海洋生物技术 抗冻蛋白.pdfVIP

Mar Biotechnol (2014) 16:502–512 DOI 10.1007/s10126-014-9567-y ORIGINAL ARTICLE Isolation and Characterization of Antifreeze Proteins from the Antarctic Marine Microalga Pyramimonas gelidicola Woongsic Jung Yunho Gwak Peter L. Davies Hak Jun Kim EonSeon Jin Received: 16 October 2012 /Accepted: 21 January 2014 /Published online: 8 March 2014 # Springer Science+Business Media New York 2014 Abstract Antifreeze proteins (AFPs) play an important role helical surfaces is neither regular nor well-conserved. This in the psychrophilic adaptation of polar organisms. AFPs might be a characteristic of AFPs used for freeze tolerance encoded by an Antarctic chlorophyte, identified as as opposed to freeze avoidance. A role for P. gelidicola AFPs Pyramimonas gelidicola, were isolated and characterized. in freeze tolerance is also consistent with their relatively low Two AFP isoforms were found from cDNAs and their de- TH values. duced molecular weights were estimated to be 26.4 kDa (Pg- 1-AFP) and 27.1 kDa (Pg-2-AFP). Both AFP cDNAs were Keywords Antifreeze proteins . Antarctic marine cloned and expressed in Escherichia coli. The purified recom- prasinophyte . Ice binding . Thermal hysteresis . In silico binant Pg-1-rAFP and Pg-2-rAFP both showed antifreeze protein modeling activity based on the measurement of thermal hysteresis (TH) and morphological changes to single ice crystals. Pg-1- rAFP shaped ice crystals into a snowflake pattern with a TH Introduction value of 0.6±0.02 °C at ~15 mg/ml. Single ice crystals in Pg- 2-rAFP showed a dendritic morphology with a TH value of Antarctica has one of the harshest environments on earth, with 0.25±0.02 °C at the same protein concentration. Based on in strong winds, frequent blizzards,