We report the genome sequence of a novel species of nontuberculous mycobacterium (NTM) belonging to the Mycobacterium terrae complex (MTC). The strain SVM_VP21 was isolated from the mouthwash sample of a healthy human adult residing in Delhi, India. The strain exhibited smooth, cream-coloured colonies on Lowenstein Jensen (LJ) medium after 2 weeks of incubation and could not be identified by Hain’s Genotype Mycobacterium CM/AS line probe assay. Whole genome sequencing (WGS) of SVM_VP21 was performed using Illumina NovaSeq 6000 platform for final identification. The whole genome sequence was assembled using Shovill. Gene annotation was performed by Rapid Annotation using Subsystem Technology (RAST) and Prokaryotic Genome Annotation Pipeline (PGAP). The genome of 4,882,418 bp exhibited 67.3

Overexpression of efflux pumps has been associated with drug resistance in Mycobacterium tuberculosis. However, their specific role in delamanid resistance remains unclear. This study aimed to identify efflux pump genes influenced by delamanid exposure through transcriptomic analysis. Cultures of M. tuberculosis H37Rv were exposed to delamanid at a sub-inhibitory concentration of ½ MIC for 24 h and transcriptomic sequencing was performed on exposed and unexposed cultures. Differentially expressed genes (DEGs) were annotated using the UniProt database. Transcriptomic analysis revealed that 23 efflux pump genes were significantly upregulated under delamanid stress, with 17/23 (73.9%) belonging to the ABC family, 4/23 (17.39%) to the RND family and 2/23 (8.69%) to the MFS superfamily. Quantitative real-time PCR (qRT-PCR) was conducted to validate the transcriptomic results for efflux pump genes following exposure of M. tuberculosis H37Rv to delamanid at ½ and ¼ MIC. qRT-PCR confirmed that 20/23 (86.95%) genes were significantly upregulated at ½ MIC, with 14/20 (70%) of these belonging to the ABC family. In contrast, only 3/23 (13%) efflux pump genes demonstrated upregulation at ¼ MIC. This study identified a set of efflux pump genes significantly upregulated in response to sub-inhibitory concentrations of delamanid and suggested that ½ MIC is a more suitable concentration than ¼ MIC for evaluating the expression of efflux pump genes in response to delamanid in M. tuberculosis.

We report the draft genome of Mycolicibacterium conceptionense, a rapidly growing nontuberculous mycobacterium, isolated from the sputum sample of a patient undergoing treatment for multidrug-resistant tuberculosis in Delhi, India. The 6,366,717-bp genome contains 6,124 coding sequences, one 5S rRNA, three 16S rRNAs, six 23S rRNAs, and 49 tRNAs.

In view of the devastating tuberculosis infections and emerging resistance against available drugs, novel 5phenyl-3-(thiophen-2-yl)-4,5-dihydro-1H-pyrazol-1-yl)(pyridin-4-yl)methanones were designed and efficiently synthesized. The in-vitro biological screening of the synthesized isoniazid-based pyrazolines reveals their remarkable antimycobacterial and antioxidant ability. The pyrazoline analog 4i containing 2,4-dimethoxy phenyl substituent exhibited the highest anti-TB action against the H37Rv Mtb strain with an MIC of 6.35 mu M, while the pyrazoline analog 4f with 2-nitrophenyl substituent demonstrates the highest radical quenching ability with an IC50 of 96.19 mu M. Through DFT execution, the quantum-mechanical properties, reactivity indices, and molecular electrostatic potential mapping of the synthesized entities were determined, showing their optimum chemical and biological compatibility. The potent antitubercular effect of the synthesized entities was further supported by docking assessment against one of the most promising antitubercular targets, InhA (PDB Id: 2X23), displaying a significant binding range of -8.3 to -9.6 kcal/mol, and favorable binding interactions with the key residues like TYR 158, THR 196. etc. The pharmaceutical compliance of the synthesized moieties was further assessed through in-silico ADMET screening, which illustrates their excellent drug likeliness and appreciable pharmacokinetic behavior.

A series of aliphatic isoniazid carbo(x/thio)amides has been synthesized by a facile one-pot green method in aqueous media by harnessing microwave irradiations. In vitro antitubercular screening of the syn-thesized compounds against Mycobacterium tuberculosis H37Rv (MTB strain) reveals promising antimy-cobacterial potential with MIC in the range of 1 to 4 mu g/ml. A computational approach comprising DFT modeling, molecular docking and ADMET analysis was executed to elucidate the chemical nature, bind -ing affinity and ADMET efficacy of the lead compounds. The docking of synthesized analogs with InhA displays weak binding interactions with the receptor and the absence of important interactions specific to direct inhibitors. However, the docking results illustrate notable binding affinity of the synthesized molecules with the KatG receptor 1SJ2 and compounds 3h and 3e showed the best docking scores (-7.2 and-6.8 kcal/mol, respectively) indicating the KatG mediated mechanism of InhA inhibition similar to isoniazid. In silico ADMET analysis of the title compounds indicate their outstanding pharmacokinetic proficiency and lower toxicity than isoniazid. The bioavailability radar further illustrates their exemplary oral availability and drug-likeliness.(c) 2023 Elsevier B.V. All rights reserved.

Globally, plant-based medicines as an alternative to chemically synthesized medicines, are growing in popularity. Abrus precatorius , L., belongs to the family Fabaceae and commonly known as Gunja in Hindi, is a multipurpose ethnomedicinal plant, traditionally used as an abortifacient, contraceptive, for treating infections, stomach ailments, worm infestation, jaundice, arthritis, snake bite, cold, fever, etc. Investigations on plant extracts of A. precatorius for assessing their phytoconstituents and ethnopharmacological properties (viz. anti-inflammatry, antioxidant, anti-fertility, neuroprotective, nephroprotective, immunomodulatory, etc.) have provided the scientific basis of its traditional usage as ethnomedicines. Thus, this plant with its diverse medicinal phytochemicals is viewed as a potential bioresource for commercial production of herbal medicines. Keeping in view, the growing importance of this medicinal plant as shown by bibliometric analysis, the present review comprehensively discusses the association between its important phytoconstituents, pharmacognostic properties and the ethnopharmacological aspects. Further, the future prospects include the aspects of ethnopharmacological activities which need further investigations for application in the health sector.

Abstract Tumor necrosis factor-alpha (TNF-α) is an inflammatory cytokine that plays a major role in immune regulation, homeostatic function, and cellular organization. The present study was undertaken to overproduce recombinant human TNF-α (rhTNF-α) in Escherichia coli (E.coli) in high cell density culture. The use of a codon-optimized gene and strong promoter-based (T7) expression system, choice of Terrific Broth (TB) as medium, and subsequent optimization of culture conditions in shake flasks resulted in production of 0.95 g/L insoluble rhTNF-α comprising upto 50% of total cellular protein (TCP) The protein yield further increased upto 1.26 g/L in 1 L TB medium batch culture in bioreactor with the controlled temperature, pH, and dissolved oxygen. In a series of chemostats operated at dilution rates of 0.2 h−1, 0.3 h−1, 0.4 h−1 and 0.5 h−1 the specific growth rate (μ) positively correlated with specific yield (Yp / x ) and a maximum yield of 164 mg/g DCW was obtained at μ = 0.4 h−1 within 4 h post-induction. A fed-batch cultivation in TB with an exponential feeding profile (μ = ∼0.4 h−1) of concentrated feed resulted in an accumulation of 5.5 g/L of rhTNF-α within 14 h of cultivation which accounted for ∼29% of TCP.

In developing countries like India, fecal pollution of surface waters is a major threat to public and environmental health. The aim of the study was to assess serological, phylogenetic and molecular diversity among aquatic Escherichia coli isolates from Yamuna river and their comparison with the animal fecal isolates. A total of 97 E. coli isolates from Yamuna river and domesticated animals were characterized by multilocus variable number tandem repeat analysis (MLVA) using four VNTR loci. The pathogenicity of these strains by serological and phylogenetic analysis was also determined. E. coli strains were differentiated into 53 distinct MLVA types with high discriminatory power, Simpson's index of 0.95 (95% CI 0.923–0.978). Cluster analysis and population modeling using minimum spanning tree suggested a possible epidemiological linkage among aquatic and fecal isolates. The study also reported the presence of highly diverse and pathogenic serotypes belonging to STEC and EPEC strains, particularly O157 and high prevalence of pathogenic phylogroups (phylogroup, B2 and D). The presence of such a high molecular heterogeneity among aquatic and fecal E. coli isolates emphasizes upon the need to develop proper fecal pollution abatement strategies for Indian natural bodies.

Cyclotides is a rapidly growing class of plant‐derived cyclic peptides exhibiting several bioactivities with potential applications in the agricultural and pharmaceutical sectors. Both natural and grafted cyclotides have shown promise in cancer therapy. Approximately 70 natural cyclotides belonging to three plant families (Fabaceae, Rubiaceae, and Violaceae) have shown cytotoxicity against several cancer cell lines. Cyclotides exhibit considerable stability against thermal and enzymatic proteolysis, owing to their unique structure with knotted topology and head to tail cyclization. Further, their small size, high stability, oral bioavailability, and tolerance to amino acid substitution in structural loops make them an ideal platform for designing peptide‐based drugs for cancer. Thus, cyclotides provide ideal scaffolds for bioactive epitope grafting and facilitating drug delivery in cancer treatment. Many anticancer linear peptides have been grafted in cysteine knotted cyclic framework of cyclotide for enhancing their cell permeability across cellular membranes, thereby improving their delivery and pharmacokinetics. The present review comprehensively discusses the distribution, toxicity, and anticancer bioactivity of natural cyclotides. Further, it systematically elaborates on the role and action of epitopes' into grafted cyclotides in targeting cancer. The review also encompasses related patents landscape study and future challenges in peptide‐based cancer therapy.

Acyclotides are plant-based, acyclic miniproteins with cystine knot motif formed by three conserved disulfide linkages and lack head to tail ligation. Acyclotides may not necessarily be less stable, even though they lack cyclic backbone, as the conserved cystine knot feature provides the required stability. Violacin A was the first acyclotide, isolated from Viola odorata in 2006. Until now, acyclotides have been reported from five dicot families (Violaceae, Rubiaceae, Cucurbitaceae, Solanaceae, Fabaceae) and one monocot family (Poaceae). In Poaceae, only acyclotides have been found whereas in dicot families both cyclotides and acyclotides have been isolated. In last 15 years, several acyclotides with antimicrobial, cytotoxic and hemolytic bioactivities have been discovered. Thus, although many naturally expressed acyclotides do exhibit bioactivities, the linearization of the cyclic peptides may result in loss of bioactivities. Although, bioactivities of acyclotides are comparable to their cyclic counterparts, the numbers of isolated acyclotides are still few. Further, those discovered, have the scope to be screened for agriculturally important activities (insecticidal, anti-helminthic, molluscicidal) and pharmaceutical properties (anticancer, anti-HIV, immuno-stimulant). The feasibility of application of acyclotides is because of their relatively less complex biological synthesis compared to cyclotides, as the cyclization step is not needed. This attribute facilitates the production of transgenic crops and/or its expression in heterologous organisms, lacking cyclization machinery. Keeping in view the bioactivities and the wide array of emerging potential applications of acyclotides, the present review discusses their distribution in plants, gene and protein structure, biosynthesis, bioactivities and mechanism of action. Further, their potential applications and future perspectives to exploit them in agriculture and pharmaceutical industries have been highlighted.

Amino acid deprivation therapy (AADT) is emerging as a promising strategy for the development of novel therapeutics against cancer. This biological therapy relies upon the differences in the metabolism of cancer and normal cells. The rapid growth of tumors results in decreased expression of certain enzymes leading to auxotrophy for some specific amino acids. These auxotrophic tumors are targeted by amino acid–depleting enzymes. The depletion of amino acid selectively inhibits tumor growth as the normal cells can synthesize amino acids by their usual machinery. The enzymes used in AADT are mostly obtained from microbes for their easy availability. Microbial l -asparaginase is already approved by FDA for the treatment of acute lymphoblastic leukemia. Arginine deiminase and methionase are under clinical trials and the therapeutic potential of lysine oxidase, glutaminase and phenylalanine ammonia lyase is also being explored. The present review provides an overview of microbial amino acid depriving enzymes. Various attributes of these enzymes like structure, mode of action, production, formulations, and targeted cancers are discussed. The challenges faced and the combat strategies to establish AADT in standard cancer armamentarium are also reviewed. Key Points • Amino acid deprivation therapy is a potential therapy for auxotrophic tumors. • Microbial enzymes are used due to their ease of manipulation and high productivity. • Enzyme properties are improved by PEGylation, encapsulation, and genetic engineering. • AADT can be employed as combinational therapy for better containment of cancer.

Acyclotides are plant-based, acyclic miniproteins with cystine knot motif formed by three conserved disulfide linkages and lack head to tail ligation. Acyclotides may not necessarily be less stable, even though they lack cyclic backbone, as the conserved cystine knot feature provides the required stability. Violacin A was the first acyclotide, isolated from Viola odorata in 2006. Until now, acyclotides have been reported from five dicot families (Violaceae, Rubiaceae, Cucurbitaceae, Solanaceae, Fabaceae) and one monocot family (Poaceae). In Poaceae, only acyclotides have been found whereas in dicot families both cyclotides and acyclotides have been isolated. In last 15 years, several acyclotides with antimicrobial, cytotoxic and hemolytic bioactivities have been discovered. Thus, although many naturally expressed acyclotides do exhibit bioactivities, the linearization of the cyclic peptides may result in loss of bioactivities. Although, bioactivities of acyclotides are comparable to their cyclic counterparts, the numbers of isolated acyclotides are still few. Further, those discovered, have the scope to be screened for agriculturally important activities (insecticidal, anti-helminthic, molluscicidal) and pharmaceutical properties (anticancer, anti-HIV, immuno-stimulant). The feasibility of application of acyclotides is because of their relatively less complex biological synthesis compared to cyclotides, as the cyclization step is not needed. This attribute facilitates the production of transgenic crops and/or its expression in heterologous organisms, lacking cyclization machinery. Keeping in view the bioactivities and the wide array of emerging potential applications of acyclotides, the present review discusses their distribution in plants, gene and protein structure, biosynthesis, bioactivities and mechanism of action. Further, their potential applications and future perspectives to exploit them in agriculture and pharmaceutical industries have been highlighted.

Antibiotic-resistant genes (ARGs) are regarded as emerging environmental pollutants and pose a serious health risk to the human population. Integrons are genetic elements that are involved in the spread of ARGs amongst bacterial species. They also act as reservoirs of these resistance traits, further contributing to the development of multi-drug resistance in several water-borne pathogens. Due to inter- and intra-species transfer, integrons are now commonly reported in important water-borne pathogens such as Vibrio , Campylobacter , Salmonella , Shigella , Escherichia coli and other opportunistic pathogens. These pathogens exhibit immense diversity in their resistance gene cassettes. The evolution of multiple novel and complex gene cassettes in integrons further suggests the selection and horizontal transfer of ARGs in multi-drug resistant bacteria. Thus, the detection and characterization of these integrons in water-borne pathogens, especially in epidemic and pandemic strains, is of the utmost importance. It will provide a framework in which health authorities can conduct improved surveillance of antibiotic resistance in our natural water bodies. Such a study will also be helpful in developing better strategies for the containment and cure of infections caused by these bacteria.

In the present study a high arginine deiminase (ADI) yielding bacterium was isolated from soil samples of Haryana, India and identified as Pseudomonas furukawaii. The specific enzyme activity was optimized to 1.420 IU/ml by OFAT and further enhanced to 2.708 IU/ml (an increase of 90.7%) with the help of statistical parametric optimization approaches using GA-ANN and GA-ANFIS. The obtained value of the coefficient of correlation (R = 0.88) for ANN and epoch error (0.12) for ANFIS, indicates the prediction accuracy and strength of these data training models. ADI production was improved significantly in simple super broth media supplemented with 1.5% fructose and 1.75% arginine at pH 7 at 37 degrees C using multilevel algorithms and evolutionary hybrid tools. The native enzyme was partially purified (ten-fold) up to a specific enzyme activity of 29.559 IU/mg.

Introduction: Rise in incidence of various cancers and growing adoption of biological therapy to avoid side effects of conventional cancer therapies is driving the growth of the cancer biotherapy market globally. One such therapy available for the treatment of certain tumors employs arginine-lowering enzymes (ALEs). Several patents have been filed in this technology domain, and many Phase I/II clinical trials of the ALEs especially arginine deiminase (ADI) are underway. Areas covered: Patents and clinical trials in the domain of ALEs for the treatment of cancer were studied with an objective to understand technology trends, targeted areas, key players, and inventors involved. Expert opinion: Amongst the various ALEs, ADI is the most promising enzyme for cancer therapy. ADI-based cancer therapy holds potential in treating liver, skin, lung, gastrointestinal, and blood cancer. ADI-PEG20 has proved to be very effective when used as a component of combination therapy in a first-line treatment. Polaris Group holds the worldwide rights for ADI-PEG20 and is the leading player in developing ADI as a therapeutic agent. Many clinical studies, especially in a combinatorial approach, are underway whose success will pave the way for ADI-PEG to the multimillion cancer market.

The present study investigates the production kinetics of recombinant human tumor necrosis factor-alpha (TNF-α) in Escherichia coli under three different nonconventional inducible expression systems, namely, salt, the thermal, and auto-inducible system in batch culture in a bioreactor. A codon-optimized synthetic gene of human TNF-α was inserted into plasmid pET-14b under T7 promoter to obtain its plasmid-based expression in E. coli. Very high specific yield (~47% of total cellular protein (TCP)) was obtained with auto-inducible expression system. The thermal and salt-induced expressions could yield up to ~30% and ~17% of TCP, respectively. Maximum specific productivities were reported to be 0.092 g/gDCW/h, 0.1 g/gDCW/h, and 0.079 g/gDCW/h for auto-inducible, thermal-inducible, and salt-inducible expression systems, respectively.

Purpose: Conventional wastewater treatment technologies are not good enough to completely remove all endocrine disrupting compounds (EDCs) from the water. Membrane separation systems have emerged as an attractive alternative to conventional clarification processes for waste and drinking water. Coupling of a membrane separation process with an enzymatic reaction has opened up new avenues to further enhance the quality of water. This review article deliberates the feasibility of implementing enzymatic membrane reactors has been deliberated. Materials and methods: A comprehensive study of conventional water treatment technologies was carried out and their shortcomings were pointed out. Research findings from the leading groups working on enzyme grafted membrane based water purification were summarized. This review also comprehends the patent documents pertinent to the technology of enzyme grafted membranes for water purification. Results: Immobilization of an enzyme on a membrane improves the performance of membrane filtration, and processes for the treatment of polluted water. Research has started exploring the potential for laccase enzymes because it can catalyze the oxidation of a wide range of substrates, structurally comparable to EDCs, by a radical-catalyzed reaction mechanism, with corresponding reduction of oxygen to water in an electron transfer process. Further, in the presence of certain mediators, the substrate range of laccases can be further enhanced to non-aromatic substrates. Conclusions: Removal of EDCs by laccase cross-linked enzyme aggregates in fixed-bed reactors or fluidized-bed reactors and laccase immobilized ultrafiltration (LIUF) membranes are proving their worth in water purification technology. The major operational issues with the use of LIUF membranes are enzyme instability in real wastewater and membrane fouling. In view of the above-stated characteristics, laccases are considered as the most promising enzyme for a greener and less expensive water purification technology.

The contamination of surface waters with multidrug resistant (MDR) coliforms is a major public health concern in developing countries. This study was aimed to evaluate the occurrence of antibiotic resistance and role of integrons in the spread of resistance genes in Escherichia coli isolated from urban waters of river Yamuna. One hundred and forty-one strains of E. coli were isolated and assessed for antibiotic resistance wherein high resistance was observed for Cefazolin. Integrons (class 1 and class 2) were detected in 32% of the isolates. Variable region of class 1 integron carried different gene cassettes, namely. dfrA17-aadA5, dfrA12-orfF-aadA2, blaOXA-1-aadA1, and unusual phage tail tape measure protein. These integron-positive isolates were further characterized by phylogrouping, serotyping, and BOX-PCR typing. Phylogroup B2 was found to be the most prevalent. Pathogenic E. coli O157 was also reported. Majority of the isolates (54%; 7/13) carrying "dfrA17-aadA5" gene cassette were clustered predominantly into a single BOX-PCR type (B8), suggesting a genetic relatedness among the isolates. This study thus depicts very high incidence of multidrug resistance and class 1 integrons in surface water of India. The prevalence of integrons in aquatic E. coli correlated well with resistance to increasing number of antibiotic classes and multiple antibiotic resistance (MAR) index at various sites. Integron-positive MDR E. coli isolates were found to be serologically and genetically diverse suggesting major role of integrons in the emergence and dissemination of resistance traits in waterborne E. coli.