Fig. 4
Functional validation of site-directed mutagenesis of EhDH domains. a Complementation plate assay of the dehydratase (DH) domains in Escherichia coli (E. coli) FabA Temperature-sensitive (Ts) mutant as conducted in Fig. 3. FabA, E. coli wild type dehydratase; Mutant, E. coli FabA Ts mutant with an empty vector; EhDH1-M, the mutant expresses the mutated DH1 domains from Emiliania huxleyi; EhDH2-M, the mutant expresses the mutated EhDH2 domains; EhDH1-M-DH2, the mutant expresses the mutated DH1 along with DH2; EhDH1-DH2-M, the mutant expresses the mutated DH2 along with DH1. b Fatty acid production following DH overexpression in wild-type E. coli. EV, E. coli Rosetta with an empty vector; EhDH1-M, E. coli Rosetta overexpressing the mutated EhDH1 domain; EhDH2-M, E. coli Rosetta overexpressing the mutated EhDH2 domain; EhDH1-M-DH2, E. coli Rosetta overexpressing the mutated DH1 along with DH2; EhDH1-DH2-M, E. coli Rosetta overexpressing the mutated DH2 along with DH1. Fatty acid abbreviations are the following: 14:0, myristic acid; 16:0, palmitic acid; 16:1, palmitoleic acid; 18:0, octadecanoic acid; 18:1, cis-vaccenic acid; SFAs saturated fatty acids, UFAs unsaturated fatty acids, Total total fatty acids. Values are reported as the means ± standard deviation of three independent biological replicates. Means with the same letters do not differ significantly. Statistical analysis of the results was conducted using one-way analysis of variance (P < 0.05)