title: Identification of a cyclooxygenase gene from the red alga Gracilaria vermiculophylla and bioconversion of arachidonic acid to PGF(2α) in engineered Escherichia coli.
abstract: Prostaglandins (PGs) are important local messenger molecules in many tissues and organs of animals including human. For applications in medicine and animal care, PGs are mostly purified from animal tissues or chemically synthesized. To generate a clean, reliable, and inexpensive source for PGs, we have now engineered expression of a suitable cyclooxygenase gene in Escherichia coli and achieved production levels of up to 2.7 mg l(-1) PGF(2α). The cyclooxygenase gene cloned from the red alga Gracilaria vermiculophylla appears to be fully functional without any eukaryotic modifications in E. coli. A crude extract of the recombinant E. coli cells is able to convert in vitro the substrate arachidonic acid (AA) to PGF(2α). Furthermore, these E. coli cells produced PGF(2α) in a medium supplemented with AA and secreted the PGF(2α) product. To our knowledge, this is the first report of the functional expression of a cyclooxygenase gene and concomitant production of PGF(2α) in E. coli. The successful microbial synthesis of PGs with reliable yields promises a novel pharmaceutical tool to produce PGF(2α) at significantly reduced prices and greater purity.
authors.affiliation: Laboratory of Plant Gene Technology, Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi-machi, Ishikawa 921-8836, Japan.
authors.forename: Hirosuke
authors.initials: H
authors.lastname: Kanamoto
authors.forename: Miho
authors.initials: M
authors.lastname: Takemura
authors.forename: Kanji
authors.initials: K
authors.lastname: Ohyama
mesh_terms: D000595:Amino Acid Sequence; D016718:Arachidonic Acid / Q000378:metabolism*; D001711:Biotransformation; D003001:Cloning, Molecular; D003470:Culture Media / Q000737:chemistry; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D015237:Dinoprost / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics; D005818:Genetic Engineering; D044724:Gracilaria / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D053858:Metabolic Networks and Pathways / Q000235:genetics; D008969:Molecular Sequence Data; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D003470:Culture Media; D018744:DNA, Plant; D016718:Arachidonic Acid; D015237:Dinoprost; D011451:Prostaglandin-Endoperoxide Synthases