Directed evolution of glyphosate oxidase and a chemiluminescence system for glyphosate detection: A comprehensive practical laboratory experiment on biotechnology

IF 1.2 4区教育学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry and Molecular Biology Education Pub Date : 2023-03-27 DOI:10.1002/bmb.21721

Chanjuan Li, Gaobing Wu, Xuezhu Fan, Siqi Guo, Yuqing Qin, Yonggang Hu, Lifang Ruan

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引用次数: 0

Abstract

This article describes a comprehensive practical laboratory method for developing an enzyme to more easily measure glyphosate levels in solution. Through this article, undergraduate students of biology majors can conduct research experiments in critical fields by utilizing various techniques, such as chemiluminescence (CL) biosensors with engineered enzymes and are guided in molecular biology laboratories. A glyphosate oxidase mutant library was constructed by DNA shuffling, and a glyphosate oxidase variant with increased glyphosate degradation activity was selected by using a high-throughput screening assay. Following protein overexpression in Escherichia coli (DE3) and purification by affinity chromatography, the glyphosate oxidase variant protein combined with luminol–H₂O₂ reaction was constructed as a new CL biosensor for detecting glyphosate in soils.

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草甘膦氧化酶定向进化及检测草甘膦的化学发光系统:生物技术综合实验室实践实验

本文介绍了一种综合实用的实验室方法，用于开发一种更容易测量溶液中草甘膦水平的酶。通过本文，生物专业本科生可以在分子生物学实验室的指导下，利用工程酶化学发光(CL)生物传感器等多种技术，在关键领域进行研究实验。通过DNA重组构建草甘膦氧化酶突变体文库，并通过高通量筛选筛选出草甘膦降解活性增加的草甘膦氧化酶突变体。通过在大肠杆菌(Escherichia coli, DE3)中过表达该蛋白并通过亲和层析纯化，构建了结合鲁米诺- h2o2反应的草甘膦氧化酶变异蛋白，作为检测土壤中草甘膦的新型CL生物传感器。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biochemistry and Molecular Biology Education 生物-生化与分子生物学

CiteScore

2.60

自引率

14.30%

发文量

审稿时长

6-12 weeks

期刊介绍： The aim of BAMBED is to enhance teacher preparation and student learning in Biochemistry, Molecular Biology, and related sciences such as Biophysics and Cell Biology, by promoting the world-wide dissemination of educational materials. BAMBED seeks and communicates articles on many topics, including: Innovative techniques in teaching and learning. New pedagogical approaches. Research in biochemistry and molecular biology education. Reviews on emerging areas of Biochemistry and Molecular Biology to provide background for the preparation of lectures, seminars, student presentations, dissertations, etc. Historical Reviews describing "Paths to Discovery". Novel and proven laboratory experiments that have both skill-building and discovery-based characteristics. Reviews of relevant textbooks, software, and websites. Descriptions of software for educational use. Descriptions of multimedia materials such as tutorials on various aspects of biochemistry and molecular biology.