Genetic Engineering of the Phosphocarrier Protein NPr of the Escherichia coli Phosphotransferase System Selectively Improves Sugar Uptake Activity

Santos, Yossef Lopez-de los; Chan, Henry Lik Yuen; Cantu, Vito A.; Rettner, Rachael; Sánchez, F.; Zhang, Zhongge; Saier, Milton H.; Soberón, Xavier

doi:10.1074/jbc.m112.345660

Cited by 4 publications

(4 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The fruB gene encodes a component of the fructose PTS, and contains an EIIA domain and a HPr‐like domain called FPr. It was also shown in E. coli and Salmonella typhimurium that FPr can substitute for HPr in ptsH mutants, and that the double mutant of HPr and FPr was not able to utilize any PTS sugars …”

Section: Resultsmentioning

confidence: 99%

“…It was also shown in E. coli and Salmonella typhimurium that FPr can substitute for HPr in ptsH mutants, [34] and that the double mutant of HPr and FPr was not able to utilize any PTS sugars. [35,36] To test the role of FruB, we deleted fruB from unevolved JK30. The resulting strain JK31 did not grow well in an M9 minimal medium containing glucose as a sole carbon source, and, in contrast to the evolved JK30E isolated containing fruB mutations, exhibited preferential utilization of xylose over glucose when grown in a M9 minimal medium containing minimal media agar plates supplemented with glucose and incubated at 37°C and large colonies were isolated after two days.…”

Section: Adaptive Laboratory Evolution Of Pts Mutants Andmentioning

confidence: 99%

See 1 more Smart Citation

Systems Metabolic Engineering of Escherichia coli Improves Coconversion of Lignocellulose‐Derived Sugars

Kim

Tremaine

Grass

et al. 2019

Biotechnology Journal

View full text Add to dashboard Cite

Currently, microbial conversion of lignocellulose‐derived glucose and xylose to biofuels is hindered by the fact that most microbes (including Escherichia coli [E. coli], Saccharomyces cerevisiae, and Zymomonas mobilis) preferentially consume glucose first and consume xylose slowly after glucose is depleted in lignocellulosic hydrolysates. In this study, E. coli strains are developed that simultaneously utilize glucose and xylose in lignocellulosic biomass hydrolysate using genome‐scale models and adaptive laboratory evolution. E. coli strains are designed and constructed that coutilize glucose and xylose and adaptively evolve them to improve glucose and xylose utilization. Whole‐genome resequencing of the evolved strains find relevant mutations in metabolic and regulatory genes and the mutations’ involvement in sugar coutilization is investigated. The developed strains show significantly improved coconversion of sugars in lignocellulosic biomass hydrolysates and provide a promising platform for producing next‐generation biofuels.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Adaptive Laboratory Evolution Of Pts Mutants Andmentioning

confidence: 99%

Systems Metabolic Engineering of Escherichia coli Improves Coconversion of Lignocellulose‐Derived Sugars

Kim

Tremaine

Grass

et al. 2019

Biotechnology Journal

View full text Add to dashboard Cite

show abstract

“…Hence, there is little or no crossphosphorylation between the protein pairs EI Ntr /NPr and EI/HPr [Rabus et al, 1999]. On the other hand, EIIA Ntr was shown to readily exchange phosphoryl groups with HPr in vitro and transfer of phosphoryl groups was also observed between NPr and EIIA of the transport PTS [Lopez-de los Santos et al, 2012;Powell et al, 1995]. Previous in vivo protein phosphorylation analysis using metabolic 32 P-labeling confirmed that transfer of phosphoryl groups from the transport-PTS to PTS Ntr is possible when cells grow on non-PTS carbohydrates [Zimmer et al, 2008].…”

Section: Discussionmentioning

confidence: 99%

Cross-Talk between the Canonical and the Nitrogen-Related Phosphotransferase Systems Modulates Synthesis of the KdpFABC Potassium Transporter in <b><i>Escherichia coli</i></b>

2015

View full text Add to dashboard Cite

Many Proteobacteria possess the regulatory nitrogen-related phosphotransferase system (PTS^Ntr), which operates in parallel to the transport PTS. PTS^Ntr is composed of the proteins EI^Ntr and NPr and the final phosphate acceptor EIIA^Ntr. Both PTSs can exchange phosphoryl groups among each other. Proteins governing K⁺ uptake represent a major target of PTS^Ntr in Escherichia coli. Nonphosphorylated EIIA^Ntr binds and stimulates the K⁺ sensor KdpD, which activates expression of the kdpFABC operon encoding a K⁺ transporter. Here we show that this regulation also operates in an ilvG⁺ strain ruling out previous concern about interference with a nonfunctional ilvG allele present in many strains. Furthermore, we analyzed phosphorylation of EIIA^Ntr. In wild-type cells EIIA^Ntr is predominantly phosphorylated, regardless of the growth stage and the utilized carbon source. However, cross-phosphorylation of EIIA^Ntr by the transport PTS becomes apparent in the absence of EI^Ntr: EIIA^Ntr is predominantly nonphosphorylated when cells grow on a PTS sugar and phosphorylated when a non-PTS carbohydrate is utilized. These differences in phosphorylation are transduced into corresponding kdpFABC transcription levels. Thus, the transport PTS may affect phosphorylation of EIIA^Ntr and accordingly modulate processes controlled by EIIA^Ntr. Our data suggest that this cross-talk becomes most relevant under conditions that would inhibit activity of EI^Ntr.

show abstract

“…Therefore, it is an ideal model system to test the proposal on the potential dual role of HPr as both HPr and NPr. A recent study showed that CPr14, a genetically engineered NPr, was proven to improve sugar uptake activity in E. coli . In this study, the three‐dimensional structure of IIA Ntr from B. pseudomallei ( Bp IIA Ntr ) has been determined.…”

Section: Introductionmentioning

confidence: 99%

New molecular interaction of IIA^Ntr and HPr from Burkholderia pseudomallei identified by X‐ray crystallography and docking studies

et al. 2013

View full text Add to dashboard Cite

The nitrogen-related phosphoenolpyruvate phosphotransferase system (PTS(Ntr) ) is involved in controlling ammonia assimilation and nitrogen fixation. The additional role of PTS(Ntr) as a regulatory link between nitrogen and carbon utilization in Escherichia coli is assumed to be closely related to molecular functions of IIA(Ntr) in potassium homeostasis. We have determined the crystal structure of IIA(Ntr) from Burkholderia pseudomallei (BpIIA(Ntr) ), which is a causative agent of melioidosis. The crystal structure of dimeric BpIIA(Ntr) determined at 3.0 Å revealed that its active sites are mutually blocked. This dimeric state is stabilized by charge and weak hydrophobic interactions. Overall monomeric structure and the active site residues, Arg51 and His67, of BpIIA(Ntr) are well conserved with those of IIA(Ntr) enzymes from E. coli and Neisseria meningitides. Interestingly, His113 of BpIIA(Ntr) , which corresponds to a key residue in another phosphoryl group relay in the mannitol-specific enzyme EIIA family (EIIA(Mtl) ), is located away from the active site due to the loop connecting β5 and α3. Combined with other differences in molecular surface properties, these structural signatures distinguish the IIA(Ntr) family from the EIIA(Mtl) family. Since, there is no gene for NPr in the chromosome of B. pseudomallei, modeling and docking studies of the BpIIA(Ntr) -BpHPr complex has been performed to support the proposal on the NPr-like activity of BpHPr. A potential dual role of BpHPr as a nonspecific phosphocarrier protein interacting with both sugar EIIAs and IIA(Ntr) in B. pseudomallei has been discussed.

show abstract

Genetic Engineering of the Phosphocarrier Protein NPr of the Escherichia coli Phosphotransferase System Selectively Improves Sugar Uptake Activity

Cited by 4 publications

References 44 publications

Systems Metabolic Engineering of Escherichia coli Improves Coconversion of Lignocellulose‐Derived Sugars

Systems Metabolic Engineering of Escherichia coli Improves Coconversion of Lignocellulose‐Derived Sugars

Cross-Talk between the Canonical and the Nitrogen-Related Phosphotransferase Systems Modulates Synthesis of the KdpFABC Potassium Transporter in <b><i>Escherichia coli</i></b>

New molecular interaction of IIA^Ntr and HPr from Burkholderia pseudomallei identified by X‐ray crystallography and docking studies

Contact Info

Product

Resources

About

Genetic Engineering of the Phosphocarrier Protein NPr of the Escherichia coli Phosphotransferase System Selectively Improves Sugar Uptake Activity

Cited by 4 publications

References 44 publications

Systems Metabolic Engineering of Escherichia coli Improves Coconversion of Lignocellulose‐Derived Sugars

Systems Metabolic Engineering of Escherichia coli Improves Coconversion of Lignocellulose‐Derived Sugars

Cross-Talk between the Canonical and the Nitrogen-Related Phosphotransferase Systems Modulates Synthesis of the KdpFABC Potassium Transporter in <b><i>Escherichia coli</i></b>

New molecular interaction of IIANtr and HPr from Burkholderia pseudomallei identified by X‐ray crystallography and docking studies

Contact Info

Product

Resources

About

New molecular interaction of IIA^Ntr and HPr from Burkholderia pseudomallei identified by X‐ray crystallography and docking studies