Bharti P. Dave

Enhanced chrysene degradation by halotolerant Achromobacter xylosoxidans using Response Surface Methodology

Degradation of chrysene, a four ring High Molecular Weight (HMW) Polycyclic Aromatic Hydrocarbon (PAH) is of intense environmental interest, being carcinogenic, teratogenic and mutagenic. Multiple PAH degrading halotolerant Achromobacter xylosoxidans was isolated from crude oil polluted saline site. Response Surface Methodology (RSM) using Central Composite Design (CCD) of Bushnell-Haas medium components was successfully employed for optimization resulting 40.79% chrysene degradation on 4th day. The interactions between variables as chrysene and glucose concentrations, pH and inoculum size on degradation were examined by RSM. Under optimum conditions, A. xylosoxidans exhibited 85.96% chrysene degradation on 5th day. The optimum values predicted by RSM were confirmed through confirmatory experiments. It was also noted that pH and glucose as co-substrate play a dynamic role in enhancement of chrysene degradation. Hence, A. xylosoxidans can be further used for subsequent microcosm and in situ experiments for its potential to remediate PAH contaminated saline and non-saline soils.

Enhanced biodegradation of total polycyclic aromatic hydrocarbons (TPAHs) by marine halotolerant Achromobacter xylosoxidans using Triton X-100 and β-cyclodextrin--a microcosm approach.

Ability of Achromobacter xylosoxidans, a chrysene degrading marine halotolerant bacterium to degrade polycyclic aromatic hydrocarbons (PAHs) using a cost effective laboratory microcosm approach, was investigated. Effect of variables as chrysene, glucose as a co-substrate, Triton X-100 as a non-ionic surfactant and β-cyclodextrin as a PAHs solubilizer was examined on degradation of low molecular weight (LMW) and high molecular weight (HMW) PAHs. A total of eleven PAHs detected from polluted saline soil were found to be degraded. Glucose, in combination with Triton X-100 and β-cyclodextrin resulted in 2.8 and 1.4-fold increase in degradation of LMW PAHs and 7.59 and 2.23-fold increase in degradation of HMW PAHs, respectively. Enhanced biodegradation of total PAHs (TPAHs) by amendments with Triton X-100 and β-cyclodextrin using Achromobacter xylosoxidans can prove to be promising approach for in situ bioremediation of marine sites contaminated with PAHs.

Application of response surface methodology for rapid chrysene biodegradation by newly isolated marine-derived fungus Cochliobolus lunatus strain CHR4D

For the first time, Cochliobolus lunatus strain CHR4D, a marine-derived ascomycete fungus isolated from historically contaminated crude oil polluted shoreline of Alang-Sosiya ship-breaking yard, at Bhavnagar coast, Gujarat has been reported showing the rapid and enhanced biodegradation of chrysene, a four ringed high molecular weight (HMW) polycyclic aromatic hydrocarbon (PAH). Mineral Salt Broth (MSB) components such as ammonium tartrate and glucose along with chrysene, pH and trace metal solution have been successfully optimized by Response Surface Methodology (RSM) using central composite design (CCD). A validated, two-step optimization protocol has yielded a substantial 93.10% chrysene degradation on the 4th day, against unoptimized 56.37% degradation on the 14th day. The results depict 1.65 fold increase in chrysene degradation and 1.40 fold increase in biomass with a considerable decrement in time. Based on the successful laboratory experiments, C. lunatus strain CHR4D can thus be predicted as a potential candidate for mycoremediation of HMW PAHs impacted environments.

Synthesis and antimicrobial activity of novel quinoline derivatives bearing pyrazoline and pyridine analogues

The present investigation is in the interest of some synthesized novel derivatives containing (5-(2-chloroquinolin-3-yl)-3-(aryl)-4,5-dihydro-1H-pyrazol-1-yl)(pyridin-4-yl)methanones (4a–o) moieties incorporated with different biological active heterocycles such as quinoline, pyrazoline and pyridine derivatives. For the determination of the compounds reported in this paper was based on IR, 1H NMR, 13C NMR and mass spectral data and same compounds were screened for their antibacterial and antifungal activity on four bacteria (Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa) and three fungi (Candida albicans, Aspergillus niger, Aspergillus clavatus) using ampicillin and griseofulvin as the standard drugs. Cytotoxicity study was carried out using MTT colorimetric assay (HeLa cell line). Among the screened compounds, 4e, 4f and 4n showed most potent antibacterial activity, while compounds 4d and 4g emerged as the most active against fungal strains. The results demonstrated that compound 4o was remarkably active against all microbial strains. From the viewpoint of SAR studies, it was observed that the presence of electron withdrawing groups remarkably enhanced the antimicrobial activity of synthesized compounds. Additionally, preliminary MTT cytotoxicity studies on HeLa cells suggested that effective antimicrobial activity of 4e–g, 4n and 4o was accompanied by low cytotoxicity.

Polycyclic aromatic hydrocarbons (PAHs) at the Gulf of Kutch, Gujarat, India: Occurrence, source apportionment, and toxicity of PAHs as an emerging issue

The present study extrapolates the assessment and characterization of a barely studied region, the Gulf of Kutch, (near Jamnagar), Gujarat, India, in terms of PAH exposure, adverse effects caused by them, and various toxicological indices showing the catastrophic effects of their elevated concentrations. ΣPAH concentration in the site ranged from 118,280 to 1,099,410ngg-1 dw, with a predominance of 2-3-ring PAHs (79.09%) as compared to 4-5- and 6-ring PAHs (20.91%). The concentrations of carcinogenic PAHs were found to be between 8120 and 160,000ngg-1 dw, with a mean of 63,810ngg-1 dw, which is much higher than normal acceptable values. The toxic equivalent quotient for 6CPAHs ranged from 150.47 to 26,330ngg-1 BaPeq, encompassing 50.63% of ΣPAH toxicity. This toxicological profile of the present study site would be of paramount importance as it offers fresh information regarding the load of legacy pollutants such as PAHs and the inputs and methods to cope with their extremely high concentrations in less explored marine habitats.

Rhodovulum algae sp. nov., isolated from an algal mat.

A reddish-brown-pigmented, phototrophic bacterium, designated strain JA877T, was isolated from a brown algae mat sample collected from Jalandhar beach, Gujarat, India. On the basis of the 16S rRNA gene sequence, strain JA877T belongs to the class Alphaproteobacteria and is closely related to the type strains Rhodovulum viride JA756T (99.0 %), Rhodovulum sulfidophilum Hansen W4T (98.9 %), Rhodovulumvisakhapatnamense JA181T (98.8 %),Rhodovulum kholense JA297T (97.5 %) and Rhodovulum salis JA746T (97.0). However, strain JA877T showed only 20-45 % relatedness with its phylogenetic neighbours and had a ∆Tm between 5.8 and 7.0 °C. The major respiratory quinone was ubiquinone-10 (Q10), and the polar lipid profile was composed of the major components phosphatidylglycerol, phosphatidylethanolamine, an unidentified phospholipid, two unidentified sulfolipids and five unidentified lipids. The major fatty acids were C18 : 1ω5c, C18 : 1ω7c/C18 : 1ω6c, C16 : 0 and C18 : 0. The DNA G+C content was 64.5 mol%. On the basis of 16S rRNA gene sequence analysis, physiological data, and chemotaxonomic and molecular differences, strain JA877T is significantly different from other species of the genus Rhodovulum and represents a novel species, for which the name Rhodovulum algae sp. nov. is proposed. The type strain is JA877T (=LMG 29228T= KCTC 42963T).

Characterization of L-asparaginase from marine-derived Aspergillus niger AKV-MKBU, its antiproliferative activity and bench scale production using industrial waste

L-asparaginase (LA), an enzyme with anticancer activities, produced by marine-derived Aspergillus niger was subjected to purification and characterization. The purified enzyme was observed to have molecular weight ∼90KDa. The enzyme retained activity over a wide range of pH, i.e. pH 4-10. The enzyme was quite stable in temperature range 20-40°C. Tween 80 and Triton X-100 were observed to enhance LA activity while inhibition of LA activity was observed in presence of heavy metals. The values for Km was found to be 0.8141 mM and Vmax was 6.228μM/mg/min. The enzyme exhibited noteworthy antiproliferative activity against various cancer cell lines tested. Successful bench scale production (in 5L bioreacator) of LA using groundnut oil cake as low cost substrate has also been carried out.

Isolation and Investigation of Biodegradation Potential of Multiple Polycyclic Aromatic Hydrocarbons (PAHs) Degrading Marine Bacteria near Bhavnagar Coast, India

Present work deals with modified isolation methods for indigenous microorganisms with a capability to use both low molecular weight (LMW) and high molecular weight (HMW)PAHs, which are pervasive recalcitrant pollutants. Methods such as biphasic enrichment, and specific isolation methods has resulted in the isolation of organisms such as Sphingomonas, Pseudomonas, Mycobacterium, Achromobacter, and Streptomyces species with efficacy to degrade majority of LMW PAHs up to 85% after four days of experiments, which is substantially rapid rate of degradation attributed by microorganisms. Moreover, the organisms had shown up to 30% degradation of HMW PAHs within the same time frame, making the isolation strategies more credible. The study thus, holds prime importance of conquering the difficulties in the isolation of multiple hydrocarbons degrading microorganisms, which can be further applied for the successful application for bioremediation of hydrocarbon impacted environments.

Description of Rhodobacter azollae sp. nov. and Rhodobacter lacus sp. nov.

Three strains (JA826T, JA912T and JA913), which were yellowish brown colour, rod to oval shaped, Gram-stain-negative, motile, phototrophic bacteria with a vesicular architecture of intracytoplasmic membranes, were isolated from different pond samples. The DNA G+C content of the three strains was between 64.6 and 65.5 mol%. The highest 16S rRNA gene sequence similarity of all three strains was with the type strains of the genus Rhodobacter sensu stricto in the family Rhodobacteraceae. Strain JA826T had highest sequence similarity with Rhodobacter maris JA276T (98.5 %), Rhodobacter viridis JA737T (97.5 %) and other members of the genus Rhodobacter (<97 %). Strain JA912T had highest sequence similarity with Rhodobacter viridis JA737T (99.6 %), Rhodobacter sediminis N1T (99.3 %), Rhodobacter capsulatus ATCC 11166T (98.8 %) and less than 97 % similarity with other members of the genus Rhodobacter. The 16S rRNA gene sequence similarity between strains JA826T and JA912T was 96.9 %. DNA-DNA hybridization showed that strains JA826T and JA912T (values among themselves and between the type strains of nearest members <44 %) did not belong to any of the nearest species of the genus Rhodobacter. However, strains JA912T and JA913 were closely related (DNA-DNA hybridization value >90 %). The genomic distinction was also supported by differences in phenotypic and chemotaxonomic characteristics in order to propose strains JA826T (=KCTC 15478T=LMG 28758T) and JA912T (=KCTC 15475T=LMG 28748T) as new species in the genus Rhodobacter sensu stricto with the names Rhodobacter lacus and Rhodobacter azollae, respectively.

Marine-Derived Fungi: Potential Candidates for Fungal Nanobiotechnology

Due to unique properties, gold and silver nanoparticles (GNPs and AgNPs, respectively) have wide applications in diverse fields like biomedicine, catalysis, imaging and photonics, solar energy conversion and nanoelectronics etc. and hence, are in great demand. Available physicochemical synthesis protocols generally face limitations like high cost, polluting nature, and also have restricted use in clinical and pharma applications. In order to overcome these limitations, biosynthesis of nanoparticles could be a promising alternative. While it has been suggested that initiatives should be taken for exploitation of marine microbial resources in the area of nanobiotechnology, despite the unique traits of marine-derived fungi, they are comparatively less explored for biosynthesis of GNPs and AgNPs. Though a few, available reports suggest marine-derived fungi as promising candidates for such purpose. Recent reports on observation of the laser speckle pattern and weak localization of light by AgNPs and GNPs biosynthesized by marine-derived fungi assert their novel application potentialities.