Heterologous expression and purification of the bicarbonate transporter BicA from Synechocystis sp. PCC 6803

Guanhong Bu, Sydney Parrish, Patrick R. Gleason, David R. Nielsen, Brent L. Nannenga

Research output: Contribution to journalArticlepeer-review

Abstract

The high-flux/low-affinity cyanobacterial bicarbonate transporter BicA is a member of sulfate permease/solute carrier 26 (SulP/SLC26) family and plays a major role in cyanobacterial inorganic carbon uptake. In order to study this important membrane protein, robust platforms for over-expression and protocols for purification are required. In this work we have optimized the expression and purification of BicA from strain Synechocystis sp. PCC 6803 (BicA6803) in Escherichia coli. It was determined that expression with C43 (DE3) Rosetta2 at 37 °C produced the highest levels of over-expressed BicA6803 relative to other strains screened, and membrane solubilization with n-dodecyl-β-D-maltopyranoside facilitated the purification of high levels of stable and homogenous BicA6803. Using these expression and purification strategies, the final yields of purified BicA were 6.5 ± 1.0 mg per liter of culture.

Original languageEnglish (US)
Article number105716
JournalProtein Expression and Purification
Volume175
DOIs
StatePublished - Nov 2020

Keywords

  • Bicarbonate transporter
  • Escherichia coli
  • Membrane protein
  • Protein overexpression
  • Protein purification
  • Synechocystis sp.PCC 6803

ASJC Scopus subject areas

  • Biotechnology

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