化工学报2016，Vol.67Issue(10)：4389-4398,10.DOI:10.11949/j.issn.0438-1157.20160355
乙二胺四乙酸二酐对草酸脱羧酶的修饰改性
Chemical modification of oxalate decarboxylase with ethylenediaminetetraacetic dianhydride
摘要
Abstract
In order to improve the stability and application performance of oxalate decarboxylase (Oxdc) in the prevention and treatment of urinary calculi, chemical modification of Oxdc with ethylenediaminetetraacetic dianhydride (EDTAD) was investigated. The results of single-factor experiment showed that the extent of modification and the recovery rate of the enzymatic activity were 71.91% and 75.42%, respectively, when the reaction time was 8 h, the molar ratio of EDTAD/Oxdc was 50:1, pH 7.0, and the temperature was 37℃. The analysis results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS) indicated that Oxdc and EDTAD have been covalently bound. The ultraviolet-visible spectrum (UV) and circular dichroic (CD) measurement showed that the structure and conformation of Oxdc were tinily altered after modification by EDTAD. The enzymology changes of Oxdc were also analyzed, the results showed the optimum pH of EDTAD-modified Oxdc was shifted to the alkaline side about 1.5 unit and it had a higher thermostability. Moreover, through modification the adsorption capacity of Oxdc onto calcium oxalate monohydrate crystals was increased by 42.42%. These results suggested that the stability and application performance of Oxdc were significantly improved under this experiment.关键词
泌尿系结石/酶/草酸脱羧酶/纯化/乙二胺四乙酸二酐/化学修饰/吸附Key words
urinary calculi/enzyme/oxalate decarboxylase/purification/ethylenediaminetetraacetic dianhydride/chemical modification/adsorption分类
生物科学引用本文复制引用
贺俊斌,林日辉,龙寒,巫佳,蔡杏华,杨颖,陈盛峰..乙二胺四乙酸二酐对草酸脱羧酶的修饰改性[J].化工学报,2016,67(10):4389-4398,10.基金项目
国家教育部留学回国人员科研启动基金项目（教外司留[2012]1707号）；广西自然科学基金项目（2014GXNSFAA118045）；广西民族大学研究生教育创新计划项目（gxun-chxs2015089）。@@@@Foundation item：supported by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, Ministry of Education ([2012]1707), the Natural Science Foundation of Guangxi (2014GXNSFAA118045) and the Innovation Project of Guangxi University for Nationalities (gxun-chxs2015089) （[2012]1707）
