Engineering Escherichia coli for the synthesis of taxadiene, a key intermediate in the biosynthesis of taxol.
Taxadiene, the key intermediate of paclitaxel (Taxol) biosynthesis, has been prepared enzymatically from isopentenyl diphosphate in cell-free extracts of Escherichia coli by overexpressing genes encoding isopentenyl diphosphate isomerase, geranylgeranyl diphosphate synthase and taxadiene synthase. In addition, by the expression of three genes encoding four enzymes on the terpene biosynthetic pathway in a single strain of E. coli, taxadiene can be conveniently synthesized in vivo, at the unoptimized yield of 1.3mg per liter of cell culture. The success of both in vitro and in vivo synthesis of taxadiene bodes well for the future production of taxoids by non-paclitaxel producing organisms through pathway engineering.
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Center for Biological NMR, Department of Chemistry, PO Box 30012, Texas A & M University, College Station, TX 77842-3012, USA.
Roessner
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Croteau
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Scott
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D000475:Alkenes / Q000032:analysis / Q000378:metabolism*; D019883:Alkyl and Aryl Transferases; D000970:Antineoplastic Agents; D019748:Carbon-Carbon Double Bond Isomerases / Q000235:genetics / Q000378:metabolism; D004224:Diterpenes / Q000032:analysis / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D051232:Geranylgeranyl-Diphosphate Geranylgeranyltransferase; D045782:Hemiterpenes; D007535:Isomerases / Q000235:genetics / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D017239:Paclitaxel / Q000096:biosynthesis*; D014166:Transferases / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
D000475:Alkenes; D000970:Antineoplastic Agents; D004224:Diterpenes; D045782:Hemiterpenes; C093125:taxa-4(5),11(12)diene; D010088:Oxidoreductases; D014166:Transferases; C109461:deoxyxylulose-5-phosphate synthase; D019883:Alkyl and Aryl Transferases; D051232:Geranylgeranyl-Diphosphate Geranylgeranyltransferase; D007535:Isomerases; C095926:taxa-4(5),11(12)-diene synthase; D019748:Carbon-Carbon Double Bond Isomerases; C021453:isopentenyldiphosphate delta-isomerase; D017239:Paclitaxel