The enzymology of the bacterial phosphoenolpyruvate-dependent sugar transport systems

1982 ◽  
Vol 46 (1) ◽  
Author(s):  
G.T. Robillard
Keyword(s):  
Sugar Transport ◽  
Transport Systems

scholarly journals The Glucuronic Acid Utilization Gene Cluster from Bacillus stearothermophilus T-6

1999 ◽  
Vol 181 (12) ◽  
pp. 3695-3704 ◽  
Author(s):  
Smadar Shulami ◽  
Orit Gat ◽  
Abraham L. Sonenshein ◽  
Yuval Shoham
Keyword(s):  
Gene Cluster ◽  
Transport System ◽  
Bacillus Stearothermophilus ◽  
Glucuronic Acid ◽  
Genomic Library ◽  
Regulatory Gene ◽  
Sugar Transport ◽  
Transcriptional Unit ◽  
Transport Systems ◽  
Potential Operator

ABSTRACT A λ-EMBL3 genomic library of Bacillus stearothermophilus T-6 was screened for hemicellulolytic activities, and five independent clones exhibiting β-xylosidase activity were isolated. The clones overlap each other and together represent a 23.5-kb chromosomal segment. The segment contains a cluster of xylan utilization genes, which are organized in at least three transcriptional units. These include the gene for the extracellular xylanase, xylanase T-6; part of an operon coding for an intracellular xylanase and a β-xylosidase; and a putative 15.5-kb-long transcriptional unit, consisting of 12 genes involved in the utilization of α-d-glucuronic acid (GlcUA). The first four genes in the potential GlcUA operon (orf1, -2, -3, and -4) code for a putative sugar transport system with characteristic components of the binding-protein-dependent transport systems. The most likely natural substrate for this transport system is aldotetraouronic acid [2-O-α-(4-O-methyl-α-d-glucuronosyl)-xylotriose] (MeGlcUAXyl3). The following two genes code for an intracellular α-glucuronidase (aguA) and a β-xylosidase (xynB). Five more genes (kdgK,kdgA, uxaC, uxuA, anduxuB) encode proteins that are homologous to enzymes involved in galacturonate and glucuronate catabolism. The gene cluster also includes a potential regulatory gene, uxuR, the product of which resembles repressors of the GntR family. The apparent transcriptional start point of the cluster was determined by primer extension analysis and is located 349 bp from the initial ATG codon. The potential operator site is a perfect 12-bp inverted repeat located downstream from the promoter between nucleotides +170 and +181. Gel retardation assays indicated that UxuR binds specifically to this sequence and that this binding is efficiently prevented in vitro by MeGlcUAXyl3, the most likely molecular inducer.

scholarly journals Utilization of Monosaccharides by Hungateiclostridium thermocellum ATCC 27405 Through Adaptive Evolution

2020 ◽  
Author(s):  
Dung Minh Ha-Tran ◽  
Trinh Thi My Nguyen ◽  
Shou-Chen Lo ◽  
Chieh-Chen Huang
Keyword(s):  
Sugar Transport ◽  
Central Carbon Metabolism ◽  
Transport Systems ◽  
Crystalline Cellulose ◽  
Rna Seq ◽  
Genome Resequencing ◽  
Adaptive Laboratory Evolution ◽  
Poor Growth ◽  
Genomic Screening ◽  
Central Carbon

Hungateiclostridium thermocellum ATCC 27405 is a promising bacterium with a robust ability to degrade lignocellulosic biomass complexes, including crystalline cellulose components, through a multienzyme cellulosomal system. In contrast, it exhibits poor growth on simple monosaccharides such as fructose and glucose. This phenomenon raises many important questions concerning its glycolytic pathways and sugar transport systems. Until now, the detailed mechanisms of H. thermocellum adaptation to growth on monosaccharides have been poorly explored. In this study, adaptive laboratory evolution was applied to train the bacterium on monosaccharides, and genome resequencing was used to detect the genes that had mutated during adaptation. RNA-seq data of the 1st-generation culture growing on either fructose or glucose revealed that several glycolytic genes in the EMP pathway were expressed at lower levels in these cells than in cellobiose-grown cells. After 8 generations of culture on fructose and glucose, the evolved H. thermocellum strains grew faster and yielded greater biomass than the nonadapted strains. Genomic screening also revealed several mutation events in the genomes of the evolved strains, especially in genes responsible for sugar transport and central carbon metabolism. Consequently, these genes could be applied as targets for further metabolic engineering to improve this bacterium for bioindustrial usage.

Regulation of sugar transport systems of Kluyveromyces marxianus: the role of carbohydrates and their catabolism

1988 ◽  
Vol 939 (3) ◽  
pp. 569-576 ◽  
Author(s):  
Adriaan W. De Bruijne ◽  
Johanna Schuddemat ◽  
Peter J.A. Van den Broek ◽  
Johnny Van Steveninck
Keyword(s):  
Kluyveromyces Marxianus ◽  
Sugar Transport ◽  
Transport Systems

scholarly journals Neutral-sugar transport by rat liver lysosomes

1990 ◽  
Vol 272 (2) ◽  
pp. 323-326 ◽  
Author(s):  
A J Jonas ◽  
P Conrad ◽  
H Jobe
Keyword(s):  
Rat Liver ◽  
Cytochalasin B ◽  
Sugar Transport ◽  
Transport Systems ◽  
Neutral Sugar ◽  
Temperature Dependent ◽  
Acetylneuraminic Acid ◽  
Liver Lysosomes ◽  
Acidic Sugars ◽  
Rat Liver Lysosomes

Transport of D-glucose was studied in Percoll-gradient-purified rat liver lysosomes. D-Glucose uptake had a Km of 22 mM and a t1/2 of approx. 30 s. D-Fucose, 2-deoxyglucose and methyl alpha-glucoside were the most effective competitors for uptake of D-glucose, although D-galactose, D-mannose, D-xylose and L-fucose also appeared to compete for uptake. L-Glucose was a poor competitor for uptake. No competition was observed with N-acetyl-D-glucosamine, N-acetyl-D-galactosamine, D-glucuronic acid, N-acetylneuraminic acid, D-glucosamine or the amino acids L-glycine, L-lysine and L-proline. Uptake was unaffected by N-ethylmaleimide, dithiothreitol, KCl, NaCl, ATP/Mg or alteration of buffer pH. D-Glucose efflux from lysosomes was temperature-dependent, with a Q10 of 2.3, and was inhibited by cytochalasin B. Counter-transport could not be demonstrated. In contrast, L-fucose uptake had a Km of 65 mM and was largely unaffected by 5 M excess of neutral D-sugars. Both uptake and efflux of L-fucose were inhibited by cytochalasin B. It appears that lysosomes possess a facilitated transport system for D-glucose and perhaps other neutral D-sugars that is discrete from transport systems for acetylated and acidic sugars.

Effects of phloretin and theophylline on 3-O-methylglucose transport by intestinal epithelial cells

1978 ◽  
Vol 234 (3) ◽  
pp. C64-C72 ◽  
Author(s):  
J. Randles ◽  
G. A. Kimmich
Keyword(s):  
Epithelial Cells ◽  
Transport System ◽  
Intestinal Epithelial Cells ◽  
Sugar Transport ◽  
Inhibitory Effect ◽  
Metabolic Effects ◽  
Transport Systems ◽  
Facilitated Diffusion ◽  
Intestinal Epithelial ◽  
Gradient Enhancement

Phloretin and theophylline each exert an immediate inhibitory effect on the Na+-independent, facilitated-diffusion transport system for sugar associated with intestinal epithelial cells. Phloretin inhibits approximately 50% more of the total Na+-independent sugar flux than theophylline. Neither agent has an immediate effect on the Na+-dependent, concentrative sugar transport system, although preincubation of the cells with phloretin causes a significant inhibition. The slowly developing effect is correlated with a decrease in cellular adenosine triphosphate (ATP) and an elevation of intracellular Na+. Other agents which elevate cell Na+ also inhibit Na+-dependent sugar influx, even if ATP levels are not depleted. On the other hand, if ATP is depleted by phloretin under conditions in which the cells do not gain Na+, the inhibitory effect on Na+-dependent sugar flux tends to disappear. The slow-onset phloretin effects are due to transinhibition of the Na+-dependent sugar carrier by cellular Na+. When the passive sugar carrier is inhibited by phloretin or theophylline, the concentrative system can establish an enhanced sugar gradient. Because of the secondary metabolic effects of phloretin, theophylline induces a greater gradient enhancement despite its more limited effect on the passive sugar-transport system. Sugar gradients as large as 20-fold are induced by theophylline, in contrast to 12-fold gradients observed in the presence of phloretin and approximately 7- to 8-fold for untreated cells. These results are discussed in terms of conceptual questions regarding the energetics of Na+-dependent transport systems.

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