In this work, we found that both the F- and V-type ATPases are expressed C. themocellum. Co-presence of V- and F-type ATPases in a bacterium is uncommon. Previously, only Enterococcus hirae was reported to utilize both types of ATPases [18]. The E. hirae

V-type ATPase differs from typical V-type ATPase in preferentially transporting Na+ [19, 20] instead of H+. In the thermophilic Clostridium fervidus, a second example of Na+-pumping V-type ATPase was reported [21]. It is reasonable to speculate that the V-type ATPase in C. thermocellum is a Na+-pumping ATPase. Most bacteria contain either F-type or V-type ATPase, among those that contain PI3K Inhibitor Library both types of ATPases, new functional variants of ATPases could be identified and their roles in bacterial physiology could be investigated. Bifunctional acetaldehyde/alcohol Mocetinostat dehydrogenase (ALDH-ADH, Cthe_0423, 96 kDa) was detected at over 880 kDa. ADHs could be classified into 3 classes based on their length: short chain ADH (approximately 250 residues) and medium chain ADH (approximately 370 residues) exist in a homotetramer form [22], but a structure of long chain ADH (over 380 amino acids and often as many as 900 amino acid residues) was not reported. The ALDH-ADH of C. thermocellum appears to be a long chain ADH and forms a homo-multimer like the ADH in Entamoeba histolytica [23]. Alcohol dehydrogenases were reported to be membrane-bound protein complexes

[24–26], it is reasonable to PXD101 supplier observe ADH in C. thermocellum membrane fraction. Complexes in lipid transport and metabolism Carboxyl transferase (CT, Cthe_0699, 56 kDa) was identified at ~220 kDa. In eubacteria, CT is part of acetyl coenzyme A carboxylase (ACC) complex, which normally consists

of biotin carboxylase (BC), biotin carboxyl carrier protein (BCCP), and CT. Typically, CT contains two subunits in a stable α2β2 form [27, 28]. But, in Streptomyces coelicolor, the ACC enzyme has Vildagliptin a subunit (590 residues) with fused BC and BCCP domains, and another subunit (530 residues) that contains the fused CT domains [29]. In archaea, ACC is a multi-subunit enzyme, with BC, BCCP and CT subunits. The archael CT subunit is also a single protein (520 residues) in a CT4 form, rather than two separate subunits, which is similar to the β subunit (CT) of the ACC from Streptomyces [30]. In C. thermocellum, CT is a 56 kDa protein, which contains two domains of carboxyl transferase, and we did not detect other ACC subunits on BN/SDS-PAGE. So the CT appears to be a sub complex of CT4 not associated with BC and BCCP. CT was also detected at over 880 kDa, which maybe due to precipitation during electrophoresis or CT formed a large complex with other subunits of ACC. Previous studies also suggested ACC may form a membrane-associated protein complex [31, 32]. Complexes in amino acid transport and metabolism Serine-Acetyl-Transferase (SAT, Cthe_1840, 33.