title: The production of the sesquiterpene β-caryophyllene in a transgenic strain of the cyanobacterium Synechocystis.
abstract: The plant secondary metabolite, β-caryophyllene, is a ubiquitous component of many plant resins that has traditionally been used in the cosmetics industry to provide a woody, spicy aroma to cosmetics and perfumes. Clinical studies have shown it to be potentially effective as an antibiotic, anesthetic, and anti-inflammatory agent. Additionally, there is significant interest in engineering phototrophic microorganisms with sesquiterpene synthase genes for the production of biofuels. Currently, the isolation of β-caryophyllene relies on purification methods from oleoresins extracted from large amounts of plant material. An engineered cyanobacterium platform that produces β-caryophyllene may provide a more sustainable and controllable means of production. To this end, the β-caryophyllene synthase gene (QHS1) from Artemisia annua was stably inserted, via double homologous recombination, into the genome of the cyanobacterium Synechocystis sp. strain PCC6803. Gene insertion into Synechocystis was confirmed through PCR assays and sequencing reactions. Transcription and expression of QHS1 were confirmed using RT-PCR, and synthesis of β-caryophyllene was confirmed in the transgenic strain using GC-FID and GC-MS analysis.
authors.affiliation: School of Biological Sciences, University of Northern Colorado, Ross Hall, Room 2480, Greeley, CO 80639, United States.
authors.forename: Robert E
authors.initials: RE
authors.lastname: Reinsvold
authors.forename: Robert E
authors.initials: RE
authors.lastname: Jinkerson
authors.forename: Randor
authors.initials: R
authors.lastname: Radakovits
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Posewitz
authors.forename: Chhandak
authors.initials: C
authors.lastname: Basu
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000235:genetics*; D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000032:analysis / Q000378:metabolism*; D031172:Artemisia annua / Q000201:enzymology / Q000235:genetics*; D018076:DNA, Complementary / Q000235:genetics; D008401:Gas Chromatography-Mass Spectrometry; D015870:Gene Expression; D017343:Genes, Plant / Q000235:genetics; D030781:Organisms, Genetically Modified; D000081223:Polycyclic Sesquiterpenes; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012717:Sesquiterpenes / Q000032:analysis / Q000378:metabolism*; D046939:Synechocystis / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D014169:Transformation, Bacterial
chemical_list: D000894:Anti-Inflammatory Agents, Non-Steroidal; D018076:DNA, Complementary; D000081223:Polycyclic Sesquiterpenes; D012717:Sesquiterpenes; C024714:caryophyllene; D019883:Alkyl and Aryl Transferases; C108877:terpene synthase