Identification of a cyclooxygenase gene from the red alga Gracilaria vermiculophylla and bioconversion of arachidonic acid to PGF2α in engineered Escherichia coli
Identification of a cyclooxygenase gene from the red alga Gracilaria vermiculophylla and bioconversion of arachidonic acid to PGF2α in engineered Escherichia coli
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DOI：
10.1007/s00253-011-3349-5
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发表时间：
2011-08-01
影响因子：
5
通讯作者：
Ohyama, Kanji
中科院分区：
文献类型：
--
作者：
Kanamoto, Hirosuke;Takemura, Miho;Ohyama, Kanji
前列腺素（Prostaglandins，PGs）是动物包括人类在内的多种组织器官中重要的局部信使分子。对于在医学和动物护理中的应用，PG主要从动物组织中纯化或化学合成。为了产生一种清洁、可靠和廉价的前列腺素来源，我们现在已经在大肠杆菌中设计了合适的环氧合酶基因的表达，并实现了高达2.7 mg l（-1）PGF（2 α）的生产水平。克隆的红江蓠环氧合酶基因在大肠杆菌中没有经过任何真核修饰，具有完整的功能。杆菌重组E.大肠杆菌细胞能够在体外将底物花生四烯酸（AA）转化为PGF（2 α）。此外，这些E.大肠杆菌细胞在补充有AA的培养基中产生PGF（2 α）并分泌PGF（2 α）产物。据我们所知，这是第一次报道环氧合酶基因在大肠杆菌中的功能性表达和PGF（2 α）的伴随产生。杆菌具有可靠产量的PG的成功微生物合成有望成为以显著降低的价格和更高的纯度生产PGF（2 α）的新型制药工具。
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(2 alpha). 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(2 alpha). Furthermore, these E. coli cells produced PGF(2 alpha) in a medium supplemented with AA and secreted the PGF(2 alpha) product. To our knowledge, this is the first report of the functional expression of a cyclooxygenase gene and concomitant production of PGF(2 alpha) in E. coli. The successful microbial synthesis of PGs with reliable yields promises a novel pharmaceutical tool to produce PGF(2 alpha) at significantly reduced prices and greater purity.