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  • Heterologous overexpression of a monotopic glucosyltransferase (MGS) induces fatty acid remodeling in Escherichia coli membranes.

Heterologous overexpression of a monotopic glucosyltransferase (MGS) induces fatty acid remodeling in Escherichia coli membranes.

Biochimica et biophysica acta (2014-04-15)
Candan Ariöz, Hansjörg Götzke, Ljubica Lindholm, Jonny Eriksson, Katarina Edwards, Daniel O Daley, Andreas Barth, Ake Wieslander
ZUSAMMENFASSUNG

The membrane protein monoglucosyldiacylglycerol synthase (MGS) from Acholeplasma laidlawii is responsible for the creation of intracellular membranes when overexpressed in Escherichia coli (E. coli). The present study investigates time dependent changes in composition and properties of E. coli membranes during 22h of MGS induction. The lipid/protein ratio increased by 38% in MGS-expressing cells compared to control cells. Time-dependent screening of lipids during this period indicated differences in fatty acid modeling. (1) Unsaturation levels remained constant for MGS cells (~62%) but significantly decreased in control cells (from 61% to 36%). (2) Cyclopropanated fatty acid content was lower in MGS producing cells while control cells had an increased cyclopropanation activity. Among all lipids, phosphatidylethanolamine (PE) was detected to be the most affected species in terms of cyclopropanation. Higher levels of unsaturation, lowered cyclopropanation levels and decreased transcription of the gene for cyclopropane fatty acid synthase (CFA) all indicate the tendency of the MGS protein to force E. coli membranes to alter its usual fatty acid composition.

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Sigma-Aldrich
L-α-Phosphatidylethanolamin aus Eigelb, Type III, ≥97% (TLC), lyophilized powder
Sigma-Aldrich
L-α-Phosphatidylethanolamine from Glycine max (soybean), Type IV, ≥98% (TLC), lyophilized powder
Sigma-Aldrich
Phosphorylase b aus Kaninchenmuskel, lyophilized powder, ≥20 units/mg protein, 2× crystallization
Sigma-Aldrich
L-α-Phosphatidylethanolamin aus Eigelb, Type III, 10 mg/mL in chloroform, ≥97%, solution
Sigma-Aldrich
Phosphorylase b aus Kaninchenmuskel, For use as a marker in SDS-PAGE