Functional analysis of genes involved in the biosynthesis of isoprene in Bacillus subtilis

Julsing, Mattijs K.; Rijpkema, Michael; Woerdenbag, Herman J.; Quax, Wim J.; Kayser, Oliver

doi:10.1007/s00253-007-0953-5

Cited by 91 publications

(85 citation statements)

References 43 publications

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“…We observed a very strong up-regulation of the phage shock protein PspA, which is thought to counteract proton leakage upon membrane damage (25), and an even stronger up-regulation of the PspA homolog LiaH. In addition, we observed upregulation of the lia response regulator LiaR, ResD, which is involved in regulation of anaerobic respiration, and IspH, which is involved in the synthesis of the lipid carrier isopentenyl phosphate and the menaquinone precursor isoprenoid (26)(27)(28)(29).…”

Section: Resultsmentioning

confidence: 63%

Daptomycin inhibits cell envelope synthesis by interfering with fluid membrane microdomains

Müller

Wenzel

Strahl

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

370

506

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Daptomycin is a highly efficient last-resort antibiotic that targets the bacterial cell membrane. Despite its clinical importance, the exact mechanism by which daptomycin kills bacteria is not fully understood. Different experiments have led to different models, including (i) blockage of cell wall synthesis, (ii) membrane pore formation, and (iii) the generation of altered membrane curvature leading to aberrant recruitment of proteins. To determine which model is correct, we carried out a comprehensive mode-of-action study using the model organism Bacillus subtilis and different assays, including proteomics, ionomics, and fluorescence light microscopy. We found that daptomycin causes a gradual decrease in membrane potential but does not form discrete membrane pores. Although we found no evidence for altered membrane curvature, we confirmed that daptomycin inhibits cell wall synthesis. Interestingly, using different fluorescent lipid probes, we showed that binding of daptomycin led to a drastic rearrangement of fluid lipid domains, affecting overall membrane fluidity. Importantly, these changes resulted in the rapid detachment of the membrane-associated lipid II synthase MurG and the phospholipid synthase PlsX. Both proteins preferentially colocalize with fluid membrane microdomains. Delocalization of these proteins presumably is a key reason why daptomycin blocks cell wall synthesis. Finally, clustering of fluid lipids by daptomycin likely causes hydrophobic mismatches between fluid and more rigid membrane areas. This mismatch can facilitate proton leakage and may explain the gradual membrane depolarization observed with daptomycin. Targeting of fluid lipid domains has not been described before for antibiotics and adds another dimension to our understanding of membrane-active antibiotics.antibiotics | daptomycin | membrane potential | cell wall biosynthesis | Bacillus subtilis

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Section: Resultsmentioning

confidence: 63%

Daptomycin inhibits cell envelope synthesis by interfering with fluid membrane microdomains

Müller

Wenzel

Strahl

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

370

506

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“…Additionally, the knockout provides evidence that isoprene production closely tracks changes in DMAPP levels. While Julsing et al did not detect a change in isoprene emission in the B. subtilis 168 IPP isomerase mutant, 31) this strain produces substantially less isoprene than the 6051 strain used here, making differences in underlying rates of biosynthesis difficult to measure.…”

Section: Discussionmentioning

confidence: 85%

Evidence of Isoprenoid Precursor Toxicity inBacillus subtilis

Sivy

Fall

Rosenstiel

2011

Bioscience, Biotechnology, and Biochemistry

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“…Saccharomyces cerevisiae (14,38,39). G3P, glyceraldehyde-3-phosphate; DXP, 1-deoxy-D-xylulose 5-phosphate; MEP, 2-C-methyl-D-erythritol 4-phosphate; CDP-ME, 4-diphosphocytidyl-2-C-methylerythritol; CDP-MEP, CDP-ME 2-phosphate; MEC, 2-C-methyl-D-erythritol-2,4-cyclodiphosphate; HMBPP, (E)-4-hydroxy-3-methylbut-2-enyl-diphosphate; AcCoA, acetyl coenzyme A (acetyl-CoA); AACoA, acetoacetyl-CoA; HMG-CoA, hydroxymethylglutaryl-CoA; MVAP, mevalonate-5-phosphate; MVAPP, mevalonate-5-diphosphate; GPP, geranyl diphosphate; FPP, farnesyl diphosphate; GGPP, geranylgeranyl diphosphate; Dxs, 1-deoxy-D-xylulose-5-phosphate synthase; IspC, 1-deoxy-D-xylulose-5-phosphate reductoisomerase; IspD, 4-diphosphocytidyl-2-C-methyl-Derythritol synthase; IspE, 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase; IspF, 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase; IspG, 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate synthase; IspH, 1-hydroxy-2-methyl-butenyl 4-diphosphate reductase; IspA, farnesyl diphosphate synthase; IspS, isoprene synthase; AtoB, acetoacetyl-CoA thiolase; HMGS, hydroxymethylglutaryl-CoA synthase; HMGR, hydroxymethylglutaryl-CoA reductase; MK, mevalonate kinase; PMK, phosphomevalonate kinase; MPD, mevalonate pyrophosphate decarboxylase; Idi, isopentenyl pyrophosphate isomerase.…”

Section: Fig 1 Pathways Of Isoprenoid Biosynthesis In E Coli B Smentioning

confidence: 99%

“…Pyruvate and G3P are condensed to form DXP by 1-deoxy-D-xylulose-5-phosphate synthase (Dxs), and subsequently, 6 enzymes, encoded by the ispC (dxr), ispD, ispE, ispF, ispG, and ispH genes, catalyze sequential reactions converting DXP to isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) (39). IPP and its isomer DMAPP can be interconverted by isopentenyl pyrophosphate isomerase, which is encoded by idi (fni) (14). IPP and DMAPP are universal precursors for the biosynthesis of isoprene and other isoprenoids, such as sterols, carotenoids, and dolichols.…”

mentioning

confidence: 99%

Enhancing Isoprene Production by Genetic Modification of the 1-Deoxy- d -Xylulose-5-Phosphate Pathway in Bacillus subtilis

Xue

Ahring

2011

Appl Environ Microbiol

109

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To enhance the production of isoprene, a volatile 5-carbon hydrocarbon, in the Gram-positive spore-forming rod-shaped bacterium Bacillus subtilis, 1-deoxy-D-xylulose-5-phosphate synthase (Dxs) and 1-deoxy-D-xylulose-5-phosphate reductoisomerase (Dxr) were overexpressed in B. subtilis DSM 10. For the strain that overexpresses Dxs, the yield of isoprene was increased 40% over that by the wild-type strain. In the Dxr overexpression strain, the level of isoprene production was unchanged. Overexpression of Dxr together with Dxs showed an isoprene production level similar to that of the Dxs overproduction strain. The effects of external factors, such as stress factors including heat (48°C), salt (0.3 M NaCl), ethanol (1%), and oxidative (0.005% H 2 O 2 ) stress, on isoprene production were further examined. Heat, salt, and H 2 O 2 induced isoprene production; ethanol inhibited isoprene production. In addition, induction and repression effects are independent of SigB, which is the general stress-responsive alternative sigma factor of Gram-positive bacteria.Isoprene, also known as 2-methyl-1,3-butadiene, is a volatile 5-carbon pure hydrocarbon. Derived from biomass through a biochemical process, isoprene can be utilized as a fossil fuel alternative and a platform chemical for the production of highvalue biobased chemicals, such as rubber, elastomers, and isoprenoid medicines (17, 39). Isoprene has several advantages over ethanol. First, it is much easier to separate from the fermentation broth, since it is present in the upper gas phase of a fermentor due to its low boiling point (34°C) and low solubility in water. Second, bacteria are more tolerant to isoprene than to ethanol (the MIC of isoprene for Bacillus subtilis strain DSM 10 is more than 10%, which is higher than the MIC of ethanol according to our unpublished data). Third, isoprene is a highly versatile molecule that can be biochemically and thermochemically re-formed into more complicated products.Isoprene is produced both by eukaryotes and by prokaryotes, including humans, plants, yeasts, and bacteria (18). Two pathways in isoprene biosynthesis have been discovered: the mevalonate (MVA) pathway and the nonmevalonate route, or the 1-deoxy-D-xylulose-5-phosphate (DXP) pathway ( Fig. 1) (13, 39). The MVA pathway is present in eukaryotes, archaea, and the cytosol of higher plants; the DXP pathway is present in most bacteria, green algae, and the chloroplasts of higher plants (18). B. subtilis, the host in the present study, uses the DXP pathway to produce isoprene ( Fig. 1) (37). The DXP pathway initiates with the glycolytic intermediates pyruvate and glyceraldehyde-3-phosphate (G3P). Pyruvate and G3P are condensed to form DXP by 1-deoxy-D-xylulose-5-phosphate synthase (Dxs), and subsequently, 6 enzymes, encoded by the ispC (dxr), ispD, ispE, ispF, ispG, and ispH genes, catalyze sequential reactions converting DXP to isopentenyl diphosphate (IPP) and dimethylallyl diphosphate (DMAPP) (39). IPP and its isomer DMAPP can be interconverted by isopentenyl pyrophosphat...

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Functional analysis of genes involved in the biosynthesis of isoprene in Bacillus subtilis

Cited by 91 publications

References 43 publications

Daptomycin inhibits cell envelope synthesis by interfering with fluid membrane microdomains

Daptomycin inhibits cell envelope synthesis by interfering with fluid membrane microdomains

Evidence of Isoprenoid Precursor Toxicity inBacillus subtilis

Enhancing Isoprene Production by Genetic Modification of the 1-Deoxy- d -Xylulose-5-Phosphate Pathway in Bacillus subtilis

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