粘质沙雷氏菌突变几丁质酶C的包涵体复性OA
Renaturation of Inclusion Bodies of Serratia marcescens Mutant Chitinase C
为筛选大肠杆菌 BL21(DE3)表达的粘质沙雷氏菌突变几丁质酶 C 包涵体复性液组成,对大肠杆菌表达的粘质沙雷氏菌突变体几丁质酶包涵体进行分离、洗涤及金属螯合层析纯化.采用分式析因设计,考察 13个因素,设定酶活性和蛋白溶解度为响应指标,以优化几丁质酶 C 包涵体的复性液并评估复性效果.结果表明:最佳复性液组成为二价阳离子 2 mmol·L-1、尿素、β-巯基乙醇 5 mmol·L-1、NaCl 与 KCl 浓度比为 250 mmol·L-1∶10 mmol·L-1、PEG 6000 质量分数 0.1%、山梨醇 0.5 mol·L-1、pH＝8.5,在此条件下,几丁质酶 C 包涵体的复性率达 77.9%.
To screen the composition of the renaturation buffer for inclusion bodies of the Serratia marcescens mutant chitinase C expressed in E.coli BL21(DE3),the inclusion bodies were isolated,washed,and purified by metal chelate affinity chromatography.Using fractional factorial design,13 factors were examined,and en-zyme activity and protein solubility were set as response indicators to optimize the refolding solution of chit-i nase C inclusion bodies and evaluate the refolding effect.The results showed that the optimal refolding solution composition was 2 mmol·L-1 divalent cations,urea,5 mmol·L-1β-mercaptoethanol,NaCl and KCl concen-tration ratio of 250 mmol·L-1∶10 mmol·L-1,0.1%mass fraction of PEG 6000,0.5 mol·L-1 sorbitol,and pH 8.5.Under these conditions,the refolding yield of chitinase C inclusion bodies reached 77.9%.
刘嘉;贺淹才
华侨大学 医学院,福建 泉州 362021华侨大学 医学院,福建 泉州 362021
生物科学
几丁质酶C包涵体复性分式析因设计
chitinase Cinclusion bodiesrenaturationfractional factorial design
《华侨大学学报（自然科学版）》 2026 (3)
309-317,9
华侨大学实验教学与管理改革项目(612-50423031)
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