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

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 subse...

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Main Authors:	Douglas Law Sie Nguong,, Lim, Yao Jun, Nor Idaya Yatim,, Sheila Nathan,, Abdul Munir Abdul Murad,, Nor Muhammad Mahadi,, Farah Diba Abu Bakar,
Format:	Article
Language:	English
Published:	Universiti Kebangsaan Malaysia 2011
Online Access:	http://journalarticle.ukm.my/2392/1/07_Douglas.pdf http://journalarticle.ukm.my/2392/ http://www.ukm.my/jsm/
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spelling	my-ukm.journal.23922016-12-14T06:31:29Z http://journalarticle.ukm.my/2392/ Characterising yeast isolates from Malaysia towards the development of alternative heterologous protein expression system Douglas Law Sie Nguong, Lim, Yao Jun Nor Idaya Yatim, Sheila Nathan, Abdul Munir Abdul Murad, Nor Muhammad Mahadi, Farah Diba Abu Bakar, 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, p1926Zeo containing the yeast 26S rDNA and Zeocin® resistance cassette. The p1926Zeo 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. Universiti Kebangsaan Malaysia 2011-04 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/2392/1/07_Douglas.pdf Douglas Law Sie Nguong, and Lim, Yao Jun and Nor Idaya Yatim, and Sheila Nathan, and Abdul Munir Abdul Murad, and Nor Muhammad Mahadi, and Farah Diba Abu Bakar, (2011) Characterising yeast isolates from Malaysia towards the development of alternative heterologous protein expression system. Sains Malaysiana, 40 (4). pp. 323-329. ISSN 0126-6039 http://www.ukm.my/jsm/
institution	Universiti Kebangsaan Malaysia
building	Perpustakaan Tun Sri Lanang Library
collection	Institutional Repository
continent	Asia
country	Malaysia
content_provider	Universiti Kebangsaan Malaysia
content_source	UKM Journal Article Repository
url_provider	http://journalarticle.ukm.my/
language	English
description	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, p1926Zeo containing the yeast 26S rDNA and Zeocin® resistance cassette. The p1926Zeo 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.
format	Article
author	Douglas Law Sie Nguong, Lim, Yao Jun Nor Idaya Yatim, Sheila Nathan, Abdul Munir Abdul Murad, Nor Muhammad Mahadi, Farah Diba Abu Bakar,
spellingShingle	Douglas Law Sie Nguong, Lim, Yao Jun Nor Idaya Yatim, Sheila Nathan, Abdul Munir Abdul Murad, Nor Muhammad Mahadi, Farah Diba Abu Bakar, Characterising yeast isolates from Malaysia towards the development of alternative heterologous protein expression system
author_facet	Douglas Law Sie Nguong, Lim, Yao Jun Nor Idaya Yatim, Sheila Nathan, Abdul Munir Abdul Murad, Nor Muhammad Mahadi, Farah Diba Abu Bakar,
author_sort	Douglas Law Sie Nguong,
title	Characterising yeast isolates from Malaysia towards the development of alternative heterologous protein expression system
title_short	Characterising yeast isolates from Malaysia towards the development of alternative heterologous protein expression system
title_full	Characterising yeast isolates from Malaysia towards the development of alternative heterologous protein expression system
title_fullStr	Characterising yeast isolates from Malaysia towards the development of alternative heterologous protein expression system
title_full_unstemmed	Characterising yeast isolates from Malaysia towards the development of alternative heterologous protein expression system
title_sort	characterising yeast isolates from malaysia towards the development of alternative heterologous protein expression system
publisher	Universiti Kebangsaan Malaysia
publishDate	2011
url	http://journalarticle.ukm.my/2392/1/07_Douglas.pdf http://journalarticle.ukm.my/2392/ http://www.ukm.my/jsm/
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score	13.211869

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

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