木基载体固定化酶及其对染料的降解  

Enzyme Immobilization on Wood-derived Cellulose Scaffolds for Degradation of Dye Molecules

作　　者：陈贵茹 张智红 宋冉 宋先亮[1] 雷建都[1] 王璐莹[1] CHEN Guiru;ZHANG Zhihong;SONG Ran;SONG Xianliang;LEI Jiandu;WANG Luying(College of Materials Science and Technology,Beijing Forestry University,Beijing 100083,China)

机构地区：[1]北京林业大学材料科学与技术学院,北京100083

出　　处：《中国生物工程杂志》2024年第10期52-63,共12页China Biotechnology

基　　金：国家重点研发计划(2019YFB1309703);北京市自然科学基金(2202034)资助项目。

摘　　要：目的:利用固定化酶技术提高可溶性酶在工业应用中存在的回收困难、环境敏感和成本高等缺点,提高酶环境稳定性以及可重复使用性,降低应用成本。方法:选取巴沙木为载体原料,脱除木质素后制备得到木基载体,分别使用柠檬酸改性法、戊二醛活化法以及二者结合处理法对木基载体进行改性并固定化辣根过氧化物酶(horse radish peroxidase,HRP);使用傅里叶红外光谱和扫描电子显微镜对原始巴沙木、木基载体(wood)、酶@木基载体(HRP@wood)进行官能团与结构研究。结果:结合处理法的木基载体具有更高的酶负载量,可达43.0 mg/g,该固定化酶材料即为HRP@wood;分别选取结晶紫、亚甲基蓝、刚果红三种染料对HRP@wood进行静态脱色性能测试,研究染料浓度、H_(2)O_(2)浓度、反应时间对染料脱色效果的影响,确定HRP@wood对于多种染料的最佳脱色条件。结论:当结晶紫的质量浓度为12 mg/L和H_(2)O_(2)浓度为2 mmol/L,亚甲基蓝的质量浓度为4 mg/L和H_(2)O_(2)浓度为0.5 mmol/L,刚果红的质量浓度为100 mg/L和H_(2)O_(2)浓度为2 mmol/L,30 min时脱色率分别可达76.9%、78.2%和82.4%;当脱色时间延长直至120 min时,亚甲基蓝的脱色率可达100%;以HRP@wood构建了一种动态脱色反应器,结果表明,该反应器具有优异的可循环使用性和脱色效果。Objective:The use of immobilized enzyme technology overcomes the disadvantages of soluble enzymes in industrial applications,such as difficult recovery,environmental sensitivity and high cost,improving enzyme environmental stability and reusability of enzymes and reducing application costs.Methods:Balsa wood was selected as the raw material for the carriers,and the wood-based carriers were prepared by removing the lignin.The wood-based carriers were modified and immobilized with horseradish peroxidase(HRP)using citric acid modification,glutaraldehyde activation,and a combination of the two treatments,respectively.The functional groups and structures of the original balsa wood,wood,and HRP@wood were investigated using Fourier infrared spectroscopy(FT-IR)and scanning electron microscopy(SEM).Conclusion:The wood-based carrier with the combination of the two treatments had a higher immobilization of the enzyme up to 43.0 mg/g,and this immobilized enzyme material was named [e-mail] dyes,crystal violet,methylene blue and Congo red,were selected to test the static decolorization performance of HRP@wood,and the optimal decolorization conditions of HRP@wood for a variety of dyes were determined by investigating the effects of dye concentration,H_2O_(2)concentration and reaction time on the decolorization effect of the dyes.Results:The dyes were decolored at up to 76.9%,78.2%,and 82.4%,respectively in the first half hour when decolorized under ideal circumstances,which included 12 mg/L crystal violet and 2 mmol/L H_2O_2,4 mg/L methylene blue and 0.5 mmol/L H_2O_2,and 100 mg/L Congo red and 2 mmol/L H_2O_2.The decolorization rate of methylene blue can reach 100%when the decolorization period was increased to 120 mins.A dynamic decolorization reactor was constructed with this HRP@wood,and the results showed that the reactor had excellent recyclability and decolorization effect.

关 键 词：木材 辣根过氧化物酶 固定化酶 染料降解 

分 类 号：TQ35[化学工程]