Characterising yeast isolates from Malaysia towards the development of alternative heterologous protein expression systems

Douglas Law Sie Nguong, Lim Yao Jun, Nor Idaya Yatim, Sheila Nathan, Abdul Munir Abd. Murad, Nor Muhammad Mahadi, Farah Diba Abu Bakar

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Yeasts with GRAS (Generally Regarded as Safe) status are commonly used as hosts for heterologous protein production. Yeasts are suitable expression hosts as they have been extensively characterised genetically. The objective of this project was to isolate yeasts from Malaysian food sources and subsequently to develop these as alternative hosts for heterologous protein production. Yeasts were isolated from Malaysian traditional fermented food namely 'tapai', 'tuak' and 'ragi' A total of 23 isolates were obtained and subjected to molecular identification by amplification and sequencing of the universally conserved ribosomal internal transcribed spacer (ITS), 26S rDNA and 18S rDNA sequences. We identified three species of yeasts, Saccharomyces cerevisiae, Hanseniaspora guilliermondii and Pichia anomala, which have a long tradition of usage in food production and have no adverse effects on humans. To test the feasibility of the yeasts as heterologous expression hosts, we have constructed an integrative vector, pl926Zeo containing the yeast 26S rDNA and Zeocin® resistance cassette. The pl926Zeo vector was linearised and transformed into both P. anomala and H. guilliermondii isolates via electroporation. Both hosts were successfully transformed at a relatively high efficiency. The transformants obtained had a growth profile similar to the respective wild type, indicating that integration of the plasmids into the host chromosome did not affect growth. These transformants were stable as they exhibited resistance to Zeocin even after 20 generations. Thus, both P. anomala and H. guilliermondii isolates exhibited the potential to be further developed as alternative heterologous protein expression hosts.

Original languageEnglish
Pages (from-to)323-329
Number of pages7
JournalSains Malaysiana
Volume40
Issue number4
Publication statusPublished - Apr 2011

Fingerprint

Malaysia
yeasts
Wickerhamomyces anomalus
Hanseniaspora
Eleusine coracana subsp. coracana
alternative hosts
fermented foods
electroporation
traditional foods
heterologous gene expression
food production
internal transcribed spacers
Saccharomyces cerevisiae
plasmids
adverse effects
chromosomes
testing

Keywords

  • Hanseniaspora guilliermondii
  • Heterologous protein
  • Pichia anomala
  • Yeast expression systems

ASJC Scopus subject areas

  • General

Cite this

Characterising yeast isolates from Malaysia towards the development of alternative heterologous protein expression systems. / Nguong, Douglas Law Sie; Jun, Lim Yao; Yatim, Nor Idaya; Nathan, Sheila; Abd. Murad, Abdul Munir; Mahadi, Nor Muhammad; Abu Bakar, Farah Diba.

In: Sains Malaysiana, Vol. 40, No. 4, 04.2011, p. 323-329.

Research output: Contribution to journalArticle

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