Heinrich Kroukamp - Academia.edu (original) (raw)

Papers by Heinrich Kroukamp

Research paper thumbnail of Strain Breeding Enhanced Heterologous Cellobiohydrolase Secretion by Saccharomyces cerevisiae in a Protein Specific Manner

Biotechnology Journal

The yeast Saccharomyces cerevisiae has a long association with alcoholic fermentation industries ... more The yeast Saccharomyces cerevisiae has a long association with alcoholic fermentation industries and has received renewed interest as a biocatalyst for second-generation bioethanol production. Rational engineering strategies are used to create yeast strains for consolidated bioprocessing of lignocellulosic biomass. Although significant progress is made in this regard with the expression of different cellulolytic activities in yeast, cellobiohydrolase (CBH) titers remain well below ideal levels. Through classical breeding, S. cerevisiae strains with up to twofold increased CBH secretion titers is obtained in strains expressing a single gene copy. An increase of up to 3.5-fold in secreted cellobiohydrolase activity is subsequently shown for strains expressing the heterologous gene on a high copy episomal vector. To our knowledge, this is the first report of classical breeding being used to enhance heterologous protein secretion and also the most significant enhancement of CBH secretion in yeast yet reported. This enhanced secretion phenotype is specific for cellobiohydrolase I secretion, indicating that reporter protein properties might be a major determining factor for efficient protein secretion in yeast. By exploring the latent potential of different S. cerevisiae strains, the authors show that the allele pool of various strains is a valuable engineering resource to enhance secretion in yeast.

Research paper thumbnail of First report of urease activity in the novel systemic fungal pathogen Emergomyces africanus: a comparison with the neurotrope Cryptococcus neoformans

Research paper thumbnail of Yeast's balancing act between ethanol and glycerol production in low-alcohol wines

Microbial biotechnology, Mar 1, 2017

Alcohol is fundamental to the character of wine, yet too much can put a wine off-balance. A wine ... more Alcohol is fundamental to the character of wine, yet too much can put a wine off-balance. A wine is regarded to be well balanced if its alcoholic strength, acidity, sweetness, fruitiness and tannin structure complement each other so that no single component dominates on the palate. Balancing a wine's positive fruit flavours with the optimal absolute and relative concentration of alcohol can be surprisingly difficult. Over the past three decades, consumers have increasingly demanded wine with richer and riper fruit flavour profiles. In response, grape and wine producers have extended harvest times to increase grape maturity and enhance the degree of fruit flavours and colour intensity. However, a higher degree of grape maturity results in increased grape sugar concentration, which in turn results in wines with elevated alcohol concentration. On average, the alcohol strength of red wines from many warm wine-producing regions globally rose by about 2% (v/v) during this period. Notw...

Research paper thumbnail of Overexpression of native PSE1 and SOD1 in Saccharomyces cerevisiae improved heterologous cellulase production

Research paper thumbnail of Cellobiohydrolase secretion by yeast: Current state and prospects for improvement

Process Biochemistry, 2013

Research paper thumbnail of Engineering Saccharomyces cerevisiae for next generation ethanol production

Journal of Chemical Technology & Biotechnology, 2013

ABSTRACT Conversion of cellulose, hemicellulose or starch to ethanol via a biological route requi... more ABSTRACT Conversion of cellulose, hemicellulose or starch to ethanol via a biological route requires enzymatic conversion of these substrates to monosaccharides that can be assimilated by a fermenting organism. Consolidation of these events in a single processing step via a cellulolytic or amylolytic microorganism(s) is a promising approach to low-cost production of fuels and chemicals. One strategy for developing a microorganism capable of such consolidated bioprocessing (CBP) involves engineering Saccharomyces cerevisiae to expresses a heterologous enzyme system enabling (hemi)cellulose or starch utilization. The fundamental principle behind consolidated bioprocessing as a microbial phenomenon has been established through the successful expression of the major (hemi)cellulolytic and amylolytic activities in S. cerevisiae. Various strains of S. cerevisiae were subsequently enabled to grow on cellobiose, amorphous and crystalline cellulose, xylan and various forms of starch through the combined expression of these activities. Furthermore, host cell engineering and adaptive evolution have yielded strains with higher levels of secreted enzymes and greater resistance to fermentation inhibitors. These breakthroughs bring the application of CBP at commercial scale ever closer. This mini-review discusses the current status of different aspects related to the engineering of S. cerevisiae for next generation ethanol production. © 2013 Society of Chemical Industry

Research paper thumbnail of Overexpression of native PSE1 and SOD1 in Saccharomyces cerevisiae improved heterologous cellulase secretion

Research paper thumbnail of The Smell of Synthetic Biology: Engineering Strategies for Aroma Compound Production in Yeast

Yeast—especially Saccharomyces cerevisiae—have long been a preferred workhorse for the production... more Yeast—especially Saccharomyces cerevisiae—have long been a preferred workhorse for the production of numerous recombinant proteins and other metabolites. S. cerevisiae is a noteworthy aroma compound producer and has also been exploited to produce foreign bioflavour compounds. In the past few years, important strides have been made in unlocking the key elements in the biochemical pathways involved in the production of many aroma compounds. The expression of these biochemical pathways in yeast often involves the manipulation of the host strain to direct the flux towards certain precursors needed for the production of the given aroma compound. This review highlights recent advances in the bioengineering of yeast—including S. cerevisiae—to produce aroma compounds and bioflavours. To capitalise on recent advances in synthetic yeast genomics, this review presents yeast as a significant producer of bioflavours in a fresh context and proposes new directions for combining engineering and biology principles to improve the yield of targeted aroma compounds.

Research paper thumbnail of Strain Breeding Enhanced Heterologous Cellobiohydrolase Secretion by Saccharomyces cerevisiae in a Protein Specific Manner

Biotechnology Journal

The yeast Saccharomyces cerevisiae has a long association with alcoholic fermentation industries ... more The yeast Saccharomyces cerevisiae has a long association with alcoholic fermentation industries and has received renewed interest as a biocatalyst for second-generation bioethanol production. Rational engineering strategies are used to create yeast strains for consolidated bioprocessing of lignocellulosic biomass. Although significant progress is made in this regard with the expression of different cellulolytic activities in yeast, cellobiohydrolase (CBH) titers remain well below ideal levels. Through classical breeding, S. cerevisiae strains with up to twofold increased CBH secretion titers is obtained in strains expressing a single gene copy. An increase of up to 3.5-fold in secreted cellobiohydrolase activity is subsequently shown for strains expressing the heterologous gene on a high copy episomal vector. To our knowledge, this is the first report of classical breeding being used to enhance heterologous protein secretion and also the most significant enhancement of CBH secretion in yeast yet reported. This enhanced secretion phenotype is specific for cellobiohydrolase I secretion, indicating that reporter protein properties might be a major determining factor for efficient protein secretion in yeast. By exploring the latent potential of different S. cerevisiae strains, the authors show that the allele pool of various strains is a valuable engineering resource to enhance secretion in yeast.

Research paper thumbnail of First report of urease activity in the novel systemic fungal pathogen Emergomyces africanus: a comparison with the neurotrope Cryptococcus neoformans

Research paper thumbnail of Yeast's balancing act between ethanol and glycerol production in low-alcohol wines

Microbial biotechnology, Mar 1, 2017

Alcohol is fundamental to the character of wine, yet too much can put a wine off-balance. A wine ... more Alcohol is fundamental to the character of wine, yet too much can put a wine off-balance. A wine is regarded to be well balanced if its alcoholic strength, acidity, sweetness, fruitiness and tannin structure complement each other so that no single component dominates on the palate. Balancing a wine's positive fruit flavours with the optimal absolute and relative concentration of alcohol can be surprisingly difficult. Over the past three decades, consumers have increasingly demanded wine with richer and riper fruit flavour profiles. In response, grape and wine producers have extended harvest times to increase grape maturity and enhance the degree of fruit flavours and colour intensity. However, a higher degree of grape maturity results in increased grape sugar concentration, which in turn results in wines with elevated alcohol concentration. On average, the alcohol strength of red wines from many warm wine-producing regions globally rose by about 2% (v/v) during this period. Notw...

Research paper thumbnail of Overexpression of native PSE1 and SOD1 in Saccharomyces cerevisiae improved heterologous cellulase production

Research paper thumbnail of Cellobiohydrolase secretion by yeast: Current state and prospects for improvement

Process Biochemistry, 2013

Research paper thumbnail of Engineering Saccharomyces cerevisiae for next generation ethanol production

Journal of Chemical Technology & Biotechnology, 2013

ABSTRACT Conversion of cellulose, hemicellulose or starch to ethanol via a biological route requi... more ABSTRACT Conversion of cellulose, hemicellulose or starch to ethanol via a biological route requires enzymatic conversion of these substrates to monosaccharides that can be assimilated by a fermenting organism. Consolidation of these events in a single processing step via a cellulolytic or amylolytic microorganism(s) is a promising approach to low-cost production of fuels and chemicals. One strategy for developing a microorganism capable of such consolidated bioprocessing (CBP) involves engineering Saccharomyces cerevisiae to expresses a heterologous enzyme system enabling (hemi)cellulose or starch utilization. The fundamental principle behind consolidated bioprocessing as a microbial phenomenon has been established through the successful expression of the major (hemi)cellulolytic and amylolytic activities in S. cerevisiae. Various strains of S. cerevisiae were subsequently enabled to grow on cellobiose, amorphous and crystalline cellulose, xylan and various forms of starch through the combined expression of these activities. Furthermore, host cell engineering and adaptive evolution have yielded strains with higher levels of secreted enzymes and greater resistance to fermentation inhibitors. These breakthroughs bring the application of CBP at commercial scale ever closer. This mini-review discusses the current status of different aspects related to the engineering of S. cerevisiae for next generation ethanol production. © 2013 Society of Chemical Industry

Research paper thumbnail of Overexpression of native PSE1 and SOD1 in Saccharomyces cerevisiae improved heterologous cellulase secretion

Research paper thumbnail of The Smell of Synthetic Biology: Engineering Strategies for Aroma Compound Production in Yeast

Yeast—especially Saccharomyces cerevisiae—have long been a preferred workhorse for the production... more Yeast—especially Saccharomyces cerevisiae—have long been a preferred workhorse for the production of numerous recombinant proteins and other metabolites. S. cerevisiae is a noteworthy aroma compound producer and has also been exploited to produce foreign bioflavour compounds. In the past few years, important strides have been made in unlocking the key elements in the biochemical pathways involved in the production of many aroma compounds. The expression of these biochemical pathways in yeast often involves the manipulation of the host strain to direct the flux towards certain precursors needed for the production of the given aroma compound. This review highlights recent advances in the bioengineering of yeast—including S. cerevisiae—to produce aroma compounds and bioflavours. To capitalise on recent advances in synthetic yeast genomics, this review presents yeast as a significant producer of bioflavours in a fresh context and proposes new directions for combining engineering and biology principles to improve the yield of targeted aroma compounds.