Bioremediation of acid mine drainage : a proteomic and genomic approach

The most significant environmental issue that has arisen in the mining industries over the last two decades is that of Acid Mine Drainage or Acid Rock Drainage (AMD​/ARD) from tailing dumps and waste mined rock piles. Bacillus subtilis and its metabolic products have been used in our studies. In this work, selective sepn. of pyrite from oxides such as quartz and calcite as well as galena (sulfide mineral) was achieved. Through microbially induced flotation, pyrite could be sepd. with 90​% efficiency from quartz and calcite leading to desulphurisation and depyritization making the process environmentally benign. Mineral specific proteins were found in extracellular proteins isolated from the cell free ext. when B. subtilis was grown in presence of minerals through SDS-​PAGE. The suppression of some genes was obsd. in the RAPD profile of the genomic DNA in presence of minerals. The cells of B. subtilis proved to be 45-​60​% efficient in the biosorption of lead metal ions at 10 and 50ppm of lead metal ion. The exopolymers extd. from B. subtilis proved to be 50-​60​% efficient in biosorption of lead metal ions. This study shows possibilities for development of eco friendly technologies for bioremediation

Bioremediation of acid mine drainage : a proteomic and genomic approach

The most significant environmental issue that has arisen in the mining industries over the last two decades is that of Acid Mine Drainage or Acid Rock Drainage (AMD​/ARD) from tailing dumps and waste mined rock piles. Bacillus subtilis and its metabolic products have been used in our studies. In this work, selective sepn. of pyrite from oxides such as quartz and calcite as well as galena (sulfide mineral) was achieved. Through microbially induced flotation, pyrite could be sepd. with 90​% efficiency from quartz and calcite leading to desulphurisation and depyritization making the process environmentally benign. Mineral specific proteins were found in extracellular proteins isolated from the cell free ext. when B. subtilis was grown in presence of minerals through SDS-​PAGE. The suppression of some genes was obsd. in the RAPD profile of the genomic DNA in presence of minerals. The cells of B. subtilis proved to be 45-​60​% efficient in the biosorption of lead metal ions at 10 and 50ppm of lead metal ion. The exopolymers extd. from B. subtilis proved to be 50-​60​% efficient in biosorption of lead metal ions. This study shows possibilities for development of eco friendly technologies for bioremediation

Isolation and Screening of Chlorella Sorokiniana for Wastewater Treatment and Biodiesel Production

Abstract- Microalgae grown on wastewater are a probable source of low cost wastewater treatment and biodiesel production. In the study, microalgae was enumerated and identified as Chlorella sorokiniana by 18S rDNA sequence which was cultivated in different wastewater for nutrient removal as well as biodiesel production were studied. The results reveal that the pH of different wastewater samples almost neutralized by microalgae, the total dissolved solids elimination ranging from 52 to 66%, the biological oxygen demand removal efficiency varied much among different wastewaters the removal rate is of 72 to 90%, the chemical oxygen demand removal ranges from 59 to 75%. The magnesium level was decreased and efficiency is about 36 to 60%, the sulphates absorption efficiency was ranging from 57 to 68% and the chloride removal efficiency was 13 to 33%. The lipid content was obtained from the algal biomass which are grown in different wastewater samples was transesterified for biodiesel production, the biodiesel was analyzed by FTIR which meets the ASTM and EU standards, hence from the current study it is evident that Chlorella sorokiniana can be effectively used for potential source for phycoremediation and biodiesel productio

In vitro flowering, somatic embryogenesis and regeneration in Boerhaavia diffusa Linn. - A medicinal plant

In vitro callogenesis was achieved from leaf explants of Boerhaavia diffusa Linn. on basal medium supplemented with different concentrations of BAP, NAA, Kn and IAA. Regeneration of shoot buds were observed on MS medium supplemented with BAP and NAA at different concentrations. In some of the replicates the shoot buds were elongated and terminated in flower buds. In vitro flowering occurred within two weeks of subculture. The highest percentage of flowering was obtained on medium supplemented with BAP (2 mg l 1). Excised shoots after transferring to rooting liquid medium (0.5-2.0 mg l 1 IAA or NAA) roots were initiated. The regenerated plantlets have been successfully established in vermiculite soil. Subsequently, they were shifted to the field conditions. Regenerated plantlets resemble the parent plants morphologically and cytologically. Histological study of the cultures showed the presence of meristemoids, somatic embryos and embryoids

Microalgae Cultivation in Different pH, Temperature and Media for Lipid Production

Lipids produced by microalgal biomass can be grouped into nonpolar lipids and polar lipids, which can be easily converted into biofuels. Microalgal samples were collected from three different ponds of Bangalore and cultured in the laboratory to find the effect of different pH, temperature and media on the production of biomass and lipids. Among these, pH-9, temperature -25 ° C and Beneck’s media was most suitable for production of biomass (35.80 g/L) and lipids from the isolated microalgae Chlorella sp. compare to Chladospora sp. (13.33 g/L). Chlorella sp. Showed 0.32 (OD) at pH-9, 0.43 (OD) at temperature-25 ° C and 2.94 (OD) in Beneck’s media. Our result revealed that nutrient supply along with measured variables affects the production of biomass and lipids in different microalgae. DOI: http://dx.doi.org/10.3126/ijls.v8i2.10227 International Journal of Life Sciences Vol.8(2): 2014; 13-1

Biodiesel Production from Oleaginous Fungi

Biodiesel involves the mixture of fatty acyl methyl/ethyl esters, produced from transesterification neutral lipids and if the origin of the source is from oleaginous micro organisms, then it is termed as micro diesel. In the present work, aiming to exploit fungi for biodiesel production, 12 fungal isolates were screened for lipid content by Sudan Black B staining method. Among 12 isolates, lipid rich five species viz, Mortierella alpina , M.ramanianna, M.vinancea, M.hyalina and M.verticella have been taken for fatty acids analysis by spectrophotometry, which revealed that the amount of free fatty acids were ranged from highest in M.alpina 35 ?moles of Oleic acid , 25 ?moles of Palmitic acid and 14 ?moles of Myristic acids to lower as much as 21 ?moles of Oleic acid , 18 ?moles of Palmitic acid and 16 ?moles of Myristic acids respectively in M.ramanianna

Strain Improvement of an Endophytic Fungal spp. for Biodiesel Production .

The Aspergillus niger JGK – 12 was subjected to mutation through physical and chemical methods (UV and EMS), and after treatment, the putative survived cultures were subjected to quantitative production of biomass and total lipid yield. Compared to control culture of Aspergillus niger JGK-12, UV treated mutant showed little enhancement in dry biomass content and total lipid production, whereas the EMS treated culture did not produce high biomass and lipid contents. The isolated lipids were then subjected to purification in which case, neutral lipids were found to be the major components accounting up to 85% of the total lipids, which infers that Aspergillus niger JGK – 12 could be a good microbial source for biodiesel production. The fractionated lipids were subjected to alkaline transesterification to produce biodiesel, which are mainly composed of fatty acid methyl esters and the compositions were found to be similar to that of regular biodiesel from plant origin. The biodiesel were further analyzed by GC-MS, HPTLC, 1H NMR, C13 NMR and FTIR techniques to confirm their purity and quality. Hence, with all the research findings, it can be concluded that Aspergillus niger JGK – 12 can be treated as a promising feedstock for biodiesel production in the futur

Evaluation of oleaginous bacteria for potential biofuel

The present work was aiming to exploit bacteria for biodiesel prodn., twenty two bacterial isolates were screened for lipid content by Sudan black B staining method, among those three isolates is lipid rich viz, F. oryzihabitans, P. aeruginosa and Morococcus sp. which have been taken for lipid extn. by Bligh and dyer method and thin layer chromatog. for detn. of fatty acid, which revealed that the amt. of lipid was highest in Morococcus sp. (0.68mL​/100mL) of Lauric acid (C12)​. Optimization of C​/N ratio the amt. of lipid was ranged highest in Morococcus sp. i.e. carbon (0.70mL​/100mL) of palmitic acid (C18)​, nitrogen (0.65mL​/100mL) of arachidonic acid (C20) and at pH 7.0 (1.96mL​/100mL) of oleic acid (C18) and in strain improvement by mutagenesis F. oryzihabitans got improved and accumulate high lipid at 10 min (1.50mL​/100mL) of linoleic acid (C16)​

Molecular Characterization of Macrophomina phaseolina, the Incitant of Coleus forskohlii Revealed by RAPD Markers

Coleus forskohlii belong to family lamiaceae is one of the commercial plants grown extensively in the country, the chemical found in the Coleus which has both medicinal application and gives great economy to the industrial organizations. Unfortunately, these plants are being highly succumbed to serious diseases like wilt and root rot caused by a fungus, hence the growers and industrialists are facing severe problem in safeguarding this crop in the field irrespective of the agro climatic conditions. Root rot disease, is one of the major diseases of Coleus forskohlii which, is caused by Macrophomina phaseolina , Pathogen variability was studied at both morphological and molecular level using cultural characteristics and Rapid Amplification of Polymorphic DNA (RAPD) analysis respectively. Totally thirty two isolates were isolated from roots of Coleus forskohlii . In RAPD 165 bands were obtained out of them 121 bands (73.3%) were polymorphic with a similarity coefficient of 0.48-0.66. Clusters analysis of RAPD data when Unweighted Pair Group Method with Arithmetic Mean (UPGMA) Tree constructed using NTSYS, it showed 6 groups. Among them two were major clusters and 4 were minor clusters with similarity coefficient 0.48-0.66. The pathogenicity of the isolates was tested on Coleus forskohlii plants. Analysis of the pathogenicity tests results revealed that the isolates grouped under two major clusters which were different from the one obtained using RAPD data. The results indicate that the data from RAPD analysis and Pathogenicity tests do not correlate with each other

Molecular phylogenetic analysis of soil bacteria

The molecular phylogeny for five strains (bearing accession numbers KR856284, KR856288, KR856287, KR856286 and KR856285) was constructed for a collection of soil bacteria isolated from different soil types. Sequences from 16s rRNA gene amplified with universal primers were used to infer evolutionary significance and were concatenated to infer a composite molecular phylogeny within the species. The isolates from each taxa formed tight species clusters in the individual gene trees aligned in BIOEDIT and constructed using MEGA7, Neighbour-Joining(NJ) and UPGMA methods were adopted as a comparative account suggesting the existence of evolutionary relationship to traditional species designations. The resulting sequence data and gene trees provide the first data set to analyses the evolutionary significance of soil bacteria from varied environments. The results showed similarity to Rhodococcus pyridinivorans, Arthrobacter koreensis, Alkaligenes faecalis, Pseudomonas pleicoglossicid and Bacillus licheniformis with bootstrap values of 100,84,100,100 and 70 respectively