Part:BBa_K5255003

MLACS1

Basic parts MLacs1 (BBa_K5255003) is a mutant of wild-type Lacs1. Long-chain acyl-coenzyme A (CoA) synthetases (LACSs) activate free fatty acids to acyl-CoA thioesters and play critical roles in fatty acid metabolism. Lacs1 can use DHA, CoA and ATP as substrates to catalyze the synthesis of DHA-CoA, which is the precursor of DHA-PC. MLacs1 sequence was obtained by mutating the active site of Lacs1 and it was introduced into Saccharomyces cerevisiae INVSC1 and E.coli BL21 (DE3). The activity of the enzyme expressed by MLacs1 was verified by Escherichia coli and the product catalyzed by MLacs1, DHA-CoA, was verified by Saccharomyces cerevisiae to ensure the synthesis of DHA-PC and the feasibility of subsequent synthesis of DHA-PC.

Figure 1 | MLacs1 generated using homology protein modeling with SWISS-MODEL. The model showed a large N-terminal and a small C-terminal

domain linked by highlighted cyan linker (Asp513-Leu518), with ligand ATP and cofactor Mg2+ residing inbetween the two domains.