Biotechnological production of the European corn borer sex pheromone in the yeast Yarrowia lipolytica. Issue 6 (18th March 2021)

• Record Type: Journal Article
• Authors: Petkevicius, Karolis; Koutsoumpeli, Eleni; Betsi, Petri Christina; Ding, Bao‐Jian; Kildegaard, Kanchana Rueksomtawin; Jensen, Hilbert; Mezo, Nora; Mazziotta, Andrea; Gabrielsson, Anders; Sinkwitz, Christina; Lorantfy, Bettina; Holkenbrink, Carina; Löfstedt, Christer; Raptopoulos, Dimitris; Konstantopoulou, Maria; Borodina, Irina
• Is Part Of: Biotechnology journal. Volume 16:Issue 6(2021)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-03-18
• Subjects: fatty acyl‐CoA desaturases -- fatty acyl‐CoA reductases -- fatty alcohols -- insect pheromones -- Yarrowia lipolytica; Biotechnology -- Periodicals; 660.605
• DOI: 10.1002/biot.202100004
• Languages: English
• ISSNs: 1860-6768

Abstract

Abstract: The European corn borer (ECB) Ostrinia nubilalis is a widespread pest of cereals, particularly maize. Mating disruption with the sex pheromone is a potentially attractive method for managing this pest; however, chemical synthesis of pheromones requires expensive starting materials and catalysts and generates hazardous waste. 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 ( Z 11‐14:OH), which can then be chemically acetylated to ( Z )‐11‐tetradecenyl acetate ( Z 11‐14:OAc), the main pheromone component of the Z‐race of O. nubilalis . First, a C14 platform strain with increased biosynthesis of myristoyl‐CoA was obtained by introducing a point mutation into the α‐subunit of fatty acid synthase, replacing isoleucine 1220 with phenylalanine (Fas2p I1220F ). The intracellular accumulation of myristic acid increased 8.4‐fold. Next, fatty acyl‐CoA desaturases (FAD) and fatty acyl‐CoA reductases (FAR) from nine different species of Lepidoptera were screened in the C14 platform strain, individually and in combinations. A titer of 29.2 ± 1.6 mg L ‐1 Z 11‐14: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, reaching 93.9 ± 11.7 mg L ‐1 in small‐scale cultivation. When the same engineered strain was cultivated in controlled 1 L bioreactors in fed‐batch mode, 188.1 ± 13.4 mg L ‐1 of Z 11‐14:OH was obtained. Fatty alcohols were extracted from the biomass and chemically acetylated to obtain Z 11‐14: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, although full precopulatory behavior was observed less often than for the chemically synthesized pheromone blend. The study paves the way for the production of ECB pheromone by fermentation. Abstract : European Corn Borer ( Ostrinia nubilalis ) is a serious pest of crops worldwide. In order to tackle infestation of this insect, mating disruption by sex pheromones could be used. In this study authors established bio‐based production of O. nubilalis sex pheromone Z11‐14:OAc by engineering yeast Y. lipolytica . Yeast‐derived pheromone modulated the behavior of O. nubilalis males. This study shows potential of sustainable production of O. nubilalis pheromone via yeast fermentation.