Modeling Neisseria meningitidis B Metabolism at Different Specific Growth Rates

被引:20
作者
Baart, Gino J. E. [1 ,2 ]
Willemsen, Marieke [1 ,2 ]
Khatami, Elnaz [1 ]
de Haan, Alex [1 ]
Zomer, Bert [1 ]
Beuvery, E. Coen [3 ]
Tramper, Johannes [2 ]
Martens, Dirk E. [2 ]
机构
[1] NVI, Unit Res & Dev, NL-3720AL Bilthoven, Netherlands
[2] Wageningen Univ, Food & Bioproc Engn Grp, NL-6700 EV Wageningen, Netherlands
[3] PAT Consultancy, NL-4132BG Vianen, Netherlands
关键词
Neisseria meningitides; growth rate dependent biomass composition; flux balance analysis; substrate and energy parameters estimation; vaccine process development;
D O I
10.1002/bit.22016
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Neisseria meningitidis is a human pathogen that can infect diverse sites within the human host. The major diseases caused by N. meningitidis are responsible for death and disability, especially in young infants. At the Netherlands Vaccine Institute (NVI) a vaccine against serogroup B organisms is currently being developed. This study describes the influence of the growth rate of N. meningitidis on its macro-molecular composition and its metabolic activity and was determined in chemostat cultures. In the applied range of growth rates, no significant changes in RNA content and protein content with growth rate were observed in N. meningitidis. The DNA content in N. meningitidis was somewhat higher at the highest applied growth rate. The phospholipid and lipopolysaccharide content in N. meningitidis changed with growth rate but no specific trends were observed. The cellular fatty acid composition and the amino acid composition did not change significantly with growth rate. Additionally, it was found that the PorA content in outer membrane vesicles was significantly lower at the highest growth rate. The metabolic fluxes at various growth rates were calculated using flux balance analysis. Errors in fluxes were calculated using Monte Carlo Simulation and the reliability of the calculated flux distribution could be indicated, which has not been reported for this type of analysis. The yield of biomass on substrate (Y-x/s) and the maintenance coefficient (m(s)) were determined as 0.44 (+/- 0.04) gg(-1) and 0.04 (+/- 0.02) gg(-1)h(-1), respectively. The growth associated energy requirement (Y-x/ATP) and the non-growth associated ATP requirement for maintenance (MATP) were estimated as 0.13 (+/- 0.04) mol mol(-1) and 0.43 (+/- 0.14) mol mol(-1) h(-1), respectively. It was found that the split ratio between the Entner-Doudoroff and the pentose phosphate pathway, the sole glucose utilizing pathways in N. meningitidis, had a minor effect on ATP formation rate but a major effect on the fluxes going through for instance the citric-acid cycle. For this reason, we presented flux ranges for underdetermined parts of metabolic network rather than presenting single flux values, which is more commonly done in literature.
引用
收藏
页码:1022 / 1035
页数:14
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