Expression of phosphofructokinase in Neisseria meningitidis.
Neisseria meningitidis serogroup B is a pathogen that can infect diverse sites within the human host. According to the N. meningitidis genomic 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 silico flux balance analysis predicted an increase of biomass yield as a result of PFK expression. However, analysis of a genetically engineered N. meningitidis strain that expressed a heterologous PFK showed that the yield of biomass on substrate decreased in comparison with a pfkA-deficient control strain, which was associated mainly with an increase in CO(2) production, whereas production of by-products was similar in the two strains. This might explain why the pfkA gene has not been obtained by horizontal gene transfer, since it is initially unfavourable for biomass yield. No large effects related to heterologous expression of pfkA were 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.
Baart
Gino J E
GJE
Wageningen University, Food and Bioprocess Engineering Group, PO Box 8129, 6700 EV Wageningen, The Netherlands.|Netherlands Vaccine Institute (NVI), Unit Research and Development, PO Box 457, 3720 AL Bilthoven, The Netherlands.
Langenhof
Marc
M
Wageningen University, Food and Bioprocess Engineering Group, PO Box 8129, 6700 EV Wageningen, The Netherlands.|Netherlands Vaccine Institute (NVI), Unit Research and Development, PO Box 457, 3720 AL Bilthoven, The Netherlands.
van de Waterbeemd
Bas
B
Netherlands Vaccine Institute (NVI), Unit Research and Development, PO Box 457, 3720 AL Bilthoven, The Netherlands.
Hamstra
Hendrik-Jan
HJ
Netherlands Vaccine Institute (NVI), Unit Research and Development, PO Box 457, 3720 AL Bilthoven, The Netherlands.
Zomer
Bert
B
Netherlands Vaccine Institute (NVI), Unit Research and Development, PO Box 457, 3720 AL Bilthoven, The Netherlands.
van der Pol
Leo A
LA
Netherlands Vaccine Institute (NVI), Unit Research and Development, PO Box 457, 3720 AL Bilthoven, The Netherlands.
Beuvery
E C
EC
PAT consultancy, Kerkstraat 66, 4132 BG Vianen, The Netherlands.
Tramper
Johannes
J
Wageningen University, Food and Bioprocess Engineering Group, PO Box 8129, 6700 EV Wageningen, The Netherlands.
Martens
Dirk E
DE
Wageningen University, Food and Bioprocess Engineering Group, PO Box 8129, 6700 EV Wageningen, The Netherlands.
D018533:Biomass; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D020869:Gene Expression Profiling; D053858:Metabolic Networks and Pathways; D038541:Neisseria meningitidis, Serogroup B / Q000145:classification / Q000201:enzymology* / Q000235:genetics; D025221:Phosphofructokinases / Q000096:biosynthesis* / Q000235:genetics; D010802:Phylogeny; D012329:RNA, Bacterial / Q000235:genetics
D012329:RNA, Bacterial; D025221:Phosphofructokinases