维生素C 对嗜酸乳杆菌GIM1.208 β-葡萄糖苷酶 激活效应及分子互作关系  

Effect of vitamin C on the activation ofβ-glucosidase in Lactobacillus acidophilus GIM1.208 and the interaction between them

作　　者：江桂兰 王翼 丁筑红 李彬彬 JIANG Guilan;WANG Yi;DING Zhuhong;LI Binbin(Guizhou Rosa Roxburghii Research Institute,Engineering Technology Research Center for Rosa Roxburghii of National Forestry and Grassland Administration,School of Liquor and Food Engineering,Guizhou University,Guiyang 550025,Guizhou,China)

机构地区：[1]贵州大学酿酒与食品工程学院国家林业和草原局刺梨工程技术研究中心贵州省刺梨研究院,贵州贵阳550025

出　　处：《微生物学通报》2023年第12期5392-5403,共12页Microbiology China

基　　金：国家自然科学基金(32160532)。

摘　　要：【背景】研究发现维生素C(vitamin C,VC)对嗜酸乳杆菌GIM1.208β-葡萄糖苷酶(β-glucosidase,BGL)具有明显激活作用。【目的】探究VC对BGL的影响特性以及二者互作关系。【方法】通过对硝基苯酚法、电化学法、Zeta电位法及核磁共振氢谱法研究VC对BGL的活性影响及互作特性。【结果】VC对BGL催化底物活性具有明显促效作用,K_(m)=1.167 mmol/L,V_(max)随着VC浓度增大而增大;VC浓度为3.5%时酶活性达到最高;在VC浓度为3.5%、温度低于30℃时,酶活性受温度影响较小,VC对BGL具有良好的稳定作用,相对酶活保持在90%以上。VC与BGL结合,两者存在弱静电作用且两者的相互作用使得VC的电氧化反应变得更加容易,同时推测VC分子中乙二醇支链片段与BGL之间存在氢键作用。【结论】VC对BGL具有明显激活作用,试验探究了两者作用力类型及作用位点,为BGL应用及天然酶激活剂的高效利用提供设计理论参考。[Background]A preliminary study showed that vitamin C(VC)had a significant activation effect on theβ-glucosidase(BGL)from Lactobacillus acidophilus GIM1.208.[Objective]To explore the effect of VC on BGL and the interaction between VC and BGL.[Methods]The 4-nitrophenol method,electrochemistry,Zeta potential,and nuclear magnetic resonance were employed to study the effect of VC on BGL activity and the interaction characteristics.[Results]VC exerted a significant effect on the catalytic activity of BGL,with K_(m)=1.167 mmol/L,and the V_(max) increased with the increase in VC concentration.BGL presented the highest activity when the VC concentration was 3.5%.At the VC concentration of 3.5%and a temperature below 30℃,the BGL activity was barely affected by temperature,and VC had a good stabilizing effect on BGL,which showed the relative activity above 90%.VC can bind to BGL,and the weak electrostatic interaction between VC and BGL facilitated the electroxidation of VC.We hypothesized that there was a hydrogen bond between the ethylene glycol branch chain in VC molecule and BGL.[Conclusion]VC can activate BGL.We studied the type and site of interaction between VC and BGL,aiming to provide a theoretical reference for the application of BGL and the efficient utilization of natural enzyme activators.

关 键 词：Β-葡萄糖苷酶 维生素C 酶活性 相互作用 动力学参数 嗜酸乳杆菌GIM1.208 

分 类 号：TS201.3[轻工技术与工程—食品科学]