In silico multi-epitope Bunyumwera virus vaccine to target virus nucleocapsid N protein.

BACKGROUND: Bunyumwera virus can cause 82% mortality in humans currently with no vaccine or drugs for treatment. We described an in silico multi-epitope vaccine targeting Bunyumwera virus nucleocapsid N-protein and predicted B and T cell epitopes for immunogenicity, allergenicity, toxicity, and conservancy. For creating the most potent immunological response possible, docking epitopes with HLA alleles are chosen to screen them. The 3D vaccination was docked with the Toll-like receptor-8 using molecular dynamic simulations. To ensure production efficiency, the vaccine sequence was further cloned in silico in a plasmid pIB2 vector. For efficacy and safety, results must be supported in vitro and in vivo. RESULTS: The vaccine was cloned to enable expression and translation in a plasmid vector pIB2. It was expected to be antigenic, non-allergenic, and have a high binding affinity with TLR-8 in silico cloning. This multi-epitope vaccination may stimulate both innate and adaptive immunity. CONCLUSION: The vaccine developed in this work was based on the nucleocapsid N-protein of the Bunyumwera virus and was created using a reverse vaccinology method. Further experimental validation is required to assess the vaccine's therapeutic effectiveness and immunogenicity.

Quercetin mitigates the deoxynivalenol mycotoxin induced apoptosis in SH-SY5Y cells by modulating the oxidative stress mediators.

Deoxynivalenol (DON) is Fusarium mycotoxin that is frequently found in many cereal-based foods, and its ingestion has a deleterious impact on human health. In this investigation, we studied the mechanism of DON-induced neurotoxicity and followed by cytoprotective efficacy of quercetin (QUE) in contradiction of DON-induced neurotoxicity through assessing the oxidative stress and apoptotic demise in the human neuronal model, i.e. SH-SY5Y cells. DON diminished the proliferation of cells in the manner of dose and time-dependent as revealed by cell viability investigations, i.e. MTT and lactate dehydrogenase assays. Additional studies, such as intracellular reactive oxygen species (ROS), lipid peroxidation (LPO), mitochondrial membrane potential (MMP), DNA damage, cell cycle, and neuronal biomarkers (amino acid decarboxylase, tyrosine hydroxylase, and brain-derived neurotrophic factor) demonstrated that DON induces apoptotic demise in neuronal cells through oxidative stress intermediaries. On another hand, pre-treatment of neuronal cells with 1 mM of quercetin (QUE) showed decent viability upon exposure to 100 µM of DON. In detailed studies demonstrated that QUE (1 mM) pre-treated cells show strong attenuation efficiency against DON-induced ROS generation, LPO, MMP loss, DNA impairment, cell cycle arrest, and down-regulation of neuronal biomarkers. The consequences of the investigation concluded that QUE mitigates the DON-induced stress viz., decreased ROS production and LPO generation, upholding MMP and DNA integrity and regulation of neuronal biomarker gene expression in SH-SY5Y cells.

Phytoassisted synthesis of magnesium oxide nanoparticles from Pterocarpus marsupium rox.b heartwood extract and its biomedical applications.

BACKGROUND: Unlike chemical techniques, the combination of metal oxide nanoparticles utilizing plant concentrate is a promising choice. The purpose of this work was to synthesize magnesium oxide nanoparticles (MgO-NPs) utilizing heartwood aqueous extract of Pterocarpus marsupium. The heartwood extract of Pterocarpus marsupium is rich in polyphenolic compounds and flavonoids that can be used as a green source for large-scale, simple, and eco-friendly production of MgO-NPs. The phytoassisted synthesis of MgO is characterized by UV-Visible spectroscopy, X-ray diffraction (XRD), dynamic light scattering (DLS), Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM) with EDS (energy dispersive X-ray spectroscopy), and transmission electron microscopy (TEM). RESULTS: The formation of MgO-NPs is confirmed by a visual color change from colorless to dark brown and they displayed a wavelength of 310 nm in UV-Spectrophotometry analysis. The crystalline nature of the obtained biosynthesized nanoparticles are revealed by X-ray diffraction analysis. SEM results revealed the synthesized magnesium oxide nanoparticles formed by this cost-effective method are spherically shaped with an average size of < 20 nm. The presence of magnesium and oxygen were confirmed by the EDS data. TEM analysis proved the spherical shape of the nanoparticles with average particle size of 13.28 nm and SAED analysis confirms the crystalline nature of MgO-NPs. FT-IR investigation confirms the existence of the active compounds required to stabilize the magnesium oxide nanoparticles with hydroxyl and carboxyl and phenolic groups that act as reducing, stabilizing, and capping agent. All the nanoparticles vary in particle sizes between 15 and 25 nm and obtained a polydispersity index value of 0.248. The zeta-potential was measured and found to be - 2.9 mV. Further, MgO-NPs were tested for antibacterial action against Staphylococcus aureus (Gram-positive bacteria) and Escherichia coli (Gram-negative bacteria) by minimum inhibitory concentration technique were found to be potent against both the bacteria. The blended nanoparticles showed good antioxidant activity examined by the DPPH radical scavenging method, showed good anti-diabetic activity determined by alpha-amylase inhibitory activity, and displayed strong anti-inflammatory activity evaluated by the albumin denaturation method. CONCLUSIONS: The investigation reports the eco-friendly, cost-effective method for synthesizing magnesium oxide nanoparticles from Pterocarpus marsupium Rox.b heartwood extract with biomedical applications.

Multi-Biofunctional Properties of Phytofabricated Selenium Nanoparticles From Carica papaya Fruit Extract: Antioxidant, Antimicrobial, Antimycotoxin, Anticancer, and Biocompatibility.

The present study focused on phytofabrication of selenium nanoparticles (SeNPs) from Carica papaya extract and exploration of their multi-biofunctional features. Total phenolics and flavonoids of C. papaya fruit extract were determined as 23.30 ± 1.88 mg gallic acid equivalents and 19.21 ± 0.44 mg quercetin equivalents per gram, respectively, which suggested that C. papaya fruit extract could be a competitive reducing and stabilizing agent during phytofabrication of nanoparticles. UV-Vis and FTIR spectroscopy showed the formation of SeNPs from sodium selenite, which could be related to the reducing and stabilizing activities of C. papaya fruit extract. The SeNPs were found to be stable with a Zeta potential of -32 mV. The average hydrodynamic size of SeNPs was found as 159 nm by dynamic light scattering. The SeNPs showed a broader XRD pattern with no sharp Bragg's peaks and found to be amorphous. SEM showed that SeNPs were spherical in shape and EDX pattern showed that SeNPs were made up of Se (71.81%), C (11.41%), and O (14.88%). The HR-TEM picture showed that SeNPs were spherical in morphology and have a size range of 101-137 nm. The SeNPs exhibited potent antioxidant activity and their EC50 values (effective concentration required to inhibit 50% of radicals) were 45.65 ± 2.01 and 43.06 ± 3.80 µg/ml in DPPH and ABTS assays, respectively. The antimicrobial action of SeNPs was found as a broad spectrum and suppressed microbial pathogens in ascending order: fungi > Gram-positive bacteria > Gram-negative bacteria. The SeNPs have been demonstrated to reduce the growth and ochratoxin A (OTA) of mycotoxigenic Aspergillus ochraceus and Penicillium verrucosum at 40 µg/ml in broth culture, which is noteworthy. The SeNPs reduced cancer cell proliferation (RAW 264.7, Caco-2, MCF-7, and IMR-32) more preferentially than normal cells (Vero), found to be highly biocompatible. Lower doses of SeNPs (up to 50 µg/ml) were shown to be less toxic and did not cause death in Danio rerio (zebrafish) embryos, implying that lower doses of SeNPs could be beneficial for biological purposes. The present study concluded that phytofabricated SeNPs have multiple biofunctional properties, including antioxidant, antimicrobial, antimycotoxin, and anticancer activities, as well as high biocompatibility.

Selecting better diagnostic kits for diagnosis of malarial parasites at point of care.

Malaria is a fatal life-threatening parasitic infection and a leading cause of morbidity and mortality. The present study was aimed to evaluate simple, inexpensive, accurate, reliable, easily available better diagnostic for rapid detection of malaria at point of care (POC). The study includes 1403 samples collected from the patients, of which 1227 were clinically suspected cases and 176 from consecutive feverish patients. Among the suspected cases only 338 samples were confirmed positive and 889 samples were negative for Plasmodium species by PCR. All the 889 samples showed negative result for plasmodium species by microscopy, Malarial Ag rapid kits but only 867 samples were confirmed negative with malarial Ab rapid kits. Of the 338 PCR positive samples, 337 samples were confirmed positive by microscopy and Malarial Ag rapid kits, but only 284 samples were confirmed positive using malarial Ab rapid kits. Overall the microscopy and the malaria antigen-based lateral flow assay exhibited similar sensitivity, specificity, PPV, NPV and efficiency, respectively, whereas the PCR assay had 100% sensitivity, specificity, PPV, NPV and efficiency. But the evolutionary data for malaria antibody lateral flow assay has 92.81% sensitivity, 94.13% specificity, 84.02% PPV, 97.52% NPV and 93.80% efficiency. The developed Malaria pf/pv antigen and antibody field-deployable kits are simple, rapid, accurate, reliable, inexpensive, user friendly, POC. In addition the kits are highly sensitive and species-specific. The pf/pv antigen kit is found to be more accurate with 99.7% sensitivity and 100% specificity than to Malaria pf/pv antibody rapid kits. Of the two rapid kits developed, Malaria pf/pv antigen kit is found be more accurate with 99.7% sensitivity and 100% specificity than to Malaria pf/pv antibody rapid kits.

Green synthesis of silver and gold nanoparticles using Stemona tuberosa Lour and screening for their catalytic activity in the degradation of toxic chemicals.

In the present study, silver and gold nanoparticles (AgNPs and AuNPs) were green synthesised using the aqueous plant extract of Stemona tuberosa Lour. When plant extract was mixed with AgNO3 and HAuCl4 solutions in separate reactions, the amalgamated solutions turned deep reddish brown and dark purple in colour after 48 h indicating the formation of AgNPs and AuNPs. UV-Visible analysis of green synthesised AgNPs and AuNPs have shown absorption maximum at 443.85 nm and 539.72 respectively after 48 h. Energy dispersive X-ray spectroscopy (EDX) analysis confirmed the presence of pure silver in the green synthesised AgNPs and pure gold in the plant-mediated AuNPs. X-ray diffractometer (XRD) data revealed the face-centred cubic nature of AgNPs. Fluorescence transmission infrared (FTIR) spectrum has shown the characteristic peaks of different phytochemicals in the plant extract which acted as stabilising or capping agents of AgNPs. Scanning electron microscopy (SEM) analysis of AgNPs and AuNPs revealed that the nanoparticles are monodispersed. Transmission electron microscopy (TEM) studies revealed that AgNPs were mostly spherical with an average size of 25 nm whereas selected area electron diffraction (SAED) analysis confirmed their crystalline nature. Both AgNPs and AuNPs of S. tuberosa Lour have shown potential catalytic activity in the presence of sodium borohydride (NaBH4) in the degradation and removal of 4-nitrophenol, methylene blue, methyl orange and methyl red.

Development of a FRET-based fluorescence aptasensor for the detection of aflatoxin B1 in contaminated food grain samples.

The present study aimed to develop an aptamer-based FRET detection strategy for the specific and sensitive detection of AFB1 in contaminated food grains. The study comprises generation of ssDNA aptamers against AFB1 by whole-cell SELEX and their application in a FRET-based platform utilizing graphene oxide (GO) and quantum dots (QDs). The generated aptamers were characterized to determine their specificity and sensitivity using indirect ELISA where AFB1-OVA was used as a coating antigen. Among the aptamers generated, the ATB1 aptamer showed good reactivity and selectivity against AFB1. This aptamer was further characterized to determine its secondary structure and KD value, which was found to be 5.9 kcal mol-1. The characterized aptamers were conjugated onto Cd/Se quantum dots to develop a fluorimetric system for the detection of aflatoxin B1 using a graphene oxide platform. The presence of graphene oxide quenches the fluorescence ability of the quantum dots due to π-π stacking interactions between the aptamer and GO. Upon target addition, the aptamer forms a complex with aflatoxin B1 thereby restoring the fluorescence intensity. The developed assay shows a linear response from 0.002 µg µl-1 to 0.2 µg µl-1 with a detection limit of 0.004 µg µl-1 for the AFB1 standard toxin and showed no cross-reactivity with other closely related mycotoxins. To validate the reliability of the developed method, several field samples spiked with AFB1 were included in this study and the results obtained were cross verified using a standard commercial AFB1 kit. In conclusion, the developed method may find good utility in routine food testing laboratories for risk assessment of AFB1.

Isolation, identification, optimization, and metabolite profiling of Streptomyces sparsus VSM-30.

Deep sea sediment samples of Bay of Bengal (Visakhapatnam) have been analyzed for actinomycetes as an elite source to screen for the production of bioactive metabolites. The actinomycetes strain VSM-30 has an exciting bioactivity profile and was isolated during our systemic screening of marine actinomycetes. It was identified as Streptomyces sparsus based on morphological, physiological, biochemical, and molecular approaches. Response surface methodology regression analysis was carried out to fit the experimental data of each response by the second-order polynomial. The results have proven right interaction among process variables at optimized values of incubation time at 12 days, pH at 8, temperature at 30 °C, concentrations of starch at 1%, and tryptone at 1% and the data have been adequately fitted into the second-order polynomial models. Under these conditions, the responses (zones of inhibition) of plant pathogenic fungi Aspergillus niger, Aspergillus flavus, Fusarium oxysporum, Fusarium solani, and Penicillium citrinum were also matched with experimental and predicted results. Chemotypic analysis of ethyl acetate extract of the strain was done using LC-Q-TOF-MS revealed the presence of bioactive compounds including tryptophan dehydrobutyrine diketopiperazine, maculosin, 7-o-demethyl albocycline, albocycline M-2, and 7-o-demethoxy-7-oxo albocycline in a negative ion mode. The ethyl acetate extract of actinobacterium has been subjected to gas chromatography and mass spectroscopy (GC-MS) revealed the presence of diverse compounds such as dotriacontane, tetracosane 11-decyl-, diheptyl phthalate, 1-hexadecanesulfonyl chloride, L-alanyl-L-tryptophan, phthalic acid ethyl pentyl ester, 4-trifluoroacetoxyhexadecane, and 1H-imidazole 4,5-dihydro-2,4-dimethyl. Hence, the ethyl acetate extract of Streptomyces sparsus VSM-30 may have antibacterial, antifungal, and antioxidant activities due to the presence of secondary metabolites in ethyl acetate extract. The study also supports marine sediment samples of Bay of Bengal, a promising marine ecosystem remained to be explored for new bioactive compounds.

Bioactive metabolites produced by Streptomyces Cheonanensis VUK-A from Coringa mangrove sediments: isolation, structure elucidation and bioactivity.

The strain VUK-A was isolated from a sediment sample of the Coringa mangrove ecosystem was identified as Streptomyces cheonanensis based on morphological, physiological, biochemical and molecular properties. Chemical investigation of the secondary metabolites of the strain Streptomyces cheonanensis VUK-A has led to the segregation of two bioactive compounds, namely 2-Methyl butyl propyl phthalate (1) and Diethyl phthalate (2) using column chromatography. The chemical structure of the active compounds was established on the basis of spectroscopic analysis, including 1H NMR and 13C NMR spectroscopies, FTIR and EIMS. The antimicrobial activity of the bioactive compounds produced by the strain was tested against a wide variety of bacteria and fungi and expressed in terms of minimum inhibitory concentration. The compounds (1&2) were active against all the bacteria tested, and the best activity of compound 1 was recorded against Proteus vulgaris (4 µg/ml). Compounds (1&2) were active against dermatophytes and fungi but compound 1 displayed high antifungal activity against Candida albicans (8 µg/ml) and Fusarium solani (16 µg/ml) compared to standard antifungal agents. The cytotoxicity of the bioactive compound 1 was tested against MDA-MB-231, OAW-42, HeLa, and MCF-7 cell lines. The highest activity of 100 µM by compound 1 was recorded against HeLa cancer cell lines. In fact, this is the first report of 2-Methyl butyl propyl phthalate from the genus Streptomyces.

Isolation and biological evaluation of N-(4-aminocyclooctyl)-3, 5-dinitrobenzamide, a new semisynthetic derivative from the Mangrove-associated actinomycete Pseudonocardia endophytica VUK-10.

The present study was aimed to isolate novel bioactive compounds from actinomycetes species isolated from mangrove habitats. With this connection, Pseudonocardia endophytica (VUK-10) was isolated using dilution plate technique and was examined for its secondary metabolite profiling. After successive purification and spectroscopic characterization viz., FTIR, mass, NMR, DEPT, HMQC, HMBC, and COSY spectroscopy, two compounds were identified including a semi synthetic derivative N-(4-aminocyclooctyl)-3, 5-dinitrobenzamide (1), obtained from the precursor of novel natural product cyclooctane-1,4-diamine (3), along with a known compound 3-((1H-indol-6-yl) methyl) hexahydropyrrolo [1, 2-a] pyrazine-1, 4-dione (2). Anti cancer activities of the characterized compounds against in vitro cancerous cell line models, MDA-MB-231, OAW-42, HeLa, and MCF-7 reveal that HELA cells are most susceptible (IC50-10 nM compound 1 and 2) followed by other studied cells. On the other hand, antibacterial and antifungal activities of the studied compounds against tested pathogens revealed that there is a significant antimicrobial activity with all the tested bacterial and fungal species. Moreover, compound 1 showed the lowest MIC values against Streptococcus mutans as 4  and 16 µg/ml for Candida albicans. In conclusion, the identified novel chemical compounds in the present study may have a potential application in anticancer therapy as well as to mitigate the bacterial and fungal pathogens thus to control the infectious diseases.

Optimization of culture conditions by response surface methodology and unstructured kinetic modeling for bioactive metabolite production by Nocardiopsis litoralis VSM-8.

Response surface methodology-based central composite design on five variables incubation time, pH, temperature, sucrose concentration, and soya peptone concentration was employed for optimization of the production of bioactive compounds by Nocardiopsis litoralis strain VSM 8. The main quadratic effects and interactions of the five variables on the production of bioactive metabolites were investigated. A second-order polynomial model produced a satisfied fit for experimental data with regard to the production of the bioactive metabolites. Regression analysis showed that high R 2 values of all the five responses are significant and adjusted R 2 values showed good agreement with the experimental and predicted values. The present model was used to evaluate the direct interaction and quadratic effects to optimize the physico-chemical parameters for the production of bioactive metabolites that inhibit the pathogenic microorganisms measured in terms of zones of inhibition (responses). Mathematical kinetic model development and estimation of kinetic parameters also showed good approximation in terms of model fitting and regression analysis.

Bioactive natural products from Pseudonocardia endophytica VUK-10.

Two proline containing cyclic dipeptides (CDPs), cyclo (L-Pro-L-Tyr) (1) and cyclo (L-Pro-L-Phe) (2) were isolated from the fermentation broth of Pseudonocardia endophytica VUK-10 originating from the Nizampatnam mangrove ecosystem on the south coast of Andhra Pradesh, India. The structures of the compounds were established by 1H NMR and 13C NMR spectroscopy, FTIR and EIMS. The antimicrobial and cytotoxic activities of the compounds were tested against a variety of medicinally and agriculturally important bacteria and fungi as well as on the MDA-MB-231, OAW-42, HeLa and MCF-7 human cell lines. Xanthomonas malvacearum was most sensitive toward 1 (MIC 4 µg/ml), whereas compound 2 had good antibacterial activity against Xanthomonas campestris (MIC 8 µg/ml). Fusarium solani was highly sensitive toward 1 (MIC 16 µg/ml). The compounds were cytotoxic against the human cell lines at micro molar concentrations; the highest activity (IC50 < 10 µM) of 1 was recorded against the MDA-MB-231 cancer cell line.

Bioactive Metabolites Produced by Pseudonocardia endophytica VUK-10 from Mangrove Sediments: Isolation, Chemical Structure Determination and Bioactivity.

Chemical investigation of the actinobacterial isolate Pseudonocardia endophytica VUK-10 has led to the segregation of two known bioactive compounds, namely 4-(2-acetamidoethyl) phenyl acetate and 4-((1, 4-dioxooctahydropyrrolo [1, 2-a] pyrazin-3-yl) methyl) phenyl acetate. The strain was isolated from a sediment sample of the Nizampatnam mangrove ecosystem, south coastal Andhra Pradesh, India. The chemical structure of the active compounds was established on the basis of spectroscopic analysis, including (1)H NMR and (13)C NMR spectroscopies, FTIR, and EIMS. The antimicrobial and cytotoxic activities of the bioactive compounds produced by the strain were tested against opportunistic and pathogenic bacteria and fungi and on MDA-MB-231, OAW, HeLa, and MCF-7 cell lines. The compounds exhibited antimicrobial activities against gram-positive and gram-negative bacteria and fungi and also showed potent cytotoxic activity against MDA-MB-231, OAW, HeLa, and MCF-7 cell lines. This is the first example for this class of bioactive compounds isolated from Pseudonocardia of mangrove origin.

Production and optimization of L-asparaginase by an actinobacterium isolated from Nizampatnam mangrove ecosystem.

The aim of the present study was to isolate and screen actinomycetes from the mangrove sediments of Nizampatnam that are potent to produce L-asparaginase, an enzyme that catalyses the hydrolysis of asparagine. A total of 31 actinomycetes strains were isolated, of which 6 strains were positive for L-asparaginase. Several physico-chemical parameters were optimized for maximizing L-asparaginase production by the potent strain identified as Pseudonocardia endophytica VUK-10. Production of L-asparaginase by the strain was high in modified Asparagine glucose salts broth (FM-4)(3.96 IU/ml) as compared to other tested media. Maltose(6.99 IU ml(-1)) and L-asparagine (7.42 IU ml(-1)) were found to be the most suitable carbon and nitrogen sources for optimum enzyme production. Maximum production of L-asparaginase was found in the culture medium with pH 8 and temperature 30 degrees C incubated for four days. This is the first report on the production of L-asparaginase by Pseudonocardia endophytica VUK-10 from Nizampatnam mangrove sediments.

Molecular characterization of Mtb-OMP decarboxylase by modeling, docking and dynamic studies.

Tuberculosis (TB), the second most deadly disease in the world is caused by Mycobacterium tuberculosis (Mtb). In the present work a unique enzyme of Mtb orotidine 5' monophosphate decarboxylase (Mtb-OMP Decase) is selected as drug target due to its indispensible role in biosynthesis of pyrimidines. The present work is focused on understanding the structural and functional aspects of Mtb-OMP Decase at molecular level. Due to absence of crystal structure, the 3D structure of Mtb-OMP Decase was predicted by MODELLER9V7 using a known structural template 3L52. Energy minimization and refinement of the developed 3D model was carried out with Gromacs 3.2.1 and the optimized homology model was validated by PROCHECK,WHAT-IF and PROSA2003. Further, the surface active site amino acids were quantified by WHAT-IF pocket. The exact binding interactions of the ligands, 6-idiouridine 5' monophosphate and its designed analogues with the receptor Mtb-OMP Decase were predicted by docking analysis with AUTODOCK 4.0. This would be helpful in understanding the blockade mechanism of OMP Decase and provide a candidate lead for the discovery of Mtb-OMP Decase inhibitors, which may bring insights into outcome new therapy to treat drug resistant Mtb.

Studies on influence of natural biowastes on cellulase production by Aspergillus niger.

The objective of this study was to determine the influence of natural biowaste substrates such as banana peel powder and coir powder at varying environmental parameters of pH (4-9) and temperature (20-50 degrees C) on the cellulase enzyme production by Aspergillus niger. The cellulase enzyme production was analyzed by measuring the amount of glucose liberated in IU ml(-1) by using the dinitrosalicylic acid assay method. The substrates were pretreated with 1% NaOH (alkaline treatment) and autoclaved. The maximum activity of the enzyme was assayed at varying pH with temperatures being constant and varying temperatures with pH being constant. The highest activity of the enzyme at varying pH was recorded at pH 6 for banana peel powder (0.068 +/- 0.002 IU ml) and coir powder (0.049 +/- 0.002 IU ml(-1)) and the maximum activity of the enzyme at varying temperature was recorded at 35 degrees C for both banana peel powder (0.072 +/- 0.001 IU ml(-1)) and coir powder (0.046 +/- 0.003 IU ml(-1)). At varying temperatures and pH the high level of enzyme production was obtained at 35 degrees C and pH 6 by using both the substrates, respectively. However among the two substrates used for the production of cellulases by Aspergillus niger banana peel powder showed maximum enzymatic activity than coir powder as substrate.

Optimization of Culturing Conditions for Improved Production of Bioactive Metabolites by Pseudonocardia sp. VUK-10

The purpose of the present study was to investigate the influence of cultural and environmental parameters affecting the growth and bioactive metabolite production of the rare strain VUK-10 of actinomycete Pseudonocardia, which exhibits a broad spectrum of in vitro antimicrobial activity against bacteria and fungi. Production of bioactive metabolites by the strain was high the in modified yeast extract-malt extract-dextrose (ISP-2) broth, as compared to other tested media. Glucose (1%) and tryptone (0.25%) were found to be the most suitable carbon and nitrogen sources, respectively, for optimum production of growth and bioactive metabolites. Maximum production of bioactive metabolites was found in the culture medium with initial pH 7 incubated with the strain for four days at 30degrees C, under shaking conditions. This is the first report on the optimization of bioactive metabolites by Pseudonocardia sp. VUK-10.