Ancestral reconstruction and consensus mutagenesis provide insight into the substrate specificity and evolution of the marine algae Δ6-desaturases — ASN Events
  1. Schedule
  2. Session Five - Poster Session A including Refreshments
  3. Ancestral reconstruction and consensus mutagenesis provide insight into the substrate specificity and evolution of the marine algae Δ6-desaturases

Ancestral reconstruction and consensus mutagenesis provide insight into the substrate specificity and evolution of the marine algae Δ6-desaturases (#139)

Dongdi Li , Thomas Vanhercke 1 , James R Petrie 1 , Surinder P Singh 1 , Colin Jackson 2
  1. CSIRO Food Futures National Research Flagship, Canberra, ACT 2601, Australia
  2. Research School of Chemistry, The Australian National University, Canberra, ACT 0200, Australia

In marine algae, the biosynthesis of these molecules begins with the catalyzed desaturation of omega-3/6-long-chain polyunsaturated fatty acids (ω3/6-LCPUFAs) by membrane bound Δ6 desaturases. In order to gain more insight into the substrate specificity of these enzymes, we have conducted consensus mutagenesis and ancestral reconstruction of the Δ6 desaturase family, focusing on the Δ6 desaturase from Micromonas pusilla (MpΔ6des). Analysis of the variants not only highlights the importance of the residues near the putative active site in determining substrate preference, it also demonstrated the contribution of the N-terminal cytochrome b5 domain to substrate specificity. In addition, the recombinant cytochrome b5 was expressed and purified for structural characterization. This result could lead to detailed understanding of the substrate binding mode of this class of enzymes.