Research
| Title： | Biochemical Properties and Substrate Specificity of Two Acyl-CoA:Lysophosphatidic Acid Acyltransferases (PtATS2a and PtATS2b) from Diatom Phaeodactylum tricornutum |
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| First author： | Jasieniecka-Gazarkiewicz, Katarzyna; Polonska, Ada; Gong, Yangmin; Banas, Antoni |
| Journal： | INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES |
| Years： | 2025 |
| DOI： | 10.3390/ijms26209936 |
| Abstract： | Microsomal fractions from yeast Delta ale1 cells harbouring the empty plasmid pYES2/CT and from yeast cells overexpressing PtATS2a (Phatr3_J11916) or PtATS2b (Phatr3_J43099) were used in the studies. When sn-1-18:1-LPA and [14C]16:0-CoA were used as exogenous substrates, both PtATS2a and PtATS2b showed the highest activity at 23 degrees C in the range of temperatures tested from 10 to 60 degrees C. Both enzymes showed the highest activity in alkaline pH. For PtATS2a, it was pH 10 while for PtATS2b, it was pH 11. At pH 6 and pH 12, the activities of both enzymes were very low. The calcium ions at concentrations of 0.05-1 mM drastically decreased the activity of both enzymes. The magnesium ions at a concentration of 0.05 mM had a little effect on the activity of both enzymes, while higher concentrations (0.5 mM and 1 mM) significantly inhibited their activity. To study the substrate specificity, seventeen different acyl-CoAs in combinations with sn-1-[14C]18:1-LPA were used. PtATS2a showed the highest preference for 18:4-CoA n-3 while PtATS2b for 18:1-CoA. The pattern of utilisation of other acyl-CoAs tested also differed between the two enzymes. The presented studies, for the first time, characterised LPAAT type enzymes from diatoms, organisms that naturally produced very-long-chain polyunsaturated fatty acids (VLC-PUFA). |
