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M.A. Vieira Lara
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7 records found
1
Journal article
(2026)
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M. Warmerdam, Y.O. Pijman, M.P. Pedersen, K. Wattel, M.A. Vieira Lara, J.T. Pronk
Ethanol is an interesting renewable feedstock for microbial biotechnology. Compared to other yeasts, Cyberlindnera jadinii has an exceptionally high biomass yield on ethanol. Simulations with a stoichiometric metabolic model identified an intramitochondrial rather than cytosolic localization of NAD+-dependent alcohol and acetaldehyde dehydrogenases as a possible explanation for this high yield. This hypothesis was tested in a comparative study with Ogataea parapolymorpha, which, like C. jadinii, contains a proton-pumping Complex I NADH dehydrogenase, but shows a lower biomass yield on ethanol. Experiments with an aox null mutant showed that the lower biomass yield of O. parapolymorpha was not caused by in vivo alcohol oxidase activity. Proteome analyses showed that the most abundant ADH isozyme in C. jadinii and O. parapolymorpha contained and lacked a predicted mitochondrial targeting sequence, respectively. The ADH protein with the highest prevalence in ethanol-limited cultures of C. jadinii was predicted to be mitochondrial and accounted for almost 4% of the detected proteome. In subcellular fractionation studies with cells grown under ethanol limitation, ADH showed 10-fold lower specific activity in the mitochondria-enriched fraction of cell homogenates of O. parapolymorpha than in the cytosolic fraction. Conversely, C. jadinii showed a 50% higher ADH activity in the mitochondria-enriched fraction. A low overall recovery of ADH in mitochondria-enriched fractions of C. jadinii was attributed to disruption of the reticulate mitochondrial structures observed in ethanol-grown cells during preparation of cell homogenates. This study demonstrates how subcellular localization of an NAD+-dependent dehydrogenase can influence respiratory energy coupling in yeasts that harbor a Complex I NADH dehydrogenase. IMPORTANCE: This study provides theoretical and experimental evidence that intracellular localization of NAD+-dependent dehydrogenases can be a key factor for respiratory energy coupling in yeasts. Based on this, alteration of the cellular localization of NAD+-dehydrogenases is identified as an interesting metabolic engineering strategy for improving respiratory energy coupling in eukaryotic cell factories.
...
Ethanol is an interesting renewable feedstock for microbial biotechnology. Compared to other yeasts, Cyberlindnera jadinii has an exceptionally high biomass yield on ethanol. Simulations with a stoichiometric metabolic model identified an intramitochondrial rather than cytosolic localization of NAD+-dependent alcohol and acetaldehyde dehydrogenases as a possible explanation for this high yield. This hypothesis was tested in a comparative study with Ogataea parapolymorpha, which, like C. jadinii, contains a proton-pumping Complex I NADH dehydrogenase, but shows a lower biomass yield on ethanol. Experiments with an aox null mutant showed that the lower biomass yield of O. parapolymorpha was not caused by in vivo alcohol oxidase activity. Proteome analyses showed that the most abundant ADH isozyme in C. jadinii and O. parapolymorpha contained and lacked a predicted mitochondrial targeting sequence, respectively. The ADH protein with the highest prevalence in ethanol-limited cultures of C. jadinii was predicted to be mitochondrial and accounted for almost 4% of the detected proteome. In subcellular fractionation studies with cells grown under ethanol limitation, ADH showed 10-fold lower specific activity in the mitochondria-enriched fraction of cell homogenates of O. parapolymorpha than in the cytosolic fraction. Conversely, C. jadinii showed a 50% higher ADH activity in the mitochondria-enriched fraction. A low overall recovery of ADH in mitochondria-enriched fractions of C. jadinii was attributed to disruption of the reticulate mitochondrial structures observed in ethanol-grown cells during preparation of cell homogenates. This study demonstrates how subcellular localization of an NAD+-dependent dehydrogenase can influence respiratory energy coupling in yeasts that harbor a Complex I NADH dehydrogenase. IMPORTANCE: This study provides theoretical and experimental evidence that intracellular localization of NAD+-dependent dehydrogenases can be a key factor for respiratory energy coupling in yeasts. Based on this, alteration of the cellular localization of NAD+-dehydrogenases is identified as an interesting metabolic engineering strategy for improving respiratory energy coupling in eukaryotic cell factories.
The high cost of cultivation media remains one of the barriers to the commercialization of cultivated meat. Current serum-free media reduce reliance on fetal bovine serum but only achieve modest cost savings. Especially at larger scales (> 20 m3), pharmaceutical-grade amino acids and recombinant proteins remain major cost drivers. Techno-economic analyses suggest that replacing purified amino acids is essential to achieve price parity with conventional meat. Microbial hydrolysates, also referred to as extracts, offer a promising alternative, as these can supply complex mixtures of amino acids and peptides at lower cost and with greater scalability than chemically defined formulations. In particular, yeast and bacterial extracts combine high protein (and therefore high amino acid) content, rapid growth, and established industrial-scale production. Comparative analyses indicate that microbial amino acid profiles broadly overlap with those of animal cells and traditional meat, but this similarity does not translate into meeting cellular amino acid demand. Consumption data indicate that key amino acids, such as glutamine, cysteine, serine and arginine, would be supplied insufficiently by microbial extracts. This mismatch highlights the need for engineering of microbial biomass and optimization of extraction methods. Extraction methods such as autolysis, enzymatic lysis, or physical disruption strongly influence nutrient release and composition, underscoring the need for standardizing microbial extract preparation. In addition, challenges remain in ensuring consistency, safety, and bioavailability of nutrients, as microbial extracts also contain nucleic acids and potential toxins. This review summarizes current knowledge on microbial hydrolysates for cultivated meat media, including biomass composition, extraction methods, and analytical tools to assess quality and performance. We identify key knowledge gaps, particularly in quantitative amino acid consumption data for relevant cell lines and performance testing in scalable cultures. Addressing these gaps could enable cost-effective media development and accelerate research toward sustainable, commercially viable cultivated meat.
...
The high cost of cultivation media remains one of the barriers to the commercialization of cultivated meat. Current serum-free media reduce reliance on fetal bovine serum but only achieve modest cost savings. Especially at larger scales (> 20 m3), pharmaceutical-grade amino acids and recombinant proteins remain major cost drivers. Techno-economic analyses suggest that replacing purified amino acids is essential to achieve price parity with conventional meat. Microbial hydrolysates, also referred to as extracts, offer a promising alternative, as these can supply complex mixtures of amino acids and peptides at lower cost and with greater scalability than chemically defined formulations. In particular, yeast and bacterial extracts combine high protein (and therefore high amino acid) content, rapid growth, and established industrial-scale production. Comparative analyses indicate that microbial amino acid profiles broadly overlap with those of animal cells and traditional meat, but this similarity does not translate into meeting cellular amino acid demand. Consumption data indicate that key amino acids, such as glutamine, cysteine, serine and arginine, would be supplied insufficiently by microbial extracts. This mismatch highlights the need for engineering of microbial biomass and optimization of extraction methods. Extraction methods such as autolysis, enzymatic lysis, or physical disruption strongly influence nutrient release and composition, underscoring the need for standardizing microbial extract preparation. In addition, challenges remain in ensuring consistency, safety, and bioavailability of nutrients, as microbial extracts also contain nucleic acids and potential toxins. This review summarizes current knowledge on microbial hydrolysates for cultivated meat media, including biomass composition, extraction methods, and analytical tools to assess quality and performance. We identify key knowledge gaps, particularly in quantitative amino acid consumption data for relevant cell lines and performance testing in scalable cultures. Addressing these gaps could enable cost-effective media development and accelerate research toward sustainable, commercially viable cultivated meat.
Journal article
(2025)
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Joel C. Fisher, Aaffien C. Reijne, Kaja Hribar, Marcel A. Vieira-Lara, Alzbeta Talarovicova, Dirk Jan Reijngoud, David D. van Niekerk, Jacky L. Snoep, Gertjan van Dijk, More authors...
Dietary restriction (DR) improves insulin sensitivity, however, it has not been tested in long-term interventions and with diet type as variable. Therefore, we exposed mice to either a low-fat (LF) or high-fat-sucrose (HFS) diet, either fed ad libitum (AL) or in a DR regimen from weaning till 2 years of age. Using an oral glucose tolerance test with [6,6-2H2]-labelled glucose, we found that DR markedly reduced plasma insulin concentrations and strongly elevated hepatic and peripheral insulin sensitivity when compared to AL cohorts. These effects of DR, however, appeared to depend on diet and age, with stable increases in hepatic and peripheral insulin sensitivities across all ages in the LF condition, while these became clearly less elevated in the HFS condition with advancing age.
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Dietary restriction (DR) improves insulin sensitivity, however, it has not been tested in long-term interventions and with diet type as variable. Therefore, we exposed mice to either a low-fat (LF) or high-fat-sucrose (HFS) diet, either fed ad libitum (AL) or in a DR regimen from weaning till 2 years of age. Using an oral glucose tolerance test with [6,6-2H2]-labelled glucose, we found that DR markedly reduced plasma insulin concentrations and strongly elevated hepatic and peripheral insulin sensitivity when compared to AL cohorts. These effects of DR, however, appeared to depend on diet and age, with stable increases in hepatic and peripheral insulin sensitivities across all ages in the LF condition, while these became clearly less elevated in the HFS condition with advancing age.
Journal article
(2024)
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Marcel A. Vieira-Lara, Marieke Warmerdam, Erik A.F. de Hulster, Marcel van den Broek, Jean Marc Daran, Jack T. Pronk
Background
Elimination of greenhouse gas emissions in industrial biotechnology requires replacement of carbohydrates by alternative carbon substrates, produced from CO2 and waste streams. Ethanol is already industrially produced from agricultural residues and waste gas and is miscible with water, self-sterilizing and energy-dense. The yeast C. jadinii can grow on ethanol and has a history in the production of single-cell protein (SCP) for feed and food applications. To address a knowledge gap in quantitative physiology of C. jadinii during growth on ethanol, this study investigates growth kinetics, growth energetics, nutritional requirements, and biomass composition of C. jadinii strains in batch, chemostat and fed-batch cultures.
Results
In aerobic, ethanol-limited chemostat cultures, C. jadinii CBS 621 exhibited a maximum biomass yield on ethanol (Y max X/S) of 0.83 gbiomass (gethanol)−1 and an estimated maintenance requirement for ATP (mATP) of 2.7 mmolATP (gbiomass)−1 h−1. Even at specific growth rates below 0.05 h−1, a stable protein content of approximately 0.54 gprotein (gbiomass)−1 was observed. At low specific growth rates, up to 17% of the proteome consisted of alcohol dehydrogenase proteins, followed by aldehyde dehydrogenases and acetyl-CoA synthetase. Of 13 C. jadinii strains evaluated, 11 displayed fast growth on ethanol (μmax > 0.4 h−1) in mineral medium without vitamins, and CBS 621 was found to be a thiamine auxotroph. The prototrophic strain C. jadinii CBS 5947 was grown on an inorganic salts medium in fed-batch cultures (10-L scale) fed with pure ethanol. Biomass concentrations in these cultures increased up to 100 gbiomass (kgbroth)−1, with a biomass yield of 0.65 gbiomass (gethanol)−1. Model-based simulation, based on quantitative parameters determined in chemostat cultures, adequately predicted biomass production. A different protein content of chemostat- and fed-batch-grown biomass (54 and 42%, respectively) may reflect the more dynamic conditions in fed-batch cultures.
Conclusions
Analysis of ethanol-grown batch, chemostat and fed-batch cultures provided a quantitative physiology baseline for fundamental and applied research on C. jadinii. Its high maximum growth rate, high energetic efficiency of ethanol dissimilation, simple nutritional requirements and high protein content, make C. jadinii a highly interesting platform for production of SCP and other products from ethanol. ...
Elimination of greenhouse gas emissions in industrial biotechnology requires replacement of carbohydrates by alternative carbon substrates, produced from CO2 and waste streams. Ethanol is already industrially produced from agricultural residues and waste gas and is miscible with water, self-sterilizing and energy-dense. The yeast C. jadinii can grow on ethanol and has a history in the production of single-cell protein (SCP) for feed and food applications. To address a knowledge gap in quantitative physiology of C. jadinii during growth on ethanol, this study investigates growth kinetics, growth energetics, nutritional requirements, and biomass composition of C. jadinii strains in batch, chemostat and fed-batch cultures.
Results
In aerobic, ethanol-limited chemostat cultures, C. jadinii CBS 621 exhibited a maximum biomass yield on ethanol (Y max X/S) of 0.83 gbiomass (gethanol)−1 and an estimated maintenance requirement for ATP (mATP) of 2.7 mmolATP (gbiomass)−1 h−1. Even at specific growth rates below 0.05 h−1, a stable protein content of approximately 0.54 gprotein (gbiomass)−1 was observed. At low specific growth rates, up to 17% of the proteome consisted of alcohol dehydrogenase proteins, followed by aldehyde dehydrogenases and acetyl-CoA synthetase. Of 13 C. jadinii strains evaluated, 11 displayed fast growth on ethanol (μmax > 0.4 h−1) in mineral medium without vitamins, and CBS 621 was found to be a thiamine auxotroph. The prototrophic strain C. jadinii CBS 5947 was grown on an inorganic salts medium in fed-batch cultures (10-L scale) fed with pure ethanol. Biomass concentrations in these cultures increased up to 100 gbiomass (kgbroth)−1, with a biomass yield of 0.65 gbiomass (gethanol)−1. Model-based simulation, based on quantitative parameters determined in chemostat cultures, adequately predicted biomass production. A different protein content of chemostat- and fed-batch-grown biomass (54 and 42%, respectively) may reflect the more dynamic conditions in fed-batch cultures.
Conclusions
Analysis of ethanol-grown batch, chemostat and fed-batch cultures provided a quantitative physiology baseline for fundamental and applied research on C. jadinii. Its high maximum growth rate, high energetic efficiency of ethanol dissimilation, simple nutritional requirements and high protein content, make C. jadinii a highly interesting platform for production of SCP and other products from ethanol. ...
Background
Elimination of greenhouse gas emissions in industrial biotechnology requires replacement of carbohydrates by alternative carbon substrates, produced from CO2 and waste streams. Ethanol is already industrially produced from agricultural residues and waste gas and is miscible with water, self-sterilizing and energy-dense. The yeast C. jadinii can grow on ethanol and has a history in the production of single-cell protein (SCP) for feed and food applications. To address a knowledge gap in quantitative physiology of C. jadinii during growth on ethanol, this study investigates growth kinetics, growth energetics, nutritional requirements, and biomass composition of C. jadinii strains in batch, chemostat and fed-batch cultures.
Results
In aerobic, ethanol-limited chemostat cultures, C. jadinii CBS 621 exhibited a maximum biomass yield on ethanol (Y max X/S) of 0.83 gbiomass (gethanol)−1 and an estimated maintenance requirement for ATP (mATP) of 2.7 mmolATP (gbiomass)−1 h−1. Even at specific growth rates below 0.05 h−1, a stable protein content of approximately 0.54 gprotein (gbiomass)−1 was observed. At low specific growth rates, up to 17% of the proteome consisted of alcohol dehydrogenase proteins, followed by aldehyde dehydrogenases and acetyl-CoA synthetase. Of 13 C. jadinii strains evaluated, 11 displayed fast growth on ethanol (μmax > 0.4 h−1) in mineral medium without vitamins, and CBS 621 was found to be a thiamine auxotroph. The prototrophic strain C. jadinii CBS 5947 was grown on an inorganic salts medium in fed-batch cultures (10-L scale) fed with pure ethanol. Biomass concentrations in these cultures increased up to 100 gbiomass (kgbroth)−1, with a biomass yield of 0.65 gbiomass (gethanol)−1. Model-based simulation, based on quantitative parameters determined in chemostat cultures, adequately predicted biomass production. A different protein content of chemostat- and fed-batch-grown biomass (54 and 42%, respectively) may reflect the more dynamic conditions in fed-batch cultures.
Conclusions
Analysis of ethanol-grown batch, chemostat and fed-batch cultures provided a quantitative physiology baseline for fundamental and applied research on C. jadinii. Its high maximum growth rate, high energetic efficiency of ethanol dissimilation, simple nutritional requirements and high protein content, make C. jadinii a highly interesting platform for production of SCP and other products from ethanol.
Elimination of greenhouse gas emissions in industrial biotechnology requires replacement of carbohydrates by alternative carbon substrates, produced from CO2 and waste streams. Ethanol is already industrially produced from agricultural residues and waste gas and is miscible with water, self-sterilizing and energy-dense. The yeast C. jadinii can grow on ethanol and has a history in the production of single-cell protein (SCP) for feed and food applications. To address a knowledge gap in quantitative physiology of C. jadinii during growth on ethanol, this study investigates growth kinetics, growth energetics, nutritional requirements, and biomass composition of C. jadinii strains in batch, chemostat and fed-batch cultures.
Results
In aerobic, ethanol-limited chemostat cultures, C. jadinii CBS 621 exhibited a maximum biomass yield on ethanol (Y max X/S) of 0.83 gbiomass (gethanol)−1 and an estimated maintenance requirement for ATP (mATP) of 2.7 mmolATP (gbiomass)−1 h−1. Even at specific growth rates below 0.05 h−1, a stable protein content of approximately 0.54 gprotein (gbiomass)−1 was observed. At low specific growth rates, up to 17% of the proteome consisted of alcohol dehydrogenase proteins, followed by aldehyde dehydrogenases and acetyl-CoA synthetase. Of 13 C. jadinii strains evaluated, 11 displayed fast growth on ethanol (μmax > 0.4 h−1) in mineral medium without vitamins, and CBS 621 was found to be a thiamine auxotroph. The prototrophic strain C. jadinii CBS 5947 was grown on an inorganic salts medium in fed-batch cultures (10-L scale) fed with pure ethanol. Biomass concentrations in these cultures increased up to 100 gbiomass (kgbroth)−1, with a biomass yield of 0.65 gbiomass (gethanol)−1. Model-based simulation, based on quantitative parameters determined in chemostat cultures, adequately predicted biomass production. A different protein content of chemostat- and fed-batch-grown biomass (54 and 42%, respectively) may reflect the more dynamic conditions in fed-batch cultures.
Conclusions
Analysis of ethanol-grown batch, chemostat and fed-batch cultures provided a quantitative physiology baseline for fundamental and applied research on C. jadinii. Its high maximum growth rate, high energetic efficiency of ethanol dissimilation, simple nutritional requirements and high protein content, make C. jadinii a highly interesting platform for production of SCP and other products from ethanol.
Ethanol is produced at industrial scale from non-agricultural feedstocks by gas fermentation, while research on other low-emission processes for ethanol production is accelerating. In view of its degree of reduction, water solubility and relatively low toxicity, ethanol is an interesting candidate to replace sugars in aerobic, zero-emission processes for yeast-based production of whole-cell protein and low-molecular-weight compounds. Currently, little information is available on specific growth rates, biomass yields and biomass composition of yeast species during growth on synthetic medium with ethanol as sole carbon source. In this study, strains of 52 Saccharomycotina yeasts were screened for their growth characteristics on ethanol. After first screening in microtiter plates, 21 fast-growing strains that were further analysed in aerobic shake-flask cultures showed specific growth rates of 0.12-0.46 h−1. Five fast-growing strains were further studied in aerobic, ethanol-limited chemostats (dilution rate 0.10 h−1). Strains of the industrial yeasts Saccharomyces cerevisiae and Kluyveromyces lactis, whose genomes lack genes for a proton-coupled Complex-I NADH dehydrogenase, both showed biomass yields of 0.6 g biomass (g ethanol)−1. Of three yeasts whose genome does contain Complex-I genes, Phaffomyces thermotolerans, showed the same biomass yield as S. cerevisiae, while Ogataea parapolymorpha and Cyberlindnera jadinii showed biomass yields of 0.67 ± 0.01 and 0.73 ± 0.00 g g−1, respectively. The biomass yield of C. jadinii, which also showed the highest protein content of the 5 yeasts tested in chemostats, corresponded to 88% of the theoretical biomass yield in a scenario where growth is limited by assimilation rather than by energy metabolism.
...
Ethanol is produced at industrial scale from non-agricultural feedstocks by gas fermentation, while research on other low-emission processes for ethanol production is accelerating. In view of its degree of reduction, water solubility and relatively low toxicity, ethanol is an interesting candidate to replace sugars in aerobic, zero-emission processes for yeast-based production of whole-cell protein and low-molecular-weight compounds. Currently, little information is available on specific growth rates, biomass yields and biomass composition of yeast species during growth on synthetic medium with ethanol as sole carbon source. In this study, strains of 52 Saccharomycotina yeasts were screened for their growth characteristics on ethanol. After first screening in microtiter plates, 21 fast-growing strains that were further analysed in aerobic shake-flask cultures showed specific growth rates of 0.12-0.46 h−1. Five fast-growing strains were further studied in aerobic, ethanol-limited chemostats (dilution rate 0.10 h−1). Strains of the industrial yeasts Saccharomyces cerevisiae and Kluyveromyces lactis, whose genomes lack genes for a proton-coupled Complex-I NADH dehydrogenase, both showed biomass yields of 0.6 g biomass (g ethanol)−1. Of three yeasts whose genome does contain Complex-I genes, Phaffomyces thermotolerans, showed the same biomass yield as S. cerevisiae, while Ogataea parapolymorpha and Cyberlindnera jadinii showed biomass yields of 0.67 ± 0.01 and 0.73 ± 0.00 g g−1, respectively. The biomass yield of C. jadinii, which also showed the highest protein content of the 5 yeasts tested in chemostats, corresponded to 88% of the theoretical biomass yield in a scenario where growth is limited by assimilation rather than by energy metabolism.
Journal article
(2024)
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Marieke Warmerdam, Marcel A. Vieira-Lara, Robert Mans, Jean Marc Daran, Jack T. Pronk
Emerging low-emission production technologies make ethanol an interesting substrate for yeast biotechnology, but information on growth rates and biomass yields of yeasts on ethanol is scarce. Strains of 52 Saccharomycotina yeasts were screened for growth on ethanol. The 21 fastest strains, among which representatives of the Phaffomycetales order were overrepresented, showed specific growth rates in ethanol-grown shake-flask cultures between 0.12 and 0.46 h−1. Seven strains were studied in aerobic, ethanol-limited chemostats (dilution rate 0.10 h−1). Saccharomyces cerevisiae and Kluyveromyces lactis, whose genomes do not encode Complex-I-type NADH dehydrogenases, showed biomass yields of 0.59 and 0.56 gbiomass gethanol−1, respectively. Different biomass yields were observed among species whose genomes do harbour Complex-I-encoding genes: Phaffomyces thermotolerans (0.58 g g−1), Pichia ethanolica (0.59 g g−1), Saturnispora dispora (0.66 g g−1), Ogataea parapolymorpha (0.67 g g−1), and Cyberlindnera jadinii (0.73 g g−1). Cyberlindnera jadinii biomass showed the highest protein content (59 ± 2%) of these yeasts. Its biomass yield corresponded to 88% of the theoretical maximum that is reached when growth is limited by assimilation rather than by energy availability. This study suggests that energy coupling of mitochondrial respiration and its regulation will become key factors for selecting and improving yeast strains for ethanol-based processes.
...
Emerging low-emission production technologies make ethanol an interesting substrate for yeast biotechnology, but information on growth rates and biomass yields of yeasts on ethanol is scarce. Strains of 52 Saccharomycotina yeasts were screened for growth on ethanol. The 21 fastest strains, among which representatives of the Phaffomycetales order were overrepresented, showed specific growth rates in ethanol-grown shake-flask cultures between 0.12 and 0.46 h−1. Seven strains were studied in aerobic, ethanol-limited chemostats (dilution rate 0.10 h−1). Saccharomyces cerevisiae and Kluyveromyces lactis, whose genomes do not encode Complex-I-type NADH dehydrogenases, showed biomass yields of 0.59 and 0.56 gbiomass gethanol−1, respectively. Different biomass yields were observed among species whose genomes do harbour Complex-I-encoding genes: Phaffomyces thermotolerans (0.58 g g−1), Pichia ethanolica (0.59 g g−1), Saturnispora dispora (0.66 g g−1), Ogataea parapolymorpha (0.67 g g−1), and Cyberlindnera jadinii (0.73 g g−1). Cyberlindnera jadinii biomass showed the highest protein content (59 ± 2%) of these yeasts. Its biomass yield corresponded to 88% of the theoretical maximum that is reached when growth is limited by assimilation rather than by energy availability. This study suggests that energy coupling of mitochondrial respiration and its regulation will become key factors for selecting and improving yeast strains for ethanol-based processes.
Journal article
(2022)
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Francine J. Boonekamp, Ewout Knibbe, Marcel A. Vieira-Lara, Melanie Wijsman, Marijke A.H. Luttik, Karen van Eunen, Maxime den Ridder, Martin Pabst, Jean Marc Daran, Pascale Daran-Lapujade
Although transplantation of single genes in yeast plays a key role in elucidating gene functionality in metazoans, technical challenges hamper humanization of full pathways and processes. Empowered by advances in synthetic biology, this study demonstrates the feasibility and implementation of full humanization of glycolysis in yeast. Single gene and full pathway transplantation revealed the remarkable conservation of glycolytic and moonlighting functions and, combined with evolutionary strategies, brought to light context-dependent responses. Human hexokinase 1 and 2, but not 4, required mutations in their catalytic or allosteric sites for functionality in yeast, whereas hexokinase 3 was unable to complement its yeast ortholog. Comparison with human tissues cultures showed preservation of turnover numbers of human glycolytic enzymes in yeast and human cell cultures. This demonstration of transplantation of an entire essential pathway paves the way for establishment of species-, tissue-, and disease-specific metazoan models.
...
Although transplantation of single genes in yeast plays a key role in elucidating gene functionality in metazoans, technical challenges hamper humanization of full pathways and processes. Empowered by advances in synthetic biology, this study demonstrates the feasibility and implementation of full humanization of glycolysis in yeast. Single gene and full pathway transplantation revealed the remarkable conservation of glycolytic and moonlighting functions and, combined with evolutionary strategies, brought to light context-dependent responses. Human hexokinase 1 and 2, but not 4, required mutations in their catalytic or allosteric sites for functionality in yeast, whereas hexokinase 3 was unable to complement its yeast ortholog. Comparison with human tissues cultures showed preservation of turnover numbers of human glycolytic enzymes in yeast and human cell cultures. This demonstration of transplantation of an entire essential pathway paves the way for establishment of species-, tissue-, and disease-specific metazoan models.