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Characterisation of Acetyl‐CoA Thiolase: The First Enzyme in the Biosynthesis of Terpenic Sex Pheromone Components in the Labial Gland of Bombus terrestris
Authors:Jana Brabcová  Zuzana Demianová  Jiří Kindl  Iva Pichová  Irena Valterová  Marie Zarevúcka
Affiliation:1. Institute of Organic Chemistry and Biochemistry AS CR, 166 10 Prague 6 (Czech Republic);2. Research Institute of Molecular Pathology (IMP), Dr. Bohr‐Gasse 7, 1030 Vienna(Austria)
Abstract:Buff‐tailed bumblebees, Bombus terrestris, use a male sex pheromone for premating communication. Its main component is a sesquiterpene, 2,3‐dihydrofarnesol. This paper reports the isolation of a thiolase (acetyl‐CoA thiolase, AACT_BT), the first enzyme involved in the biosynthetic pathway leading to formation of isoprenoids in the B. terrestris male sex pheromone. Characterisation of AACT_BT might contribute to a better understanding of pheromonogenesis in the labial gland of B. terrestris males. The protein was purified to apparent homogeneity by column chromatography with subsequent stepwise treatment. AACT_BT showed optimum acetyltransferase activity at pH 7.1 and was strongly inhibited by iodoacetamide. The enzyme migrated as a band with an apparent mass of 42.9 kDa on SDS‐PAGE. MS analysis of an AACT_BT tryptic digest revealed high homology to representatives of the thiolase family. AACT_BT has 96 % amino acid sequence identity with the previously reported Bombus impatiens thiolase.
Keywords:acetyl‐CoA thiolase  biosynthesis  Bombus terrestris  labial gland  pheromones  terpenoids
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