Mullan Alan, Quinn John P, McGrath John W
School of Biology and Biochemistry and QUESTOR Centre, The Queen's University of Belfast, Medical Biology Centre, Ireland.
Anal Biochem. 2002 Sep 15;308(2):294-9. doi: 10.1016/s0003-2697(02)00249-x.
Studies of polyphosphate (polyP) metabolism in microorganisms have been hampered by the lack of a convenient method for the assay in cell extracts of the activity of polyphosphate kinase (PPK), the enzyme principally responsible for microbial polyP biosynthesis. We report the development of such an assay, based on the well-established metachromatic reaction, with toluidine blue, of the polyP formed during the PPK-catalyzed reaction. The method was successfully used in the characterization of PPK activity in crude extracts of an environmental Burkholderia cepacia isolate. The development of a protocol for the physical recovery of polyP from solution is also reported.
微生物中多聚磷酸盐（polyP）代谢的研究一直受到阻碍，因为缺乏一种方便的方法来测定细胞提取物中多聚磷酸盐激酶（PPK）的活性，该酶是微生物多聚磷酸盐生物合成的主要负责酶。我们报告了这样一种测定方法的开发，该方法基于已确立的用甲苯胺蓝对PPK催化反应过程中形成的多聚磷酸盐进行的异色反应。该方法成功用于表征环境洋葱伯克霍尔德菌分离株粗提物中的PPK活性。还报告了从溶液中物理回收多聚磷酸盐的方案的开发。
