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Concetta Compagno - One of the best experts on this subject based on the ideXlab platform.
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Heterologous Protein Production in Zygosaccharomyces bailii: physiological effects and fermentative strategies.
FEMS yeast research, 2005Co-Authors: Ileana Vigentini, Annamaria Merico, Danilo Porro, Paola Branduardi, Luca Brambilla, Concetta CompagnoAbstract:The optimisation and scale-up of a specific Protein Production process have to take into account cultivation conditions as well as cell physiology of growth and the influence of foreign Protein expression on host cell metabolism. The ability of Zygosaccharomyces bailii to tolerate high sugar concentrations as well as high temperatures and acidic environments renders this "non-conventional" yeast suitable for the development of biotechnological processes like Heterologous Protein Production. This work addresses the Production of human interleukin-1beta by a recombinant Z. bailii strain. We found that the Heterologous Protein Production causes some modifications of the Z. bailii carbon metabolism, leading to a reduced biomass yield. The other important factor is the dependence of the recombinant IL-1beta Production/secretion on the growth rate. Among the cultivation strategies studied, the most appropriate in terms of Production and productivity was the fed-batch mode.
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Heterologous Protein Production in Zygosaccharomyces bailii: physiological effects and fermentative strategies.
Fems Yeast Research, 2004Co-Authors: Ileana Vigentini, Annamaria Merico, Danilo Porro, Paola Branduardi, Luca Brambilla, Concetta CompagnoAbstract:The optimisation and scale-up of a specific Protein Production process have to take into account cultivation conditions as well as cell physiology of growth and the influence of foreign Protein expression on host cell metabolism. The ability of Zygosaccharomyces bailii to tolerate high sugar concentrations as well as high temperatures and acidic environments renders this “non-conventional” yeast suitable for the development of biotechnological processes like Heterologous Protein Production. This work addresses the Production of human interleukin-1β by a recombinant Z. bailii strain. We found that the Heterologous Protein Production causes some modifications of the Z. bailii carbon metabolism, leading to a reduced biomass yield. The other important factor is the dependence of the recombinant IL-1β Production/secretion on the growth rate. Among the cultivation strategies studied, the most appropriate in terms of Production and productivity was the fed-batch mode.
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How physiological and cultural conditions influence Heterologous Protein Production in Kluyveromyces lactis.
Journal of biotechnology, 2004Co-Authors: Annamaria Merico, Ileana Vigentini, Daniele Capitanio, Bianca Maria Ranzi, Concetta CompagnoAbstract:The optimization and scale-up of a specific Protein Production process have to take into account cultural conditions as well as cell physiology of growth and influence of foreign Protein expression on host cell metabolism. Growth on cheap substrates, efficient secretion ability and a weaker tendency to hypermannosilate Proteins than S. cerevisiae, make K. lactis an excellent and well-accepted host for Heterologous Protein Production, even for human use. A fairly good Heterologous glucoamylase yield and the setting of the optimal conditions to produce it were obtained expressing the Arxula adeninivorans glucoamylase in a strain of K. lactis and its isogenic mutant, which seems to have higher secretion ability. We performed batch cultures of both strains to analyze the influence of different physiological and environmental parameters on glucoamylase Production/secretion. Interestingly, the maintenance of pH in the range of neutrality causes the consumption of a larger amount of carbon source, a longer time of Production and a better stability of the active form of the enzyme, thus increasing biomass and glucoamylase Production. Furthermore, the enrichment of the culture medium adds up to the action of pH control, forcing the mutant Production/secretion to higher levels.
Luca Brambilla - One of the best experts on this subject based on the ideXlab platform.
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Induction by Hypoxia of Heterologous-Protein Production with the KlPDC1 Promoter in Yeasts
Applied and environmental microbiology, 2006Co-Authors: Andrea Camattari, Danilo Porro, Paola Branduardi, Michele M. Bianchi, Luca BrambillaAbstract:The control of promoter activity by oxygen availability appears to be an intriguing system for Heterologous Protein Production. In fact, during cell growth in a bioreactor, an oxygen shortage is easily obtained simply by interrupting the air supply. The purpose of our work was to explore the possible use of hypoxic induction of the KlPDC1 promoter to direct Heterologous gene expression in yeast. In the present study, an expression system based on the KlPDC1 promoter was developed and characterized. Several Heterologous Proteins, differing in size, origin, localization, and posttranslational modification, were successfully expressed in Kluyveromyces lactis under the control of the wild type or a modified promoter sequence, with a Production ratio between 4 and more than 100. Yields were further optimized by a more accurate control of hypoxic physiological conditions. Production of as high as 180 mg/liter of human interleukin-1β was obtained, representing the highest value obtained with yeasts in a lab-scale bioreactor to date. Moreover, the transferability of our system to related yeasts was assessed. The lacZ gene from Escherichia coli was cloned downstream of the KlPDC1 promoter in order to get β-galactosidase activity in response to induction of the promoter. A centromeric vector harboring this expression cassette was introduced in Saccharomyces cerevisiae and in Zygosaccharomyces bailii, and effects of hypoxic induction were measured and compared to those already observed in K. lactis cells. Interestingly, we found that the induction still worked in Z. bailii; thus, this promotor constitutes a possible inducible system for this new nonconventional host.
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Heterologous Protein Production in Zygosaccharomyces bailii: physiological effects and fermentative strategies.
FEMS yeast research, 2005Co-Authors: Ileana Vigentini, Annamaria Merico, Danilo Porro, Paola Branduardi, Luca Brambilla, Concetta CompagnoAbstract:The optimisation and scale-up of a specific Protein Production process have to take into account cultivation conditions as well as cell physiology of growth and the influence of foreign Protein expression on host cell metabolism. The ability of Zygosaccharomyces bailii to tolerate high sugar concentrations as well as high temperatures and acidic environments renders this "non-conventional" yeast suitable for the development of biotechnological processes like Heterologous Protein Production. This work addresses the Production of human interleukin-1beta by a recombinant Z. bailii strain. We found that the Heterologous Protein Production causes some modifications of the Z. bailii carbon metabolism, leading to a reduced biomass yield. The other important factor is the dependence of the recombinant IL-1beta Production/secretion on the growth rate. Among the cultivation strategies studied, the most appropriate in terms of Production and productivity was the fed-batch mode.
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Heterologous Protein Production in Zygosaccharomyces bailii: physiological effects and fermentative strategies.
Fems Yeast Research, 2004Co-Authors: Ileana Vigentini, Annamaria Merico, Danilo Porro, Paola Branduardi, Luca Brambilla, Concetta CompagnoAbstract:The optimisation and scale-up of a specific Protein Production process have to take into account cultivation conditions as well as cell physiology of growth and the influence of foreign Protein expression on host cell metabolism. The ability of Zygosaccharomyces bailii to tolerate high sugar concentrations as well as high temperatures and acidic environments renders this “non-conventional” yeast suitable for the development of biotechnological processes like Heterologous Protein Production. This work addresses the Production of human interleukin-1β by a recombinant Z. bailii strain. We found that the Heterologous Protein Production causes some modifications of the Z. bailii carbon metabolism, leading to a reduced biomass yield. The other important factor is the dependence of the recombinant IL-1β Production/secretion on the growth rate. Among the cultivation strategies studied, the most appropriate in terms of Production and productivity was the fed-batch mode.
Lieve Van Mellaert - One of the best experts on this subject based on the ideXlab platform.
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Metabolic impact assessment for Heterologous Protein Production in Streptomyces lividans based on genome-scale metabolic network modeling.
Mathematical biosciences, 2013Co-Authors: Ivan Lule, Pieter-jan D'huys, Lieve Van Mellaert, Jozef Anné, Kristel Bernaerts, Jan Van ImpeAbstract:The metabolic impact exerted on a microorganism due to Heterologous Protein Production is still poorly understood in Streptomyces lividans. In this present paper, based on exometabolomic data, a proposed genome-scale metabolic network model is used to assess this metabolic impact in S. lividans. Constraint-based modeling results obtained in this work revealed that the metabolic impact due to Heterologous Protein Production is widely distributed in the genome of S. lividans, causing both slow substrate assimilation and a shift in active pathways. Exchange fluxes that are critical for model performance have been identified for metabolites of mouse tumor necrosis factor, histidine, valine and lysine, as well as biomass. Our results unravel the interaction of Heterologous Protein Production with intracellular metabolism of S. lividans, thus, a possible basis for further studies in relieving the metabolic burden via metabolic or bioprocess engineering.
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On the influence of overexpression of phosphoenolpyruvate carboxykinase in Streptomyces lividans on growth and Production of human tumour necrosis factor-alpha
Applied microbiology and biotechnology, 2012Co-Authors: Ivan Lule, Pieter-jan D'huys, Lieve Van Mellaert, Kristel Bernaerts, Jan Van Impe, Bárbara Maldonado, Jozef AnnéAbstract:Streptomyces lividans has shown potential as an expression system for Heterologous Proteins. Overexpression of proteic factors important for Heterologous Protein Production is a valuable approach to improve yields of such Proteins. Comparative transcriptomic analysis revealed that several genes were differentially expressed in strains involved in Heterologous Protein Production. For instance, the gene-encoding phosphoenolpyruvate carboxykinase (pepck) showed a significant twofold change in recombinant S. lividans producing human tumour necrosis factor-alpha (hTNF-α). The effect of pepck overexpression on S. lividans TK24 and its hTNF-α producing recombinant was thus investigated in bench-top fermenters. Results obtained revealed that pepck overexpression resulted into a twofold increase in specific PEPCK activity during growth. This overexpression is correlated with slower growth rate, reduced excretion of pyruvate and less alkalinisation of the growth medium when compared with the control strain. After 26 h of fermentation, hTNF-α yields were enhanced (up to 1.7-fold) in the pepck-overexpressing S. lividans TK24, demonstrating that this metabolic engineering approach is indeed promising for Heterologous Protein Production.
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MiniReviewStreptomyces lividans as host for Heterologous Protein Production
Fems Microbiology Letters, 1993Co-Authors: Jozef Anné, Lieve Van MellaertAbstract:Streptomycetes are Gram-positive soil bacteria with a differentiated morphology. They are considered interesting candidates for the Production of Heterologous Proteins for several reasons, including their efficient secretion mechanism by which the secreted Proteins are localized into the culture supernatant. In view of this potential, this review article describes different aspects of gene expression and regulation in Streptomyces, and summarizes and discusses results obtained using Streptomyces lividans as host for secretion of heterologus Proteins of prokaryotic and eukaryotic origin.
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Streptomyces lividans as host for Heterologous Protein Production.
FEMS microbiology letters, 1993Co-Authors: Jozef Anné, Lieve Van MellaertAbstract:Streptomycetes are Gram-positive soil bacteria with a differentiated morphology. They are considered interesting candidates for the Production of Heterologous Proteins for several reasons, including their efficient secretion mechanism by which the secreted Proteins are localized into the culture supernatant. In view of this potential, this review article describes different aspects of gene expression and regulation in Streptomyces, and summarizes and discusses results obtained using Streptomyces lividans as host for secretion of heterologus Proteins of prokaryotic and eukaryotic origin.
Ileana Vigentini - One of the best experts on this subject based on the ideXlab platform.
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Heterologous Protein Production in Zygosaccharomyces bailii: physiological effects and fermentative strategies.
FEMS yeast research, 2005Co-Authors: Ileana Vigentini, Annamaria Merico, Danilo Porro, Paola Branduardi, Luca Brambilla, Concetta CompagnoAbstract:The optimisation and scale-up of a specific Protein Production process have to take into account cultivation conditions as well as cell physiology of growth and the influence of foreign Protein expression on host cell metabolism. The ability of Zygosaccharomyces bailii to tolerate high sugar concentrations as well as high temperatures and acidic environments renders this "non-conventional" yeast suitable for the development of biotechnological processes like Heterologous Protein Production. This work addresses the Production of human interleukin-1beta by a recombinant Z. bailii strain. We found that the Heterologous Protein Production causes some modifications of the Z. bailii carbon metabolism, leading to a reduced biomass yield. The other important factor is the dependence of the recombinant IL-1beta Production/secretion on the growth rate. Among the cultivation strategies studied, the most appropriate in terms of Production and productivity was the fed-batch mode.
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Heterologous Protein Production in Zygosaccharomyces bailii: physiological effects and fermentative strategies.
Fems Yeast Research, 2004Co-Authors: Ileana Vigentini, Annamaria Merico, Danilo Porro, Paola Branduardi, Luca Brambilla, Concetta CompagnoAbstract:The optimisation and scale-up of a specific Protein Production process have to take into account cultivation conditions as well as cell physiology of growth and the influence of foreign Protein expression on host cell metabolism. The ability of Zygosaccharomyces bailii to tolerate high sugar concentrations as well as high temperatures and acidic environments renders this “non-conventional” yeast suitable for the development of biotechnological processes like Heterologous Protein Production. This work addresses the Production of human interleukin-1β by a recombinant Z. bailii strain. We found that the Heterologous Protein Production causes some modifications of the Z. bailii carbon metabolism, leading to a reduced biomass yield. The other important factor is the dependence of the recombinant IL-1β Production/secretion on the growth rate. Among the cultivation strategies studied, the most appropriate in terms of Production and productivity was the fed-batch mode.
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How physiological and cultural conditions influence Heterologous Protein Production in Kluyveromyces lactis.
Journal of biotechnology, 2004Co-Authors: Annamaria Merico, Ileana Vigentini, Daniele Capitanio, Bianca Maria Ranzi, Concetta CompagnoAbstract:The optimization and scale-up of a specific Protein Production process have to take into account cultural conditions as well as cell physiology of growth and influence of foreign Protein expression on host cell metabolism. Growth on cheap substrates, efficient secretion ability and a weaker tendency to hypermannosilate Proteins than S. cerevisiae, make K. lactis an excellent and well-accepted host for Heterologous Protein Production, even for human use. A fairly good Heterologous glucoamylase yield and the setting of the optimal conditions to produce it were obtained expressing the Arxula adeninivorans glucoamylase in a strain of K. lactis and its isogenic mutant, which seems to have higher secretion ability. We performed batch cultures of both strains to analyze the influence of different physiological and environmental parameters on glucoamylase Production/secretion. Interestingly, the maintenance of pH in the range of neutrality causes the consumption of a larger amount of carbon source, a longer time of Production and a better stability of the active form of the enzyme, thus increasing biomass and glucoamylase Production. Furthermore, the enrichment of the culture medium adds up to the action of pH control, forcing the mutant Production/secretion to higher levels.
Paola Branduardi - One of the best experts on this subject based on the ideXlab platform.
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Induction by Hypoxia of Heterologous-Protein Production with the KlPDC1 Promoter in Yeasts
Applied and environmental microbiology, 2006Co-Authors: Andrea Camattari, Danilo Porro, Paola Branduardi, Michele M. Bianchi, Luca BrambillaAbstract:The control of promoter activity by oxygen availability appears to be an intriguing system for Heterologous Protein Production. In fact, during cell growth in a bioreactor, an oxygen shortage is easily obtained simply by interrupting the air supply. The purpose of our work was to explore the possible use of hypoxic induction of the KlPDC1 promoter to direct Heterologous gene expression in yeast. In the present study, an expression system based on the KlPDC1 promoter was developed and characterized. Several Heterologous Proteins, differing in size, origin, localization, and posttranslational modification, were successfully expressed in Kluyveromyces lactis under the control of the wild type or a modified promoter sequence, with a Production ratio between 4 and more than 100. Yields were further optimized by a more accurate control of hypoxic physiological conditions. Production of as high as 180 mg/liter of human interleukin-1β was obtained, representing the highest value obtained with yeasts in a lab-scale bioreactor to date. Moreover, the transferability of our system to related yeasts was assessed. The lacZ gene from Escherichia coli was cloned downstream of the KlPDC1 promoter in order to get β-galactosidase activity in response to induction of the promoter. A centromeric vector harboring this expression cassette was introduced in Saccharomyces cerevisiae and in Zygosaccharomyces bailii, and effects of hypoxic induction were measured and compared to those already observed in K. lactis cells. Interestingly, we found that the induction still worked in Z. bailii; thus, this promotor constitutes a possible inducible system for this new nonconventional host.
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Heterologous Protein Production in Zygosaccharomyces bailii: physiological effects and fermentative strategies.
FEMS yeast research, 2005Co-Authors: Ileana Vigentini, Annamaria Merico, Danilo Porro, Paola Branduardi, Luca Brambilla, Concetta CompagnoAbstract:The optimisation and scale-up of a specific Protein Production process have to take into account cultivation conditions as well as cell physiology of growth and the influence of foreign Protein expression on host cell metabolism. The ability of Zygosaccharomyces bailii to tolerate high sugar concentrations as well as high temperatures and acidic environments renders this "non-conventional" yeast suitable for the development of biotechnological processes like Heterologous Protein Production. This work addresses the Production of human interleukin-1beta by a recombinant Z. bailii strain. We found that the Heterologous Protein Production causes some modifications of the Z. bailii carbon metabolism, leading to a reduced biomass yield. The other important factor is the dependence of the recombinant IL-1beta Production/secretion on the growth rate. Among the cultivation strategies studied, the most appropriate in terms of Production and productivity was the fed-batch mode.
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Heterologous Protein Production in Zygosaccharomyces bailii: physiological effects and fermentative strategies.
Fems Yeast Research, 2004Co-Authors: Ileana Vigentini, Annamaria Merico, Danilo Porro, Paola Branduardi, Luca Brambilla, Concetta CompagnoAbstract:The optimisation and scale-up of a specific Protein Production process have to take into account cultivation conditions as well as cell physiology of growth and the influence of foreign Protein expression on host cell metabolism. The ability of Zygosaccharomyces bailii to tolerate high sugar concentrations as well as high temperatures and acidic environments renders this “non-conventional” yeast suitable for the development of biotechnological processes like Heterologous Protein Production. This work addresses the Production of human interleukin-1β by a recombinant Z. bailii strain. We found that the Heterologous Protein Production causes some modifications of the Z. bailii carbon metabolism, leading to a reduced biomass yield. The other important factor is the dependence of the recombinant IL-1β Production/secretion on the growth rate. Among the cultivation strategies studied, the most appropriate in terms of Production and productivity was the fed-batch mode.