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

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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.

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Acknowledgments

This research was funded by the Ministry of Science, Technology and Innovation (MOSTI), Malaysia under the research grants UKM-MGI-NBD-0015-2007 and 07-05-MGI-GMB014. We acknowledge support given by the Australian Antarctic Division and the Malaysian Antarctic Research Programme (MARP) of the Academy of Science, Malaysia.

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Authors and Affiliations

  1. School of Biosciences and Biotechnology, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600, Bangi, Selangor, Malaysia

    Noor Haza Fazlin Hashim, Izwan Bharudin, Douglas Law Sie Nguong, Farah Diba Abu Bakar, Sheila Nathan, Amir Rabu & Abdul Munir Abdul Murad

  2. Department of Life Science and Biotechnology, Kansai University, 3-3-35 Yamate-cho, Suita, Osaka, 564-8680, Japan

    Sakura Higa & Hidehisa Kawahara

  3. Malaysia Genome Institute, 43000, Kajang, Selangor, Malaysia

    Sheila Nathan & Nor Muhammad Mahadi

  4. Department of Bioprocess Engineering, Faculty of Chemical and Natural Resources Engineering, Universiti Teknologi Malaysia, 81310, Skudai, Johor, Malaysia

    Rosli Md. Illias

  5. School of Biological Sciences, Universiti Sains Malaysia, 11800, Penang, Malaysia

    Nazalan Najimudin

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  1. Noor Haza Fazlin Hashim
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  11. Nor Muhammad Mahadi
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  12. Abdul Munir Abdul Murad
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Correspondence to Abdul Munir Abdul Murad.

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Communicated by F. Robb.

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Hashim, N.H.F., Bharudin, I., Nguong, D.L.S. et al. Characterization of Afp1, an antifreeze protein from the psychrophilic yeast Glaciozyma antarctica PI12. Extremophiles 17, 63–73 (2013). https://doi.org/10.1007/s00792-012-0494-4

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  • DOI: https://doi.org/10.1007/s00792-012-0494-4

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