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Cloning, Purification, Crystallization and Preliminary X-Ray Diffraction Studies of periplasmic Glucose Binding Protein of <i>Pseudomonas putida</i>CSV86

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Advances in Bioscience and Biotechnology 2015年第3期6卷 164-171页

作者： Suman Pandey Arnab Modak Prashant S. Phale Prasenjit Bhaumik Department of Biosciences and Bioengineering Indian Institute of Technology Bombay Mumbai India

Biochemical data and genomic analysis indicate the involvement of a putative ABC transporter for glucose transport in Pseudomonas putida CSV86. The periplasmic solute binding proteins are known to confer substrate specificity to the ABC transporters by binding specifically to the substrate and transferring them to their cognate inner membrane transport assembly. periplasmic glucose binding protein from Pseudomonas putida CSV86 (ppGBP) was found to be glucose specific. The gene encoding ppGBP was cloned. Recombinant ppGBP was overexpressed and purified to homogeneity. The purified recombinant protein showed glucose binding activity of 752 pmol/mg of protein and was crystallized as a complex with glucose. The crystal diffracted to 1.7 Å resolution using home X-ray source. Preliminary analysis of diffraction data showed that the crystals belonged to space group P21212 with unit-cell parameters a = 102.56, b = 119.2, c = 66.65 Å and α = β = γ = 90°. Matthews coefficient calculation showed the presence of two molecules in the asymmetric unit with solvent content of 45.7%.

关键词： periplasmic Glucose Binding Protein Pseudomonas Crystallization

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Nickel uptake and intracellular localization in Cupriavidus pauculus KPS 201, native to ultramafic ecosystem

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Advances in Bioscience and Biotechnology 2010年第4期1卷 276-280页

作者： Arundhati Pal A. K. Paul 不详

The nickel-resistant bacterium, Cupriavidus pauculus KPS 201 was isolated from the rhizosphere of Rinorea bengalensis (Wall.) O. K. endemic to metal-percolated ultramafic ecosystem of Andaman, India. This study investigates nature of Ni resistance, growth associated uptake and localization of Ni in cellular compartments of KPS 201. Growth kinetics of C. pauculus KPS 201 exhibited a typical inducible Ni resistance in Ni-supplemented (1.0-10.0 mM) Tris-minimal medium. The Ni-induced cells showed a high degree of Ni resistance and accumulated a maximum of 29.3 μM Ni/g protein after 48 h of growth in 5 mM Ni. The accumulated Ni was preferentially retained (90.6%) in the periplasm and was associated with the expression of two periplasmic proteins (74 and 66 kDa) under Ni-induced condition. Inducible nickel resistance in C. pauculus KPS 201 may possibly be due to extracytoplasmic binding and accumulation coupled with expression of specific periplasmic proteins. These findings will provide an insight in understanding metal-microbe interaction in geogenous environments and their exploitation in bioremediation of heavy metal pollutants.

关键词： Cupriavidus pauculus Inducible Ni Resistance Intracellular Uptake periplasmic Proteins Ultramafic Soil

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