얼음 결합 단백질을 이용한 유용 미세조류의 동결보존

Abstract

Recently, useful marine microalgae are applying to diverse bio-industry. It could be enough energy for demands of increasing energy continuously, because it is eco-friendly and unicellular organism. The cryopreservation technology is the best method of maintaining them for long-terms. Ice-Binding Protein (IBP) is able to increase the cell viability by preventing ice recrystallization. Therefore, we proved the effect of IBP by applying 2 kinds of IBP (LeIBP, FfIBP) to 3 kinds of useful microalgae (Isochrysis galbana, Isochrysis galbana green, Chlamydomonas coccoides). We used 10% dimethylsulfoxide (DMSO) which has the lowest toxicity compared with other 10% CPAs (Ethylene glycol, Glycerol, Methanol) on cytotoxicity tests as cryopreservation agents (CPAs) and combined with IBPs. In freezing method, two step slow freezing were used and then plunged into liquid nitrogen (-196 ℃). After that, the samples were rapidly thawed and cultured in each optimal condition. The viability was measured every two day during 18 days. Results show that viability of samples had LeIBP and FfIBP were very high. However, only Isochrysis galbana green growth was fast, especially sample combined with DMSO and 0.05 mg/mL LeIBP. In other two species, existence of IBP increase its viability and growth rate after thaw compared to other samples. Usually LeIBP samples showed pattern of fast growth and after that, FfIBP samples grew fast. Moreover, the green algae cryopreservation is best because there was the fast growth of Chlamydomonas coccoides.