Department of Pharmacy

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Evaluation of biphenyl- and polychlorinated-biphenyl (PCB) degrading Rhodococcus sp. MAPN-1 on growth of Morus alba by pot study
(Taylor & Francis, 2020-06) Paul, Atish Tulshiram; Jha, Prabhat Nath
This study focused on isolation of bacteria with biphenyl/polychlorinated biphenyl (PCB) degrading ability from the rhizosphere of Morus alba (mulberry plant). Repetitive enrichment of rhizospheric soil samples with biphenyl resulted in the isolation of Rhodococcus sp. MAPN-1, identified by 16S rRNA gene sequence analysis. The bacterium showed growth on five different aromatic compounds (naphthalene, salicylic acid, benzoic acid, dibenzofuran and anthracene). Benzoic acid was detected as the major metabolite during biphenyl degradation using high-performance thin-layer chromatography (HPTLC) with Rf 0.42 at 254 nm. Further GC-MS/MS study showed 95% and 15% degradation of biphenyl and dichlorobiphenyl, respectively. A pot study was conducted to evaluate the effect of presence of biphenyl on M. alba and the role of biphenyl degrader Rhodococcus sp. MAPN-1 in relation to phytoremediation. Morus alba twigs in biphenyl spiked soil (100 mg/kg and 300 mg/kg) inoculated with Rhodococcus sp. MAPN-1 showed growth, whereas, growth of plants (control) was adversely affected in biphenyl-spiked uninoculated soil. It is the first report of isolation of Rhodococcus sp. MAPN-1 from the rhizosphere of Morus alba, its capability to degrade biphenyl, thereby showing a positive effect on the plant growth grown in biphenyl spiked soil.
Integrated metabolomic, molecular, and morphological insights into the degradation of polychlorinated biphenyls (PCB) by priestia megaterium mapb-27
(ACS, 2025-10) Paul, Atish Tulshiram; Jha, Prabhat Nath
Polychlorinated biphenyls (PCBs) are persistent organic pollutants that cause profound deleterious effects on the environment and human health. Exposure to PCBs and biphenyl can induce changes in cellular metabolite levels. However, metabolic responses to utilize and adapt to PCBs are not well understood. Therefore, this study meticulously examined the PCB degradation potential, gene expression, and metabolic responses of Priestia megaterium MAPB-27 exposed to biphenyl. MAPB-27 showed growth and chemotaxis toward PCB degradation intermediates such as biphenyl, dihydroxy biphenyl, benzoate, and catechol. We employed GC-MS/MS to elucidate disparities in the main metabolic pathways in the biphenyl-exposed MAPB-27 through variations in metabolite composition and PCB biodegradation, while Field-emission scanning electron microscopy (FESEM) was used to study cell morphology. GC-MS/MS analysis highlighted the degradation of trichlorobiphenyl, tetrachlorobiphenyl, pentachlorobiphenyl, and hexachlorobiphenyl by P. megaterium MAPB-27, exhibiting 92.5, 62.9, 3.7, and 2.4%, respectively. GC-MS/MS analysis identified 4-dihydroxy-2-oxo-valerate, benzoic acid, and 2,3-dihydroxybenzoic acid as the major degradative metabolites in MAPB-27. MAPB-27 extract also contains metabolites with a wide range of direct industrial applications, such as poly(3-hydroxybutyrate) (3-hydroxybutyrate), a biobased organic acid (3-hydroxypropionoic acid), and antibacterial and antifungal compounds (phenyllactic acid, 4-hydroxyphenyllactic acid, and β-sitosterol). Glyoxylate and dicarboxylate metabolism and fatty acid biosynthesis were observed to be the active metabolisms in MAPB-27 grown in biphenyl-supplemented Minimal Medium. Overall, the results of this study provided important insights into microbial adaptation to biphenyl and the biodegradation of PCB. Thus, the P. megaterium MAPB-27 strain can be used for the development of efficient PCB biodegradation strategies and for the exploration of industrial applications.
Metagenomic analysis for taxonomic and functional potential of Polyaromatic hydrocarbons (PAHs) and Polychlorinated biphenyl (PCB) degrading bacterial communities in steel industrial soil
(PLOS, 2022-04) Paul, Atish Tulshiram; Jha, Prabhat Nath
Iron and steel industries are the major contributors to persistent organic pollutants (POPs). The microbial community present at such sites has the potential to remediate these contaminants. The present study highlights the metabolic potential of the resident bacterial community of PAHs and PCB contaminated soil nearby Bhilai steel plant, Chhattisgarh (India). The GC-MS/MS analysis of soil samples MGB-2 (sludge) and MGB-3 (dry soil) resulted in identification of different classes of POPs including PAHs {benzo[a]anthracene (nd; 17.69%), fluorene (15.89%, nd), pyrene (nd; 18.7%), benzo(b)fluoranthene (3.03%, nd), benzo(k)fluoranthene (11.29%; nd), perylene (5.23%; nd)} and PCBs (PCB-15, PCB-95, and PCB-136). Whole-genome metagenomic analysis by Oxford Nanopore GridION Technology revealed predominance of domain bacteria (97.4%; 97.5%) followed by eukaryote (1.4%; 1.5%), archaea (1.2%; 0.9%) and virus (0.02%; 0.04%) in MGB-2 and MGB-3 respectively. Proteobacteria (44.3%; 50.0%) to be the prominent phylum followed by Actinobacteria (22.1%; 19.5%) in MBG-2 and MBG-3, respectively. However, Eukaryota microbial communities showed a predominance of phylum Ascomycota (20.5%; 23.6%), Streptophyta (18.5%, 17.0%) and unclassified (derived from Eukaryota) (12.1%; 12.2%) in MGB-2 and MGB-3. The sample MGB-3 was richer in macronutrients (C, N, P), supporting high microbial diversity than MGB-2. The presence of reads for biphenyl degradation, dioxin degradation, PAH degradation pathways can be further correlated with the presence of PCB and PAH as detected in the MGB-2 and MGB-3 samples. Further, taxonomic vis-à-vis functional analysis identified Burkholderia, Bradyrhizobium, Mycobacterium, and Rhodopseudomonas as the keystone degrader of PAH and PCB. Overall, our results revealed the importance of metagenomic and physicochemical analysis of the contaminated site, which improves the understanding of metabolic potential and adaptation of bacteria growing under POP contaminated environments.
PCB-77 biodegradation potential of biosurfactant producing bacterial isolates recovered from contaminated soil
(Frontiers, 2022-09) Paul, Atish Tulshiram; Jha, Prabhat Nath
Polychlorinated biphenyls (PCBs) are persistent organic pollutants widely distributed in the environment and possess deleterious health effects. The main objective of the study was to obtain bacterial isolates from PCB-contaminated soil for enhanced biodegradation of PCB-77. Selective enrichment resulted in the isolation of 33 strains of PCB-contaminated soil nearby Bhilai steel plant, Chhattisgarh, India. Based on the prominent growth using biphenyl as the sole carbon source and the confirmation of its degradation by GC-MS/MS analysis, four isolates were selected for further study. The isolates identified by 16S rRNA gene sequencing were Pseudomonas aeruginosa MAPB-2, Pseudomonas plecoglossicida MAPB-6, Brucella anthropi MAPB-9, and Priestia megaterium MAPB-27. The isolate MAPB-9 showed a degradation of 66.15% biphenyl, while MAPB-2, MAPB-6, and MAPB-27 showed a degradation of 62.06, 57.02, and 56.55%, respectively in 48 h. Additionally, the degradation ability of these strains was enhanced with addition of co-metabolite glucose (0.2%) in the culture medium. Addition of glucose showed 100% degradation of biphenyl by MAPB-9, in 48 h, while MAPB-6, MAPB-2, and MAPB-27 showed 97.1, 67.5, and 53.3% degradation, respectively as analyzed by GC-MS/MS. Furthermore, in the presence of inducer, PCB-77 was found to be 59.89, 30.49, 27.19, and 4.43% degraded by MAPB-6, MAPB-9, MAPB-2, and MAPB-27, respectively in 7 d. The production of biosurfactants that aid in biodegradation process were observed in all the isolates. This was confirmed by ATR-FTIR analysis that showed the presence of major functional groups (CH2, CH3, CH, = CH2, C–O–C, C-O) of the biosurfactant. The biosurfactants were further identified by HPTLC and GC-MS/MS analysis. Present study is the first to report PCB-77 degradation potential of Pseudomonas aeruginosa, B. anthropi, Pseudomonas plecoglossicida, and Priestia megaterium. Similarly, this is the first report on Pseudomonas plecoglossicida and Priestia megaterium for PCB biodegradation. Our results suggest that the above isolates can be used for the biodegradation of biphenyl and PCB-77 in PCB-contaminated soil.
QbD-driven formulation development and evaluation of topical hydrogel containing ketoconazole loaded cubosomes
(Elsevier, 2021-02) Jha, Prabhat Nath; Singhvi, Gautam
The proposed study aimed to develop topical hydrogel containing ketoconazole loaded cubosomes with lower surfactant concentrations using the ‘Quality by Design’ (QbD) approach. Risk assessment was performed, followed by screening and optimization of formulations by 32 factorial design using Design-Expert® software. Keeping the combination of constituents similar to that of the optimized batches as predicted post conduct of ‘Design of Experiment’ (DoE) studies, scale-up batches were prepared. The 32 factorial design model successfully predicted the composition of the optimized formulation within the confidence limits. In vitro drug release study was performed and analyzed by various mathematical models. Ex vivo permeation study was investigated using goat ear skin. These ketoconazole loaded cubosomes showed a release pattern similar to the Korsmeyer-Peppas model experiencing Fickian diffusion having 67% cumulative ketoconazole release within 24 h. Ex vivo permeation study of hydrogel containing ketoconazole loaded cubosomes revealed a sustained release pattern through the goat ear skin with around 92.73 % release within 24 h. Scale-up studies also gave the confirmatory results for the post characterization studies, whereby the particle size of ketoconazole loaded cubosomes was 198 nm with 45% ketoconazole entrapment efficiency. This hydrogel containing ketoconazole loaded cubosomes can be used for topical drug delivery.
Unfolding the Potential of Pyrrole- and Indole-Based Allylidene Hydrazine Carboximidamides as Antimicrobial Agents
(ACS, 2025-01) Jadhav, Hemant R.; Jha, Prabhat Nath
Antimicrobial drug resistance is a significant global health challenge, causing hundreds of thousands of deaths annually and severely impacting healthcare systems worldwide. Several reported antimicrobial compounds have a guanidine motif, as the positive charge on guanidine promotes cell lysis. Therefore, pyrrole- and indole-based allylidene hydrazine carboximidamide derivatives with guanidine motifs are proposed as antimicrobial agents that mimic cationic antimicrobial peptides (CAMPs). A total of 72 derivatives having pyrrol-2-yl-phenyl allylidene hydrazine carboximidamide and indol-3-yl-phenyl allylidene hydrazine carboximidamide scaffolds were assessed for their inhibitory potential against a panel of Gram-positive and Gram-negative bacteria. Analogs 1j, 1k, 1s, 2j, 2q, 4a, 4c, 4h, 5b, 6a, and 6d exhibited potent broad-spectrum antimicrobial activity better than the standard antibiotics. Also, these compounds showed no cytotoxicity up to 3-fold of the minimum inhibitory concentration, and structure–activity relationship was established. Further, the most active compound, 6a, showed a strong biofilm disruption, acted on the bacterial membrane, and lysed it. The further development of these compounds as novel antimicrobial agents is warranted.