The Experts below are selected from a list of 315 Experts worldwide ranked by ideXlab platform
Sonjaverena Albers - One of the best experts on this subject based on the ideXlab platform.
-
identification of xylr the activator of arabinose xylose inducible regulon in sulfolobus acidocaldarius and its application for homologous protein expression
Frontiers in Microbiology, 2020Co-Authors: Nienke Van Der Kolk, Alexander Wagner, Michaela Wagner, Bianca Wasmer, Bettina Siebers, Sonjaverena AlbersAbstract:The thermophilic archaeon Sulfolobus acidocaldarius can use different carbon sources for growth, including the Pentoses D-xylose and L-arabinose. In this study, we identified the activator XylR (saci_2116) responsible for the transcriptional regulation of the Pentose transporter and Pentose metabolizing genes in S. acidocaldarius. A xylR deletion mutant showed growth retardation on D-xylose/L-arabinose containing media and the lack of transcription of the respective ABC transporter. In contrast to so far used promoters for expression in S. acidocaldarius, the xylR responsive promoters have a very low background activity. Finally, two XylR dependent promoters next to the long-established maltose inducible promotor were used to construct a high-throughput expression vector system for S. acidocaldarius to efficiently clone and express proteins in S. acidocaldarius.
-
sulfolobus acidocaldarius transports Pentoses via a carbohydrate uptake transporter 2 cut2 type abc transporter and metabolizes them through the aldolase independent weimberg pathway
Applied and Environmental Microbiology, 2017Co-Authors: Michaela Wagner, Nienke Van Der Kolk, Bettina Siebers, Lu Shen, Andreas Albersmeier, Sujin Kim, Jaeho Cha, Christopher Brasen, Jorn Kalinowski, Sonjaverena AlbersAbstract:ABSTRACT Sulfolobus spp. possess a great metabolic versatility and grow heterotrophically on various carbon sources, such as different sugars and peptides. Known sugar transporters in Archaea predominantly belong to ABC transport systems. Although several ABC transporters for sugar uptake have been characterized in the crenarchaeon Sulfolobus solfataricus, only one homologue of these transporters, the maltose/maltooligomer transporter, could be identified in the closely related Sulfolobus acidocaldarius. Comparison of the transcriptome of S. acidocaldarius MW001 grown on peptides alone and peptides in the presence of d-xylose allowed for the identification of the ABC transporter for d-xylose and l-arabinose transport and the gaining of deeper insights into Pentose catabolism under the respective growth conditions. The d-xylose/l-arabinose substrate binding protein (SBP) (Saci_2122) of the ABC transporter is unique in Archaea and shares more similarity to bacterial SBPs of the carbohydrate uptake transporter-2 (CUT2) family than to any characterized archaeal one. The identified Pentose transporter is the first CUT2 family ABC transporter analyzed in the domain of Archaea. Single-gene deletion mutants of the ABC transporter subunits exemplified the importance of the transport system for d-xylose and l-arabinose uptake. Next to the transporter operon, enzymes of the aldolase-independent Pentose catabolism branch were found to be upregulated in N-Z-Amine and d-xylose medium. The α-ketoglutarate semialdehyde dehydrogenase (KGSADH; Saci_1938) seemed not to be essential for growth on Pentoses. However, the deletion mutant of the 2-keto-3-deoxyarabinoate/xylonate dehydratase (KDXD [also known as KDAD]; Saci_1939) was no longer able to catabolize d-xylose or l-arabinose, suggesting the absence of the aldolase-dependent branch in S. acidocaldarius. IMPORTANCE Thermoacidophilic microorganisms are emerging model organisms for biotechnological applications, as their optimal growth conditions resemble conditions used in certain biotechnologies such as industrial plant waste degradation. Because of its high genome stability, Sulfolobus acidocaldarius is especially suited as a platform organism for such applications. For use in (ligno)cellulose degradation, it was important to understand Pentose uptake and metabolism in S. acidocaldarius. This study revealed that only the aldolase-independent Weimberg pathway is required for growth of S. acidocaldarius MW001 on d-xylose and l-arabinose. Moreover, S. acidocaldarius employs a CUT2 ABC transporter for Pentose uptake, which is more similar to bacterial than to archaeal ABC transporters. The identification of Pentose-inducible promoters will expedite the metabolic engineering of S. acidocaldarius for its development into a platform organism for (ligno)cellulose degradation.
Michaela Wagner - One of the best experts on this subject based on the ideXlab platform.
-
identification of xylr the activator of arabinose xylose inducible regulon in sulfolobus acidocaldarius and its application for homologous protein expression
Frontiers in Microbiology, 2020Co-Authors: Nienke Van Der Kolk, Alexander Wagner, Michaela Wagner, Bianca Wasmer, Bettina Siebers, Sonjaverena AlbersAbstract:The thermophilic archaeon Sulfolobus acidocaldarius can use different carbon sources for growth, including the Pentoses D-xylose and L-arabinose. In this study, we identified the activator XylR (saci_2116) responsible for the transcriptional regulation of the Pentose transporter and Pentose metabolizing genes in S. acidocaldarius. A xylR deletion mutant showed growth retardation on D-xylose/L-arabinose containing media and the lack of transcription of the respective ABC transporter. In contrast to so far used promoters for expression in S. acidocaldarius, the xylR responsive promoters have a very low background activity. Finally, two XylR dependent promoters next to the long-established maltose inducible promotor were used to construct a high-throughput expression vector system for S. acidocaldarius to efficiently clone and express proteins in S. acidocaldarius.
-
sulfolobus acidocaldarius transports Pentoses via a carbohydrate uptake transporter 2 cut2 type abc transporter and metabolizes them through the aldolase independent weimberg pathway
Applied and Environmental Microbiology, 2017Co-Authors: Michaela Wagner, Nienke Van Der Kolk, Bettina Siebers, Lu Shen, Andreas Albersmeier, Sujin Kim, Jaeho Cha, Christopher Brasen, Jorn Kalinowski, Sonjaverena AlbersAbstract:ABSTRACT Sulfolobus spp. possess a great metabolic versatility and grow heterotrophically on various carbon sources, such as different sugars and peptides. Known sugar transporters in Archaea predominantly belong to ABC transport systems. Although several ABC transporters for sugar uptake have been characterized in the crenarchaeon Sulfolobus solfataricus, only one homologue of these transporters, the maltose/maltooligomer transporter, could be identified in the closely related Sulfolobus acidocaldarius. Comparison of the transcriptome of S. acidocaldarius MW001 grown on peptides alone and peptides in the presence of d-xylose allowed for the identification of the ABC transporter for d-xylose and l-arabinose transport and the gaining of deeper insights into Pentose catabolism under the respective growth conditions. The d-xylose/l-arabinose substrate binding protein (SBP) (Saci_2122) of the ABC transporter is unique in Archaea and shares more similarity to bacterial SBPs of the carbohydrate uptake transporter-2 (CUT2) family than to any characterized archaeal one. The identified Pentose transporter is the first CUT2 family ABC transporter analyzed in the domain of Archaea. Single-gene deletion mutants of the ABC transporter subunits exemplified the importance of the transport system for d-xylose and l-arabinose uptake. Next to the transporter operon, enzymes of the aldolase-independent Pentose catabolism branch were found to be upregulated in N-Z-Amine and d-xylose medium. The α-ketoglutarate semialdehyde dehydrogenase (KGSADH; Saci_1938) seemed not to be essential for growth on Pentoses. However, the deletion mutant of the 2-keto-3-deoxyarabinoate/xylonate dehydratase (KDXD [also known as KDAD]; Saci_1939) was no longer able to catabolize d-xylose or l-arabinose, suggesting the absence of the aldolase-dependent branch in S. acidocaldarius. IMPORTANCE Thermoacidophilic microorganisms are emerging model organisms for biotechnological applications, as their optimal growth conditions resemble conditions used in certain biotechnologies such as industrial plant waste degradation. Because of its high genome stability, Sulfolobus acidocaldarius is especially suited as a platform organism for such applications. For use in (ligno)cellulose degradation, it was important to understand Pentose uptake and metabolism in S. acidocaldarius. This study revealed that only the aldolase-independent Weimberg pathway is required for growth of S. acidocaldarius MW001 on d-xylose and l-arabinose. Moreover, S. acidocaldarius employs a CUT2 ABC transporter for Pentose uptake, which is more similar to bacterial than to archaeal ABC transporters. The identification of Pentose-inducible promoters will expedite the metabolic engineering of S. acidocaldarius for its development into a platform organism for (ligno)cellulose degradation.
Nienke Van Der Kolk - One of the best experts on this subject based on the ideXlab platform.
-
identification of xylr the activator of arabinose xylose inducible regulon in sulfolobus acidocaldarius and its application for homologous protein expression
Frontiers in Microbiology, 2020Co-Authors: Nienke Van Der Kolk, Alexander Wagner, Michaela Wagner, Bianca Wasmer, Bettina Siebers, Sonjaverena AlbersAbstract:The thermophilic archaeon Sulfolobus acidocaldarius can use different carbon sources for growth, including the Pentoses D-xylose and L-arabinose. In this study, we identified the activator XylR (saci_2116) responsible for the transcriptional regulation of the Pentose transporter and Pentose metabolizing genes in S. acidocaldarius. A xylR deletion mutant showed growth retardation on D-xylose/L-arabinose containing media and the lack of transcription of the respective ABC transporter. In contrast to so far used promoters for expression in S. acidocaldarius, the xylR responsive promoters have a very low background activity. Finally, two XylR dependent promoters next to the long-established maltose inducible promotor were used to construct a high-throughput expression vector system for S. acidocaldarius to efficiently clone and express proteins in S. acidocaldarius.
-
sulfolobus acidocaldarius transports Pentoses via a carbohydrate uptake transporter 2 cut2 type abc transporter and metabolizes them through the aldolase independent weimberg pathway
Applied and Environmental Microbiology, 2017Co-Authors: Michaela Wagner, Nienke Van Der Kolk, Bettina Siebers, Lu Shen, Andreas Albersmeier, Sujin Kim, Jaeho Cha, Christopher Brasen, Jorn Kalinowski, Sonjaverena AlbersAbstract:ABSTRACT Sulfolobus spp. possess a great metabolic versatility and grow heterotrophically on various carbon sources, such as different sugars and peptides. Known sugar transporters in Archaea predominantly belong to ABC transport systems. Although several ABC transporters for sugar uptake have been characterized in the crenarchaeon Sulfolobus solfataricus, only one homologue of these transporters, the maltose/maltooligomer transporter, could be identified in the closely related Sulfolobus acidocaldarius. Comparison of the transcriptome of S. acidocaldarius MW001 grown on peptides alone and peptides in the presence of d-xylose allowed for the identification of the ABC transporter for d-xylose and l-arabinose transport and the gaining of deeper insights into Pentose catabolism under the respective growth conditions. The d-xylose/l-arabinose substrate binding protein (SBP) (Saci_2122) of the ABC transporter is unique in Archaea and shares more similarity to bacterial SBPs of the carbohydrate uptake transporter-2 (CUT2) family than to any characterized archaeal one. The identified Pentose transporter is the first CUT2 family ABC transporter analyzed in the domain of Archaea. Single-gene deletion mutants of the ABC transporter subunits exemplified the importance of the transport system for d-xylose and l-arabinose uptake. Next to the transporter operon, enzymes of the aldolase-independent Pentose catabolism branch were found to be upregulated in N-Z-Amine and d-xylose medium. The α-ketoglutarate semialdehyde dehydrogenase (KGSADH; Saci_1938) seemed not to be essential for growth on Pentoses. However, the deletion mutant of the 2-keto-3-deoxyarabinoate/xylonate dehydratase (KDXD [also known as KDAD]; Saci_1939) was no longer able to catabolize d-xylose or l-arabinose, suggesting the absence of the aldolase-dependent branch in S. acidocaldarius. IMPORTANCE Thermoacidophilic microorganisms are emerging model organisms for biotechnological applications, as their optimal growth conditions resemble conditions used in certain biotechnologies such as industrial plant waste degradation. Because of its high genome stability, Sulfolobus acidocaldarius is especially suited as a platform organism for such applications. For use in (ligno)cellulose degradation, it was important to understand Pentose uptake and metabolism in S. acidocaldarius. This study revealed that only the aldolase-independent Weimberg pathway is required for growth of S. acidocaldarius MW001 on d-xylose and l-arabinose. Moreover, S. acidocaldarius employs a CUT2 ABC transporter for Pentose uptake, which is more similar to bacterial than to archaeal ABC transporters. The identification of Pentose-inducible promoters will expedite the metabolic engineering of S. acidocaldarius for its development into a platform organism for (ligno)cellulose degradation.
Bettina Siebers - One of the best experts on this subject based on the ideXlab platform.
-
identification of xylr the activator of arabinose xylose inducible regulon in sulfolobus acidocaldarius and its application for homologous protein expression
Frontiers in Microbiology, 2020Co-Authors: Nienke Van Der Kolk, Alexander Wagner, Michaela Wagner, Bianca Wasmer, Bettina Siebers, Sonjaverena AlbersAbstract:The thermophilic archaeon Sulfolobus acidocaldarius can use different carbon sources for growth, including the Pentoses D-xylose and L-arabinose. In this study, we identified the activator XylR (saci_2116) responsible for the transcriptional regulation of the Pentose transporter and Pentose metabolizing genes in S. acidocaldarius. A xylR deletion mutant showed growth retardation on D-xylose/L-arabinose containing media and the lack of transcription of the respective ABC transporter. In contrast to so far used promoters for expression in S. acidocaldarius, the xylR responsive promoters have a very low background activity. Finally, two XylR dependent promoters next to the long-established maltose inducible promotor were used to construct a high-throughput expression vector system for S. acidocaldarius to efficiently clone and express proteins in S. acidocaldarius.
-
sulfolobus acidocaldarius transports Pentoses via a carbohydrate uptake transporter 2 cut2 type abc transporter and metabolizes them through the aldolase independent weimberg pathway
Applied and Environmental Microbiology, 2017Co-Authors: Michaela Wagner, Nienke Van Der Kolk, Bettina Siebers, Lu Shen, Andreas Albersmeier, Sujin Kim, Jaeho Cha, Christopher Brasen, Jorn Kalinowski, Sonjaverena AlbersAbstract:ABSTRACT Sulfolobus spp. possess a great metabolic versatility and grow heterotrophically on various carbon sources, such as different sugars and peptides. Known sugar transporters in Archaea predominantly belong to ABC transport systems. Although several ABC transporters for sugar uptake have been characterized in the crenarchaeon Sulfolobus solfataricus, only one homologue of these transporters, the maltose/maltooligomer transporter, could be identified in the closely related Sulfolobus acidocaldarius. Comparison of the transcriptome of S. acidocaldarius MW001 grown on peptides alone and peptides in the presence of d-xylose allowed for the identification of the ABC transporter for d-xylose and l-arabinose transport and the gaining of deeper insights into Pentose catabolism under the respective growth conditions. The d-xylose/l-arabinose substrate binding protein (SBP) (Saci_2122) of the ABC transporter is unique in Archaea and shares more similarity to bacterial SBPs of the carbohydrate uptake transporter-2 (CUT2) family than to any characterized archaeal one. The identified Pentose transporter is the first CUT2 family ABC transporter analyzed in the domain of Archaea. Single-gene deletion mutants of the ABC transporter subunits exemplified the importance of the transport system for d-xylose and l-arabinose uptake. Next to the transporter operon, enzymes of the aldolase-independent Pentose catabolism branch were found to be upregulated in N-Z-Amine and d-xylose medium. The α-ketoglutarate semialdehyde dehydrogenase (KGSADH; Saci_1938) seemed not to be essential for growth on Pentoses. However, the deletion mutant of the 2-keto-3-deoxyarabinoate/xylonate dehydratase (KDXD [also known as KDAD]; Saci_1939) was no longer able to catabolize d-xylose or l-arabinose, suggesting the absence of the aldolase-dependent branch in S. acidocaldarius. IMPORTANCE Thermoacidophilic microorganisms are emerging model organisms for biotechnological applications, as their optimal growth conditions resemble conditions used in certain biotechnologies such as industrial plant waste degradation. Because of its high genome stability, Sulfolobus acidocaldarius is especially suited as a platform organism for such applications. For use in (ligno)cellulose degradation, it was important to understand Pentose uptake and metabolism in S. acidocaldarius. This study revealed that only the aldolase-independent Weimberg pathway is required for growth of S. acidocaldarius MW001 on d-xylose and l-arabinose. Moreover, S. acidocaldarius employs a CUT2 ABC transporter for Pentose uptake, which is more similar to bacterial than to archaeal ABC transporters. The identification of Pentose-inducible promoters will expedite the metabolic engineering of S. acidocaldarius for its development into a platform organism for (ligno)cellulose degradation.
Huimin Zhao - One of the best experts on this subject based on the ideXlab platform.
-
discovery and characterization of novel d xylose specific transporters from neurospora crassa and pichia stipitis
Molecular BioSystems, 2010Co-Authors: Jing Du, Sijin Li, Huimin ZhaoAbstract:Saccharomyces cerevisiae is considered one of the most promising organisms for ethanol production from lignocellulosic feedstock. Unfortunately, Pentose sugars, which comprise up to 30% of lignocellulose, cannot be utilized by wild type S. cerevisiae. Heterologous pathways were introduced into S. cerevisiae to enable utilization of D-xylose, the most abundant Pentose sugar. However, the resulting recombinant S. cerevisiae strains exhibited a slow growth rate and poor sugar utilization efficiency when grown on D-xylose as the sole carbon source. D-xylose uptake is the first step of D-xylose utilization. D-xylose can only enter yeast cells through hexose transporters, which have two orders of magnitude lower affinity towards D-xylose compared to hexoses. It was also shown that inefficient Pentose uptake is the limiting step in some D-xylose metabolizing yeast strains. Here we report the cloning and characterization of two novel D-xylose-specific transporters from Neurospora crassa and Pichia stipitis. These two transporters were identified from a total of 18 putative Pentose transporters. They were functionally expressed and properly localized in S. cerevisiae as indicated by HPLC analysis and fluorescence confocal microscopy, respectively. Kinetic parameters of the D-xylose-specific transporters were determined using a 14C-labeled sugar uptake assay. Use of Pentose-specific transporters should improve D-xylose consumption and ethanol production in fast D-xylose assimilating strains, thereby lowering the cost of lignocellulosic ethanol production.