Presently the world is facing the problem of scarcity of fossil fuel as it is a non-renewable source of energy. Biodiesel can be an alternative energy source having the advantage of being renewable as well as environment friendly. Enzymes are better catalysts for the production biodiesel as enzymes are more stable and their production is more convenient and safer. Microalgae Scenedesmus sp. was used as a feedstock for biodiesel production. Lipase producing bacteria was isolated from the pulp and paper mill and identified as Microbacterium sp. using 16S rDNA sequencing method. Lipase enzyme was purified by sequential methods of ammonium sulphate precipitation and Sephadex G-100 gel column chromatography. The molecular weight of purified enzyme was 40 kDa on SDS-PAGE. This purification procedure resulted in 2.1fold purification of lipase with a 20.8 % final yield. The purified lipase exhibited maximal hydrolytic activity at a temperature of 50 0 C and a pH of 8.5. The Km of lipase was 3.2 mM and the Vmax 50 μmol/min/mg. Lipase activity was stimulated by Triton X-100 and SDS and inhibited by Tween 20 and Tween 80. Biodiesel was prepared through sodium hydroxide, potassium hydroxide and Lipase (Celite and charcoal bound as substrate) catalyzed transesterification process, which enabled a yield of 72.5%, 90% (95.1%, and 15.5%) respectively determined by gas chromatography/mass spectrometry (GC/MS) analysis.
Present study involves the simple, rapid, non-toxic and in vitro method of extracellular silver nanoparticles synthesis using Entomopathogenic fungus (Beauveria bassiana). The development of silver nanoparticle in fungal supernatant was confirmed by the absorbance peak at 450 nm in UV-Vis spectrophotometer. Further, presence of AgNPs and its crystal lattice was confirmed by EDS and XRD, respectively. TEM micrograph confirmed the presence of differently shaped (triangular, circular, hexagonal) nanoparticles with size ranging from 10 to 50 nm. Variable shape and size of fungal assisted AgNps was also confirmed in SEM study. The optimal pH and temperature for biosynthesis of nanoparticles was found to be 6.0 and 25 °C, respectively. The continuous effects of AgNPs against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus in time dependent manner was confirmed in growth kinetic studies. During 36 h of growth study, maximum reduction in O.D 560 was found in E. coli (67.2%) followed by P. aeruginosa (63.3%) and S. aureus (56.8%) at 30 °C. The MIC values of fungal assisted AgNPs against E. coli, P. aeruginosa and S. aureus was found to be 2.5, 3 and 4.5 ppm, respectively. The MIC values of Ciprofloxacin was observed to be 0.5, 0.5 and 0.7 ppm, whereas MICs of AgNPs + Ciprofloxacin showed 0.4, 0.4, 0.5 ppm against E. coli, P. aeruginosa and S. aureus, respectively, clearly highlighting the synergistic effect of AgNPs in combination with Ciprofloxacin. In the view of challenges for developing antimicrobial nanoparticles of variable shape and size by various other methods, tuning nanoparticles synthesis via fungi can be a wonderful approach to resolve existing hurdles.
A chemolithotrophic bacterium enriched in the chemostat in presence of sodium bicarbonate as sole carbon source was identified as Serratia sp. by 16S rRNA sequencing. Carbon dioxide sequestering capacity of bacterium was detected by carbonic anhydrase enzyme and ribulose-1, 5- bisphosphate carboxylase/oxygenase (RuBisCO). The purified carbonic anhydrase showed molecular weight of 29 kDa. Molecular weight of RuBisCO was 550 kDa as determined by fast protein liquid chromatography (FPLC), however, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed presence of two subunits whose molecular weights were 56 and 14 kDa. The Western blot analysis of the crude protein and purified sample cross reacted with RuBisCO large-subunit polypeptides antibodies showed strong band pattern at molecular weight around 56 kDa regions. Whole cell soluble proteins of Serratia sp. grown under autotrophic and heterotrophic conditions were resolved by two-dimensional gel electrophoresis and MALDI-TOF/MS for differential expression of proteins. In proteomic analysis of 63 protein spots, 48 spots were significantly up-regulated in the autotrophically grown cells; seven enzymes showed its utilization in autotrophic carbon fixation pathways and other metabolic activities of bacterium including lipid metabolisms indicated sequestration potency of carbon dioxide and production of biomaterials.
Bacterial community of palaeoproterozoic metasediments was enriched in the chemostat in the presence of different concentrations of NaHCO3. Six bacterial isolates were isolated from the chemostat on nutrient agar plates on the basis of distinct morphology. Denaturing gradient gel electrophoresis (DGGE) proved the presence of six operational taxonomic units (OTUs) at 50 and 100 mM NaHCO3. The OTU was reduced to three and one at enrichment concentration of 150 and 200 mM NaHCO3 respectively. These six isolates were tested for sequestration of carbon dioxide by (14)C metabolic labeling of NaH(14)CO3. Among the six isolates, one of the bacterium showed better potency to fix radiolabeled NaH(14)CO3. The isolate (ISTD04) was identified as Serratia sp. by 16S ribosomal RNA (16S rRNA) sequence analysis and was found to be same as the DGGE OTU sequence at 200-mM NaHCO3 concentration. The bacterium was tested for product formation in form of calcite crystals in presence of 5 % CO2. Scanning electron microscopy (SEM) of product formed by the bacterium revealed defined faceted rhombohedral structure which resembled calcite and vaterite phases of the crystal. Formation of calcium carbonate crystals was further confirmed by Fourier transform infrared (FTIR) spectroscopy as carbonate group showing strong vibration at 1,456 cm(-1). Major calcite phase diffraction peaks were determined by X-ray diffraction (XRD) analysis, and energy-dispersive X-ray (EDX) analysis showed the presence of CaO (72 %) and carbon (18 %). Bacterium use bicarbonate as carbon source for their growth as well as by-product formation in form of calcite shows carbon circulation and storage.
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