Advancing small-molecule drug discovery by encoded dual-display technologies.

DNA-encoded chemical library technology (DECL or DEL) has become an important pillar for small-molecule drug discovery. The technology rapidly identifies small-molecule hits for relevant target proteins at low cost and with a high success rate, including ligands for targeted protein degradation (TPD). More recently, the setup of DNA- or peptide nucleic acid (PNA)-encoded chemical libraries based on the simultaneous display of ligand pairs, termed dual-display, allows for more sophisticated applications which will be reviewed herein. Both stable and dynamic dual-display DEL technologies enable innovative affinity-based selection modalities, even on and in cells. Novel methods for a seamless conversion between single- and double-stranded library formats allow for even more versatility. We present the first candidates emerging from dual-display technologies and discuss the future potential of dual-display for drug discovery.

Unbiased and biased chemometric analysis of LC-MS data from human urine following coffee intake.

We carried out a human volunteer study with 14 participants, eight of whom were asked to consume one cup of coffee at four different time points. Urine samples were collected at eight time points and analyzed by HPLC-MS analysis. The LC-MS data were subjected to unsupervised multivariate statistical analysis (principal component analysis) followed by supervised multivariate analysis (linear discriminant analysis). In an unbiased approach, in the absence of data preselection and filtering, the most important features explaining differences between coffee consumers and the control group observed showed variations in endogenous human hormonal steroid metabolites as well as xanthine derivatives. Only after a biased data treatment data revealed differences between the sample groups based on literature reported chlorogenic acid metabolites resulting directly from coffee intake. Such analysis could confirm the presence of 21 previously reported chlorogenic acid plasma metabolites as urinary metabolites. The application of tandem MS molecular networking revealed the presence of five bioavailable chlorogenic acid derivatives in urine previously not reported, including both quinic acid lactone and dimethoxy caffeoyl esters. Selected cinnamic acids were quantified in urine.

Technical note: Realization and uncertainty analysis for an adjustable 3D structured breast phantom in digital breast tomosynthesis.

BACKGROUND: Projection imaging phantoms are often optimized for 2-dimensional image characteristics in homogeneous backgrounds. Therefore, evaluation of image quality in tomosynthesis (DBT) lacks accepted and established phantoms. PURPOSE: We describe a 3D breast phantom with a structured, variable background. The phantom is an adaptable and advanced version of the L1 phantom by Cockmartin et al. Phantom design and its use for quality assurance measurements for DBT devices are described. Four phantoms were compared to assess the objectivity. METHODS: The container size was increased to a diameter of 24 cm and a total height of 53.5 mm. Spiculated masses were replaced by five additional non-spiculated masses for higher granularity in threshold diameter resolution. These patterns are adjustable to the imaging device. The masses were printed in one session with a base layer using two-component 3D printing. New materials compared to the L1 phantom improved the attenuation difference between the lesion models and the background. Four phantoms were built and intra-human observer, inter-human observer and inter-phantom variations were determined. The latter assess the reproducibility of the phantom production. Coefficients of variance (V) were calculated for all three variations. RESULTS: The difference of the attenuation coefficients between the lesion models and the background was 0.20 cm-1 (with W/Al at 32 kV, equivalent to 19-20 keV effective energy) compared to 0.21 cm-1 for 50/50 glandular/adipose breast tissue and cancerous lesions. PMMA equivalent thickness of the phantom was 47.0 mm for the Siemens Mammomat Revelation. For the masses, the V i n t r a $V_{intra}$ for the intra-observer variation was 0.248, the averaged inter-observer variation, V ¯ i n t e r $\overline{V}_{inter}$ was 0.383. V p h a n t o m $V_{phantom}$ for phantom variance was 0.321. For the micro-calcifications, V i n t r a $V_{intra}$ was 0.0429, V ¯ i n t e r = $\overline{V}_{inter}=$ 0.0731 and V p h a n t o m = $V_{phantom}=$ 0.0759. CONCLUSIONS: Position, orientation and shape of the masses are reproducible and attenuation differences appropriate. The phantom presented proved to be a candidate test object for quality control.

Investigating the magnetic and magnetocaloric behaviors of LiSm(PO3)4.

We report a detailed study on the magnetic behaviors and magnetocaloric (MC) effect of a single crystal of lithium samarium tetraphosphate, LiSm(PO3)4. The analyses of temperature-dependent magnetization data have revealed magnetic ordering established with decreasing temperature below T p, where T p is the minimum of a dM/dT vs. T curve and varies as a linear function of the applied field H. The Curie temperature has been extrapolated from T p(H) data, as H → 0, to be about 0.51 K. The establishment of magnetic-ordering causes a substantial change in the heat capacity C p. Above T p, the crystal exhibits paramagnetic behavior. Using the Curie-Weiss (CW) law and Arrott plots, we have found the crystal to have a CW temperature θ CW ≈ -36 K, and short-range magnetic order associated with a coexistence of antiferromagnetic and ferromagnetic interactions ascribed to the couplings of magnetic dipoles and octupoles at the Γ7 and Γ8 states. An assessment of the MC effect has shown increases in value of the absolute magnetic-entropy change (|ΔS m|) and adiabatic-temperature change (ΔT ad) when lowering the temperature to 2 K, and increasing the magnetic-field H magnitude. Around 2 K, the maximum value of |ΔS m| is about 3.6 J kg-1 K-1 for the field H = 50 kOe, and ΔT ad is about 5.8 K for H = 20 kOe, with the relative cooling power (RCP) of ∼82.5 J kg-1. In spite of a low MC effect in comparison to Li(Gd,Tb,Ho)(PO3)4, the absence of magnetic hysteresis reflects that LiSm(PO3)4 is also a candidate for low-temperature MC applications below 25 K.

Opportunities and Challenges to Profile mRNA Modifications in Escherichia†coli.

mRNA methylation is an important regulator of many physiological processes in eukaryotes but has not been studied in depth in prokaryotes. Working with bacterial mRNA is challenging because it lacks a poly(A)-tail. However, methods for detecting RNA modifications, both sequencing and mass spectrometry, rely on efficient preparation of mRNA. Here, we compared size-dependent separation by electrophoresis and rRNA depletion for enrichment of Escherichia coli mRNA. The purification success was monitored by qRT-PCR and RNA sequencing. Neither method allowed complete removal of rRNA. Nevertheless, we were able to quantitatively analyze several modified nucleosides in the different RNA types. We found evidence for stress dependent RNA modification reprofiling in rRNA, but also several modified nucleosides in the mRNA enriched fractions showed significant changes.

LC-MS based metabolomic approach for the efficient identification and relative quantification of bioavailable cocoa phenolics in human urine.

This study was designed to investigate the rate and extent of urinary excretion of cocoa phenolic metabolites after human intake using metabolomics approach. In this context, a feeding trial was conducted where urine samples were collected at different time points over 48-h period. Several biomarkers were highlighted in LC-MS based chemometrics using principal component (PCA) and partial least squares discriminant analysis (PLS-DA), which revealed the presence of both epicatechin and gut microbial phenyl-γ-valerolactones (PVLs) conjugated analogues. The presences of these metabolites segregated and grouped the samples based on cocoa and non-cocoa ingestion. Furthermore, semi quantification of major bioavailable metabolites was performed to determine the interindividual differences and assess the relative bioavailability of cocoa compounds in the human body. Our approach presented here is unique in displaying a combination of LC-MS based chemometrics visualization strategies, which revealed and identified significant biomarkers that could reduce the problems associated with data screening complexity.

On the relevance of modulation transfer function measurements in digital mammography quality control.

Purpose: The relevance of presampling modulation transfer function (MTF) measurements in digital mammography (DM) quality control (QC) is examined. Two studies are presented: a case study on the impact of a reduction in MTF on the technical image quality score and analysis of the robustness of routine QC MTF measurements. Approach: In the first study, two needle computed radiography (CR) plates with identical sensitivities were used with differences in the 50% point of the MTF ( f MTF 0.5 ) larger than the limiting value in the European guidelines ( > 10 % change between successive measurements). Technical image quality was assessed via threshold gold thickness of the CDMAM phantom and threshold microcalcification diameter of the L1 structured phantom. For the second study, presampling MTF results from 595 half-yearly QC tests of 55 DM systems (16 types, six manufacturers) were analyzed for changes from the baseline value and changes in f MTF 0.5 between successive tests. Results: A reduction of 20% in f MTF 0.5 of the two CR plates was observed. There was a tendency to a lower score for task-based metrics, but none were significant. Averaging over 55 systems, the absolute relative change in f MTF 0.5 between consecutive tests (with 95% confidence interval) was 3% (2.5% to 3.4%). Analysis of the maximum relative change from baseline revealed changes of up to - 10 % for one a-Se based system and - 15 % for a group of CsI-based systems. Conclusions: A limit of 10% is a relevant action level for investigation. If exceeded, then the impact on performance has to be verified with extra metrics.

Elongation factor P is required for EIIGlc translation in Corynebacterium glutamicum due to an essential polyproline motif.

Translating ribosomes require elongation factor P (EF-P) to incorporate consecutive prolines (XPPX) into nascent peptide chains. The proteome of Corynebacterium glutamicum ATCC 13032 contains a total of 1,468 XPPX motifs, many of which are found in proteins involved in primary and secondary metabolism. We show here that synthesis of EIIGlc , the glucose-specific permease of the phosphoenolpyruvate (PEP): sugar phosphotransferase system (PTS) encoded by ptsG, is strongly dependent on EF-P, as an efp deletion mutant grows poorly on glucose as sole carbon source. The amount of EIIGlc is strongly reduced in this mutant, which consequently results in a lower rate of glucose uptake. Strikingly, the XPPX motif is essential for the activity of EIIGlc , and substitution of the prolines leads to inactivation of the protein. Finally, translation of GntR2, a transcriptional activator of ptsG, is also dependent on EF-P. However, its reduced amount in the efp mutant can be compensated for by other regulators. These results reveal for the first time a translational bottleneck involving production of the major glucose transporter EIIGlc , which has implications for future strain engineering strategies.

LC-MS/MS based molecular networking approach for the identification of cocoa phenolic metabolites in human urine.

Dietary phenolic compounds are often transformed by gut microbiota prior to absorption. This transformation may modify their structures, producing novel gut flora metabolites associated with numerous health benefits. Traditional mass spectrometry (MS) based approaches for assessing dietary exposure of cocotea (cocoa, coffee and tea) products provided very little information about the modification and fate of dietary phenolics after ingestion, mainly due to limitation of complex sample nature and their data analyses. Mass spectrometry techniques are well-suited to a high-throughput characterization of natural products, however, analyzing MS based data of complex biological matrix is still considered a challenge. In order to overcome such limitations and simplify the analysis of complex MS data, a cocotea based human trial was conducted where MS based molecular networking approach was implemented. To demonstrate the utility of this approach in one of the specific cocotea diets, we have applied it to a diverse collection of human (n = 15) urine samples, who consumed cocoa rich in polyphenols over a 48-h period. This approach illustrated the power of the new strategy, allowing the rapid identification of new analogues of cocoa metabolites after human consumption. Analysis of human urine samples after cocoa consumption revealed (by assignment of unknown metabolites based on the network similarities) that monomeric flavanols are mainly absorbed and transformed directly into their glucuronide and sulfated moieties. Subsequently, the hydroxy and methoxy phenyl-g-velerolactone as well as their smaller metabolites (such as hydroxyphenyl valeric acids, hydroxy and methoxy phenyl propionic acids and their derivates) are indicative of bacterial metabolism of cocoa major flavanols. For the first time, our study exemplifies and highlight the implementation of MS based molecular networking approach in illustrating the tracking of various structural motifs of ingested cocoa phenolics in human based study.

NAIL-MS reveals the repair of 2-methylthiocytidine by AlkB in E. coli.

RNAs contain post-transcriptional modifications, which fulfill a variety of functions in translation, secondary structure stabilization and cellular stress survival. Here, 2-methylthiocytidine (ms2C) is identified in tRNA of E. coli and P. aeruginosa using NAIL-MS (nucleic acid isotope labeling coupled mass spectrometry) in combination with genetic screening experiments. ms2C is only found in 2-thiocytidine (s2C) containing tRNAs, namely tRNAArgCCG, tRNAArgICG, tRNAArgUCU and tRNASerGCU at low abundances. ms2C is not formed by commonly known tRNA methyltransferases. Instead, we observe its formation in vitro and in vivo during exposure to methylating agents. More than half of the s2C containing tRNA can be methylated to carry ms2C. With a pulse-chase NAIL-MS experiment, the repair mechanism by AlkB dependent sulfur demethylation is demonstrated in vivo. Overall, we describe ms2C as a bacterial tRNA modification and damage product. Its repair by AlkB and other pathways is demonstrated in vivo by our powerful NAIL-MS approach.

Model and human observer reproducibility for detection of microcalcification clusters in digital breast tomosynthesis images of three-dimensionally structured test object.

We compare the reproducibility of the human observers and a channelized Hotelling observer (CHO), when reading digital breast tomosynthesis (DBT) images of a physical phantom containing a breast simulating structured background and calcification clusters at three dose levels. The phantom is scanned 217 times on a Siemens Inspiration DBT system. Volumes of interest, with and without the calcification targets, are extracted and the human observers' percentage of correct (PC) scores is evaluated using a four-alternative forced choice method. A two-layer CHO is developed using the human observer results. The first layer consists of a localizing CHO that identifies the most conspicuous calcifications using two Laguerre-Gauss channels. Then a CHO with eight Gabor channels estimates the PC score for the calcification cluster. Observer reproducibility is estimated by bootstrapping, and the standard deviation (SD) is used as a figure of merit. The CHO closely approximated the human observer results for all the three dose levels with a correlation of > 0.97 . For the larger calcification cluster sizes, both observers have similar reproducibility, whereas the CHO is more reproducible for the smaller calcifications, with a maximum of 5.5 SD against 13.1 SD for the human observers. The developed CHO is a good candidate for automated reading of the calcification clusters of the structured phantom, with better reproducibility than the human readers for small calcifications.

Systematic approach to a channelized Hotelling model observer implementation for a physical phantom containing mass-like lesions: Application to digital breast tomosynthesis.

PURPOSE: to develop a channelized model observer (CHO) that matches human reader (HR) scoring of a physical phantom containing breast simulating structure and mass lesion-like targets for use in quality control of digital breast tomosynthesis (DBT) imaging systems. METHODS: A total of 108 DBT scans of the phantom was acquired using a Siemens Inspiration DBT system. The detectability of mass-like targets was evaluated by human readers using a 4-alternative forced choice (4-AFC) method. The percentage correct (PC) values were then used as the benchmark for CHO tuning, again using a 4-AFC method. Three different channel functions were considered: Gabor, Laguerre-Gauss and Difference of Gaussian. With regard to the observer template, various methods for generating the expected signal were studied along with the influence of the number of training images used to form the covariance matrix for the observer template. Impact of bias in the training process on the observer template was evaluated next, as well as HR and CHO reproducibility. RESULTS: HR performance was most closely matched by 8 Gabor channels with tuned phase, orientation and frequency, using an observer template generated from training image data. Just 24 DBT image stacks were required to give robust CHO performance with 0% bias, although a bias of up to 33% in the training images also gave acceptable performance. CHO and HR reproducibility were similar (on average 3.2 PC versus 3.4 PC). CONCLUSIONS: The CHO algorithm developed matches human reader performance and is therefore a promising candidate for automated readout of phantom studies.

Inter-laboratory comparison of channelized hotelling observer computation.

PURPOSE: The task-based assessment of image quality using model observers is increasingly used for the assessment of different imaging modalities. However, the performance computation of model observers needs standardization as well as a well-established trust in its implementation methodology and uncertainty estimation. The purpose of this work was to determine the degree of equivalence of the channelized Hotelling observer performance and uncertainty estimation using an intercomparison exercise. MATERIALS AND METHODS: Image samples to estimate model observer performance for detection tasks were generated from two-dimensional CT image slices of a uniform water phantom. A common set of images was sent to participating laboratories to perform and document the following tasks: (a) estimate the detectability index of a well-defined CHO and its uncertainty in three conditions involving different sized targets all at the same dose, and (b) apply this CHO to an image set where ground truth was unknown to participants (lower image dose). In addition, and on an optional basis, we asked the participating laboratories to (c) estimate the performance of real human observers from a psychophysical experiment of their choice. Each of the 13 participating laboratories was confidentially assigned a participant number and image sets could be downloaded through a secure server. Results were distributed with each participant recognizable by its number and then each laboratory was able to modify their results with justification as model observer calculation are not yet a routine and potentially error prone. RESULTS: Detectability index increased with signal size for all participants and was very consistent for 6 mm sized target while showing higher variability for 8 and 10 mm sized target. There was one order of magnitude between the lowest and the largest uncertainty estimation. CONCLUSIONS: This intercomparison helped define the state of the art of model observer performance computation and with thirteen participants, reflects openness and trust within the medical imaging community. The performance of a CHO with explicitly defined channels and a relatively large number of test images was consistently estimated by all participants. In contrast, the paper demonstrates that there is no agreement on estimating the variance of detectability in the training and testing setting.

Environment of the Eu3+ Ion within Nanocrystalline Eu-Doped BaAl2O4: Correlation of X-ray Diffraction, Mössbauer Spectroscopy, X-ray Absorption Spectroscopy, and Photoluminescence Investigations.

Powder samples of pure BaAl2O4 and doped with 4.9 atom % Eu in relation to Ba were prepared by a hydrothermal route. The samples were characterized by X-ray diffraction, 151Eu Mössbauer spectroscopy, synchrotron-based X-ray absorption spectroscopy at the Ba L3- and Eu L3-edges, and photoluminescence measurements. Diffraction lines were broadened, indicating that the samples were nanocrystallline. The samples possessed a hexagonal crystal structure, space group P63. 151Eu Mössbauer spectroscopy revealed the presence of Eu in the 3+ oxidation state. The same information on the Eu oxidation state was also obtained by the Eu L3-edge X-ray absorption near-edge structure of the doped sample. Extended X-ray absorption fine structure showed an Eu3+ ion substituted for Ba2+ on the Ba2 site in the BaAl2O4 host structure, with charge compensation by an interstitial O in the vicinity of the Ba2 site. That was confirmed by a Rietveld structure refinement for the Eu-doped BaAl2O4 sample. Analysis of the diffraction line broadening for the prepared samples was performed simultaneously with the structure refinement. Both the dopant Eu3+ and the interstitial O acted as defects in the host BaAl2O4 lattice, which increased the lattice strain from 0.02% for pure BaAl2O4 to 0.17% for the Eu-doped sample. Crystallite sizes in the samples increased with Eu doping from 32 nm for pure BaAl2O4 to 36 nm for Eu-doped BaAl2O4. This could likely be related to the increase in the diffusion rate of the cations in the sample when a part of the Ba2+ cation content was exchanged with smaller Eu3+ cations. The Eu-doped BaAl2O4 sample exhibited red photoluminescence under excitation with λexc = 308 nm. The observed emission spectrum indicated that Eu3+ ions occupied the Ba site with lower symmetry in the doped sample.

Lattice Enthalpies of Lanthanide Orthoferrites LnFeO3.

Lattice enthalpies Δ(L)H(θ) of lanthanide orthoferrites, LnFeO(3) have been determined by the Born-Haber cycle and compared with those calculated by an empirical equation. Enthalpies of formation of LnFeO(3) from two different sources have been employed: from oxides (Ln(2)O(3), Fe(2)O(3)), for 12 LnFeO(3), and from elements, for 8 LnFeO(3), but the differences in Δ(L)H(θ) are very small. The Born-Haber cycle in both routes results in close values of Δ(L)H(θ) to those obtained by the empirical equation of Glasser and Jenkins. A correspondence in dimension and magnitude has been found between the partial derivative of the lattice enthalpies to the molar volumes and an upper limit of the shear moduli of the lanthanide orthoferrites.

Transcription of malP is subject to phosphotransferase system-dependent regulation in Corynebacterium glutamicum.

The Gram-positive Corynebacterium glutamicum co-metabolizes most carbon sources such as the phosphotransferase system (PTS) sugar glucose and the non-PTS sugar maltose. Maltose is taken up via the ABC-transporter MusEFGK2I, and is further metabolized to glucose phosphate by amylomaltase MalQ, maltodextrin phosphorylase MalP, glucokinase Glk and phosophoglucomutase Pgm. Surprisingly, growth of C. glutamicum strains lacking the general PTS components EI or HPr was strongly impaired on the non-PTS sugar maltose. Complementation experiments showed that a functional PTS phosphorelay is required for optimal growth of C. glutamicum on maltose, implying its involvement in the control of maltose metabolism and/or uptake. To identify the target of this PTS-dependent control, transport measurements with 14C-labelled maltose, Northern blot analyses and enzyme assays were performed. The activities of the maltose transporter and enzymes MalQ, Pgm and GlK were not decreased in PTS-deficient C. glutamicum strains, which was corroborated by comparable transcript amounts of musE, musK and musG, as well as of malQ, in C. glutamicum ΔptsH and WT. By contrast, MalP activity was significantly reduced and only residual amounts of malP transcripts were detected in C. glutamicum ΔptsH when compared to WT. Promoter activity assays with the malP promoter in C. glutamicum ΔptsH and WT confirmed that malP transcription is reduced in the PTS-deficient strain. Taken together, we show here for what is to the best of our knowledge the first time a regulatory function of the PTS in C. glutamicum and identify malP transcription as its target.

Nephelauxetic effect and 〈r(k)〉4f radial integrals of Tm³âº in crystals.

Bonding and covalency parameters have been evaluated from the nephelauxetic ratios ßk=Fk (crystal)/Fk (free ion), with k=2, 4, 6, for 24 halide and chalcogenide crystals containing Tm(3+) ions. The radial expectation values for 4f electrons 〈r(k)〉4f of Tm(3+) ion in certain complex oxides, fluorides, and a sulfide have been determined by means of experimental Slater parameter shifts ΔFk relative to the Fk values for the free ion Tm IV. The 〈r(k)〉1f values derived in the dielectric screening model have been compared with those computed by different types of 4f wave functions as well as with other estimates.

Lattice enthalpies, polarizabilities and shear moduli of lanthanide orthophosphates LnPO4.

Lattice energies ΔLHθ of lanthanide orthophosphates, LnPO4 (Ln=Ce-Lu, excluding Pm) have been determined from the Born-Haber cycle and compared with those calculated by other methods. The Born-Haber cycle results in close values of ΔLHθ to those obtained after an empirical equation proposed by Glasser and Jenkins. It has been found that: (i) the partial derivative of the lattice enthalpies to the molar volumes corresponds by dimension and magnitude to the shear moduli of these crystals; (ii) these moduli differ for the monazite- and xenotime-type structures of LnPO4. Molar polarizabilities have been calculated for three LnPO4 with monazite structure, Ln=Ce, Nd, Sm, and for three LnPO4 with xenotime structure, Ln=Tb, Dy, Yb.

Preparation, characterization and catalytic activity of NiOx and NiOx/ZrO2 for oxidation of phenol in aqueous solution.

In the present study bulk NiO(x) and supported NiO(x)/ZrO(2) catalysts have been prepared. The bulk NiO(x) was synthesized using the precipitation-oxidation method with reverse order of precipitation while deposition-precipitation technique was used for the preparation of zirconia supported catalyst. The as-prepared samples were characterized by means of XRD, HRTEM, SAED, IR-spectroscopy, and chemical analyses. It was found that under the applied synthesis procedure nanosized and highly dispersed oxide materials with high active oxygen content were obtained. The catalytic activity and selectivity of these oxide catalysts have been studied in reaction of low-temperature oxidation of phenol in aqueous phase. The effects of several parameters such as catalyst amount, temperature and solution pH on the degradation efficiency of the process were also investigated. Experimental results demonstrated that phenol could be completely degraded under all reaction conditions, except at pH = 12.

Phosphotransferase system-mediated glucose uptake is repressed in phosphoglucoisomerase-deficient Corynebacterium glutamicum strains.

Corynebacterium glutamicum is 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 the pgi gene, encoding the phosphoglucoisomerase, has been applied for the improvement of C. glutamicum amino 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 the pgi gene in the type strain C. glutamicum ATCC 13032. The resulting strain, C. glutamicum Δpgi, 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 in C. glutamicum Δpgi. Furthermore, Northern blot analyses revealed that expression of ptsG, which encodes the glucose-specific EII permease of the PTS, was abolished in this mutant. Applying our findings, we optimized l-lysine production in the model strain C. glutamicum DM1729 by deletion of pgi and overexpression of plasmid-encoded ptsG. l-Lysine yields and productivity with C. glutamicum Δpgi(pBB1-ptsG) were significantly higher than those with C. glutamicum Δpgi(pBB1). These results show that ptsG overexpression is required to overcome the repressed activity of PTS-mediated glucose uptake in pgi-deficient C. glutamicum strains, thus enabling efficient as well as fast l-lysine production.