재조합 Pichia pastoris의 유가식 배양을 통한 남극세균 Flavobacterium frigoris PS1 유래 결빙방지단백질의 생산

Abstract

Antifreeze proteins (AFP) inhibit ice growth to permit the survival of polar organisms in the cold environments. The recombinant AFP from an Antarctic bacterium, Flavobacterium frigoris PS1, FfIBP (Flavobacterium frigoris icebinding protein), was produced using Pichia pastoris expression system. The optimum fermentation temperature (30℃) and pH (5) for FfIBP production were determined using a fedbatch culture system. The maximal cell density and purified FfIBP were 112 g/L and 70 mg/L, respectively. The thermal hysteresis (TH) activity (0.85) of FfIBP obtained using a glycerol-methanol fed-batch culture system was 2-fold higher than that of the LeIBP (Leucosporidium ice-binding protein). This work allows for large-scale production of FfIBP, which could be extended to further application studies using recombinant AFPs.