Biosynthesis of cobamides: methods for the detection, analysis and production of cobamides and biosynthetic intermediates

Deery, Evelyne, Lawrence, Andrew D. and Warren, Martin J. (2022) Biosynthesis of cobamides: methods for the detection, analysis and production of cobamides and biosynthetic intermediates. In, Neil, E. and Marsh, G. (eds.) Methods in Enzymology: Coenzyme B12 Enzymes Part A. Elsevier Ltd., pp. 3-23. (doi:10.1016/bs.mie.2022.01.013).

Abstract

Vitamin B₁₂, cobalamin, belongs to the broader cobamide family whose members are characterized by the presence of a cobalt-containing corrinoid ring. The ability to detect, isolate and characterize cobamides and their biosynthetic intermediates is an important prerequisite when attempting to study the synthesis of this remarkable group of compounds that play diverse roles across the three kingdoms of life. The synthesis of cobamides is restricted to only certain prokaryotes and their structural complexity entails an equally complex synthesis orchestrated through a multi-step biochemical pathway. In this chapter, we have outlined methods that we have found extremely helpful in the characterization of the biochemical pathway, including a plate microbiological assay, a corrinoid affinity extraction method, LCMS characterization and a multigene cloning strategy.

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Contributors

Author: Andrew D. Lawrence

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