Proline dehydrogenase from Thermus thermophilus does not discriminate between FAD and FMN as cofactor

Flavoenzymes are versatile biocatalysts containing either FAD or FMN as cofactor. FAD often binds to a Rossmann fold, while FMN prefers a TIM-barrel or flavodoxin-like fold. Proline dehydrogenase is denoted as an exception: it possesses a TIM barrel-like fold while binding FAD. Using a riboflavin auxotrophic Escherichia coli strain and maltose-binding protein as solubility tag, we produced the apoprotein of Thermus thermophilus ProDH (MBP-TtProDH). Remarkably, reconstitution with FAD or FMN revealed that MBP-TtProDH has no preference for either of the two prosthetic groups. Kinetic parameters of both holo forms are similar, as are the dissociation constants for FAD and FMN release. Furthermore, we show that the holo form of MBP-TtProDH, as produced in E. coli TOP10 cells, contains about three times more FMN than FAD. In line with this flavin content, the crystal structure of TtProDH variant ¿ABC, which lacks helices aA, aB and aC, shows no electron density for an AMP moiety of the cofactor. To the best of our knowledge, this is the first example of a flavoenzyme that does not discriminate between FAD and FMN as cofactor. Therefore, classification of TtProDH as an FAD-binding enzyme should be reconsidered

Influence of winter waves on the longitudinal growth of the Punta Banda Estuary sandbar, Todos Santos Bay, Baja California, Mexico

Punta Banda Estuary, located within Todos Santos Bay, on the northwest coast of the Baja California Peninsula, is separated from the bay by a single sandbar, approximately 8 km long. From 1972 to 2003 this sandbar showed a longitudinal net growth of 420 m, which decreased the width of the estuary mouth by more than 400 m. Despite the bar's rapid growth it has been suggested that the estuary mouth remains open because in spring, summer and autumn the longshore sand transport is northeastward, but during winter the longshore sand transport should be toward the southwest, away from the mouth, establishing a balance between the amount of sediment transported both ways. In this study we analyze wave data acquired from November 2004 to March 2005 on the bay side of the sandbar, and for the first time provide the magnitude and direction of the longshore sand transport during winter. The results show that during this season the dominant waves approach the bay from the west-northwest and the sandbar from the west, inducing a net longshore sand transport of 817 m3 day–1 to the northeast. This result contradicts the hypothesis of a longshore sand transport balance, and shows that throughout a typical year the net longshore sand transport is to the northeast. The fact that the estuary mouth has not closed up should be attributed to other causes, such as extraordinary amounts of rain during the rainy season or inequalities between the ebb and flood tidal current velocities to the estuary.