Figure 1.
Putative GDP-β-D-virenose biosynthesis pathway.
1. F6P, fructose-6-phosphate; PMI, phosphomannose isomerase 2. M6P, mannose-6-phosphate; PMM, phosphomannomutase 3. M1P, mannose-1-phosphate, GMP, GDP-mannose pyrophosphorylase 4. GMD, GDP-mannose 4,6-dehydratase; NADP+ nicotinamide adenine dinucleotide phosphate 5. GFS, fucose synthase 6. Ado-Met, S-adenosyl methionine.
Table 1.
Predicted C. burnetii proteins catalyzing formation of GDP-D-mannose.
Figure 2.
C. burnetii CBU0671 (GMP) restores K30 expression in the E. coli cpsB (manC) mutant strain CWG152.
Immunoblot analysis with K30-specific antiserum of proteinase K treated whole cell lysates of 1. wild type E. coli CWG44, E. coli cpsB mutant strain CWG152, 3. E. coli CWG152/pCN603-1 (E. coli cpsB) induced, 4. E. coli CWG152/pCN603-1 (E. coli cpsB) not induced, 5. E. coli CWG152/pBAD (vector control), 6. E. coli CWG152/pCN606-2 (C. burnetii CBU0671) induced, 7. E. coli CWG152/pCN606-2 (C. burnetii CBU0671) not induced.
Figure 3.
C. burnetii CBU0294 (PMM) restores a smooth LPS chemotype in P. aeruginosa PAO1 O5 algC mutant.
SDS-PAGE and silver stain of proteinase K treated whole cell lysates of 1. wild-type P. aeruginosa PAO1, 2. P. aeruginosa algC mutant, 3. P. aeruginosa algC::tet/pLPS188 (P. aeruginosa algC), 4. P. aeruginosa algC::tet/pUCP20 (vector control), 5. P. aeruginosa algC::tet/pCN620.
Figure 4.
Mechaelis-Menten diagram depicting C. burnetii CBU0294 (PMM) kinetic parameters.
Reactions were carried out with D-mannose-1-P as the fixed substrate. Data points were fitted using Microsoft XLfit model 601. Inset: Cooresponding Lineweaver-Burk Plot, regression line calculated by least squares.
Table 2.
Kinetic parameters for C. burnetii CBU0671 and CBU0294.
Table 3.
Enzymatic activity of C. burnetii CBU0671 and CBU0294 in bacterial crude extracts.
Table 4.
Bacterial strains and plasmids used in this study.