Honors Theses

Life Finds a Way: Optimization of the Scalable Purification of a Recombinant Lanthanide Ion Selective Protein from Methylobacterium extorquens

Advisor

Justin Biffinger, Ph.D.

Department

Chemistry

Publication Date

4-22-2026

Document Type

Honors Thesis

Abstract

Over recent decades, global demands for lanthanides (Lns) have rapidly grown due to their irreplaceable functions in various consumer electronic devices, including smartphones, computers, tablets, and more. Despite these circumstances, conventional Ln harvesting methods, largely via industrial mining, frequently fail to meet commercial and environmental demands. Lanmodulin (LanM), a novel Ln binding protein first isolated from Methylobacterium extorquens (M. extoorquens), offers a potential solution to these issues. LanM has previously demonstrated selective sequestration of Lns from Ln-containing stocks thus making it a viable candidate for high-yield Ln recycling. Accordingly, this study focused on the development of methods to optimize the scalable production of purified LanM as a first step toward developing efficient biomining strategies for Lns. To accomplish these goals, this study utilized lactose-inducible E. coli transformed with a plasmid encoded for a recombinant form of LanM containing a 6×Histidine (6xHis) protein binding tag connected to the native LanM protein through a Tobacco Etch Virus (TEV) protease cleavage sequence. Cells containing the plasmid of interest were subjected to autoinduction via expression in lactose-containing media. Following the lysis of the cultured cells, the 6xHis-tagged LanM was isolated using nickel-nitrilotriacetic acid (Ni-NTA) affinity chromatography and the 6xHis-tag was subsequently removed through cleavage of the TEV protease cleavage sequence by a TEV protease. The viability of autoinduction and purification conditions was assessed through quantification of the yield of purified LanM via the Bradford colorimetric assay.

Permission Statement

This item is protected by copyright law (Title 17, U.S. Code) and may only be used for noncommercial, educational, and scholarly purposes.

Keywords

Undergraduate research

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