AcsA–AcsB: The core of the cellulose synthase complex from Gluconacetobacter hansenii ATCC23769

• Purification of cellulose synthase from Gluconacetobacter hansenii by two methods.
• Identification of the post-translational processing site of cellulose synthase.
• Determination of the steady state kinetic parameters for cellulose synthase.
• Exploration of the role of accessory proteins AcsC, AcsD, and ccpAx.

The gram-negative bacterium, Gluconacetobacter hansenii, produces cellulose of exceptionally high crystallinity in comparison to the cellulose of higher plants. This bacterial cellulose is synthesized and extruded into the extracellular medium by the cellulose synthase complex (CSC). The catalytic component of this complex is encoded by the gene AcsAB. However, several other genes are known to encode proteins critical to cellulose synthesis and are likely components of the bacterial CSC. We have purified an active heterodimer AcsA–AcsB from G. hansenii ATCC23769 to homogeneity by two different methods. With the purified protein, we have determined how it is post-translationally processed, forming the active heterodimer AcsA–AcsB. Additionally, we have performed steady-state kinetic studies on the AcsA–AcsB complex. Finally through mutagenesis studies, we have explored the roles of the postulated CSC proteins AcsC, AcsD, and CcpAx.