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Biotechnological production of the European corn borer sex pheromone in the yeast Yarrowia lipolytica
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  • Karolis Petkevicius,
  • Eleni Koutsoumpeli,
  • Petri Betsi,
  • Bao-Jian Ding,
  • Kanchana Rueksomtawin Kildegaard,
  • Hilbert Jensen,
  • Nora Mezo,
  • Andrea Mazziotta,
  • Anders Gabrielsson,
  • Christina Sinkwitz,
  • Bettina Lorantfy,
  • Carina Holkenbrink,
  • Christer Löfstedt,
  • Dimitris Raptopoulos,
  • Maria Konstantopoulou,
  • Irina Borodina
Karolis Petkevicius
Technical University of Denmark

Corresponding Author:[email protected]

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Eleni Koutsoumpeli
National Centre for Scientific Research-Demokritos
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Petri Betsi
National Centre for Scientific Research-Demokritos
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Bao-Jian Ding
Lund University
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Kanchana Rueksomtawin Kildegaard
BioPhero ApS
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Hilbert Jensen
BioPhero ApS
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Nora Mezo
BioPhero ApS
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Andrea Mazziotta
BioPhero ApS
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Anders Gabrielsson
BioPhero ApS
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Christina Sinkwitz
BioPhero ApS
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Bettina Lorantfy
BioPhero ApS
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Carina Holkenbrink
BioPhero ApS
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Christer Löfstedt
Lund University
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Dimitris Raptopoulos
Novagrica Hellas S.A
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Maria Konstantopoulou
National Centre for Scientific Research-Demokritos
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Irina Borodina
Technical University of Denmark
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Abstract

The European corn borer (ECB) Ostrinia nubilalis is a widespread pest of cereals. Mating disruption with the sex pheromone is a potentially attractive method for managing this pest. The goal of this study was to develop a biotechnological method for the production of ECB sex pheromone. Our approach was to engineer the oleaginous yeast Yarrowia lipolytica to produce (Z)-11-tetradecenol (Z11-14:OH), which can be chemically acetylated to (Z)-11-tetradecenyl acetate (Z11-14:OAc), the main pheromone component of the Z-race of O. nubilalis. Fatty acyl-CoA desaturases (FAD) and fatty acyl-CoA reductases (FAR) from nine different species of Lepidoptera were screened individually and in combinations. A titer of 29.2±1.6 mg/L Z11-14:OH was reached in small-scale cultivation with an optimal combination of a FAD (Lbo_PPTQ) from Lobesia botrana and FAR (HarFAR) from Helicoverpa armigera. When the second copies of FAD and FAR genes were introduced, the titer improved 2.1-fold. The native FAS1 gene’s overexpression led to a further 1.5-fold titer increase. When the same engineered strain was cultivated in controlled 1 L bioreactors in fed-batch mode, 188.1±13.4 mg/L of Z11-14:OH was obtained. Fatty alcohols were chemically acetylated to obtain Z11-14:OAc. Electroantennogram experiments showed that males of the Z-race of O. nubilalis were responsive to biologically-derived pheromone blend. Behavioral bioassays in a wind tunnel revealed attraction of male O. nubilalis at a level similar to that of the chemically synthesized pheromone used as a control, although full precopulatory behavior was observed less often. The study paves the way for the production of ECB pheromone by fermentation.
05 Jan 2021Submitted to Biotechnology Journal
14 Jan 2021Submission Checks Completed
14 Jan 2021Assigned to Editor
14 Jan 2021Reviewer(s) Assigned
01 Feb 2021Editorial Decision: Revise Minor
15 Feb 20211st Revision Received
16 Feb 2021Submission Checks Completed
16 Feb 2021Assigned to Editor
16 Feb 2021Reviewer(s) Assigned
19 Feb 2021Editorial Decision: Accept