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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
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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.

Peer review status:ACCEPTED

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