Reconstruction of the archaeal isoprenoid ether lipid biosynthesis pathway in Escherichia coli through digeranylgeranylglyceryl phosphate

One of the main differences between bacteria and archaea concerns their membrane composition. Whereas bacterial membranes are made up of glycerol-3-phosphate ester lipids, archaeal membranes are composed of glycerol-1-phosphate ether lipids. Here, we report the construction of a stable hybrid heterochiral membrane through lipid engineering of the bacterium Escherichia coli. By boosting isoprenoid biosynthesis and heterologous expression of archaeal ether lipid biosynthesis genes, we obtained a viable E. coli strain of which the membranes contain archaeal lipids with the expected stereochemistry. It has been found that the archaeal lipid biosynthesis enzymes are relatively promiscuous with respect to their glycerol phosphate backbone and that E. coli has the unexpected potential to generate glycerol-1-phosphate. The unprecedented level of 20–30% archaeal lipids in a bacterial cell has allowed for analyzing the effect on the mixed-membrane cell’s phenotype. Interestingly, growth rates are unchanged, whereas the robustness of cells with a hybrid heterochiral membrane appeared slightly increased. The implications of these findings for evolutionary scenarios are discussed.

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Draft genome analysis, poly-phasic study and lipid biosynthesis pathway of Scenedesmus sp. SVMIICT1

Bioresource Technology ◽

10.1016/j.biortech.2021.125809 ◽

2021 ◽

Vol 341 ◽

pp. 125809 ◽

Cited By ~ 1

Author(s):

Rajesh Kona ◽

S Venkata Mohan

Keyword(s):

Genome Analysis ◽

Draft Genome ◽

Lipid Biosynthesis ◽

Biosynthesis Pathway

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Ether lipid biosynthesis: alkyl-dihydroxyacetonephosphate synthase protein deficiency leads to reduced dihydroxyacetonephosphate acyltransferase activities

The Journal of Lipid Research ◽

10.1016/s0022-2275(20)32423-8 ◽

1999 ◽

Vol 40 (11) ◽

pp. 1998-2003 ◽

Cited By ~ 1

Author(s):

E.C.J.M. de Vet ◽

L. Ijlst ◽

W. Oostheim ◽

C. Dekker ◽

H.W. Moser ◽

...

Keyword(s):

Ether Lipid ◽

Lipid Biosynthesis ◽

Protein Deficiency

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Two Arabidopsis thaliana dihydrodipicolinate synthases, DHDPS1 and DHDPS2, are unequally redundant

Functional Plant Biology ◽

10.1071/fp12169 ◽

2012 ◽

Vol 39 (12) ◽

pp. 1058 ◽

Cited By ~ 9

Author(s):

Susan Jones-Held ◽

Luciana Pimenta Ambrozevicius ◽

Michael Campbell ◽

Bradley Drumheller ◽

Emily Harrington ◽

...

Keyword(s):

Escherichia Coli ◽

Arabidopsis Thaliana ◽

Growth Inhibition ◽

Segregation Analysis ◽

Narrow Range ◽

Insertion Mutant ◽

Biosynthesis Pathway ◽

Wild Type ◽

Exogenous Feeding ◽

Growth Inhibitory

In Arabidopsis thalinana (L.) Heynh., DHDPS1 and DHDPS2 encode orthologous dihydrodipicolinate synthases (DHDPS), the first enzyme of the lysine (Lys) biosynthesis pathway. A TDNA insertion mutant of dhdps2 was previously reported to be viable and to accumulate free threonine (Thr). Analysis of additional TDNA insertion lines showed that dhdps1 and dhdps2 mutants are both viable and that whereas dhdps2 mutants accumulate Thr, dhdps1 plants do not. Thr-accumulation was complemented by heterologous expression of Escherichia coli DapA, indicating that the phenotype is due to reduced DHDPS activity in dhdps2. DHDPS1 contributes ~30% towards the total DHDPS activity in leaves of young plants and DHDPS2 contributes 70%; therefore, the threshold of activity resulting in Thr accumulation lies within this narrow range. dhdps1–dhdps2 double mutants could not be isolated, even after exogenous feeding with Lys. Segregation analysis indicated that gametes lacking functional DHDPS genes are defective, as are embryos. Plants carrying only a single DHDPS2 gene do not accumulate Thr, but they show a gametophytic defect that is partially rescued by Lys application. Despite the accumulation of Thr, dhdps2 seedlings are no more sensitive than wild-type plants to growth inhibition by Lys or the Lys precursor diaminopimelate. They also are not rescued by methionine at growth-inhibitory Lys concentrations. Exogenous application of Lys and methionine to dhdps2 mutants did not reduce the accumulation of Thr.

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Utilization of exogenous fatty acids for complex lipid biosynthesis and its effect on de novo fatty acid formation in Escherichia coli K-12

Biochemistry ◽

10.1021/bi00725a027 ◽

1973 ◽

Vol 12 (1) ◽

pp. 164-171 ◽

Cited By ~ 40

Author(s):

David F. Silbert ◽

Thomas M. Ulbright ◽

J. L. Honegger

Keyword(s):

Escherichia Coli ◽

Fatty Acids ◽

Fatty Acid ◽

De Novo ◽

Lipid Biosynthesis ◽

Acid Formation ◽

Complex Lipid ◽

K 12

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Inhibitors of ether lipid biosynthesis

Biochemical and Biophysical Research Communications ◽

10.1016/0006-291x(81)91857-x ◽

1981 ◽

Vol 101 (3) ◽

pp. 1064-1070 ◽

Cited By ~ 2

Author(s):

Susan Hixson ◽

Richard Wolfenden

Keyword(s):

Ether Lipid ◽

Lipid Biosynthesis

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TDP-l-Megosamine Biosynthesis Pathway Elucidation and Megalomicin A Production in Escherichia coli

Applied and Environmental Microbiology ◽

10.1128/aem.03083-09 ◽

2010 ◽

Vol 76 (12) ◽

pp. 3869-3877 ◽

Cited By ~ 11

Author(s):

Mariana Useglio ◽

Salvador Peirú ◽

Eduardo Rodríguez ◽

Guillermo R. Labadie ◽

John R. Carney ◽

...

Keyword(s):

Escherichia Coli ◽

Heterologous Expression ◽

Gene Cluster ◽

Mass Spectrometric ◽

Mass Spectrometric Analysis ◽

Spectrometric Analysis ◽

Biosynthesis Pathway ◽

Biosynthetic Genes

ABSTRACT In vivo reconstitution of the TDP-l-megosamine pathway from the megalomicin gene cluster of Micromonospora megalomicea was accomplished by the heterologous expression of its biosynthetic genes in Escherichia coli. Mass spectrometric analysis of the TDP-sugar intermediates produced from operons containing different sets of genes showed that the production of TDP-l-megosamine from TDP-4-keto-6-deoxy-d-glucose requires only five biosynthetic steps, catalyzed by MegBVI, MegDII, MegDIII, MegDIV, and MegDV. Bioconversion studies demonstrated that the sugar transferase MegDI, along with the helper protein MegDVI, catalyzes the transfer of l-megosamine to either erythromycin C or erythromycin D, suggesting two possible routes for the production of megalomicin A. Analysis in vivo of the hydroxylation step by MegK indicated that erythromycin C is the intermediate of megalomicin A biosynthesis.

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