Biochemical Characterization of Acyl-CoA: Lysophosphatidylcholine Acyltransferase (LPCAT) Enzyme from the Seeds of Salvia hispanica

Mol Biotechnol. 2021 Oct;63(10):963-972. doi: 10.1007/s12033-021-00354-3. Epub 2021 Jun 15.

Abstract

Salvia hispanica (chia) is the highest reported terrestrial plant source of alpha-linolenic acid (ALA, ~ 65%), an ω-3 polyunsaturated fatty acid with numerous health benefits. The molecular basis of high ALA accumulation in chia is yet to be understood. We have identified lysophosphatidylcholine acyltransferase (LPCAT) gene from the developing seed transcriptome data of chia and carried out its biochemical characterization through heterologous expression in Saccharomyces cerevisiae. Expression profiling showed that the enzyme was active throughout the seed development, indicating a pivotal role in oil biosynthesis. The enzyme could utilize both saturated and unsaturated lysophosphatidylcholine substrates at the same rate, to synthesize phosphatidylcholine (PC). The enzyme also exhibited lysophosphatidic acid acyltransferase (LPAAT) activity, by preferring lysophosphatidic acid substrate. Substrate specificity studies showed that the enzyme preferred both monounsaturated and polyunsaturated fatty acyl CoAs over saturated CoAs. This activity may play a key role in enriching the PC fraction with polyunsaturated fatty acids (PUFAs). PUFAs present on PC can be transferred to oil through the action of other acyltransferases. Our results describe a new LPCAT enzyme that can be used to biotechnologically alter oilseed crops to incorporate more PUFA in its seed oil.

Keywords: Acyl CoA; PUFA; Phosphatidylcholine; Salvia hispanica; ShLPCAT.

MeSH terms

  • 1-Acylglycerophosphocholine O-Acyltransferase / genetics*
  • 1-Acylglycerophosphocholine O-Acyltransferase / metabolism*
  • Cloning, Molecular / methods*
  • Fatty Acids, Unsaturated / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation, Plant
  • Phosphatidylcholines / metabolism
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development
  • Salvia hispanica / enzymology
  • Salvia hispanica / genetics
  • Salvia hispanica / growth & development*
  • Seeds / enzymology
  • Seeds / genetics
  • Seeds / growth & development
  • Substrate Specificity

Substances

  • Fatty Acids, Unsaturated
  • Phosphatidylcholines
  • Plant Proteins
  • 1-Acylglycerophosphocholine O-Acyltransferase