海洋科学2025，Vol.49Issue(4)：66-75,10.DOI:10.11759/hykx20241210002
创伤弧菌膦酸乙醛水解酶PhnX的重组表达、活性分析及晶体生长
Recombinant expression,enzyme activity,and crystallization of the Vibrio vulnificus phosphonoacetaldehyde hydrolase(PhnX)
摘要
Abstract
Phosphorus(P)is an essential nutrient for life.P limitation often occurs in the oceans;marine microor-ganisms can utilize inorganic phosphate(Pi)and organophosphorus compounds such as phosphonates as a P-acquisition strategy.2-Aminoethylphosphonate is the most common phosphonate,and phosphonoacetaldehyde hydrolase(PhnX)is a key enzyme in converting phosphonate to Pi.However,the enzymatic properties of PhnX in marine bacteria have been rarely studied.In this effort,we successfully expressed and purified PhnX from Vibrio vulnificus(Vv PhnX),which demonstrated significant enzymatic activity.Gel filtration experiments indicated Vv PhnX to exist as a stable and soluble homomeric enzyme with a monomer molecular mass of～29.5 kDa.Thermal shift experiments indicated the melting temperature to be～37℃.After incubation at 20-70℃for 1 h,the residual activity of Vv PhnX declined with rising temperature.After incubation at 30℃for 1 h,the enzyme activity declined by～30%.At 40℃,the activity dropped by～80%and was nearly lost after 1 h at≥50℃;these results indicated the enzyme to have moderate thermal stability.The well-shaped crystals of the Vv PhnX protein were obtained,and they were suitable for X-ray analysis.Overall,this research provides valuable insights into the biological function of PhnX and its role in the marine P cycle.关键词
海洋磷循环/有机磷代谢/2-氨基乙基膦酸盐/膦酸乙醛水解酶PhnX/创伤弧菌Key words
marine phosphorus cycle/organic phosphorus metabolism/2-Aminoethylphosphonate/phosphonoacetaldehyde hydrolase PhnX/Vibrio vulnificus分类
生物科学引用本文复制引用
韩梦雪,刘长水,马庆军..创伤弧菌膦酸乙醛水解酶PhnX的重组表达、活性分析及晶体生长[J].海洋科学,2025,49(4):66-75,10.基金项目
青岛市创新创业领军人才项目(No.18-1-2-12-zhc) （No.18-1-2-12-zhc）
国家自然科学基金青年项目(41806203) Qingdao Innovation Leadership Project,No.18-1-2-12-zhc （41806203）
National Natural Science Foundation of China,No.41806203 （）
