Enhancing Production of Bio-Isoprene Using Hybrid MVA Pathway and Isoprene Synthase in E. coli

Abstract

The depleting petroleum reserve, increasingly severe energy crisis, and global climate change are reigniting enthusiasm for seeking sustainable technologies to replace petroleum as a source of fuel and chemicals. In this paper, the efficiency of the MVA pathway on isoprene production has been improved as follows: firstly, in order to increase MVA production, the source of the “upper pathway” which contains HMG-CoA synthase, acetyl-CoA acetyltransferase and HMG-CoA reductase to covert acetyl-CoA into MVA has been changed from Saccharomyces cerevisiae to Enterococcus faecalis; secondly, to further enhance the production of MVA and isoprene, a alanine 110 of the mvaS gene has been mutated to a glycine. The final genetic strain YJM25 containing the optimized MVA pathway and isoprene synthase from Populus alba can accumulate isoprene up to 6.3 g/L after 40 h of fed-batch cultivation.

Scalable production of mechanically tunable block polymers from sugar

“…2A). The strain expressing the L. casei enzymes produced 50% more mevalonate than other engineered E. coli strains described in previous work (30)(31)(32).…”

A synthetic biochemistry molecular purge valve module that maintains redox balance

“…Isoprene is a platform chemical for a variety of products, but it is mostly employed in the production of synthetic rubber [24][25][26] . The isoprenoid pathway also provides precursors for over 25,000 known biomolecules including drugs such as taxol and potential biofuels 27 .…”

“…The isoprenoid pathway also provides precursors for over 25,000 known biomolecules including drugs such as taxol and potential biofuels 27 . There have been a number of efforts to produce isoprene in microorganisms 24,26,28,29 and the best reported yield is 28% from glucose 24 . We recently showed that a synthetic biochemistry system could produce isoprene in 495% yield from pyruvate as long as high-energy cofactors were added 30 .…”

Isoprenoids Production from Lipid-Extracted Microalgal Biomass Residues Using the Engineered <em>E. coli</em>

“…Engineered E. coli strain, YJM25 was used during isoprene fermentation [20], YJM29 was utilized in α-pinene biosynthesis [21] and FHR-2 (E. coli BL21 (DE3) (pACYDuet-1-mvaE-mvaS-GPPS2-QH6, pTrcHis2B-ERG8-ERG12-ERG19-IDI1) was applied for β-pinene production [22]. The fermentation procedure was carried out as reported in the previous study [23] with some modifications.…”

“…Optical density (OD) of the bacteria was measured with a spectrophotometer (Cary-50, Varian Inc. USA) at a wavelength of 600 nm. The isoprene, α-pinene and β-pinene production were analyzed as described earlier [20][21][22] by a gas chromatograph (GC) equipped with a flame ionization detector (FID) and a HP-1 column (30 m×0.25 mm×0.25 μm, Agilent). The concentration of target production was calculated with peak area on the bases of a standard curve.…”