Cloning of a palmitoyl-acyl carrier protein thioesterase from oil palm

Biochem Soc Trans. 2000 Dec;28(6):619-22.

Abstract

A palmitoyl-acyl carrier protein (ACP) thioesterase cDNA clone was isolated from an oil palm cDNA library. The cDNA was expressed in Escherichia coli as a glutathione S-transferase fusion protein and a crude bacterial extract was assayed for acyl-CoA-hydrolysing activity. The recombinant enzyme was able to hydrolyse medium- and long-chain acyl-CoAs. Northern-blot analysis showed a high level of gene expression in leaf, flower and 15-, 17- and 18-week mesocarp tissues. Low-level gene expression was detected in germinated seedlings and 8- and 12-week mesocarp tissues, but no transcript was detected in any kernel tissues. Southern-blot analysis indicated the presence of a single gene and we have also isolated a genomic clone using the cDNA as a probe. Two genomic fragments were subcloned and a 7 kb contiguous stretch of the oil palm genome was sequenced. Comparison of this sequence with the cDNA sequence identified a putative 93 amino acid transit peptide, most of which is missing from the cDNA. The coding region of the gene consisted of seven exons and six introns.

MeSH terms

  • Cloning, Molecular
  • Escherichia coli / genetics
  • Genomic Library
  • Kinetics
  • Plant Oils
  • Plants, Edible / enzymology*
  • Plants, Edible / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / metabolism
  • Thiolester Hydrolases / genetics
  • Thiolester Hydrolases / metabolism*

Substances

  • Plant Oils
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Thiolester Hydrolases
  • oleoyl-(acyl-carrier-protein) hydrolase