Expression of phosphofructokinase in Neisseria meningitidis

Abstract

Neisseria meningitidisserogroup B is a pathogen that can infect diverse sites within the human host. According to theN. meningitidisgenomic information and experimental observations, glucose can be completely catabolized through the Entner–Doudoroff pathway and the pentose phosphate pathway. The Embden–Meyerhof–Parnas pathway is not functional, because the gene for phosphofructokinase (PFK) is not present. The phylogenetic distribution of PFK indicates that in most obligate aerobic organisms, PFK is lacking. We conclude that this is because of the limited contribution of PFK to the energy supply in aerobically grown organisms in comparison with the energy generated through oxidative phosphorylation. Under anaerobic or microaerobic conditions, the available energy is limiting, and PFK provides an advantage, which explains the presence of PFK in many (facultatively) anaerobic organisms. In accordance with this,in silicoflux balance analysis predicted an increase of biomass yield as a result of PFK expression. However, analysis of a genetically engineeredN. meningitidisstrain that expressed a heterologous PFK showed that the yield of biomass on substrate decreased in comparison with apfkA-deficient control strain, which was associated mainly with an increase in CO2production, whereas production of by-products was similar in the two strains. This might explain why thepfkAgene has not been obtained by horizontal gene transfer, since it is initially unfavourable for biomass yield. No large effects related to heterologous expression ofpfkAwere observed in the transcriptome. Although our results suggest that introduction of PFK does not contribute to a more efficient strain in terms of biomass yield, achievement of a robust, optimal metabolic network that enables a higher growth rate or a higher biomass yield might be possible after adaptive evolution of the strain, which remains to be investigated.