scholarly journals Identification and application of a different glucose uptake system that functions as an alternative to the phosphotransferase system in Corynebacterium glutamicum

2011 ◽  
Vol 90 (4) ◽  
pp. 1443-1451 ◽  
Author(s):  
Masato Ikeda ◽  
Yuta Mizuno ◽  
Shin-ichi Awane ◽  
Masahiro Hayashi ◽  
Satoshi Mitsuhashi ◽  
...  
Keyword(s):  
Glucose Uptake ◽  
Corynebacterium Glutamicum ◽  
Uptake System ◽  
Phosphotransferase System

scholarly journals Evidence for the Presence of an Alternative Glucose Transport System in Clostridium beijerinckii NCIMB 8052 and the Solvent-Hyperproducing Mutant BA101

2005 ◽  
Vol 71 (6) ◽  
pp. 3384-3387 ◽  
Author(s):  
Jieun Lee ◽  
Wilfrid J. Mitchell ◽  
Martin Tangney ◽  
H. P. Blaschek
Keyword(s):  
Glucose Uptake ◽  
Transport System ◽  
Exponential Growth ◽  
Clostridium Beijerinckii ◽  
Uptake System ◽  
Wild Type ◽  
Phosphotransferase System ◽  
Substrate Analogs ◽  
Glucose Transport System ◽  
Energy Inhibitors

ABSTRACT The effects of substrate analogs and energy inhibitors on glucose uptake and phosphorylation by Clostridium beijerinckii provide evidence for the operation of two uptake systems: a previously characterized phosphoenolpyruvate-dependent phosphotransferase system (PTS) and a non-PTS system probably energized by the transmembrane proton gradient. In both wild-type C. beijerinckii NCIMB 8052 and the butanol-hyperproducing mutant BA101, PTS activity declined at the end of exponential growth, while glucokinase activity increased in the later stages of fermentation. The non-PTS uptake system, together with enhanced glucokinase activity, may provide an explanation for the ability of the mutant to utilize glucose more effectively during fermentation despite the fact that it is partially defective in PTS activity.

scholarly journals Phosphotransferase System-Mediated Glucose Uptake Is Repressed in Phosphoglucoisomerase-Deficient Corynebacterium glutamicum Strains

2013 ◽  
Vol 79 (8) ◽  
pp. 2588-2595 ◽  
Author(s):  
Steffen N. Lindner ◽  
Dimitar P. Petrov ◽  
Christian T. Hagmann ◽  
Alexander Henrich ◽  
Reinhard Krämer ◽  
...  
Keyword(s):  
Amino Acid ◽  
Glucose Uptake ◽  
Corynebacterium Glutamicum ◽  
Pentose Phosphate ◽  
Phosphotransferase System ◽  
Negative Effects ◽  
Gene Encoding ◽  
Lysine Production ◽  
Content Type ◽  
Model Strain

ABSTRACTCorynebacterium glutamicumis particularly known for its industrial application in the production of amino acids. Amino acid overproduction comes along with a high NADPH demand, which is covered mainly by the oxidative part of the pentose phosphate pathway (PPP). In previous studies, the complete redirection of the carbon flux toward the PPP by chromosomal inactivation of thepgigene, encoding the phosphoglucoisomerase, has been applied for the improvement ofC. glutamicumamino acid production strains, but this was accompanied by severe negative effects on the growth characteristics. To investigate these effects in a genetically defined background, we deleted thepgigene in the type strainC. glutamicumATCC 13032. The resulting strain,C. glutamicumΔpgi, lacked detectable phosphoglucoisomerase activity and grew poorly with glucose as the sole substrate. Apart from the already reported inhibition of the PPP by NADPH accumulation, we detected a drastic reduction of the phosphotransferase system (PTS)-mediated glucose uptake inC. glutamicumΔpgi. Furthermore, Northern blot analyses revealed that expression ofptsG, which encodes the glucose-specific EII permease of the PTS, was abolished in this mutant. Applying our findings, we optimizedl-lysine production in the model strainC. glutamicumDM1729 by deletion ofpgiand overexpression of plasmid-encodedptsG.l-Lysine yields and productivity withC. glutamicumΔpgi(pBB1-ptsG) were significantly higher than those withC. glutamicumΔpgi(pBB1). These results show thatptsGoverexpression is required to overcome the repressed activity of PTS-mediated glucose uptake inpgi-deficientC. glutamicumstrains, thus enabling efficient as well as fastl-lysine production.

scholarly journals Phosphotransferase System-Independent Glucose Utilization in Corynebacterium glutamicum by Inositol Permeases and Glucokinases

2011 ◽  
Vol 77 (11) ◽  
pp. 3571-3581 ◽  
Author(s):  
Steffen N. Lindner ◽  
Gerd M. Seibold ◽  
Alexander Henrich ◽  
Reinhard Krämer ◽  
Volker F. Wendisch
Keyword(s):  
Glucose Uptake ◽  
Corynebacterium Glutamicum ◽  
Glucose Utilization ◽  
Growth Defect ◽  
Phosphotransferase System ◽  
Glucokinase Gene ◽  
Lysine Production ◽  
Content Type ◽  
Suppressor Mutations ◽  
Glucose Phosphorylation

ABSTRACTPhosphoenolpyruvate-dependent glucose phosphorylation via the phosphotransferase system (PTS) is the major path of glucose uptake inCorynebacterium glutamicum, but some growth from glucose is retained in the absence of the PTS. The growth defect of a deletion mutant lacking the general PTS component HPr in glucose medium could be overcome by suppressor mutations leading to the high expression of inositol utilization genes or by the addition of inositol to the growth medium if a glucokinase is overproduced simultaneously. PTS-independent glucose uptake was shown to require at least one of the inositol transporters IolT1 and IolT2 as a mutant lacking IolT1, IolT2, and the PTS component HPr could not grow with glucose as the sole carbon source. Efficient glucose utilization in the absence of the PTS necessitated the overexpression of a glucokinase gene in addition to eitheriolT1oriolT2. IolT1 and IolT2 are low-affinity glucose permeases withKsvalues of 2.8 and 1.9 mM, respectively. As glucose uptake and phosphorylation via the PTS differs from glucose uptake via IolT1 or IolT2 and phosphorylation via glucokinase by the requirement for phosphoenolpyruvate, the roles of the two pathways forl-lysine production were tested. Thel-lysine yield byC. glutamicumDM1729, a rationally engineeredl-lysine-producing strain, was lower than that by its PTS-deficient derivate DM1729Δhpr, which, however, showed low production rates. The combined overexpression ofiolT1oriolT2withppgK, the gene for PolyP/ATP-dependent glucokinase, in DM1729Δhprenabledl-lysine production as fast as that by the parent strain DM1729 but with 10 to 20% higherl-lysine yield.

scholarly journals Characterization of simultaneous uptake of xylose and glucose in Caldicellulosiruptor kronotskyensis for optimal hydrogen production

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Thitiwut Vongkampang ◽  
Krishnan Sreenivas ◽  
Jonathan Engvall ◽  
Carl Grey ◽  
Ed W. J. van Niel
Keyword(s):  
Glucose Uptake ◽  
Growth Patterns ◽  
Sugar Uptake ◽  
Uptake System ◽  
Sugar Mixtures ◽  
Xylose Uptake ◽  
Growth Profiles ◽  
High Concentrations ◽  
Xylose Transporter

Abstract Background Caldicellulosiruptor kronotskyensis has gained interest for its ability to grow on various lignocellulosic biomass. The aim of this study was to investigate the growth profiles of C. kronotskyensis in the presence of mixtures of glucose–xylose. Recently, we characterized a diauxic-like pattern for C. saccharolyticus on lignocellulosic sugar mixtures. In this study, we aimed to investigate further whether C. kronotskyensis has adapted to uptake glucose in the disaccharide form (cellobiose) rather than the monosaccharide (glucose). Results Interestingly, growth of C. kronotskyensis on glucose and xylose mixtures did not display diauxic-like growth patterns. Closer investigation revealed that, in contrast to C. saccharolyticus, C. kronotskyensis does not possess a second uptake system for glucose. Both C. saccharolyticus and C. kronotskyensis share the characteristics of preferring xylose over glucose. Growth on xylose was twice as fast (μmax = 0.57 h−1) as on glucose (μmax = 0.28 h−1). A study of the sugar uptake was made with different glucose–xylose ratios to find a kinetic relationship between the two sugars for transport into the cell. High concentrations of glucose inhibited xylose uptake and vice versa. The inhibition constants were estimated to be KI,glu = 0.01 cmol L−1 and KI,xyl = 0.001 cmol L−1, hence glucose uptake was more severely inhibited by xylose uptake. Bioinformatics analysis could not exclude that C. kronotskyensis possesses more than one transporter for glucose. As a next step it was investigated whether glucose uptake by C. kronotskyensis improved in the form of cellobiose. Indeed, cellobiose is taken up faster than glucose; nevertheless, the growth rate on each sugar remained similar. Conclusions C. kronotskyensis possesses a xylose transporter that might take up glucose at an inferior rate even in the absence of xylose. Alternatively, glucose can be taken up in the form of cellobiose, but growth performance is still inferior to growth on xylose. Therefore, we propose that the catabolism of C. kronotskyensis has adapted more strongly to pentose rather than hexose, thereby having obtained a specific survival edge in thermophilic lignocellulosic degradation communities.

scholarly journals Metabolic Fluxes in Corynebacterium glutamicum during Lysine Production with Sucrose as Carbon Source

2004 ◽  
Vol 70 (12) ◽  
pp. 7277-7287 ◽  
Author(s):  
Christoph Wittmann ◽  
Patrick Kiefer ◽  
Oskar Zelder
Keyword(s):  
Carbon Source ◽  
Corynebacterium Glutamicum ◽  
Metabolic Flux ◽  
Tca Cycle ◽  
Pentose Phosphate ◽  
Flux Analysis ◽  
Phosphotransferase System ◽  
Lysine Production ◽  
Metabolic Fluxes ◽  
Fructose 1,6 Bisphosphate

ABSTRACT Metabolic fluxes in the central metabolism were determined for lysine-producing Corynebacterium glutamicum ATCC 21526 with sucrose as a carbon source, providing an insight into molasses-based industrial production processes with this organism. For this purpose, 13C metabolic flux analysis with parallel studies on [1-13CFru]sucrose, [1-13CGlc]sucrose, and [13C6 Fru]sucrose was carried out. C. glutamicum directed 27.4% of sucrose toward extracellular lysine. The strain exhibited a relatively high flux of 55.7% (normalized to an uptake flux of hexose units of 100%) through the pentose phosphate pathway (PPP). The glucose monomer of sucrose was completely channeled into the PPP. After transient efflux, the fructose residue was mainly taken up by the fructose-specific phosphotransferase system (PTS) and entered glycolysis at the level of fructose-1,6-bisphosphate. Glucose-6-phosphate isomerase operated in the gluconeogenetic direction from fructose-6-phosphate to glucose-6-phosphate and supplied additional carbon (7.2%) from the fructose part of the substrate toward the PPP. This involved supply of fructose-6-phosphate from the fructose part of sucrose either by PTSMan or by fructose-1,6-bisphosphatase. C. glutamicum further exhibited a high tricarboxylic acid (TCA) cycle flux of 78.2%. Isocitrate dehydrogenase therefore significantly contributed to the total NADPH supply of 190%. The demands for lysine (110%) and anabolism (32%) were lower than the supply, resulting in an apparent NADPH excess. The high TCA cycle flux and the significant secretion of dihydroxyacetone and glycerol display interesting targets to be approached by genetic engineers for optimization of the strain investigated.

scholarly journals Glucose Uptake in Clostridium beijerinckii NCIMB 8052 and the Solvent-Hyperproducing Mutant BA101

2001 ◽  
Vol 67 (11) ◽  
pp. 5025-5031 ◽  
Author(s):  
Jieun Lee ◽  
H. P. Blaschek
Keyword(s):  
Glucose Uptake ◽  
Clostridium Beijerinckii ◽  
Phosphotransferase System ◽  
Stages Of Growth ◽  
Cell Extracts ◽  
Glucose Phosphorylation ◽  
Phosphorylation Rate

ABSTRACT Glucose uptake and accumulation by Clostridium beijerinckii BA101, a butanol hyperproducing mutant, were examined during various stages of growth. Glucose uptake in C. beijerinckii BA101 was repressed 20% by 2-deoxyglucose and 25% by mannose, while glucose uptake in C. beijerinckii8052 was repressed 52 and 28% by these sugars, respectively. We confirmed the presence of a phosphoenolpyruvate (PEP)-dependent phosphotransferase system (PTS) associated with cell extracts ofC. beijerinckii BA101 by glucose phosphorylation by PEP. The PTS activity associated with C. beijerinckii BA101 was 50% of that observed for C. beijerinckii 8052.C. beijerinckii BA101 also demonstrated lower PTS activity for fructose and glucitol. Glucose phosphorylation by cell extracts derived from both C. beijerinckii BA101 and 8052 was also dependent on the presence of ATP, a finding consistent with the presence of glucokinase activity in C. beijerinckii extracts. ATP-dependent glucose phosphorylation was predominant during the solventogenic stage, when PEP-dependent glucose phosphorylation was dramatically repressed. A nearly twofold-greater ATP-dependent phosphorylation rate was observed for solventogenic stage C. beijerinckii BA101 than for solventogenic stage C. beijerinckii 8052. These results suggest that C. beijerinckii BA101 is defective in PTS activity and that C. beijerinckii BA101 compensates for this defect with enhanced glucokinase activity, resulting in an ability to transport and utilize glucose during the solventogenic stage.

scholarly journals Proteomic and Transcriptomic Elucidation of the MutantRalstonia eutrophaG+1 with Regard to Glucose Utilization

2011 ◽  
Vol 77 (6) ◽  
pp. 2058-2070 ◽  
Author(s):  
Matthias Raberg ◽  
Katja Peplinski ◽  
Silvia Heiss ◽  
Armin Ehrenreich ◽  
Birgit Voigt ◽  
...  
Keyword(s):  
Glucose Uptake ◽  
Ralstonia Eutropha ◽  
Stationary Growth Phase ◽  
Insertion Mutation ◽  
Secondary Effects ◽  
Phosphotransferase System ◽  
Stationary Growth ◽  
Increase Glucose Uptake ◽  
Mutant Cells ◽  
Intracellular Glucose

ABSTRACTBy taking advantage of the available genome sequence ofRalstonia eutrophaH16, glucose uptake in the UV-generated glucose-utilizing mutantR. eutrophaG+1 was investigated by transcriptomic and proteomic analyses. Data revealed clear evidence that glucose is transported by a usuallyN-acetylglucosamine-specific phosphotransferase system (PTS)-type transport system, which in this mutant is probably overexpressed due to a derepression of the encodingnagoperon by an identified insertion mutation in gene H16_A0310 (nagR). Furthermore, a missense mutation innagE(membrane component EIICB), which yields a substitution of an alanine by threonine in NagE and may additionally increase glucose uptake, was identified. Phosphorylation of glucose is subsequently mediated by NagF (cytosolic PTS component EIIA-HPr-EI) or glucokinase (GlK), respectively. The inability of the defined deletion mutantR. eutrophaG+1 ΔnagFECto utilize glucose strongly confirms this finding. In addition, secondary effects of glucose, which is now intracellularly available as a carbon source, on the metabolism of the mutant cells in the stationary growth phase occurred: intracellular glucose degradation is stimulated by the stronger expression of enzymes involved in the 2-keto-3-deoxygluconate 6-phosphate (KDPG) pathway and in subsequent reactions yielding pyruvate. The intermediate phosphoenolpyruvate (PEP) in turn supports further glucose uptake by the Nag PTS. Pyruvate is then decarboxylated by the pyruvate dehydrogenase multienzyme complex to acetyl coenzyme A (acetyl-CoA), which is directed to poly(3-hydroxybutyrate). The polyester is then synthesized to a greater extent, as also indicated by the upregulation of various enzymes of poly-β-hydroxybutyrate (PHB) metabolism. The larger amounts of NADPH required for PHB synthesis are delivered by significantly increased quantities of proton-translocating NAD(P) transhydrogenases. The current study successfully combined transcriptomic and proteomic investigations to unravel the phenotype of this hitherto-undefined glucose-utilizing mutant.

Increased glucose utilization and cell growth of Corynebacterium glutamicum by modifying the glucose-specific phosphotransferase system (PTSGlc) genes

2016 ◽  
Vol 62 (12) ◽  
pp. 983-992 ◽  
Author(s):  
Jianzhong Xu ◽  
Junlan Zhang ◽  
Dongdong Liu ◽  
Weiguo Zhang
Keyword(s):  
Growth Rate ◽  
Cell Growth ◽  
Corynebacterium Glutamicum ◽  
Production Rate ◽  
Glucose Consumption ◽  
Chemical Mutagenesis ◽  
Transport Systems ◽  
Phosphotransferase System ◽  
Lysine Production ◽  
Consumption Growth

The phosphoenolpyruvate:glucose phosphotransferase system (PTSGlc) is the major pathway of glucose uptake in Corynebacterium glutamicum. This study investigated glucose consumption rate, cell growth, and metabolite changes resulting from modification of PTSGlc. The classical l-lysine producer C. glutamicum XQ-8 exhibited low glucose consumption, cell growth, and l-lysine production rates, whereas these parameters were significantly increased during cultivating on glucose plus maltose, through inactivation of SugR, or by overexpression of PTSGlc genes. XQ-8sugR::cat/pDXW-8-ptsI exhibited the highest increase in glucose consumption, growth rate, and l-lysine production, followed by XQ-8sugR::cat/pDXW-8-ptsG. However, overexpression of ptsH had little effect on the above-mentioned factors. Although co-overexpression of ptsGHI led to the highest glucose consumption, growth rate, and final l-lysine production; the l-lysine production rate was lower than that of XQ-8sugR::cat/pDXW-8-ptsIH. In fed-batch fermentation, XQ-8sugR::cat/pDXW-8-ptsIH had a higher growth rate of 0.54 h−1 to a dry cell mass of 66 g·L−1 after 16 h, and had a higher l-lysine production rate of 159.2 g·L−1 after 36 h. These results indicate that modification of the sugar transport systems improves amino acid production, especially for mutants obtained by repeated physical and (or) chemical mutagenesis. However, modification of these systems needs to be performed on a case-by-case basis.

Sign in / Sign up

Export Citation Format

Share Document