Efficacy of Antifreeze Proteins from Clupea harangues and Anarhichas minor on Gas Hydrate Inhibition via Cell Surface Display

Abstract The expression of anti-freeze proteins (AFPs) on the Escherichia coli using the cell surface display (CSD) strategy was investigated to determine the feasibility of the method as a potential application of kinetic hydrate inhibitors. Monomers to hexamers of type II and III AFPs were displayed on the surfaces of bacterial cells. The kinetic inhibition performance of the AFPs on the synthetic natural gas hydrate formation was tested. The displayed AFPs exhibited higher KHI effectiveness with the increase in the number of peptides attached to the cells. In addition, best KHI performance of the AFPs could be achieved with higher dosages of the cultures when the concentration of the inducer is optimized, regardless of AFP type. Only a small portion of AFPs in the cell cultures exhibited considerable KHI performance, which implies that the CSD could be an effective strategy, reducing the cost for the production and purification of the AFPs.