Engineered isoprenoid pathway enhances astaxanthin production in Escherichia coli.
The isoprenoid pathway is a versatile biosynthetic network leading to over 23,000 compounds. Similar to other biosynthetic pathways, the production of isoprenoids in microorganisms is controlled by the supply of precursors, among other factors. To engineer a host that has the capability to supply geranylgeranyl diphosphate (GGPP), a common precursor of isoprenoids, we cloned and overexpressed isopentenyl diphosphate (IPP) isomerase (encoded by idi) from Escherichia coli and GGPP synthase (encoded by gps) from the archaebacterium Archaeoglobus fulgidus. The latter was shown to be a multifunctional enzyme converting dimethylallyl diphosphate (DMAPP) to GGPP. These two genes and the gene cluster (crtBIYZW) of the marine bacterium Agrobacterium aurantiacum were introduced into E. coli to produce astaxanthin, an orange pigment and antioxidant. This metabolically engineered strain produces astaxanthin 50 times higher than values reported before. To determine the rate-controlling steps in GGPP production, the IDI-GPS pathway was compared with another construct containing idi, ispA (encoding farnesyl diphosphate (FPP) synthase in E. coli), and crtE (encoding GGPP synthase from Erwinia uredovora). Results show that the conversion from FPP to GGPP is the first bottleneck, followed sequentially by IPP isomerization and FPP synthesis. Removal of these bottlenecks results in an E. coli strain providing sufficient precursors for in vivo synthesis of isoprenoids.
Wang
C W
CW
Department of Chemical Engineering, University of California, Los Angeles, California 90095-1592, USA.
Oh
M K
MK
Liao
J C
JC
D019883:Alkyl and Aryl Transferases / Q000235:genetics / Q000378:metabolism; D001105:Archaea / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D001709:Biotechnology; D019748:Carbon-Carbon Double Bond Isomerases / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D051231:Farnesyltranstransferase; D015870:Gene Expression; D005818:Genetic Engineering; D045782:Hemiterpenes; D007700:Kinetics; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D024341:Xanthophylls; D019207:beta Carotene / Q000031:analogs & derivatives* / Q000096:biosynthesis
D017931:DNA Primers; D045782:Hemiterpenes; D011106:Polyisoprenyl Phosphates; D024341:Xanthophylls; D019207:beta Carotene; C005948:astaxanthine; D019883:Alkyl and Aryl Transferases; D051231:Farnesyltranstransferase; D019748:Carbon-Carbon Double Bond Isomerases; C021453:isopentenyldiphosphate delta-isomerase