title: Overexpression of hydroperoxide lyase, peroxygenase and epoxide hydrolase in tobacco for the biotechnological production of flavours and polymer precursors.
abstract: Plants produce short-chain aldehydes and hydroxy fatty acids, which are important industrial materials, through the lipoxygenase pathway. Based on the information that lipoxygenase activity is up-regulated in tobacco leaves upon infection with tobacco mosaic virus (TMV), we introduced a melon hydroperoxide lyase (CmHPL) gene, a tomato peroxygenase (SlPXG) gene and a potato epoxide hydrolase (StEH) into tobacco leaves using a TMV-based viral vector system to afford aldehyde and hydroxy fatty acid production. Ten days after infiltration, tobacco leaves infiltrated with CmHPL displayed high enzyme activities of 9-LOX and 9-HPL, which could efficiently transform linoleic acid into C(9) aldehydes. Protein extracts prepared from 1 g of CmHPL-infiltrated tobacco leaves (fresh weight) in combination with protein extracts prepared from 1 g of control vector-infiltrated tobacco leaves (as an additional 9-LOX source) produced 758 ± 75 μg total C(9) aldehydes in 30 min. The yield of C(9) aldehydes from linoleic acid was 60%. Besides, leaves infiltrated with SlPXG and StEH showed considerable enzyme activities of 9-LOX/PXG and 9-LOX/EH, respectively, enabling the production of 9,12,13-trihydroxy-10(E)-octadecenoic acid from linoleic acid. Protein extracts prepared from 1 g of SlPXG-infiltrated tobacco leaves (fresh weight) in combination with protein extracts prepared from 1 g of StEH-infiltrated tobacco leaves produced 1738 ± 27 μg total 9,12,13-trihydroxy-10(E)-octadecenoic acid isomers in 30 min. The yield of trihydroxyoctadecenoic acids from linoleic acid was 58%. C(9) aldehydes and trihydroxy fatty acids could likely be produced on a larger scale using this expression system with many advantages including easy handling, time-saving and low production cost.
authors.affiliation: Biotechnology of Natural Products, Technische Universität München, Freising, Germany.
authors.forename: Fong-Chin
authors.initials: FC
authors.lastname: Huang
authors.forename: Wilfried
authors.initials: W
authors.lastname: Schwab
mesh_terms: D000446:Aldehyde-Lyases / Q000378:metabolism*; D000447:Aldehydes / Q000378:metabolism*; D019662:Cucurbitaceae / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D004851:Epoxide Hydrolases / Q000378:metabolism*; D005818:Genetic Engineering; D019787:Linoleic Acid / Q000378:metabolism; D018551:Solanum lycopersicum / Q000235:genetics; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000201:enzymology; D030821:Plants, Genetically Modified / Q000378:metabolism; D011108:Polymers; D011198:Solanum tuberosum / Q000235:genetics; D014026:Nicotiana / Q000201:enzymology*
chemical_list: D000447:Aldehydes; D011108:Polymers; D003577:Cytochrome P-450 Enzyme System; D019787:Linoleic Acid; D006899:Mixed Function Oxygenases; C022333:peroxygenase; D004851:Epoxide Hydrolases; D000446:Aldehyde-Lyases; C054540:hydroperoxide lyase