Details

Autor(en) / Beteiligte

• Hashim, Noor Haza Fazlin
• Bharudin, Izwan
• Nguong, Douglas Law Sie
• Higa, Sakura
• Bakar, Farah Diba Abu
• Nathan, Sheila
• Rabu, Amir
• Kawahara, Hidehisa
• Illias, Rosli Md
• Najimudin, Nazalan
• Mahadi, Nor Muhammad
• Murad, Abdul Munir Abdul

Titel

Characterization of Afp1, an antifreeze protein from the psychrophilic yeast Glaciozyma antarctica PI12

Ist Teil von

• Extremophiles : life under extreme conditions, 2013-01, Vol.17 (1), p.63-73

Ort / Verlag

Germany

Erscheinungsjahr

2013

Link zum Volltext

Quelle

MEDLINE

Beschreibungen/Notizen

• The psychrophilic yeast Glaciozyma antarctica demonstrated high antifreeze activity in its culture filtrate. The culture filtrate exhibited both thermal hysteresis (TH) and ice recrystallization inhibition (RI) properties. The TH of 0.1 °C was comparable to that previously reported for bacteria and fungi. A genome sequence survey of the G. antarctica genome identified a novel antifreeze protein gene. The cDNA encoded a 177 amino acid protein with 30 % similarity to a fungal antifreeze protein from Typhula ishikariensis. The expression levels of AFP1 were quantified via real time-quantitative polymerase chain reaction (RT-qPCR), and the highest expression levels were detected within 6 h of growth at -12 °C. The cDNA of the antifreeze protein was cloned into an Escherichia coli expression system. Expression of recombinant Afp1 in E. coli resulted in the formation of inclusion bodies that were subsequently denatured by treatment with urea and allowed to refold in vitro. Activity assays of the recombinant Afp1 confirmed the antifreeze protein properties with a high TH value of 0.08 °C.