High-throughput detection and quantification of vitamin B12 in microbiome isolates using Escherichia coli

Hencel, K.; Sullivan, M.; Akay, A.

doi:10.1101/2025.04.08.647760

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bioRxiv · 10.1101/2025.04.08.647760

High-throughput detection and quantification of vitamin B12 in microbiome isolates using Escherichia coli

Hencel, K.·Sullivan, M.·Akay, A.

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Abstract

Vitamin B12 is an essential micronutrient produced only by prokaryotes, and animals must acquire it from their diet. Vitamin B12 is critical for the synthesis of methionine and propionyl-CoA metabolism. In humans, vitamin B12 deficiency has been linked to many disorders, including infertility and developmental abnormalities. The growing trend towards plant-based diets and the ageing populations increase the risk of vitamin B12 deficiency, and therefore, there is an increasing interest in understanding vitamin B12 biology. Accurate approaches for detecting and quantifying vitamin B12 are essential in studying its complex biology, from its biogenesis in Bacteria and Archaea to its effects in complex organisms. Here, we present an approach using the commonly available E. coli methionine auxotroph strain B834 (DE3) and a multi-well spectrophotometer to detect and quantify vitamin B12 from biological samples at picomolar concentrations. We further show that our quantification method for vitamin B12 is sufficient to reveal important differences in the production of vitamin B12 from vitamin B12-synthesising bacteria commonly found in the microbiome of wild Caenorhabditis elegans isolates. Our results establish a high-throughput and simple assay platform for detecting and quantifying vitamin B12 using the E. coli B834 (DE3) strain.

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Hencel, K., Sullivan, M., Akay, A.. 2025-04-08. High-throughput detection and quantification of vitamin B12 in microbiome isolates using Escherichia coli. https://doi.org/10.1101/2025.04.08.647760

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