Part:BBa_K1122676
IPTG inducible pdc and adhB
This parts enables expression of pyruvate decarboxylase (pdc) and alcohol dehydrogenase B (adhB) using IPTG induction. Those enzymes are involved in conversion of pyruvate to ethanol via an acetaldehyde intermediate. The pdc and adhB sequences come from Zymomonas mobilis however, they are codon optimised for E. coli.
Usage and Biology
Microbial production of ethanol is of great importance due to its possible application as a biofuel. Increasing ethanol yields in bacteria is potentially beneficial as those are able to utilise a wider variety of renewable, biomass-derived carbon sources compared to standard ethanol producer: Sacharomyces cerevisiae. Production of ethanol in E. coli was achieved by expression of Zymomonas mobilis pyruvate decarboxylase (Pdc) and alcohol dehydrogenase B (AdhB)(Ingram et al.,.1987) . Reactions catalysed by those enzymes (Figure A) enable conversion of pyruvate to ethanol.
Figure A. Ethanol is generated from pyruvate (fed into the pathway from glycolysis) via an Acetaldehyde intermediate.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1109
Illegal AgeI site found at 2315 - 1000COMPATIBLE WITH RFC[1000]
Expression alalysis
Construct submitted to the Registry is able to provide good levels of Pdc and AdhB expression (Figure B)
Figure B. Two distinct bands present in second and third lane correspond to Pdc and AdhB. They are absent in induced vector only sample (last lane)
Ethanol production
Ethanol production was analysed in 24h aerobic culture and in 72h anaerobic culture (Figure C)
Figure C. Ethanol production using BBa_K1122676
References
INGRAM, L. O., CONWAY, T., CLARK, D. P., SEWELL, G. W. & PRESTON, J. F. 1987. GENETIC-ENGINEERING OF ETHANOL-PRODUCTION IN ESCHERICHIA-COLI. Applied and Environmental Microbiology, 53, 2420-2425.
//cds/enzyme
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