LAUSR

Abstract Antifreeze proteins (AFPs) are characterized by their ability to protect organisms from subfreezing temperatures. They constitute a class of promising candidates as environmentally kinetic hydrate inhibitors (KHIs). In this study, the effectiveness of an insect cell expressed novel monomeric streptavidin fusion protein version of Rhagium mordax RmAFP1 antifreeze protein (mSA-RmAFP1), and four amino acids (histidine, lysine, tyrosine and proline), on CH4 hydrate nucleation, growth and decomposition was investigated using a rocking cell apparatus, then compared with the commercial inhibitors Polyvinylpyrrolidone (PVP) and Luvicap Bio. It was found that CH4 hydrate nucleation and growth exhibited good repeatable results under experimental conditions. The results showed that 2250 ppm mSA-RmAFP1 can inhibit CH4 hydrate nucleation as effectively as PVP at the same concentration. The histidine, lysine, tyrosine and proline exhibited weak inhibition effect on CH4 hydrate nucleation. The mSA-RmAFP1 decreased CH4 hydrate growth rate and production in the fresh and memory solutions. The CH4 hydrate formed in the solutions containing various tested KHIs present slightly lower onset decomposition temperatures than the non-inhibited system under experimental conditions. The promising performance of the insect cell expressed mSA-RmAFP1 could promote the further development of green hydrate inhibitors. The production of this protein through insect cell line fermentation provides a platform for the future production and optimization of AFPs for hydrate inhibition.