Difference between revisions of "Part:BBa K1122676"
 
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<partinfo>BBa_K1122676 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K1122676 SequenceAndFeatures</partinfo>
  
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===Expression alalysis===
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Construct submitted to the Registry is able to provide good levels of Pdc and AdhB expression (Figure B)
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[[File:sds_page_non_fusion.png]]
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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)
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===Ethanol production===
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Ethanol production was analysed in 24h aerobic culture and in 72h anaerobic culture (Figure C)
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[[File:ethanol_production_non_fusion.png]]
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Figure C. Ethanol production using BBa_K1122676
  
 
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Latest revision as of 19:27, 4 October 2013

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.

Bioethanol introduction 1.jpg

Figure A. Ethanol is generated from pyruvate (fed into the pathway from glycolysis) via an Acetaldehyde intermediate.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1109
    Illegal AgeI site found at 2315
  • 1000
    COMPATIBLE WITH RFC[1000]


Expression alalysis

Construct submitted to the Registry is able to provide good levels of Pdc and AdhB expression (Figure B)

Sds page non fusion.png

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)

Ethanol production non fusion.png

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.