8-2.6), as was also the iunH2 gene in 23K (1.2).