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Abstract Dichloroacetic acid (DCA), an organohalide that present in environmental sample and biological systems, got high attention for its therapeutic potential as the inhibitor of pyruvate dehydrogenase kinase (PDK), elevated in obesity, diabetes, heart disease and cancer. Herein, we developed a Cobalt conjugated carbon quantum dots (N-CQDs/Co) that selectively detect DCA by fluorescence “turn-on” mechanism. Utilizing TEM, DLS, UV–vis and fluorescence spectroscopy, the mechanism has been thoroughly elucidated and is attributed to disaggregation induced enhancement (DIE). The limit of detection of the N-CQDs/Co complex is 8.7 µM. The structural characteristics and size of the N-CQDs and N-CQDS/Co complex have been verified using FT-IR, XPS, HRTEM, DLS, EDX have been performed. Additionally, the complex is used to specifically find DCA in the human cell line and in zebrafish.Journal instruction requires a city for affiliations; however, these are missing in affiliation [4]. Please verify if the provided city is correct and amend if necessary.Kharagpur is the city. The address is okay.

Abstract Selective isolation of soil Actinobacteria was undertaken to isolate a new class of antibiotics and bioactive molecules. A Streptomyces sp. PSAA01 (= MTCC 13,157), isolated from soil of Eastern Himalaya foothill was cultivated on a large scale for the production of the antimicrobial SM02. It has been found that the maximum amount of SM02 produced while PSAA01 was grown in ISP-2 medium (pH 7.0) for 7 days at 30 °C in shaking (180 rpm) condition. A significant zone of inhibition against Staphylococcus aureus MTCC 96 has been found with the crude cell-free culture media (50 µL) of 7 days grown PSAA01. After the purification and chemical structural characterization, we found that SM02 is a new antimicrobial having 746 dalton molecular weight. The compound SM02 contains pyrimidine moiety in it and is produced by a species of Streptomyces and thus we have named this antibiotic pyrimidomycin. The antimicrobial spectrum of pyrimidomycin has been found to be restricted in Gram-positive organisms with a MIC of 12 µg/mL. SM02 was found active against Mycobacterium sp. and also multi-drug resistant Gram-positive bacteria with similar potency and found to disrupt the bacterial cell wall. Pyrimidomycin also showed significant impairment in the biofilm formation by S. aureus. Furthermore, pyrimidomycin showed synergy with the most used antibiotic like ampicillin, vancomycin and chloramphenicol. Pyrimidomycin did not have cytotoxicity towards human cell lines indicating its limited activity within bacteria.

Abstract The genus Lentzea is a rare group of actinobacteria having potential for the exploration of bioactive compounds. Despite its proven ability to produce compounds with medical relevance, Lentzea genome analysis remains unexplored. Here we show a detailed understanding of the genetic features, biosynthetic gene clusters (BGCs), and genetic clusters for carbohydrate-active enzymes present in the Lentzea genome. Our analysis determines the genes for core proteins, non-ribosomal peptide synthetase condensation domain, and polyketide synthases-ketide synthase domain. The antiSMASH-based sequence analysis identifies 692 BGCs among which 8% are identical to the BGCs that produce geosmin, citrulassin, achromosin (lassopeptide), vancosamine, anabaenopeptin NZ857/nostamide A, alkylresorcinol, BE-54017, and bezastatin. The remaining BGCs code for advanced category antimicrobials like calcium-dependent, glycosylated, terpenoids, lipopeptides, thiopeptide, lanthipeptide, lassopeptide, lingual antimicrobial peptide and lantibiotics together with antiviral, antibacterial, antifungal, antiparasitic, anticancer agents. About 28% of the BGCs, that codes for bioactive secondary metabolites, are exclusive in Lentzea and could lead to new compound discoveries. We also find 7121 genes that code for carbohydrate-degrading enzymes which could essentially convert a wide range of polymeric carbohydrates. Genome mining of such genus is very much useful to give scientific leads for experimental validation in the discovery of new-generation bioactive molecules of biotechnological importance.

Abstract The present study aims at the production optimization, purification, and characterization of a potent broad-spectrum antimicrobial compound (AMC) produced by Aspergillus fumigatus nHF-01 (GenBank Ac. No. MN190286). The culture conditions were optimized for a higher amount of AMC. The AMC was solvent extracted and characterized by UV–Vis, FT–IR, ESI–MS, and 1H-NMR spectroscopy. The MIC, MBC and mode of action were determined against a set of Gram-positive and Gram-negative human pathogenic bacteria. Its antibiofilm, synergistic and cytotoxic effects were also tested. The putative target site of action was evaluated through in silico molecular docking study. The stain A. fumigatus nHF-01 produced the maximum AMC (5-butyl-2-pyridine carboxylic acid) in 2% MEB (w/v) and 4% YE (w/v) at pH 6.0 and 20 °C temperature with 100 rpm agitation for ten days. It caused complete lethality of the Gram-positive and Gram-negative human pathogenic bacteria at a 129 µg/mL dose by rupture and entire dissolution of cell integrity. It showed moderate antibiofilm activity and had a synergistic activity with streptomycin and additive effects with ciprofloxacin and vancomycin. It targets a respiratory enzyme, Quinol-Fumarate Reductase (1l0v), with the highest binding affinities. It had cytotoxicity against human lung carcinoma A549 cell line and was stable up to 100 °C. Thus, the study revealed that the strain A. fumigatus nHF-01 produces a potent broad-spectrum AMC 5-butyl-2-pyridine carboxylic acid that could be used against human food and topical pathogenic bacteria. This is the first report of such a compound produced from the A. fumigatus.

Polar microbiology remains as the most fascinating area of research which mainly focuses on exploration of psychrophilic organisms for having their cold-active enzymes of biotechnological potential. In this study, we have explored a culturable bacterial community and isolated 27 bacterial isolates with a different morphology from an unexplored site of Arctic region, for the possibility of identifying various active biomolecules. Screening of various isolates in a culture dependent manner helped us to identify strains capable of producing extracellular enzymes. The optimal growth parameters of most of the isolates are ranges between 18-22°C temperature, 3-5 days of incubation, 6-9 pH, and 3-5% (w/v) NaCl in LB media. It has also been found that among these isolates, 63% are able to produce lipase, 17% amylase, 7% xylanase and 7% isolates have responded for phosphatase activity but there are no isolates found for gelatinase and cellulase production ability. In addition, few isolates can also produce secretory protease, urease, β-galactosidase, etc. 16SrRNA gene sequence-based phylogeny revealed that the isolates belong to the genera of Psychrobacter, Planococcus, Halomonas, Arthrobacter, Oceanisphaera, Marinbacter, Pseudomonas, Algoriphagus. Strikingly, none of the Arctic isolates showed resistance towards commonly used antibiotics which indicates that the unexplored habitat is devoid of antibiotic exposure and so does the rise of antimicrobial resistance. The structure-function relationship of the isolated bioactive compounds from these isolates are the major focus of future research.

Abstract Purulia is a malaria-prone district in West Bengal, India, with approximately half of the blocks defined as malaria endemic. We analyzed the malaria case in each block of the Purulia district from January 1, 2016, to December 31, 2020. As per the API, 20 blocks of Purulia were assigned to four different categories (0–3) and mapped using ArcGIS software. An exponential decay model was fitted to forecast the trend of malaria cases for each block of Purulia (2021–2025). There was a sharp decrease in total malaria cases and API from 2016 to 2020 due to the mass distribution of LLINs. The majority of cases (72.63%) were found in ≥ 15-year age group. Males were more prone to malaria (60.09%). Malaria was highly prevalent among Scheduled Tribes (48.44%). Six blocks were reported in Category 3 (high risk) and none in Category 0 (no risk) in 2016, while no blocks were determined to be in Category 3, and three blocks were in Category 0 in 2020. The exponential decay model prediction is oriented towards gaining malaria-free status in thirteen blocks of Purulia by 2025. This study will incite the government to uphold and strengthen the current efforts to meet the malaria elimination goals.

No structural data have been available for RNA polymerase holoenzymes or transcription initiation complexes that contain extracytoplasmic σ factors. Here the authors report the crystal structures of transcription initiation complexes comprising Mycobacterium tuberculosis RNA polymerase, extracytoplasmic σ factor σL and promoter DNA.

Lactic acid bacteria (LAB) play a very vital role in food production, preservation, and as probiotic agents. Some of these species can colonize and survive longer in the gastrointestinal tract (GIT), where their presence is crucially helpful to promote human health. LAB has also been used as a safe and efficient incubator to produce proteins of interest. With the advent of genetic engineering, recombinant LAB have been effectively employed as vectors for delivering therapeutic molecules to mucosal tissues of the oral, nasal, and vaginal tracks and for shuttling therapeutics for diabetes, cancer, viral infections, and several gastrointestinal infections. The most important tool needed to develop genetically engineered LABs to produce proteins of interest is a plasmid-based gene expression system. To date, a handful of constitutive and inducible vectors for LAB have been developed, but their limited availability, host specificity, instability, and low carrying capacity have narrowed their spectrum of applications. The current review discusses the plasmid-based vectors that have been developed so far for LAB; their functionality, potency, and constraints; and further highlights the need for a new, more stable, and effective gene expression platform for LAB.

Metal organic frameworks are an emerging class of materials with a broad range of electronic properties with diverse applications such as sensors, catalysts, and permeable membranes. Here, three isostructural wide bandgap (WB) zeolitic imidazole frameworks (ZIFs) are synthesized having metal ion sites occupied by Zn (ZIF-8, Eg = 5.3 eV), Co (ZIF-67, Eg = 4.3 eV), and both Zn and Co (50%–50% mixture). The conductivity mechanism in these WB ZIFs involves the Mott variable range hopping of charge carriers from one metal site to the other. The hopping probability in the mixture is governed by the constitute having a lesser activation energy. Thus, it reveals that the incorporation of a different metal ion (Co in place of Zn) in the lattice forms a parallel low resistance path through hopping at Co sites and hence reduces the sensor response as well as selectivity toward ammonia. This parallel resistance path of the Co channel does not get affected by ammonia since it is found that ammonia has high affinity toward Zn ions and not toward Co ions. Thus, the incorporation of new metal ions hinders the hopping charge transport mechanisms in ZIFs.

Abstract A chemosensor, 3-aminophenol-based rhodamine conjugate (ARC) has been developed for visualisation of diethylchlorophosphate (DCP), mimic of a chemical warfare agent, in Catfish brain. The simple detection of DCP by “turn-on” fluorescence property of the chemosensor makes it unique for easy and rapid in vivo and in vitro detection of DCP with the detection limit of 5.6 nM.

Triton X-100 (TX-100), a useful non-ionic surfactant, reduced the methicillin resistance in Staphylococcus aureus significantly. Many S. aureus proteins were expressed in the presence of TX-100. SarA, one of the TX-100-induced proteins, acts as a global virulence regulator in S. aureus. To understand the effects of TX-100 on the structure, and function of SarA, a recombinant S. aureus SarA (rSarA) and its derivative (C9W) have been investigated in the presence of varying concentrations of this surfactant using various probes. Our data have revealed that both rSarA and C9W bind to the cognate DNA with nearly similar affinity in the absence of TX-100. Interestingly, their DNA binding activities have been significantly increased in the presence of pre-micellar concentration of TX-100. The increase of TX-100 concentrations to micellar or post-micellar concentration did not greatly enhance their activities further. TX-100 molecules have altered the secondary and tertiary structures of both proteins to some extents. Size of the rSarA-TX-100 complex appears to be intermediate to those of rSarA and TX-100. Additional analyses show a relatively moderate interaction between C9W and TX-100. Binding of TX-100 to C9W has, however, occurred by a cooperative pathway particularly at micellar and higher concentrations of this surfactant. Taken together, TX-100-induced structural alteration of rSarA and C9W might be responsible for their increased DNA binding activity. As TX-100 has stabilized the somewhat weaker SarA-DNA complex effectively, it could be used to study its structure in the future.

SarA, a Staphylococcus aureus-specific dimeric protein, modulates the expression of numerous proteins including various virulence factors. Interestingly, S. aureus synthesizes multiple SarA paralogs seemingly for optimizing the expression of its virulence factors. To understand the domain structure/flexibility and the folding/unfolding mechanism of the SarA protein family, we have studied a recombinant SarA (designated rSarA) using various in vitro probes. Limited proteolysis of rSarA and the subsequent analysis of the resulting protein fragments suggested it to be a single-domain protein with a long, flexible C-terminal end. rSarA was unfolded by different mechanisms in the presence of different chemical and physical denaturants. While urea-induced unfolding of rSarA occurred successively via the formation of a dimeric and a monomeric intermediate, GdnCl-induced unfolding of this protein proceeded through the production of two dimeric intermediates. The surface hydrophobicity and the structures of the intermediates were not identical and also differed significantly from those of native rSarA. Of the intermediates, the GdnCl-generated intermediates not only possessed a molten globule-like structure but also exhibited resistance to dissociation during their unfolding. Compared to the native rSarA, the intermediate that was originated at lower GdnCl concentration carried a compact shape, whereas, other intermediates owned a swelled shape. The chemical-induced unfolding, unlike thermal unfolding of rSarA, was completely reversible in nature.

We report that bacterial RNA polymerase (RNAP) is the functional cellular target of the depsipeptide antibiotic salinamide A (Sal), and we report that Sal inhibits RNAP through a novel binding site and mechanism. We show that Sal inhibits RNA synthesis in cells and that mutations that confer Sal-resistance map to RNAP genes. We show that Sal interacts with the RNAP active-center ‘bridge-helix cap’ comprising the ‘bridge-helix N-terminal hinge’, ‘F-loop’, and ‘link region’. We show that Sal inhibits nucleotide addition in transcription initiation and elongation. We present a crystal structure that defines interactions between Sal and RNAP and effects of Sal on RNAP conformation. We propose that Sal functions by binding to the RNAP bridge-helix cap and preventing conformational changes of the bridge-helix N-terminal hinge necessary for nucleotide addition. The results provide a target for antibacterial drug discovery and a reagent to probe conformation and function of the bridge-helix N-terminal hinge.

Bacteriophage Φ11 uses Staphylococcus aureus as its host and, like lambdoid phages, harbors three homologous operators in between its two divergently oriented repressor genes. None of the repressors of Φ11, however, showed binding to all three operators, even at high concentrations. To understand why the DNA binding mechanism of Φ11 repressors does not match that of lambdoid phage repressors, we studied the N-terminal domain of the Φ11 lysogenic repressor, as it harbors a putative helix-turn-helix motif. Our data revealed that the secondary and tertiary structures of the N-terminal domain were different from those of the full-length repressor. Nonetheless, the N-terminal domain was able to dimerize and bind to the operators similar to the intact repressor. In addition, the operator base specificity, binding stoichiometry, and binding mechanism of this domain were nearly identical to those of the whole repressor. The binding affinities of the repressor and its N-terminal domain were reduced to a similar extent when the temperature was increased to 42°C. Both proteins also adequately dislodged a RNA polymerase from a Φ11 DNA fragment carrying two operators and a promoter. Unlike the intact repressor, the binding of the N-terminal domain to two adjacent operator sites was not cooperative in nature. Taken together, we suggest that the dimerization and DNA binding abilities of the N-terminal domain of the Φ11 repressor are distinct from those of the DNA binding domains of other phage repressors.