Expression of an exogenous isopentenyl diphosphate isomerase gene enhances isoprenoid biosynthesis in Escherichia coli

Abstract

Escherichia coli expressing the Erwinia carotenoid biosynthesis genes, crtE, crtB, crtI and crtY, form yellow-coloured colonies due to the presence of beta-carotene. This host was used as a visible marker for evaluating regulatory systems operating in isoprenoid biosynthesis of E. coli. cDNAs enhancing carotenoid levels were isolated from the yeast Phaffia rhodozyma and the green alga Haematococcus pluvialis. Nucleotide sequence analysis indicated that they coded for proteins similar to isopentenyl diphosphate (IPP) isomerase of the yeast Saccharomyces cerevisiae. Determination of enzymic activity confirmed the identity of the gene products as IPP isomerases. The corresponding gene was isolated from the genomic library of S. cerevisiae based on its nucleotide sequence, and was confirmed to have the same effect as the above two IPP isomerase genes when introduced into the E. coli transformant accumulating beta-carotene. In the three E. coli strains carrying the individual exogenous IPP isomerase genes, the increases in carotenoid levels are comparable to the increases in IPP isomerase enzyme activity with reference to control strains possessing the endogenous gene alone. These results imply that IPP isomerase forms an influential step in isoprenoid biosynthesis of the prokaryote E. coli, with potential for the efficient production of industrially useful isoprenoids by metabolic engineering.

The Arabidopsis thaliana Type I Isopentenyl Diphosphate Isomerases Are Targeted to Multiple Subcellular Compartments and Have Overlapping Functions in Isoprenoid Biosynthesis

“…both IPP and DMAPP (Eisenreich et al, 2004), previous work has suggested that IDI activity in the plastid is necessary for maximal rates of synthesis of isoprenoids such as carotenoids (Albrecht and Sandmann, 1994;Page et al, 2004). Furthermore, it has also been shown that in E. coli cells that were engineered to synthesize carotenoids from DMAPP and IPP produced via the MEP pathway, IDI activity was a limiting factor in carotenoid biosynthesis (Kajiwara et al, 1997;Cunningham and Gantt, 2000). That exogenously supplied mevalonate can complement a block in 1-deoxyxylulose 5-phosphate synthase activity and chlorophyll biosynthesis (presumably following the transport of IPP to the plastid) also suggests the presence of IDI activity in this compartment (Esté vez et al, 2000;Nagata et al, 2002).…”

Differential expression of two isopentenyl pyrophosphate isomerases and enhanced carotenoid accumulation in a unicellular chlorophyte

“…We previously had taken advantage of this phenotype in the identification and functional analysis of a ␤-carotene hydroxylase from Arabidopsis (11). Also, a similar construct made with the gene equivalent to ipiHp2, and an isomerase gene from the yeast Phaffia rhodozyma, led to enhanced carotenoid accumulation in E. coli (32). In the mutant Car-3, which accumulates carotenoids at an earlier stage than wild type, a higher IPP isomerase content might be expected.…”

Molecular biogeochemistry of modern and ancient marine microbes

“…One gene in the PR photosystem cluster was dispensable under our conditions: the idi gene that encodes IPP ␦-isomerase, an activity already present in E. coli (17), as is the FPP synthase, catalyzing the next two steps in the pathway (23). The presence of the idi gene in the cluster likely enables retinal production in the native organism because isomerization of IPP to dimethylallyl pyrophosphate can be a rate-limiting step in ␤-carotene biosynthesis (24,25).…”

“…As expected for the regular isoprenoid side-chain, C 22 was the least abundant homologue. C 24 tetracyclic terpane was also detected in many samples. As the biological source(s) of cheilanthanes and C 24 tetracyclic terpane are not known , little paleobiological interpretation can be made of their presence.…”