Robinson N A, Goss N H, Wood H G
Biochem Int. 1984 Jun;8(6):757-69.
Polyphosphate kinase, which catalyzes the synthesis of polyphosphate from ATP, has been partially purified from Propionibacterium shermanii. The reaction is unusual in that addition of basic protein causes the enzyme to precipitate and the insoluble form has optimal activity. The synthesized [32P]polyphosphate is non-covalently bound to the precipitated material and was isolated from the complex by proteolysis. The gel electrophoresis procedure of Maxam and Gilbert was adapted to sizing polyphosphates. When polyphosphate was treated with alkali, polyphosphates ranging from 1-100 phosphate residues were obtained as individual bands. The untreated enzymatically synthesized polyphosphate migrated as a species in excess of 200 phosphate moieties.
已从谢氏丙酸杆菌中部分纯化出催化由ATP合成多聚磷酸盐的多聚磷酸盐激酶。该反应不同寻常之处在于，添加碱性蛋白质会导致酶沉淀，且不溶性形式具有最佳活性。合成的[32P]多聚磷酸盐与沉淀物质非共价结合，并通过蛋白水解从复合物中分离出来。对Maxam和Gilbert的凝胶电泳方法进行了改进，以对多聚磷酸盐进行大小测定。用碱处理多聚磷酸盐时，可得到含有1至100个磷酸残基的多聚磷酸盐作为单独的条带。未经处理的酶促合成多聚磷酸盐迁移时呈现出超过200个磷酸部分的形态。
