Glutathione Biosynthesis in Bacteria by Bifunctional GshF Is Driven by a Modular Structure Featuring a Novel Hybrid ATP-Grasp Fold

Stout, Jan; Vos, Dirk De; Vergauwen, Bjorn; Savvides, Savvas N.

doi:10.1016/j.jmb.2011.12.046

Cited by 22 publications

(20 citation statements)

References 32 publications

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“…GshF is commonly found among mammalian pathogens (44) and could have a role in mitigating DNA damage caused by general oxidative stress (43). Mutations to yybT and gshF share the common distinction of being responsive to stress-induced damage, and earlier studies have clearly shown that insertion of the DAP-Ca 2ϩ complex into the membrane leads to a loss of ions (such as K ϩ ) and functional integrity, leading to a significant increase in oxidative stress on the cell (45).…”

Section: I440smentioning

confidence: 99%

Adaptation of Enterococcus faecalis to Daptomycin Reveals an Ordered Progression to Resistance

Miller

Kong

Tran³

et al. 2013

Antimicrob Agents Chemother

106

189

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e With increasing numbers of hospital-acquired antibiotic resistant infections each year and staggering health care costs, there is a clear need for new antimicrobial agents, as well as novel strategies to extend their clinical efficacy. While genomic studies have provided a wealth of information about the alleles associated with adaptation to antibiotics, they do not provide essential information about the relative importance of genomic changes, their order of appearance, or potential epistatic relationships between adaptive changes. Here we used quantitative experimental evolution of a single polymorphic population in continuous culture with whole-genome sequencing and allelic frequency measurements to study daptomycin (DAP) resistance in the vancomycinresistant clinical pathogen Enterococcus faecalis S613. Importantly, we sustained both planktonic and nonplanktonic (i.e., biofilm) populations in coculture as the concentration of antibiotic was raised, facilitating the development of more ecological complexity than is typically observed in laboratory evolution. Quantitative experimental evolution revealed a clear order and hierarchy of genetic changes leading to resistance, the signaling and metabolic pathways responsible, and the relative importance of these mutations to the evolution of DAP resistance. Despite the relative simplicity of this ex vivo approach compared to the ecological complexity of the human body, we showed that experimental evolution allows for rapid identification of clinically relevant adaptive molecular pathways and new targets for drug design in pathogens.

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Section: I440smentioning

confidence: 99%

Adaptation of Enterococcus faecalis to Daptomycin Reveals an Ordered Progression to Resistance

Miller

Kong

Tran³

et al. 2013

Antimicrob Agents Chemother

106

189

View full text Add to dashboard Cite

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“…However, the high local concentration of enzymes, when encapsulated, will drive oligomerization of the enzyme cargo which might be advantageous for multiunit enzymes to enhance enzymatic activities compared to bulk solution, as was reported for Escherichia coli hydrogenase 1. 19 Although the two sequential reactions of the GSH biosynthetic pathway can be catalyzed by a single protomer of GshF, intersubunit communication is suggested to influence the function in the dimer of GshF, 50 which is expected to be enhanced after encapsulation and may contribute to the slightly increased k cat values observed. The P22-GshF s showed a slightly higher activity than P22-GshF l , which again suggests that extra time for enzyme maturation before packaging is not always required to achieve high activity of encapsulated enzymes.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Synthetic Virus-like Particles for Glutathione Biosynthesis

et al. 2020

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Protein-based nanocompartments found in nature have inspired the development of functional nanomaterials for a range of applications including delivery of catalytic activities with therapeutic effects. As glutathione (GSH) plays a vital role in metabolic adaptation and many diseases are associated with its deficiency, supplementation of GSH biosynthetic activity might be a potential therapeutic when delivered directly to the disease site. Here, we report the successful design and production of active nanoreactors capable of catalyzing the partial or complete pathway for GSH biosynthesis, which was realized by encapsulating essential enzymes of the pathway inside the virus-like particle (VLP) derived from the bacteriophage P22. These nanoreactors are the first examples of nanocages specifically designed for the biosynthesis of oligomeric biomolecules. A dense packing of enzymes is achieved within the cavities of the nanoreactors, which allows us to study enzyme behavior, in a crowded and confined environment, including enzymatic kinetics and protein stability. In addition, the biomedical utility of the nanoreactors in protection against oxidative stress was confirmed using an in vitro cell culture model. Given that P22 VLP capsid was suggested as a potential liver-tropic nanocarrier in vivo, it will be promising to test the efficacy of these GSH nanoreactors as a novel treatment for GSH-deficient hepatic diseases.

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“…Until recently, bifunctional GSH synthetase (GshF) was reported to simultaneously catalyze the 2-step ATPdependent biosynthesis of GSH in those strains because GSH biosynthesis generally requires the consecutive reaction by γ-GCS and GS (Vergauwen et al, 2006;Ge et al, 2012). Bifunctional GSH synthetase contains an N-terminal γ-GCS-like domain followed by a typical ATP-grasp GS-like domain, but this GS-like domain has no sequence homology with any known GS (Stout et al, 2012). Based on sequence alignment, more than 20 microbial species, most of which are gram-positive bacteria such as L. plantarum and S. thermophilus, have been found to contain GshF, of which most have not been studied by cloning and expression (Li et al, 2011).…”

Section: Functional Analysis and Heterologous Expression Of Bifunctional Glutathione Synthetase From Lactobacillusmentioning

confidence: 99%

Functional analysis and heterologous expression of bifunctional glutathione synthetase from Lactobacillus

Xiong

Kong

Wang

et al. 2018

Journal of Dairy Science

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Bifunctional glutathione synthetase (GshF) has recently been reported to simultaneously catalyze the 2-step ATP-dependent biosynthesis of reduced glutathione (GSH). In this work, 19 putative gshF were mined from the complete sequenced genome of 20 representative Lactobacillus species. To functionally analyze these putative GshF, GshF from Lactobacillus plantarum and Lactobacillus casei were selected and successfully expressed in Escherichia coli. Compared with the control without expressing GshF, GSH titers were enhanced significantly in E. coli with overexpression of GshF, demonstrating that putative GshF from Lactobacillus have functional activities on GSH biosynthesis. Moreover, with the expression of GshF from L. plantarum in E. coli as a paradigm, GSH yield (286.5 μM) was strongly improved by 177.9% with optimized induced conditions and precursor concentration compared with the control under unoptimized conditions. Transcriptional analysis showed that key genes of endogenous GSH metabolism and precursor biosynthesis were remarkably suppressed by GshF expression, indicating that the increase of GSH titer was attributed to heterologous expression of GshF. Overall, our results suggested that gshF is enriched in Lactobacillus and that heterologous expression of GshF is an efficient strategy for improving GSH biosynthesis.

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Glutathione Biosynthesis in Bacteria by Bifunctional GshF Is Driven by a Modular Structure Featuring a Novel Hybrid ATP-Grasp Fold

Cited by 22 publications

References 32 publications

Adaptation of Enterococcus faecalis to Daptomycin Reveals an Ordered Progression to Resistance

Adaptation of Enterococcus faecalis to Daptomycin Reveals an Ordered Progression to Resistance

Synthetic Virus-like Particles for Glutathione Biosynthesis

Functional analysis and heterologous expression of bifunctional glutathione synthetase from Lactobacillus

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