Polyamine‐independent growth and biofilm formation, and functional spermidine/spermine N‐acetyltransferases in Staphylococcus aureus and Enterococcus faecalis

Li, Bin; Maezato, Yukari; Kim, Sok Ho; Kurihara, Shin; Liang, Jue; Michael, Anthony J.

doi:10.1111/mmi.14145

Cited by 34 publications

(28 citation statements)

References 80 publications

(107 reference statements)

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“…Therefore, the in vitro substrate specificity showed spm is preferred over spd, which is similar to what has been observed in several previously characterized SSAT enzymes. 10,11,18 Previous studies with the human and mouse SSAT proteins showed that Y140 was critical for activity and it was proposed to act as a general acid during the enzymatic reaction. 6,15 Based on these results, Sugiyama et al mutated the structurally equivalent EcSpeG Y133 residue to phenylalanine in the recombinant protein and tested its activity.…”

Section: Resultsmentioning

confidence: 99%

Criticality of a conserved tyrosine residue in the SpeG protein from Escherichia coli

Dang

Lim

et al. 2021

Protein Science

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The SpeG spermidine/spermine N-acetyltransferase (SSAT) from Escherichia coli belongs to the Gcn5-related N-acetyltransferase (GNAT) superfamily of proteins. In vitro characterization of this enzyme shows it acetylates the polyamines spermine and spermidine, with a preference toward spermine. This enzyme has a conserved tyrosine residue (Y135) that is found in all SSAT proteins and many GNAT functional subfamilies. It is located near acetyl coenzyme A in the active center of these proteins and has been suggested to act as a general acid in a general acid/base chemical mechanism. In contrast, a previous study showed this residue was not critical for E. coli SpeG enzymatic activity when mutated to phenylalanine. This result was quite different from previous studies with a comparable residue in the human and mouse SSAT proteins, which also acetylate spermine and spermidine. Therefore, we constructed several mutants of the E. coli SpeG Y135 residue and tested their enzymatic activity. We found this conserved residue was indeed critical for E. coli SpeG enzyme activity and may behave similarly in other SSAT proteins.

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Section: Resultsmentioning

confidence: 99%

Criticality of a conserved tyrosine residue in the SpeG protein from Escherichia coli

Dang

Lim

et al. 2021

Protein Science

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“…Polyamines, such as spermidine and norspermidine, also have a dual role. Compounds that structurally mimic norspermidine has been demonstrated to effectively inhibit the biofilm formation of Bacillus subtilis and S. aureus [ 156 ], while both spermidine and norspermidine will also induce biofilm formation in some cases [ 157 , 158 , 159 ]. Besides, four D-amino acids produce by bacteria, including D-tyrosine, D-leucine, D-tryptophan, and D-methionine, can destroy existing biofilms and prevent biofilm forming [ 160 , 161 ].…”

Section: Dispersal Moleculesmentioning

confidence: 99%

Targeting Biofilms Therapy: Current Research Strategies and Development Hurdles

2020

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Biofilms are aggregate of microorganisms in which cells are frequently embedded within a self-produced matrix of extracellular polymeric substance (EPS) and adhere to each other and/or to a surface. The development of biofilm affords pathogens significantly increased tolerances to antibiotics and antimicrobials. Up to 80% of human bacterial infections are biofilm-associated. Dispersal of biofilms can turn microbial cells into their more vulnerable planktonic phenotype and improve the therapeutic effect of antimicrobials. In this review, we focus on multiple therapeutic strategies that are currently being developed to target important structural and functional characteristics and drug resistance mechanisms of biofilms. We thoroughly discuss the current biofilm targeting strategies from four major aspects—targeting EPS, dispersal molecules, targeting quorum sensing, and targeting dormant cells. We explain each aspect with examples and discuss the main hurdles in the development of biofilm dispersal agents in order to provide a rationale for multi-targeted therapy strategies that target the complicated biofilms. Biofilm dispersal is a promising research direction to treat biofilm-associated infections in the future, and more in vivo experiments should be performed to ensure the efficacy of these therapeutic agents before being used in clinic.

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“…Why might S. aureus and C. diphtheriae use the more complex amino acidcontaining structures rather than a putrescinecontaining one? It is well known that S. aureus is a putrescine auxotroph (57,58), and BLASTP analysis of C. diphtheriae strain proteomes indicates that no known putrescine synthesizing enzymes (52) are encoded in their genomes, in contrast to the putrescine prototrophs R. pickettii (59) and L. pneumophila.…”

Section: Putrescine Prototrophy and Putrescinedependent Rhizoferrin Bmentioning

confidence: 99%

Alternative pathways utilize or circumvent putrescine for biosynthesis of putrescine-containing rhizoferrin

Deng

Kim

et al. 2021

Journal of Biological Chemistry

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The siderophore rhizoferrin ( N 1 , N 4 -dicitrylputrescine) is produced in fungi and bacteria to scavenge iron. Putrescine-producing bacterium Ralstonia pickettii synthesizes rhizoferrin and encodes a single nonribosomal peptide synthetase-independent siderophore (NIS) synthetase. From biosynthetic logic, we hypothesized that this single enzyme is sufficient for rhizoferrin biosynthesis. We confirmed this by expression of R. pickettii NIS synthetase in Escherichia coli , resulting in rhizoferrin production. This was further confirmed in vitro using the recombinant NIS synthetase, synthesizing rhizoferrin from putrescine and citrate. Heterologous expression of homologous lbtA from Legionella pneumophila , required for rhizoferrin biosynthesis in that species, produced siderophore activity in E. coli . Rhizoferrin is also synthesized by Francisella tularensis and Francisella novicida , but unlike R. pickettii or L. pneumophila , Francisella species lack putrescine biosynthetic pathways because of genomic decay. Francisella encodes a NIS synthetase FslA/FigA and an ornithine decarboxylase homolog FslC/FigC, required for rhizoferrin biosynthesis. Ornithine decarboxylase produces putrescine from ornithine, but we show here in vitro that FigA synthesizes N -citrylornithine, and FigC is an N -citrylornithine decarboxylase that together synthesize rhizoferrin without using putrescine. We co-expressed F. novicida figA and figC in E. coli and produced rhizoferrin. A 2.1 Å X-ray crystal structure of the FigC N -citrylornithine decarboxylase reveals how the larger substrate is accommodated and how active site residues have changed to recognize N -citrylornithine. FigC belongs to a new subfamily of alanine racemase-fold PLP-dependent decarboxylases that are not involved in polyamine biosynthesis. These data reveal a natural product biosynthetic workaround that evolved to bypass a missing precursor and re-establish it in the final structure.

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Polyamine‐independent growth and biofilm formation, and functional spermidine/spermine N‐acetyltransferases in Staphylococcus aureus and Enterococcus faecalis

Cited by 34 publications

References 80 publications

Criticality of a conserved tyrosine residue in the SpeG protein from Escherichia coli

Criticality of a conserved tyrosine residue in the SpeG protein from Escherichia coli

Targeting Biofilms Therapy: Current Research Strategies and Development Hurdles

Alternative pathways utilize or circumvent putrescine for biosynthesis of putrescine-containing rhizoferrin

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