生物学杂志

生物学杂志 ›› 2023, Vol. 40 ›› Issue (5): 69-.doi: 10.3969/j.issn.2095-1736.2023.05.069

• 研究报告 • 上一篇    下一篇

固定化表面展示锰过氧化物酶对甲基橙的吸附特性

王艳君1，2， 黄佳瑶1， 陈少煌1， 赵鹏飞1   

  1. 1. 福建技术师范学院 食品与生物工程学院， 福清 350300；
    2. 食品软塑包装技术福建省高校工程研究中心， 福清 350300
  • 出版日期:2023-10-18 发布日期:2023-10-17
  • 作者简介:王艳君，博士，副教授，研究方向为环境微生物，E-mail:507842885@qq.com
  • 基金资助:
    国家自然科学基金项目（21547005）； 福建省自然科学基金项目（2019J01893）； 福建省新世纪优秀人才支持计划（闽教科2016-23）； 福建省教育厅中青年教师教育科研项目（JAS180651）

Immobilized surfaces demonstrate the adsorption properties of manganese peroxidase on methyl orange

王艳君1，2， 黄佳瑶1， 陈少煌1， 赵鹏飞1   

  1. 1. School of Food and Bioengineering, Fujian Polytechnic Normal University, Fuqing 350300, China;
    2. Fujian Universities and Colleges Engineering Research Center of Soft Plastic Packaging Technology for Food,
    Fuqing 350300, China
  • Online:2023-10-18 Published:2023-10-17

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摘要： 将酵母细胞表面展示载体pYD1-MnP经LiAC法转化至酿酒酵母（Saccharomyces cerevisiae）EBY100，经SDS-PAGE及免疫荧光检测表明，MnP蛋白已成功锚定于酵母细胞表面，体外诱导转化子产锰过氧化物酶活性。为偶氮染料废水的降解提供优良的微生物资源。经2,6-二甲氧基苯酚(DMP)法测定酶活并通过选取不同培养时间、培养温度、pH、碳源、氮源、碳氮源的浓度等因素对酶活影响进行测定，并以海藻酸钠将游离酶进行固定化。锚定成功的转化子PM-2体外诱导产锰过氧化物酶的最适培养时间为60 h、诱导产酶最适温度为40 ℃；最适pH值为4.5，最适碳源为半乳糖，最适氮源为YNB，碳源浓度为0.02 g/mL，氮源浓度为0.0072 g/mL；经海藻酸钠固定化后对甲基橙模拟废水进行吸附性能研究，吸附动力学表明，吸附过程符合准二级方程；与Freundlich等温吸附方程拟合效果更佳。锚定成功的转化子PM-2具有产锰过氧化物酶活性的能力，可在偶氮染料的生物降解过程中发挥作用。

关键词: 表面展示, 锰过氧化物酶, 免疫荧光, 固定化, 甲基橙

Abstract: The yeast cell surface display vector pyd1 MnP was transformed into EBY100 by LiAC method. The MnP protein was successfully anchored on the surface of yeast cells by SDS-PAGE and immunofluorescence, and the transformant was induced to produce manganese peroxidase activity in vitro. It can provide excellent microbial resources for the degradation of azo dye wastewater. The enzyme activity was determined by 2,6-dimethoxyphenol (DMP) method, and the effects of different culture time, temperature, pH, carbon source, nitrogen source and concentration of carbon and nitrogen source on enzyme activity were determined. The enzyme was immobilized with sodium alginate. The optimal culture time and temperature of the immobilized transformant PM-2 were 60 h and 40 ℃. The optimal pH was 4.5, the optimal carbon source was galactose, the optimal nitrogen source was YNB, the carbon source concentration was 0.02 g/mL, and the nitrogen source concentration was 0.0072 g/mL. The adsorption performance of methyl orange simulated wastewater was studied by sodium alginate immobilization. The adsorption process accords with the quasi second order equation, and the fitting effect is better with Freundlich isotherm adsorption equation. The successfully anchored transformant PM-2 had the ability to produce manganese peroxidase, which can play a role in the biodegradation of azo dyes.

Key words: surface display, manganese peroxidase, immunofluorescence, immobilization, methyl orange

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