Strengthening microbial cell factories for efficient production of bioactive molecules.

High demand of bioactive molecules (food additives, antibiotics, plant growth enhancers, cosmetics, pigments and other commercial products) is the prime need for the betterment of human life where the applicability of the synthetic chemical product is on the saturation due to associated toxicity and ornamentations. It has been noticed that the discovery and productivity of such molecules in natural scenarios are limited due to low cellular yields as well as less optimized conventional methods. In this respect, microbial cell factories timely fulfilling the requirement of synthesizing bioactive molecules by improving production yield and screening more promising structural homologues of the native molecule. Where the robustness of the microbial host can be potentially achieved by taking advantage of cell engineering approaches such as tuning functional and adjustable factors, metabolic balancing, adapting cellular transcription machinery, applying high throughput OMICs tools, stability of genotype/phenotype, organelle optimizations, genome editing (CRISPER/Cas mediated system) and also by developing accurate model systems via machine-learning tools. In this article, we provide an overview from traditional to recent trends and the application of newly developed technologies, for strengthening the systemic approaches and providing future directions for enhancing the robustness of microbial cell factories to speed up the production of biomolecules for commercial purposes.

Onosma L. as a source of anticancer agents: phytochemistry to mechanistic insight.

Onosma (O.) is a genus of perennial flowering plants in the family Boraginaceae with approximately 250 species widely dispersed in temperate, tropical, and subtropical areas. It is traditionally used to treat rheumatism, fever, asthma, stomach irritation, and inflammatory ailments. The bioactive constituents present in the genus O. include benzoquinones, naphthazarins, alkaloids, phenolic, naphthoquinones, and flavonoids whereas shikonins and onosmins are the most significant. The review compiled contemporary research on O. L., including its distribution, morphology, traditional applications, phytochemistry, ethnopharmacology, and toxicology. This review also highlights a few critical challenges and possible future directions for O. L. research. Modern research has demonstrated a wide range of pharmacological effects of different species of O. L., including anti-diabetic, anticancer, anti-inflammatory, and cardiovascular protective. However, the studies on the O. genus are still not fully explored, therefore, researchers need to discover novel products with their toxicity studies, molecular mechanism, and associated side effects. Future exploration of potent constituents from this genus and clinical trials are required to explore its pharmacological importance.

Recent advances in electrochemical-based sensors amplified with carbon-based nanomaterials (CNMs) for sensing pharmaceutical and food pollutants.

Foodborne-related infections due to additives and pollutants pose a considerable task for food processing enterprises. Therefore, the competent, cost-effective, and quick investigation of nutrition additives and contaminants is essential to reduce the threat of public fitness problems. The electrochemical sensor (ECS) shows facile and potent analytical approaches desirable for food protection and quality inspection over traditional methods. The consequence of a broad display of nanomaterials has paved the path for their relevance in designing high-performance ECSs appliances for medical diagnostics and conditions and food protection. This review article has discussed the importance of electrochemical-based sensors amplified with carbon-based nanomaterials (CNMs). Initially, we have demonstrated the types of pharmaceutical and food/agriculture pollutants (such as pesticides, heavy metals, antibiotics and other medical drugs) present in water. Subsequently, we have compiled the information on electrochemical techniques (such as voltammetric and electrochemical impedance spectroscopy) and their crucial parameters for detecting pollutants. Further, the applications of CNMs for sensing pharmaceutical and food pollutants have been demonstrated in detail. Finally, the topic has been concluded with existing challenges and future prospects.

Recent advances of carbon-based nanomaterials (CBNMs) for wastewater treatment: Synthesis and application.

Carbon-based nanomaterials (CBNMs) have attracted significant alert due to the affluent science underpinning their implementations associated with a novel mixture of high aspect proportions, greater thermal and electrical performance, outstanding optical features, and high exterior area. CBNMs not only bear assurance in a broad range of implementations in medication, nano and microelectronics, and ecological remedies but may also be utilized in practical laboratory determinations. More specifically, CBNMs perform as an outstanding adsorbent in terminating heavy metal ions (HMI) from wastewater. There is presently a deficiency of powerful threat inspection instruments owing to their complex detection and related deficit in the health risk database. Therefore, our present review concentrates on spreading CBNMs to release pollutants from wastewater. The article wraps the effect of these contaminants and photocatalytic strategies towards treating these mixtures in wastewater, along with their restrictions and challenges, convincing resolutions, and possibilities of these approaches.

Recent developments in microbial degradation of polypropylene: Integrated approaches towards a sustainable environment.

Fossil-fuel-based plastics have many enticing properties, but their production has resulted in significant environmental issues that require immediate attention. Despite the fact that these polymers are manmade, some bacteria can degrade and metabolise them, suggesting that biotechnologies based on the principle of plastic biodegradation could be beneficial. Among different types of plastics, polypropylene (PP), either having low or high density, is one of the most consumed plastics (18.85%). Their debasement under natural conditions is somewhat tricky. Still, their debasement under natural conditions is rather difficult slightly. However, different scientists have still made efforts by employing other microbes such as bacteria, fungi, and guts bacteria of larvae of insects to bio-deteriorate the PP plastic. Pre-irradiation techniques (ultraviolet and gamma irradiations), compatibilizers, and bio-additives (natural fibers, starch, and polylactic acid) have been found to impact percent bio-deterioration of different PP derivatives stronglythe various. The fungal and bacterial study showed that PP macro/microplastic might serve as an energy source and sole carbon during bio-degradation. Generally, gravimetric method or physical characterization techniques such as FTIR, XRD, SEM, etc., are utilized to affirm the bio-degradation of PP plastics-based materials. However, these techniques are not enough to warrant the bio-deterioration of PP. In this regard, a new technique approach that measures the amount of carbon dioxide emitted during bacterial or fungus degradation has also been discussed. In addition, further exploration is needed on novel isolates from plastisphere environments, sub-atomic strategies to describe plastic-debasing microorganisms and improve enzymatic action strategies, and omics-based innovations to speed up plastic waste bio-deterioration.

Role of Silver Nanoparticle-Doped 2-Aminodiphenylamine Polymeric Material in the Detection of Dopamine (DA) with Uric Acid Interference.

A viable electrochemical approach for the detection of dopamine (DA) in uric acid (UA) utilizing a silver nanoparticle-doped 2-aminodiphenylamine (AgNPs-2ADPA) electrode was invented. The electrochemical performance of DA showed that the incorporated electrode displayed outstanding electrocatalytic performance to the electrochemical oxidation of DA. In our study, the AgNPs-2ADPA exhibits remarkable catalytic activity, retaining high current value and resilience when employed as a working electrode component for electrocatalytic detection of DA. We have also utilized the bare and polymeric-2ADPA in DA detection for a comparison study. This method offers a facile route with extraordinary sensitivity, selectivity, and strength for the voltammetric detection of DA, even in the presence of UA and ascorbic acid (AA) as interferents, that can be employed for pharmaceutical and biological specimens.

Novel synthesis methods and applications of MXene-based nanomaterials (MBNs) for hazardous pollutants degradation: Future perspectives.

MXenes are a quickly growing and extended group of two-dimensional (2D) substances that have earned unbelievable analysis credits for various application areas within different manufacturing areas. Due to novel essential architectural and physicochemical properties shows good properties, such as elevated exterior area, living adaptability, strong electrochemistry, and great hydrophilicity. Given the fast progress within the structure and synthesis of MBNs for water treatment, quick updates on this research field are required to remove toxic substances, such as production approaches and characterization methods for the advantages and constraints of MXenes for pollutant degradation. MXenes are determined as a proposed road toward atmosphere-clean-up machinery to identify and decrease a pattern of hazardous resistant pollutants from environmental forms. Here, in this review article, we have been focused on describing the overview, novel synthesis methods, and characteristics of the MXene-based nanomaterials (MBNs) in the field for removing hazardous contaminants from environmental conditions. In the last, the utilizations of MBNs in water sanitization, organic solvent filtration, antibiotics degradation, pesticide degradation, heavy metals degradation, ions removal, bacterial pathogens degradation, along with the conclusion, challenges, and prospects in this field, have been discussed.

Key ingredients and recycling strategy of personal protective equipment (PPE): Towards sustainable solution for the COVID-19 like pandemics.

The COVID-19 pandemic has intensified the complications of plastic trash management and disposal. The current situation of living in fear of transmission of the COVID-19 virus has further transformed our behavioural models, such as regularly using personal protective equipment (PPE) kits and single-use applications for day to day needs etc. It has been estimated that with the passage of the coronavirus epidemic every month, there is expected use of 200 billion pieces of single-use facemasks and gloves. PPE are well established now as life-saving items for medicinal specialists to stay safe through the COVID-19 pandemic. Different processes such as glycolysis, hydrogenation, aminolysis, hydrolysis, pyrolysis, and gasification are now working on finding advanced technologies to transfer waste PPE into value-added products. Here, in this article, we have discussed the recycling strategies of PPE, important components (such as medical gloves, gowns, masks & respirators and other face and eye protection) and the raw materials used in PPE kits. Further, the value addition methods to recycling the PPE kits, chemical & apparatus used in recycling and recycling components into value-added products. Finally, the biorenewable materials in PPE for textiles components have been discussed along with concluded remarks.

Synthesis and selection of de novo proteins that bind and impede cellular functions of an essential mycobacterial protein.

Recent advances in nonrational and part-rational approaches to de novo peptide/protein design have shown increasing potential for development of novel peptides and proteins of therapeutic use. We demonstrated earlier the usefulness of one such approach recently developed by us, called "codon shuffling," in creating stand-alone de novo protein libraries from which bioactive proteins could be isolated. Here, we report the synthesis and selection of codon-shuffled de novo proteins that bind to a selected Mycobacterium tuberculosis protein target, the histone-like protein HupB, believed to be essential for mycobacterial growth. Using a versatile bacterial two-hybrid system that entailed utilization of HupB and various codon-shuffled protein libraries as bait and prey, respectively, we were able to identify proteins that bound strongly to HupB. The observed interaction was also confirmed using an in vitro assay. One of the protein binders was expressed in Mycobacterium smegmatis and was shown to appreciably affect growth in the exponential phase, a period wherein HupB is selectively expressed. Furthermore, the transcription profile of hupB gene showed a significant reduction in the transcript quantity in mycobacterial strains expressing the protein binder. Electron microscopy of the affected mycobacteria elaborated on the extent of cell damage and hinted towards a cell division malfunction. It is our belief that a closer inspection of the obtained de novo proteins may bring about the generation of small-molecule analogs, peptidomimetics, or indeed the proteins themselves as realistic leads for drug candidates. Furthermore, our strategy is adaptable for large-scale targeting of the essential protein pool of Mycobacterium tuberculosis and other pathogens.

Nucleoside diphosphate kinase from Mycobacterium tuberculosis cleaves single strand DNA within the human c-myc promoter in an enzyme-catalyzed reaction.

The reason for secretion of nucleoside diphosphate kinase (NdK), an enzyme involved in maintaining the cellular pool of nucleoside triphosphates in both prokaryotes and eukaryotes, by Mycobacterium tuberculosis is intriguing. We recently observed that NdK from M.tuberculosis (mNdK) localizes within nuclei of HeLa and COS-1 cells and also nicks chromosomal DNA in situ (A. K. Saini, K. Maithal, P. Chand, S. Chowdhury, R. Vohra, A. Goyal, G. P. Dubey, P. Chopra, R. Chandra, A. K. Tyagi, Y. Singh and V. Tandon (2004) J. Biol. Chem., 279, 50142-50149). In the current study, using a molecular beacon approach, we demonstrate that the mNdK catalyzes the cleavage of single strand DNA. It displays Michaelis-Menten kinetics with a kcat/K(M) of 9.65 (+/-0.88) x 10(6) M(-1) s(-1). High affinity (K(d) approximately K(M) of approximately 66 nM) and sequence-specific binding to the sense strand of the nuclease hypersensitive region in the c-myc promoter was observed. This is the first study demonstrating that the cleavage reaction is also enzyme-catalyzed in addition to the enzymatic kinase activity of multifunctional NdK. Using our approach, we demonstrate that GDP competitively inhibits the nuclease activity with a K(I) of approximately 1.9 mM. Recent evidence implicates mNdK as a potent virulence factor in tuberculosis owing to its DNase-like activity. In this context, our results demonstrate a molecular mechanism that could be the basis for assessing in situ DNA damage by secretory mNdK.

Role of Mycobacterium tuberculosis Ser/Thr kinase PknF: implications in glucose transport and cell division.

Protein kinases have a diverse array of functions in bacterial physiology, with a distinct role in the regulation of development, stress responses, and pathogenicity. pknF, one of the 11 kinases of Mycobacterium tuberculosis, encodes an autophosphorylating, transmembrane serine/threonine protein kinase, which is absent in the fast-growing, nonpathogenic Mycobacterium smegmatis. Herein, we investigate the physiological role of PknF using an antisense strategy with M. tuberculosis and expressing PknF and its kinase mutant (K41M) in M. smegmatis. Expression of PknF in M. smegmatis led to reduction in the growth rate and shortening and swelling of cells with constrictions. Interestingly, an antisense strain of M. tuberculosis expressing a low level of PknF displayed fast growth and a deformed cell morphology compared to the wild-type strain. Electron microscopy showed that most of the cells of the antisense strain were of a smaller size with an aberrant septum. Furthermore, nutrient transport analysis of these strains was conducted using 3H-labeled and 14C-labeled substrates. A significant increase in the uptake of D-glucose but not of glycerol, leucine, or oleic acid was observed in the antisense strain compared to the wild-type strain. The results suggest that PknF plays a direct/indirect role in the regulation of glucose transport, cell growth, and septum formation in M. tuberculosis.

Nuclear localization and in situ DNA damage by Mycobacterium tuberculosis nucleoside-diphosphate kinase.

Nucleoside-diphosphate kinase of Mycobacterium tuberculosis (mNdK) is a secretory protein, but the rationale behind secreting an enzyme involved in the maintenance of cellular pool of nucleoside triphosphates is not clearly understood. To elucidate the biological significance of mNdK secretion, we expressed mNdK fused to green fluorescent protein in HeLa and COS-1 cells. Interestingly, mNdK was detected in the nuclei of HeLa and COS-1 cells. Incubation of mNdK with nuclei isolated from HeLa and COS-1 cells led to in situ damage of chromosomal DNA. Surface plasmon resonance studies demonstrated that mNdK binds supercoiled plasmid DNA lacking apurinic/apyrimidinic sites with a dissociation constant of 30 +/- 3.2 mum. Plasmid cleavage by mNdK was found to be dependent on the specific divalent metal ion and inhibited by a metal ion chelator. Moreover, the metal ion-dependent DNA cleavage by mNdK was mediated by superoxide radicals as detected by electron paramagnetic resonance. The cleavage reaction was inhibited under nitrogen atmosphere confirming the necessity of molecular oxygen for DNA cleavage. In view of the findings that mNdK is secreted by intracellular mycobacteria and damages the nuclear DNA, it can be postulated that mNdK may cause cell death that could help in the dissemination of the pathogen.