2025年 02期

Effects of Evolution and Heterologous Expression of Oxalate Decarboxylase on Oxalate Degradation Ability of Limosilactobacillus reuteri

摘要(Abstract):

为了开发能够治疗或辅助治疗高草酸尿症的益生菌,采用基于深度学习的方法对枯草芽孢杆菌的草酸脱羧酶(OXDC)进行蛋白质进化,并在罗伊氏乳杆菌Limosilactobacillus reuteri Q35中异源表达野生型OXDC和2个预测催化常数更大的突变型OXDC-M5和OXDC-M10,比较基因工程菌株在生长体系和全细胞转化体系中降解草酸盐的能力。结果表明:突变型、野生型OXDC的表达均能显著提高罗伊氏乳杆菌的草酸铵降解能力,且OXDC-M10表达菌株L_3的降解能力最强;生长体系中培养24 h,菌株L_3的草酸盐降解率为45%,比对照菌株的高52.03%,比野生型OXDC表达菌株L_1的高12.78%;转化体系中培养4 h,菌株L_3的草酸盐降解率为46.56%,比对照菌株的高23.24%,比野生型OXDC表达菌株L_1的高21.25%,利用基因工程和蛋白质工程相结合的方法可显著提高罗伊氏乳杆菌Q35的草酸盐降解能力。

关键词(KeyWords):蛋白质进化;草酸盐降解;异源表达;罗伊氏乳杆菌;草酸脱羧酶

基金项目(Foundation):国家自然科学基金项目(32302050);; 山东省自然科学基金项目(ZR2022QC004)

作者(Author): 牛冬玉,孔令慧,刘向勇,秦加阳

DOI: 10.13349/j.cnki.jdxbn.20250307.001

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