Prasanna Kumar Panda

Biosynthesis of silver nanoparticle from leaf extract of Desmodium gangeticum (L.) DC. and its biomedical potential

An aqueous leaf extract of Desmodium gangeticum was employed to synthesize silver nano particles. Rapid formation of stable silver nanoparticles were observed on exposure of the aqueous leaf extract with solution of silver nitrate. The silver nanoparticles were characterized by UV-visible spectroscopy, scanning electron microscopy, energy dispersive X-ray analysis and transmission electron microscopy (TEM), and Fourier Transform Infra-Red spectroscopy (FTIR) UV-visible spectroscopy, scanning electron microscopy (SEM), energy dispersive X-ray analysis (EDAX), transmission electron microscopy (TEM), and Fourier Transform Infra-Red spectroscopy (FTIR). UV-visible spectrum of the aqueous medium peaked at 450 nm corresponding to the plasmon absorbance of silver nanoparticles. SEM analysis revealed the spherical shape of the particles with sizes ranging from 18 to 39 nm and the EDAX spectrum confirmed the presence of silver along with other elements in the plant metabolite. Further, these biologically synthesized nanoparticles were found to be highly toxic against pathogenic bacteria Escherichia coli, thus implying significance of the present study in production of biomedical products.

Phylogenomic Study of Lipid Genes Involved in Microalgal Biofuel Production—Candidate Gene Mining and Metabolic Pathway Analyses

Optimizing microalgal biofuel production using metabolic engineering tools requires an in-depth understanding of the structure-function relationship of genes involved in lipid biosynthetic pathway. In the present study, genome-wide identification and characterization of 398 putative genes involved in lipid biosynthesis in Arabidopsis thaliana Chlamydomonas reinhardtii, Volvox carteri, Ostreococcus lucimarinus, Ostreococcus tauri and Cyanidioschyzon merolae was undertaken on the basis of their conserved motif/domain organization and phylogenetic profile. The results indicated that the core lipid metabolic pathways in all the species are carried out by a comparable number of orthologous proteins. Although the fundamental gene organizations were observed to be invariantly conserved between microalgae and Arabidopsis genome, with increased order of genome complexity there seems to be an association with more number of genes involved in triacylglycerol (TAG) biosynthesis and catabolism. Further, phylogenomic analysis of the genes provided insights into the molecular evolution of lipid biosynthetic pathway in microalgae and confirm the close evolutionary proximity between the Streptophyte and Chlorophyte lineages. Together, these studies will improve our understanding of the global lipid metabolic pathway and contribute to the engineering of regulatory networks of algal strains for higher accumulation of oil.

Screening of Fresh Water Microalgae from Eastern Region of India for Sustainable Biodiesel Production

Study of six different freshwater microalgae, collected from Odisha, eastern region of India, has been carried out to find out their potential for biodiesel production. The growth, total lipid, and fatty acid composition of six microalgal strains were determined. Chlorella sp. IMMTCC-2, which exhibited high lipid content with considerable amount of unsaturated fatty acids, was selected for culture in a self-designed photobioreactor in order to study the scale-up possibilities. The result shows significant increase in lipid accumulation from logarithmic phase to stationary phase in the photobioreactor, i.e., from 12.4 to 28.3%. Analyses of the present results suggest that Chlorella sp. IMMTCC-2 is appropriate for biodiesel production.

Controlled Synthesis of Gold Nanoparticles Using Aspergillus terreus IF0 and Its Antibacterial Potential against Gram Negative Pathogenic Bacteria

Biosynthesis of monodispersed nanoparticles, along with determination of potential responsible biomolecules, is the major bottleneck in the area of bionanotechnology research. The present study focuses on an ecofriendly, ambient temperature protocol for size controlled synthesis of gold nanoparticles, using the fungus Aspergillus terreus IF0. Gold nanoparticles were formed immediately, with the addition of chloroauric acid to the aqueous fungal extract. Synthesized nanoparticles were characterized by UV-Vis spectroscopy, TEM-EDX, and XRD analysis. Particle diameter and dispersity of nanoparticles were controlled by varying the pH of the fungal extract. At pH 10, the average size of the synthesized particles was in the range of 10–19 nm. Dialysis to obtain high and low molecular weight fraction followed by FTIR analysis revealed that biomolecules larger than 12 kDa and having –CH, –NH, and –SH functional groups were responsible for bioreduction and stabilization. In addition, the synthesized gold nanoparticles were found to be selectively bactericidal against the pathogenic gram negative bacteria, Escherichia coli.

In Search of Actionable Targets for Agrigenomics and Microalgal Biofuel Production: Sequence-Structural Diversity Studies on Algal and Higher Plants with a Focus on GPAT Protein

The triacylglycerol (TAG) pathway provides several targets for genetic engineering to optimize microalgal lipid productivity. GPAT (glycerol-3-phosphate acyltransferase) is a crucial enzyme that catalyzes the initial step of TAG biosynthesis. Despite many recent biochemical studies, a comprehensive sequence-structure analysis of GPAT across diverse lipid-yielding organisms is lacking. Hence, we performed a comparative genomic analysis of plastid-located GPAT proteins from 7 microalgae and 3 higher plants species. The close evolutionary relationship observed between red algae/diatoms and green algae/plant lineages in the phylogenetic tree were further corroborated by motif and gene structure analysis. The predicted molecular weight, amino acid composition, Instability Index, and hydropathicity profile gave an overall representation of the biochemical features of GPAT protein across the species under study. Furthermore, homology models of GPAT from Chlamydomonas reinhardtii, Arabidopsis thaliana, and Glycine max provided deep insights into the protein architecture and substrate binding sites. Despite low sequence identity found between algal and plant GPATs, the developed models exhibited strikingly conserved topology consisting of 14α helices and 9β sheets arranged in two domains. However, subtle variations in amino acids of fatty acyl binding site were identified that might influence the substrate selectivity of GPAT. Together, the results will provide useful resources to understand the functional and evolutionary relationship of GPAT and potentially benefit in development of engineered enzyme for augmenting algal biofuel production.

Biogenic unmodified gold nanoparticles for selective and quantitative detection of cerium using UV-vis spectroscopy and photon correlation spectroscopy (DLS).

The ability of self-functionalized biogenic GNPs towards highly selective colorimetric detection of rare earth element cerium is being reported for the first time. GNPs underwent rapid aggregation on addition of cerium indicated by red shift of SPR peak followed by complete precipitation. Hereby, this concept of co-ordination of cerium ions onto the GNP surface has been utilized for detection of cerium. The remarkable capacity of GNPs to sensitively detect Ce without proves beneficial compared to previous reports of colorimetric sensing. MDL was 15 and 35 ppm by DLS and UV-vis spectroscopy respectively, suggesting DLS to be highly sensitive and a practical alternative in ultrasensitive detection studies. The sensing system showed a good linear fit favouring feasible detection of cerium in range of 2-50 ppm. Similar studies further showed the superior selectivity of biogenic GNPs compared to chemically synthesized counterparts. The sensing system favours on-site analysis as it overcomes need of complex instrumentation, lengthy protocols and surface modification of GNP.

Way forward to achieve sustainable and cost-effective biofuel production from microalgae: a review

Driven by policies aimed at enhancing energy security and mitigating greenhouse gas emissions, the production and use of biofuels have significantly increased in recent years. Microalgae owing to its multiple advantages which include high lipid content, sustainable biomass production, effective land and water utilization are the most potential biofuel feedstock that can provide drop-in fossil fuel replacements without stimulating competition for agricultural resources and are considered to be more environmentally benign than the first- and second-generation biofuel feedstocks. However, there are many existing technical and scientific impediments that are yet to be resolved. Keeping this in view, the present review provides a concise account of the microalgal species known to accumulate high levels of lipid and describes the main factors that should be taken into consideration while selecting suitable algal strains for mass cultivation. The underlining advantages and limitations of raceway pond and photobioreactor cultivation systems are also examined. The recent advances in genetic engineering of microalgae to improve biomass and lipid productivity are then highlighted, which include the ongoing debate over the biosafety issues pertinent to the use of genetically modified algae. Furthermore, a wide range of high-value products that can be co-produced from microalgae have been discussed. The review concludes with a comprehensive summary of the major techno-economic constraints to commercialization of algal-derived biofuels along with promising methods for overcoming these challenges in order to produce cost-competitive and environmentally sustainable biofuel.

Genome-wide identification and evolutionary analysis of algal LPAT genes involved in TAG biosynthesis using bioinformatic approaches

Lysophosphatidyl acyltransferase (LPAT) is one of the major triacylglycerol synthesis enzymes, controlling the metabolic flow of lysophosphatidic acid to phosphatidic acid. Experimental studies in Arabidopsis have shown that LPAT activity is exhibited primarily by three distinct isoforms, namely the plastid-located LPAT1, the endoplasmic reticulum-located LPAT2, and the soluble isoform of LPAT (solLPAT). In this study, 24 putative genes representing all LPAT isoforms were identified from the analysis of 11 complete genomes including green algae, red algae, diatoms and higher plants. We observed LPAT1 and solLPAT genes to be ubiquitously present in nearly all genomes examined, whereas LPAT2 genes to have evolved more recently in the plant lineage. Phylogenetic analysis indicated that LPAT1, LPAT2 and solLPAT have convergently evolved through separate evolutionary paths and belong to three different gene families, which was further evidenced by their wide divergence at gene structure and sequence level. The genome distribution supports the hypothesis that each gene encoding a LPAT is not duplicated. Mapping of exon–intron structure of LPAT genes to the domain structure of proteins across different algal and plant species indicates that exon shuffling plays no role in the evolution of LPAT genes. Besides the previously defined motifs, several conserved consensus sequences were discovered which could be useful to distinguish different LPAT isoforms. Taken together, this study will enable the generation of experimental approximations to better understand the functional role of algal LPAT in lipid accumulation.

Homology modeling and docking studies of FabH (β-ketoacyl-ACP synthase III) enzyme involved in type II fatty acid biosynthesis of Chlorella variabilis: a potential algal feedstock for biofuel production

The concept of using microalgae as an alternative renewable source of biofuel has gained much importance in recent years. However, its commercial feasibility is still an area of concern for researchers. Unraveling the fatty acid metabolic pathway and understanding structural features of various key enzymes regulating the process will provide valuable insights to target microalgae for augmented oil content. FabH (β-ketoacyl-acyl carrier protein synthase; KAS III) is a condensing enzyme catalyzing the initial elongation step of type II fatty acid biosynthetic process and acyl carrier protein (ACP) facilitates the shuttling of the fatty acyl intermediates to the active site of the respective enzymes in the pathway. In the present study, a reliable three-dimensional structure of FabH from Chlorella variabilis, an oleaginous green microalga was modeled and subsequently the key residues involved in substrate binding were determined by employing protein–protein docking and molecular dynamics (MD) simulation protocols. The FabH-ACP complex having the lowest docking energy score showed the binding of ACP to the electropositive FabH surface with strong hydrogen bond interactions. The MD simulation results indicated that the substrate-complexed FabH adopted a more stable conformation than the free enzyme. Further, the FabH structure retained its stability throughout the simulation although noticeable displacements were observed in the loop regions. Molecular simulation studies suggested the importance of crucial hydrogen bonding of the conserved Arg91 of FabH with Glu53 and Asp56 of ACP for exhibiting high affinity between the enzyme and substrate. The molecular modeling results are consistent with available experimental results on the flexibility of FabH and the present study provides first in silico insights into the structural and dynamical aspect of catalytic mechanism of FabH, which could be used for further site-specific mutagenic experiments to develop engineered high oil-yielding microalgal strains for biofuel production.

BBProF: An Asynchronous Application Server for Rapid Identification of Proteins Associated with Bacterial Bioleaching

Recovery of metal value, especially from low-grade ores and overburden minerals using acidophilic bacteria through the process of bioleaching is an environmentally benign and commercially scalable biotechnology. In recent years, while the “OMICS” landscape has been witnessing extensive application of computational tools to understand and interpret global biological sequence data, a dedicated bioinformatic server for analysis of bacterial information in the context of its bioleaching ability is not available. We have developed an on-line Bacterial Bioleaching Protein Finder (BBProF) System, which rapidly identifies novel proteins involved in a bacterial bioleaching process and also performs phylogenetic analysis of 16S rRNA genes. BBProF uses the features of Asynchronous Java Script and XML (AJAX) to provide an efficient and fast user experience with minimal requirement of network bandwidth. In the input module the server accepts any bacterial or archaeal complete genome sequence in RAW format and provides a list of proteins involved in the microbial leaching process. BBProF web server is integrated with the European Bioinformatics Institute (EBI) web services such as BLAST for homology search and InterProScan for functional characterization of output protein sequences. Studying evolutionary relationship of bacterial strains of interest using Muscle and ClustalW2 phylogeny web services from EBI is another key feature of our server, where 16S rRNA gene sequences are considered as input through a JQUERY interface along with the sequences present in the BBProF database library. Complete genome sequences of 24 bioleaching microorganism characterized by genomic and physiological study in the laboratory and their respective 16S rRNA gene sequences were stored in the database of the BBProF library. To our knowledge BBProF is the first integrated bioinformatic web server that demonstrates its utility in identifying potential bioleaching bacteria. We hope that the server facilitate ongoing comparative genomic studies on of bioleaching microorganisms and also assist in identification and design of novel microbial consortia that are optimally efficient bioleaching agents.