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ToolsHencel, Katarzyna, Sullivan, Matthew J. and Akay, Alper (2025) A simple, fast and inexpensive approach using E. coli to detect and estimate vitamin B12 content in microbial extracts. Biology Open, 14 (9). ISSN 2046-6390
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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 ageing populations increases 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 estimate the levels of vitamin B12 from biological samples at picomolar concentrations. We further show that our method 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 estimating vitamin B12 levels using the E. coli B834 (DE3) strain.
| Item Type: | Article |
|---|---|
| Additional Information: | Data and resource availability: All data generated or analysed during this study are included in this published article and its supplementary information files. Funding information: This work was supported by a UK Research and Innovation Future Leaders Fellowship [MR/S033769/1 and MR/X024261/1] awarded to A.A. from the Medical Research Council, a Springboard Award from the Academy of Medical Sciences [SBF009\1005] and a Royal Society research grant [RGS\R1\231151] awarded to M.J.S. K.H. was funded by the University of East Anglia doctoral training programme. Open Access funding provided by University of East Anglia School of Biological Sciences. Deposited in PMC for immediate release. |
| Uncontrolled Keywords: | c. elegans,caenorhabditis elegans,cembio,cobalamin,e. coli,mete,vitamin b,biochemistry, genetics and molecular biology(all),agricultural and biological sciences(all) ,/dk/atira/pure/subjectarea/asjc/1300 |
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| UEA Research Groups: | Faculty of Medicine and Health Sciences > Research Groups > Pathogen Biology Group Faculty of Science > Research Groups > Molecular Microbiology Faculty of Science > Research Groups > Cells and Tissues |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 17 Nov 2025 15:30 |
| Last Modified: | 23 Nov 2025 07:30 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/101033 |
| DOI: | 10.1242/bio.062017 |
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