METABOLITE ANALYSIS OF CLOSTRIDIUM THERMOCELLUM USING CAPILLARY ELECTROPHORESIS BASED TECHNIQUES

Clostridium thermocellum is a thermophilic bacterium that converts biomass to ethanol directly; however, high sensitivity of this bacterium toward ethanol limits its commercial utility. To elucidate the effect of ethanol on the growth of this bacterium a metabolite analysis of C. thermocellum was performed. The hypothesis of the project was that exogenous ethanol alters the metabolite profile of C. thermocellum. For metabolite analysis, capillary electrophoresis-electrospray ionization-mass spectrometry method (CE-ESI-MS) was developed due to highly polar and charged nature of metabolites. To increase the sensitivity of CE-ESI-MS, several parameters at the ESI interface were optimized. The application of 50% isopropanol as a sheath liquid increased sensitivity for metabolite analysis dramatically. Trimethylamine acetate (pH 10) was used as background electrolyte (BGE) due to its ability to separate the structural isomers of glucose phosphate. For metabolite sample preparation, novel methods for quenching and CE compatible metabolite extraction protocols were developed. Newly developed protocols were applied to metabolite analysis of wild type (WT) and ethanol adapted (EA) strains of C. thermocellum grown in batch cultures. Significant differences were found in key intracellular metabolites such as NAD+ and pyruvic acid. Intracellular concentrations of NAD+ were low in EA cells compared to WT cells and pyruvic acid was only detected in EA cells. To further understand the effect of ethanol on metabolite fluxes, WT and EA cells were grown in increasing concentrations of ethanol and the metabolite profile for each ethanol treatment was obtained. Significant changes were found in intracellular metabolite concentrations. Metabolic data showed that the glycolysis process in WT cells was obstructed due to exogenous ethanol which was evident from accumulation of G6P. On the other hand, no such accumulation of G6P was observed in the EA strain; however pyruvate began to accumulate in EA strain. These changes in intracellular metabolite concentrations due to perturbation of exogenous ethanol supported the hypothesis. Also, this investigation revealed a correlation between ethanol and metabolite profile changes and was able to explain a possible mechanism of growth inhibition of C. thermocellum which will certainly help genetic engineers to develop superior strains of C. thermocellum for commercial cellulosic ethanol production

Effect of Sb deficiency on the thermoelectric properties of Zn4Sb3

We have investigated the effect of Sb-deficiency on the thermoelectric figure of merit (zT) of Zn4Sb3 prepared by solid state reaction route. At high temperatures, the Seebeck coefficient (S) and electrical conductivity ({\sigma}) increase with increase in Sb deficiency whereas the thermal conductivity (\k{appa}) decreases giving rise to an increase in the overall zT value. The observations suggest that creation of vacancies could be an effective route in improving the thermoelectric properties of Zn4Sb3 system. This coupled to nanostructuring strategy could lead to the ultimate maximum value of zT in this system for high temperature thermoelectric applications

Currents and Radiation from the large DD Black Hole Membrane

It has recently been demonstrated that black hole dynamics in a large number of dimensions $D$ reduces to the dynamics of a codimension one membrane propagating in flat space. In this paper we define a stress tensor and charge current on this membrane and explicitly determine these currents at low orders in the expansion in $\frac{1}{D}$. We demonstrate that dynamical membrane equations of motion derived in earlier work are simply conservation equations for our stress tensor and charge current. Through the paper we focus on solutions of the membrane equations which vary on a time scale of order unity. Even though the charge current and stress tensor are not parametrically small in such solutions, we show that the radiation sourced by the corresponding membrane currents is generically of order $\frac{1}{D^D}$. In this regime it follows that the `near horizon' membrane degrees of freedom are decoupled from asymptotic flat space at every perturbative order in the $\frac{1}{D}$ expansion. We also define an entropy current on the membrane and use the Hawking area theorem to demonstrate that the divergence of the entropy current is point wise non negative. We view this result as a local form of the second law of thermodynamics for membrane motion.Comment: 104 pages plus 69 pages appendix, 1 figure, Minor correction

Molecular modeling, simulation and docking of Rv1250 protein from Mycobacterium tuberculosis

Computational prediction and protein structure modeling have come to the aid of various biological problems in determining the structure of proteins. These technologies have revolutionized the biological world of research, allowing scientists and researchers to gain insights into their biological questions and design experimental research much more efficiently. Pathogenic Mycobacterium spp. is known to stay alive within the macrophages of its host. Mycobacterium tuberculosis is an acid-fast bacterium that is the most common cause of tuberculosis and is considered to be the main cause of resistance of tuberculosis as a leading health issue. The genome of Mycobacterium tuberculosis contains more than 4,000 genes, of which the majority are of unknown function. An attempt has been made to computationally model and dock one of its proteins, Rv1250 (MTV006.22), which is considered as an apparent drug-transporter, integral membrane protein, and member of major facilitator superfamily (MFS). The most widely used techniques, i.e., homology modeling, molecular docking, and molecular dynamics (MD) simulation in the field of structural bioinformatics, have been used in the present work to study the behavior of Rv1250 protein from M. tuberculosis. The structure of unknown TB protein, i.e., Rv1250 was retrived using homology modeling with the help of I-TASSER server. Further, one of the sites responsible for infection was identified and docking was done by using the specific Isoniazid ligand which is an inhibitor of this protein. Finally, the stability of protein model and analysis of stable and static interaction between protein and ligand molecular dynamic simulation was performed at 100 ns The designing of novel Rv1250 enzyme inhibitors is likely achievable with the use of proposed predicted model, which could be helpful in preventing the pathogenesis caused by M. tuberculosis. Finally, the MD simulation was done to evaluate the stability of the ligand for the specific protein