Biological and structural characterization of the Mycobacterium smegmatis nitroreductase NfnB, and its role in benzothiazinone resistance

Manina, Giulia; Bellinzoni, M.; Pasca, Maria Rosalia; Neres, João; Milano, Anna; Ribeiro, Ana Luisa de Jesus Lopes; Buroni, Silvia; Škovierová, Henrieta; Dianišková, Petronela; Mikušová, Katarı́na; Marák, Jozef; Makarov, Vadim; Giganti, David; Haouz, Ahmed; Lucarelli, Anna; Degiacomi, Giulia; Piazza, Aurora; Chiarelli, Laurent R.; Rossi, Edda De; Salina, Elena G.; Cole, Stewart T.; Alzari, Pedro M.; Riccardi, Giovanna

doi:10.1111/j.1365-2958.2010.07277.x

Cited by 74 publications

(124 citation statements)

References 41 publications

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“…Our group also identified and characterized a novel resistance mechanism to benzothiazinones in M. smegmatis (Manina et al, 2010a), and this mechanism was confirmed also for dinitrobenzamides (M.R. Pasca, personal communication).…”

Section: Introductionmentioning

confidence: 57%

“…In this context, another novel resistance mechanism to BTZs was described in M. smegmatis (Manina et al, 2010a). The over-expression of the nitroreductase NfnB leads to the inactivation of the drug by reduction of a critical nitro group to an amino group (Figure 3).…”

Section: Benzothiazinonesmentioning

confidence: 96%

“…The sequence analysis revealed no mutations in the nfnB gene, but a point mutation in MSMEG_6503. This mutation led to the substitution of leucine at position 137 by proline, leading to the hypothesis that the resistance of this mutant could be due to a defective repressor (MSMEG_6503), possibly causing over-expression of NfnB and, consequently, the reduction of the BTZ nitro-molecule to its less active amino-derivative (Manina et al, 2010a).…”

Section: Benzothiazinonesmentioning

confidence: 99%

“…One of these mutants presented a mutation in the hypothetical repressor binding site (Manina et al, 2010a).…”

Section: Benzothiazinonesmentioning

confidence: 99%

“…The direct involvement of NfnB in the inactivation of the lead compound BTZ043 was demonstrated by enzymology, microbiological assays and gene knockout experiments. The crystal structure of NfnB and docking analysis of NfnBbenzothiazinones have been performed in order to understand their interaction and the mechanism of nitroreduction (Manina et al, 2010a). Although M. tuberculosis seems to lack nitroreductases able to inactivate these drugs, our findings are valuable for the design of new benzothiazinones, which may be more effective in vivo.…”

Section: Introductionmentioning

confidence: 99%

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Fighting Against Resistant Strains: The Case of Benzothiazinones and Dinitrobenzamides

Buroni¹,

Riccardi²,

Pasca³

2012

Understanding Tuberculosis - New Approaches to Fighting Against Drug Resistance

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Section: Introductionmentioning

confidence: 57%

Section: Benzothiazinonesmentioning

confidence: 96%

Section: Benzothiazinonesmentioning

confidence: 99%

“…One of these mutants presented a mutation in the hypothetical repressor binding site (Manina et al, 2010a).…”

Section: Benzothiazinonesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Fighting Against Resistant Strains: The Case of Benzothiazinones and Dinitrobenzamides

Buroni¹,

Riccardi²,

Pasca³

2012

Understanding Tuberculosis - New Approaches to Fighting Against Drug Resistance

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Anionπ Interactions in Flavoproteins

Estarellas

Frontera

Quiñonero

et al. 2011

Chemistry An Asian Journal

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The importance of anion À p interactions [1] has been widely demonstrated by a great deal of theoretical and experimental investigations. [2] These interaction are gaining significant recognition, and their pivotal role in many key chemical and biological processes is becoming increasingly appreciated. [3] The design of highly selective anion receptors and channels represent important advances in this new field of supramolecular chemistry. [4] Recently, our group [5] has reported clear evidence of anion À p interactions in the active site of urate oxidase, causing inhibition of the enzymatic activity, and thereby demonstrating the crucial role of this noncovalent interaction in a biological system. In addition, Rotello and co-workers [6] have published pioneering computational and experimental model studies that establish the importance of donor-atom À p bonding (lone-pair À p interactions) on the recognition and function of flavins.Herein, we report the results of a systematic search for anion À p interactions in flavin-dependent enzymes. The structure of riboflavin (Scheme 1) consists of an isoalloxazine heterocyclic ring system, which is responsible for its redox activity. The ribitol side-chain attached to N10 can be phosphorylated in the case of FMN or attached through a diphosphate linkage to adenosine in FAD. The highly conjugated isoalloxazine tricyclic ring system is an excellent electron acceptor and therefore it should be well-suited for establishing anionÀp interactions, as confirmed by its positive quadrupole moment (Q zz = 4.42 B). This result is clearly confirmed by the electrostatic potential surface (EPS), which has been computed for a model of flavin (1) where the sidechain at N10 has been replaced by a hydrogen atom (Figure 1). A blue region can be clearly observed over the pyrimidinic ring. We have performed a search in the Protein Data Base (PDB) for anion À p interactions in proteins containing either FMN or FAD moieties. This search has been carried out manually examining all PDB entries one by one, which is a very arduous task. As a result, we have found a significant number of structures exhibiting anionÀp interactions between either FAD or FMN and Cl À , Br À , N 3 À , SCN À , SO 4 2À , and phosphate anions (see the Supporting Information). We performed a theoretical study on the p-binding ability of 1 toward several anions by performing high-level ab initio calculations (RI-MP2/CBS level of theory, CBS = complete basis set) of several complexes and computing the interaction energies (Table 1). In all cases, the interaction energies were large and negative, thereby indicating favorable interactions.As mentioned above, the search in the PDB provided evidence of anion À p interactions in a large number of proteins (51 hits). The total number of proteins in the PDB containing either FAD or FMN moieties in the structure was 1594.

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Synthesis of Pharmaceutically Relevant Arylamines Enabled by a Nitroreductase from Bacillus tequilensis

Russo,

Luján,

Fraaije

et al. 2024

ChemBioChem

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Arylamines are essential building blocks for the manufacture of valuable pharmaceuticals, pigments and dyes. However, their current industrial production involves the use of chemocatalytic procedures with a significant environmental impact. As a result, flavin‐dependent nitroreductases (NRs) have received increasing attention as sustainable catalysts for more ecofriendly synthesis of arylamines. In this study, we assessed a novel NR from Bacillus tequilensis, named BtNR, for the synthesis of pharmaceutically relevant arylamines, including valuable synthons used in the manufacture of blockbuster drugs such as vismodegib, sonidegib, linezolid and sildenafil. After optimizing the enzymatic reaction conditions, high conversion of nitroaromatics to arylamines (up to 97%) and good product yields (up to 56%) were achieved. Our results indicate that BtNR has a broad substrate scope, including bulky nitro benzenes, nitro pyrazoles and nitro pyridines. Hence, BtNR is an interesting biocatalyst for the synthesis of pharmaceutically relevant amine‐functionalized aromatics, providing an attractive alternative to traditional chemical synthesis methodologies.

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Biological and structural characterization of the Mycobacterium smegmatis nitroreductase NfnB, and its role in benzothiazinone resistance

Cited by 74 publications

References 41 publications

Fighting Against Resistant Strains: The Case of Benzothiazinones and Dinitrobenzamides

Fighting Against Resistant Strains: The Case of Benzothiazinones and Dinitrobenzamides

Anionπ Interactions in Flavoproteins

Synthesis of Pharmaceutically Relevant Arylamines Enabled by a Nitroreductase from Bacillus tequilensis

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