分子生物学
IVD分子诊断
细胞培养与分析
蛋白研究
细胞因子
重组蛋白
抗体
高通量测序建库
病原检测UCF系列
生物医药
工具酶
抑制剂激活剂与常用试剂
仪器
耗材

Rapid identification of magnesium ascorbyl phosphate utilizing phosphatase through a chromogenic change-coupled activity assay

Wang Yuli, Hu Wenyue, Deng Zixin, He Xinyi

Journal:APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

IF:4.81

DOI:10.1007/s00253-021-11229-7

PMID:33754168

Published:2021-03-22

research field:肿瘤学分子生物学血液学药物化学靶向蛋白降解

Abstract

Abstract In this study, we report a chromogenic reaction between magnesium ascorbyl phosphate (MAP) and ferric chloride to generate a Brown-Red clathrate, while the Treated MAP by phosphatases forms Colorless (BRTC) product with ferric chloride. The BRTC was indicative of phosphatase activity-mediated excision of phosphorous group from MAP and utilized to screen phosphatases from bacterial cell lysates. From ten tested strains, BRTC was observed in the cell lysate of Salmonella enterica subsp. enterica serovar Cerro 87. BRTC was again employed to track phosphatase activity of the resuspensions of the ammonium sulfate graded precipitations of the cell lysate. Two phosphatases, PhoN and YcdX, were identified by LC-MS/MS analysis in the protein fraction giving most obvious BRTC phenotype and validated by examination of in vitro activity of the purified proteins. Key points BRTC is labelling-free, naked-eye visible, and independent of any facilities. BRTC can directly screen phosphatases from microbial cell lysates. Using BRTC system, two phosphatases were identified in Salmonella enterica subsp. enterica serovar Cerro 87. Graphical abstract

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