Abstract
Ice-binding protein (IBPs) protect cells from cryo-injury during cryopreservation by inhibiting ice recrystallization (IR), which is a main cause of cell death. In the present study, we employed two IBPs, one, designated LeIBP from Arctic yeast, and the other, designated FfIBP from Antarctic sea ice bacterium, in the cryopreservation of three economically valuable marine microalgae, Isochrysis galbana, Pavlova viridis, and Chlamydomonas coccoides. Both of the IBPs showed IR inhibition in f/2 medium containing 10% DMSO, indicating that they retain their function in freezing media. Microalgal cells were frozen in 10% DMSO with or without IBP. Post-thaw viability exhibited that the supplementation of IBPs increased the viability of all cryopreserved cells. LeIBP was effective in P. viridis and C. coccoides, while FfIBP was in I. galbana. The cryopreservative effect was more drastic with P. viridis when 0.05 mg/mL LeIBP was used. These results clearly demonstrate that IBPs could improve the viability of cryopreserved microalgal cells.
Highlights
Ice-binding proteins (IBPs) are a class of protein that has an affinity to ice
Effect of CPAs on Unfrozen Marine Microalgae f/2 medium (Supplementary Table S1): dimethyl sulfoxide, DMSO; ethylene glycol, EG; glycerol, In order to select an appropriate CPA for I. galbana, P. viridis, and C. coccoides, based on the
C. coccoides tolerableindicated to three CPAs, the being culture most shouldtolerable be by far less than 0.1%
Summary
Ice-binding proteins (IBPs) are a class of protein that has an affinity to ice. Some IBPs, for example fish antifreeze proteins (AFPs) are a biological antifreeze, which bind to the ice surface and subsequently inhibit further growth of the ice crystal [1,2]. Some IBPs, for example fish antifreeze proteins (AFPs) are a biological antifreeze, which bind to the ice surface and subsequently inhibit further growth of the ice crystal [1,2] This behavior of IBP eventually lowers the freezing point of the solution, and creates a gap between freezing and melting points [3]. Psychrophilic organisms inhabiting cold environments, such as alpine, Arctic, and Antarctica, experience relatively wide temperature fluctuations. These temperature fluctuations cause smaller ice grains to combine one another to form larger ones, which is thermodynamically favorable [11,12].
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