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

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

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)63-73
Number of pages11
JournalExtremophiles
Volume17
Issue number1
DOIs
Publication statusPublished - 2013

Fingerprint

Antifreeze Proteins
Yeasts
Hot Temperature
Complementary DNA
Genome
Escherichia coli
Fungal Proteins
Inclusion Bodies
Ice
Urea
Real-Time Polymerase Chain Reaction
Fungi
Bacteria
Amino Acids
Growth
Genes
Proteins

Keywords

  • Antifreeze protein
  • Glaciozyma antarctica PI12
  • Psychrophilic yeast
  • Recrystallization inhibition
  • Thermal hysteresis

ASJC Scopus subject areas

  • Molecular Medicine
  • Microbiology

Cite this

Characterization of Afp1, an antifreeze protein from the psychrophilic yeast Glaciozyma antarctica PI12. / Hashim, Noor Haza Fazlin; Bharudin, Izwan; Nguong, Douglas Law Sie; Higa, Sakura; Abu Bakar, Farah Diba; Nathan, Sheila; Rabu, Amir; Kawahara, Hidehisa; Illias, Rosli Md; Najimudin, Nazalan; Mahadi, Nor Muhammad; Abd. Murad, Abdul Munir.

In: Extremophiles, Vol. 17, No. 1, 2013, p. 63-73.

Research output: Contribution to journalArticle

Hashim, Noor Haza Fazlin ; Bharudin, Izwan ; Nguong, Douglas Law Sie ; Higa, Sakura ; Abu Bakar, Farah Diba ; Nathan, Sheila ; Rabu, Amir ; Kawahara, Hidehisa ; Illias, Rosli Md ; Najimudin, Nazalan ; Mahadi, Nor Muhammad ; Abd. Murad, Abdul Munir. / Characterization of Afp1, an antifreeze protein from the psychrophilic yeast Glaciozyma antarctica PI12. In: Extremophiles. 2013 ; Vol. 17, No. 1. pp. 63-73.
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