Isak S. Pretorius

ARC Centre of Excellence in Synthetic Biology

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Florian F. Bauer

Stellenbosch University

Ian T. Paulsen

Australian Genome Research Facility

Jan H. Swiegers

Carlsberg Group (Denmark)

Marius G. Lambrechts

Stellenbosch University

Paul J. Chambers

Australian Wine Research Institute

Niël van Wyk

ARC Centre of Excellence in Synthetic Biology

Ricardo R. Cordero Otero

Universidad Rovira i Virgili

Anthony R. Borneman

Australian Wine Research Institute

Heinrich Kroukamp

ARC Centre of Excellence in Synthetic Biology

Willem H. van Zyl

Stellenbosch University

Cooperative Institutions

Stellenbosch University

Australian Wine Research Institute

ARC Centre of Excellence in Synthetic Biology

Macquarie University

University of Adelaide

KU Leuven

Hochschule Geisenheim University

Consejo Superior de Investigaciones Científicas

Ghent University

University of the Free State

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The Saccharomyces cerevisiae alcohol acetyl transferase Atf1p is localized in lipid particles

Yeast (2004)

Kevin J. Verstrepen Stijn D. M. Van Laere Jo Vercammen Guy Derdelinckx Jean‐Pierre Dufour

Abstract The yeast alcohol acetyl transferase I, Atf1p, is responsible for the major part of volatile acetate ester production in fermenting Saccharomyces cerevisiae cells. Some of these esters, such as ethyl acetate and isoamyl acetate, are important for the fruity flavours of wine, beer and other fermented beverages. In order to reveal the subcellular localization of Atf1p and further unravel the possible physiological role of this protein, ATF1::GFP fusion constructs were overexpressed in brewer's yeast. The transformant strain showed a significant increase in acetate ester formation, similar to that of an ATF1 overexpression strain, indicating that the Atf1p–GFP fusion protein was active. UV fluorescence microscopy revealed that the fusion protein was localized in small, sphere‐like organelles. These organelles could be selectively stained by the fluorescent dye Nile red, indicating that they contained high amounts of neutral lipids and/or sterols, a specific characteristic of yeast lipid particles. Purification of lipid particles from wild type and ATF1 deletion cells confirmed that the Atf1p–GFP fusion protein was located in these organelles. Furthermore, a clear alcohol acetyl transferase activity could be measured in the purified lipid particles of both wild type and transformed cells. The localization of Atf1p in lipid particles may indicate that Atf1p has a specific role in the lipid and/or sterol metabolism that takes place in these particles. Copyright © 2004 John Wiley & Sons, Ltd.

Organelle

Lipid droplet

Nile red

10.1002/yea.1100

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Citations (77)

Identification and physical characterization of yeast glucoamylase structural genes

MGG Molecular & General Genetics (1986)

Isak S. Pretorius Thomas Chow Julius Marmur

Identification

Human genetics

Structural gene

10.1007/bf00330381

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Citations (56)

CELLODEXTRINASE GENE IN SACCHAROMYCES CEREVISIAE

Pierre van Rensburg Willem H. van Zyl Isak S. Pretorius

SUMMARY A recombinant strain of Saccharomyces cerevtslae secreting bacterial cellodextrinase was constructed. The Rummococcus flavefaclens cellodextrinase gene (celA) was inserted between a yeast expression-secretion cassette and yeast gene terminator, and cloned into a yeast-centromeric shuttle vector. Enzyme assays revealed growth-associated production of biologically active cellodextrinase by S. cerewstae transformants.

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Terminator (solar)

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The Tailoring of designer grapevines in microbial starter strains for a market-directed and quality-focused wine industry

Taylor & Francis eBooks (2006)

Isak S. Pretorius Eveline Bartowsky Miguel de Barros Lopes Florian F. Bauer Maret du Toit

Isak S. Pretorius, Eveline J. Bartowsky, Miguel de Barros Lopes, Florian F. Bauer, Maret du Toit, Pierre van Rensburg and Melane` A. Vivier

Starter

Grape wine

Winery

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Citations (7)

Designing wine yeast for the future

Elsevier eBooks (2014)

Isak S. Pretorius Christopher D. Curtin Paul J. Chambers

10.1016/b978-1-78242-015-6.00009-8

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Citations (7)

Transcriptional Control of Glucoamylase Synthesis in Vegetatively Growing and Sporulating Saccharomyces Species

Molecular and Cellular Biology (1986)

Isak S. Pretorius Daniela Modena Marco Vanoni Sasha Englard Julius Marmur

Three unlinked, homologous genes, STA1, STA2, and STA3, encode the extracellular glycosylated glucoamylase isozymes I, II, and III, respectively, in Saccharomyces species. S. cerevisiae, which is sta0 (absence of functional STA genes in haploids), does carry a glucoamylase gene, delta sta, expressed only during sporulation (W. J. Colonna and P. T. Magee, J. Bacteriol. 134:844-853, 1978; I. Yamashita and S. Fukui, Mol. Cell. Biol. 5:3069-3073, 1985). In this study we examined some of the physiological and genetic factors that affect glucoamylase expression. It was found that STA2 strains grown in synthetic medium produce glucoamylase only in the presence of either Maltrin M365 (a mixture of maltooligosaccharides) or starch. Maximal levels of glucoamylase activity were found in cells grown in rich medium supplemented with glycerol plus ethanol, starch, or Maltrin. When various sugars served as carbon sources they all supported glucoamylase synthesis, although at reduced levels. In any given growth medium glucoamylase isozyme II synthesis was modulated by functionality of the mitochondria. Synthesis of glucoamylase is continuous throughout the growth phases, with maximal secretion taking place in the early stationary phase. In the various regimens, the differences in enzyme accumulation are accounted for by differences in the levels of glucoamylase mRNA. Both glucoamylase mRNA and enzyme activity were drastically and coordinately inhibited in MATa/MAT alpha diploids and by the presence of the regulatory gene STA10. Both effects were partially overcome when the STA2 gene was present on a multicopy plasmid. The STA2 mRNA and glucoamylase were coinduced in sporulating STA2/STA2 diploids. A smaller, coinduced RNA species was also detected by Northern blotting with a STA2 probe. The same mRNA species was detected in sporulating sta0 diploids and is likely to encode the sporulation-specific glucoamylase.

10.1128/mcb.6.9.3034-3041.1986

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Citations (48)

Applications of Yeast Synthetic Biology Geared towards the Production of Biopharmaceuticals

Genes (2018)

Roy Walker Isak S. Pretorius

Engineered yeast are an important production platform for the biosynthesis of high-value compounds with medical applications. Recent years have witnessed several new developments in this area, largely spurred by advances in the field of synthetic biology and the elucidation of natural metabolic pathways. This minireview presents an overview of synthetic biology applications for the heterologous biosynthesis of biopharmaceuticals in yeast and demonstrates the power and potential of yeast cell factories by highlighting several recent examples. In addition, an outline of emerging trends in this rapidly-developing area is discussed, hinting upon the potential state-of-the-art in the years ahead.

Synthetic Biology

Metabolic Engineering

Heterologous

10.3390/genes9070340

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Citations (47)

Co-expression of a Phanerochaete chrysosporium cellobiohydrolase gene and a Butyrivibrio fibrisolvens endo-?-1,4-glucanase gene in Saccharomyces cerevisiae

Current Genetics (1996)

Pierre van Rensburg Willem H. van Zyl Isak S. Pretorius

Phanerochaete

Chrysosporium

URA3

10.1007/s002940050128

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Citations (46)

Development of bactericidal wine yeast starter cultures

Yeast (2001)

Isak S. Pretorius Maret du Toit C. du Toit Vincenzo S. D'Aguanno

Starter

Source

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Genomic insights into the evolution of industrial yeast speciesBrettanomyces bruxellensis

FEMS Yeast Research (2014)

Christopher D. Curtin Isak S. Pretorius

Brettanomyces bruxellensis, like its wine yeast counterpart Saccharomyces cerevisiae, is intrinsically linked with industrial fermentations. In wine, B. bruxellensis is generally considered to contribute negative influences on wine quality, whereas for some styles of beer, it is an essential contributor. More recently, it has shown some potential for bioethanol production. Our relatively poor understanding of B. bruxellensis biology, at least when compared with S. cerevisiae, is partly due to a lack of laboratory tools. As it is a nonmodel organism, efforts to develop methods for sporulation and transformation have been sporadic and largely unsuccessful. Recent genome sequencing efforts are now providing B. bruxellensis researchers unprecedented access to gene catalogues, the possibility of performing transcriptomic studies and new insights into evolutionary drivers. This review summarises these findings, emphasises the rich data sets already available yet largely unexplored and looks over the horizon at what might be learnt soon through comprehensive population genomics of B. bruxellensis and related species. Genomic insights into the evolution of Brettanomyces bruxellensis.

Population genomics

10.1111/1567-1364.12198

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Citations (38)