The transcription of genes for ED pathway in Zymomonas mobilis significantly increased under anaerobic ethanol-producing conditions to facilitate energy conservation [34]. In R. eutropha under the PHA biosynthesis condition, we observed a decreasing trend in expression of the genes in ED pathway find protocol and TCA cycle. The activity of ED pathway and TCA cycle during the PHA production phase is probably attributable to pre-existing as well as newly synthesized enzymes with the reduced transcription. The probably decreased flux
of central metabolisms were supported by our recent metabolomics analysis of R. eutropha H16 that detected lower intracellular concentrations of many sugar phosphates in the PHA production phase than in the growth phase on fructose [23]. It can be assumed that the decreased
metabolic activity appeared to be enough to maintain cellular viability and P(3HB) synthesis in a condition not associated with cell growth, as seen in Corynebacterium glutamicum in a glutamate-producing condition [35]. de novo Opaganib nmr Fatty acid synthesis and β-oxidation In R. eutropha H16, accA1, accA2, accB, accC1, accC2, accC3, and accD have been annotated as genes of the acetyl-CoA carboxylase (ACC) subunits. Based on a consideration of the general quaternary structure of ACC and the expression levels of these genes, the major ACC in this strain probably consisted of AccA1 (H16_A1223) as the carboxyl transferase subunit α (CTα), AccD (H16_A2611) as the carboxyl transferase subunit β (CTβ), AccB and AccC2 (H16_A3171-A3172) as the biotin carboxyl carrier protein (BCCP) and biotin carboxylase (BC), respectively. The expression levels of these genes were high in the growth phase, and then slightly decreased in the PHA production phase (Figure 4). accC1 (H16_A0184, BC-BCCP) and H16_A0177 (CTαβ) may be another pair of ACC or
the related carboxylase, because these had weak and similar expression behaviors to each other. The expression levels of accA2 (H16_A2142, BC-BCCP) and accC3 (H16_A3290, BC-BCCP-CTαβ) were negligible throughout cultivation on fructose. The genes fabHDG-acpP-fabF (H16_A2569-A2565), fabZ (H16_A2044), and fabI1 (H16_A2410), which are involved Florfenicol in de novo fatty acid biosynthesis, were highly expressed in the growth phase, but many of the genes still had rather high expression levels in the PHA production phase. Figure 4 Transcription levels of genes involved in fatty acid biosynthesis and β-oxidation in R. eutropha H16 at growth phase F16, PHA production phase F26, and stationary phase F36 on fructose. With respect to β-oxidation enzymes, selected genes of which specific name has been assigned, or RPKM value are larger than 1,000 at least one of the three phases are shown. The log2-transformed RPKM values are visualized using the rainbow color scale in the figure. Genes with the P value above the threshold (P > 0.05) are underlined.
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