Multiple Rounds of In Vivo Random Mutagenesis and Selection in Vibrio natriegens Result in Substantial Increases in REE Binding Capacity

Medin, Sean; Dressel, Anastacia; Specht, David A.; Sheppard, Timothy J.; Holycross, Megan E.; Reid, Matthew C.; Gazel, Esteban; Wu, Mingming; Barstow, Buz

doi:10.1021/acssynbio.3c00484

Title: Multiple Rounds of In Vivo Random Mutagenesis and Selection in Vibrio natriegens Result in Substantial Increases in REE Binding Capacity

Journal Article · 06 December 2023 · ACS Synthetic Biology

DOI: https://doi.org/10.1021/acssynbio.3c00484 · OSTI ID:2228762

^[1]; Dressel, Anastacia ^[1];

^[1]; Sheppard, Timothy J. ^[1]; Holycross, Megan E. ^[1];

^[1]; Gazel, Esteban ^[1];

^[1];

^[1]

Cornell Univ., Ithaca, NY (United States)

Rare earth elements (REE) are essential ingredients in many modern technologies, yet their purification remains either environmentally harmful or economically unviable. Adsorption, or biosorption, of REE onto bacterial cell membranes offers a sustainable alternative to traditional solvent extraction methods. But in order for biosorption-based REE purification to compete economically, the capacity and specificity of biosorption sites must be enhanced. Although there have been some recent advances in characterizing the genetics of REE-biosorption, the variety and complexity of bacterial membrane surface sites make targeted genetic engineering difficult. Here, we propose using multiple rounds of in vivo random mutagenesis induced by the MP6 plasmid combined with plate-throughput REE-biosorption screening to improve a microbe’s capacity and selectivity for biosorbing REE. We engineered a strain of Vibrio natriegens capable of biosorbing 210% more dysprosium compared to the wild-type and produced selectivity improvements of up to 50% between the lightest (lanthanum) and heaviest (lutetium) REE. We believe that mutations we observed in ABC transporters as well as a nonessential protein in the BAM outer membrane β-barrel protein insertion complex likely contribute to some but almost certainly not all of the biosorption changes we observed. Given the ease of finding significant biosorption mutants, these results highlight just how many genes likely contribute to biosorption as well as the power of random mutagenesis in identifying genes of interest and optimizing a biological system for a task.

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Research Organization:: Harvard Medical School, Boston, MA (United States)

Sponsoring Organization:: USDOE Advanced Research Projects Agency - Energy (ARPA-E)

Grant/Contract Number:: AR0001341

OSTI ID:: 2228762

Alternate ID(s):: OSTI ID: 2242482

Journal Information:: ACS Synthetic Biology, Vol. 12, Issue 12; ISSN 2161-5063

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
biosorption
rare earth elements
random mutagenesis
bacterial membrane
Vibrio natriegens

Title: Multiple Rounds of In Vivo Random Mutagenesis and Selection in Vibrio natriegens Result in Substantial Increases in REE Binding Capacity

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