Staphylopine, pseudopaline, and yersinopine dehydrogenases: A structural and kinetic analysis of a new functional class of opine dehydrogenase

McFarlane, Jeffrey S.; Davis, C.L.; Lamb, Audrey L.

doi:10.1074/jbc.ra118.002007

Cited by 34 publications

(69 citation statements)

References 42 publications

(50 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The structure of CntM have been solved in a binary/tertiary complex with NADPH or NADPH and xNA, revealing the residues that are involved in the complex formation and building the active site [8,32]. First focusing on the NADPH binding site of SaCntM (Figure 2A) we observed that the phosphate group of NADPH is sandwiched by the side chains of two positively charged residues (R33 and K39) that are rather conserved in the CntM family.…”

Section: Resultsmentioning

confidence: 99%

“…In this context, nicotianamine-like metallophores have been identified in some bacteria as playing an important role in metal acquisition strategies. Staphylopine, pseudopaline and yersinopine are the three examples currently known and recently identified in S. aureus [5], P. aeruginosa [6,7] and Y. pestis [8] respectively. The biosynthesis of these nicotianamine-like metallophores occurs in two or three steps depending on the species.…”

Section: Introductionmentioning

confidence: 99%

“…Differences in the staphylopine, pseudopaline, and yersinopine biosynthetic pathways. Adapted from [5,6,8]. …”

Section: Introductionmentioning

confidence: 99%

“…Opine dehydrogenases are also involved in the biosynthesis of nicotianamine-like metallophores in bacteria and their substrate specificity results in the production of diverse metallophores. The activity of these opine dehydrogenases from S. aureus , P. aeruginosa and Y. pestis (respectively called SaCntM, PaCntM and YpCntM) has been described: SaCntM uses NADPH and pyruvate with xNA to produce staphylopine [5], YpCntM uses pyruvate and NADPH with yNA to produce yersinopine [8], and PaCntM uses NAD(P)H and α-ketoglutarate with yNA to produce pseudopaline [6,7]. Therefore, substrate specificity (and eventually stereospecificity in the case of xNA vs yNA) of CntM leads to the production of diverse opine-type metallophores.…”

Section: Introductionmentioning

confidence: 99%

“…Similarly, the exporter of staphylopine was found to be important for fitness in abscesses, even before staphylopine discovery [27]. Concerning Y. pestis , no data indicates a link between yersinopine production and virulence at this time and the discovery of this metallophore is so far restricted to in vitro studies [8].…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Simple rules govern the diversity of bacterial nicotianamine-like metallophores

Laffont

Brutesco

Hajjar

et al. 2019

Preprint

View full text Add to dashboard Cite

3In metal-scarce environments, some pathogenic bacteria produce opine-type metallophores 1 4 mainly to face the host's nutritional immunity. This is the case of staphylopine, pseudopaline and 1 5 yersinopine, identified in Staphylococcus aureus, Pseudomonas aeruginosa and Yersinia pestis 1 6respectively. These metallophores are synthesized by two (CntLM) or three enzymes (CntKLM), 1 7 CntM catalyzing the last step of biosynthesis using diverse substrates (pyruvate or α -ketoglutarate), 1 8 pathway intermediates (xNA or yNA) and cofactors (NADH or NADPH), depending on the species. 1 9Here, we explored substrate specificity of CntM by combining bioinformatics and structural analysis 2 0 with chemical synthesis and enzymatic studies. We found that NAD(P)H selectivity was mainly due to 2 1 the amino acid at position 33 (S. aureus numbering) which ensures a preferential binding to NADPH 2 2 when it is an arginine. Moreover, whereas CntM from P. aeruginosa preferentially uses yNA over 2 3 xNA, the staphylococcal enzyme is not stereospecific. Most importantly, selectivity towards α -2 4 ketoacids is largely governed by a single residue at position 150 of CntM (S. aureus numbering): an 2 5 aspartate at this position ensures selectivity towards pyruvate whereas an alanine leads to the 2 6 consumption of both pyruvate and α -ketoglutarate. Modifying this residue in P. aeruginosa led to a 2 7complete reversal of selectivity. Thus, opine-type metallophore diversity is mainly mediated by the 2 8absence/presence of a cntK gene encoding a histidine racemase, and the presence of an 2 9 aspartate/alanine at position 150 of CntM. These two simple rules predict the production of a fourth 3 0 metallophore by Paenibacillus mucilaginosus, which was confirmed in vitro and called bacillopaline. 3 1 3 2 3 3 3 5metals required for their growth. This is particularly the case for pathogenic bacteria that have to 3 6 confront the host's immune system. Indeed, the so-called "nutritional immunity" induces an additional 3 7 metal limitation by sequestering iron, zinc or manganese to prevent bacterial growth [1-4]. To face 3 8 this metal restriction, bacteria have developed metallophores to recover metals. In this context, 3 9 nicotianamine-like metallophores have been identified in some bacteria as playing an important role in 4 0 metal acquisition strategies. Staphylopine, pseudopaline and yersinopine are the three examples 4 1 2 currently known and recently identified in S. aureus [5], P. aeruginosa [6,7] and Y. pestis [8] 4 2respectively. The biosynthesis of these nicotianamine-like metallophores occurs in two or three steps 4 3 depending on the species. When it is present, as in S. aureus, CntK, a histidine racemase, transforms 4 4

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

“…Differences in the staphylopine, pseudopaline, and yersinopine biosynthetic pathways. Adapted from [5,6,8]. …”

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Simple rules govern the diversity of bacterial nicotianamine-like metallophores

Laffont

Brutesco

Hajjar

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

Opine dehydrogenases, an underexplored enzyme family for the enzymatic synthesis of chiral amines

Telek

Molnár

Vértessy

et al. 2023

Biotech & Bioengineering

View full text Add to dashboard Cite

Opines and opine‐type chemicals are valuable natural products with diverse biochemical roles, and potential synthetic building blocks of bioactive compounds. Their synthesis involves reductive amination of ketoacids with amino acids. This transformation has high synthetic potential in producing enantiopure secondary amines. Nature has evolved opine dehydrogenases for this chemistry. To date, only one enzyme has been used as biocatalyst, however, analysis of the available sequence space suggests more enzymes to be exploited in synthetic organic chemistry. This review summarizes the current knowledge of this underexplored enzyme class, highlights key molecular, structural, and catalytic features with the aim to provide a comprehensive general description of opine dehydrogenases, thereby supporting future enzyme discovery and protein engineering studies.

show abstract

Heterologous Expression and Partial Characterization of a Putative Opine Dehydrogenase from a Metagenomic Sequence of Desulfohalobium retbaense

et al. 2023

View full text Add to dashboard Cite

The aim of this research was to prove the function of the putative opine dehydrogenase from Desulfohalobium retbaense and to characterize the enzyme in terms of functional and kinetic parameters. A putative opine dehydrogenase was identified from a metagenomic library by a sequence‐based technique search of the metagenomic library, and afterward was successfully heterologously produced in Escherichia coli. In order to examine its potential for applications in the synthesis of secondary amines, first the substrate specificity of the enzyme towards different amino donors and amino acceptors was determined. The highest affinity was observed towards small amino acids, preferentially L‐alanine, and when it comes to α‐keto acids, pyruvate proved to be a preferential amino acceptor. The highest activity was observed at pH 6.5 in the absence of salts. The enzyme showed remarkable stability in a wide range of experimental conditions, such as broad pH stability (from 6.0‐11.0 after 30 min incubation in buffers at a certain pH), stability in the presence of NaCl up to 3.0 M for 24 h, it retained 80% of the initial activity after 1 h incubation at 45 °C, and 65% of the initial activity after 24 h incubation in 30% dimethyl sulfoxide.

show abstract

Staphylopine, pseudopaline, and yersinopine dehydrogenases: A structural and kinetic analysis of a new functional class of opine dehydrogenase

Cited by 34 publications

References 42 publications

Simple rules govern the diversity of bacterial nicotianamine-like metallophores

Simple rules govern the diversity of bacterial nicotianamine-like metallophores

Opine dehydrogenases, an underexplored enzyme family for the enzymatic synthesis of chiral amines

Heterologous Expression and Partial Characterization of a Putative Opine Dehydrogenase from a Metagenomic Sequence of Desulfohalobium retbaense

Contact Info

Product

Resources

About