Development of the photoheterotroph rhodobacter for functional membrane protein expression

Brandon S. Curtis, Philip D. Laible, Deborah K. Hanson, Wayne R. Curtis

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

This paper presents progress on the development of Rhodobacter sphaeroides as a platform for high-level expression of functional membrane proteins. The high capacity of Rhodobacter for membrane protein expression stems from the dense intracellular membranes synthesized by this organism under anaerobic photoheterotrophic conditions. Using plasmids with promoters from the light-harvesting pigments, inducible heterologous expression is accomplished simultaneously with membrane formation when the organism experiences reduced oxygen tension and low-light conditions. Unique growth characteristics of this organism are presented: although anaerobic growth ceases in the dark, anaerobic photoheterotrophic growth is as fast as aerobic growth and results in nearly 3-times greater biomass yield as a result of photosynthetic ATP production. Growth yield (on carbon basis) is also several-fold higher on defined media as compared to complex media - which is of particular interest for the application of isotopic labeling for membrane protein structural studies. Although transformed lines display dramatically reduced growth under low light conditions, this can be overcome with high light, even with 'knockout' strains where the dominant light harvesting protein (LH2) has been deleted to facilitate greater capacity for functional heterologous expression. Preliminary growth testing in a thin film photobioreactor combined with mass-balance rationalized fed-batch operation are shown to facilitate photoheterotrophic growth to nearly 10 grams DW/L (an order of magnitude higher than current practice in flask culture). Preliminary results demonstrate high-level membrane protein expression.

Original languageEnglish (US)
Title of host publicationAIChE100 - 2008 AIChE Annual Meeting, Conference Proceedings
StatePublished - 2008
Event2008 AIChE Annual Meeting, AIChE 100 - Philadelphia, PA, United States
Duration: Nov 16 2008Nov 21 2008

Other

Other2008 AIChE Annual Meeting, AIChE 100
CountryUnited States
CityPhiladelphia, PA
Period11/16/0811/21/08

Fingerprint

Membrane Proteins
Proteins
Membranes
Photobioreactors
Adenosinetriphosphate
Pigments
Labeling
Biomass
Plasmids
Carbon
Adenosine Triphosphate
Oxygen
Thin films
Testing

All Science Journal Classification (ASJC) codes

  • Chemical Engineering(all)
  • Chemistry(all)

Cite this

Curtis, B. S., Laible, P. D., Hanson, D. K., & Curtis, W. R. (2008). Development of the photoheterotroph rhodobacter for functional membrane protein expression. In AIChE100 - 2008 AIChE Annual Meeting, Conference Proceedings
Curtis, Brandon S. ; Laible, Philip D. ; Hanson, Deborah K. ; Curtis, Wayne R. / Development of the photoheterotroph rhodobacter for functional membrane protein expression. AIChE100 - 2008 AIChE Annual Meeting, Conference Proceedings. 2008.
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Curtis, BS, Laible, PD, Hanson, DK & Curtis, WR 2008, Development of the photoheterotroph rhodobacter for functional membrane protein expression. in AIChE100 - 2008 AIChE Annual Meeting, Conference Proceedings. 2008 AIChE Annual Meeting, AIChE 100, Philadelphia, PA, United States, 11/16/08.

Development of the photoheterotroph rhodobacter for functional membrane protein expression. / Curtis, Brandon S.; Laible, Philip D.; Hanson, Deborah K.; Curtis, Wayne R.

AIChE100 - 2008 AIChE Annual Meeting, Conference Proceedings. 2008.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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Curtis BS, Laible PD, Hanson DK, Curtis WR. Development of the photoheterotroph rhodobacter for functional membrane protein expression. In AIChE100 - 2008 AIChE Annual Meeting, Conference Proceedings. 2008