草酸脱羧酶的修饰及其对草酸钙的吸附研究
Research on the adsorption of the modified oxalate decarboxylase on calcium oxalate
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摘要: 为了提高草酸脱羧酶(OXDC)的适用性及其对草酸钙的吸附性能,本实验采用乙二胺四乙酸二酐(EDTAD)对草酸脱羧酶进行了化学修饰,通过高效液相色谱-质谱联用仪(UPLC-MS)测定其修饰位点,并研究和对比了酶在修饰前后对草酸钙的吸附差异,采用动力学和热力学吸附模型对吸附实验的结果进行拟合分析。实验结果表明,EDTAD成功修饰在草酸脱羧酶上,修饰位点位于肽N端的α-氨基上;且修饰酶(EDTAD-OXDC)对草酸钙的吸附作用,在酶初始浓度为5 mg/m L,p H为7.0,温度为37℃的条件下,比原酶(OXDC)提高了53.37%。结论:经EDTAD修饰后的草酸脱羧酶,其稳定性和吸附性能均有所提高。Abstract: In order to improve the applicability and adsorption property of oxalate decarboxylase ( OXDC) on calcium oxalate.EDTAD was used to modify the OXDC, modified site was determined by UPLC-MS, and the adsorption of the enzymes on calcium oxalate ( Ca Ox) were studied and compared. The kinetics and thermodynamics adsorption models were used to fit and analysis the experimental data. Results showed that EDTAD was successfully grafted onto OXDC covalently as an absorption group, the modified sites were located in the α-NH2 of the peptide N side, and the adsorption property of the modified enzyme ( EDTAD-OXDC) on calcium oxalate was 53.37% higher than that of OXDC in initial enzyme concentration of 5 mg/m L, p H7.0, temperature of 37 ℃. Conclusion: which inclicated the stability and adsorption properties of DXDC was improved by grafting with EDTAD.