Identification and Purification of Membrane-Bound NAD (P) + - Dependent D-Glucose Dehydrogenase (nGDH) and Thiosulfate Dehydrogenase (TDH) from Cattle Manure Inhabitant Sulphur-Oxidizing Pseudomonas PRK786

Novel membrane bound-enzymes such as NAD (P) +- dependent D-glucose dehydrogenase (nGDH) and thiosulfate dehydrogenase (TDH) were identified and separated from cattle manure compost inhabitant sulphur oxidizing Pseudomonas PRK 786. The unique bacterial strain belongs to chemolithotrophic oxidizer due to that presence of dual characters - A) oxidation of glucose B) oxidation of inorganic sulphur compounds. In membrane preparation, the optimum level (membrane of Pseudomonas PRK 786) of sonication parameters is 40 % duty cycle; power - 10; total time - 30 min. 7.2?g of phospholipids were estimated in1ml of cured membrane sample. The total activity of NAD (P) +- dependent D-glucose dehydrogenase and thiosulfate dehydrogenase were assayed at three levels (Cured preparation, Heat precipitation and Desalting) the results 28.93 ?mol min-1, 20.25 ?mol min-1, 35.69 ?mol min-1, 103.54 ?mol min-1, 86.28?mol min-1 & 150.37 ?mol min-1 respectively. An effect of catalytic processes was assayed with and without substrates, enzymes and electron acceptor. 80% of proteins were precipitated at 500C, ~80 kDs and ~48 kDs protein bands were identified by SDS-PAGE and two different peaks were observed by HPLC at retention time -1.390 min and retention time -2.170.Its clearly indicates purified sample contains of nGDH and TDH.