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: Fong-Chin Huang, Wilfried Schwab
• Journal: Plant biotechnology journal (Plant Biotechnol J) Vol. 10 Issue 9 Pg. 1099-109 (Dec 2012) ISSN: 1467-7652 [Electronic] England
• PMID: 22967031 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2012 The Authors Plant Biotechnology Journal © 2012 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd.

Chemical References

• Aldehydes
• Polymers
• Cytochrome P-450 Enzyme System
• Linoleic Acid
• Mixed Function Oxygenases
• peroxygenase
• Epoxide Hydrolases
• Aldehyde-Lyases
• hydroperoxide lyase

Topics

• Aldehyde-Lyases (metabolism)
• Aldehydes (metabolism)
• Cucurbitaceae (genetics)
• Cytochrome P-450 Enzyme System (metabolism)
• Epoxide Hydrolases (metabolism)
• Genetic Engineering
• Linoleic Acid (metabolism)
• Solanum lycopersicum (genetics)
• Mixed Function Oxygenases (metabolism)
• Molecular Sequence Data
• Plant Leaves (enzymology)
• Plants, Genetically Modified (metabolism)
• Polymers
• Solanum tuberosum (genetics)
• Tobacco (enzymology)