Methylotrophic bacteria with cobalamin-dependent mutases in primary metabolism as potential strains for vitamin B12 production

Antonie Van Leeuwenhoek. 2023 Mar;116(3):207-220. doi: 10.1007/s10482-022-01795-9. Epub 2022 Nov 17.


Several bacterial species are known for their ability to synthesize vitamin B12 but biotechnological vitamin B12 production today is restricted to Pseudomonas denitrificans and Propionibacterium freudenreichii. Nevertheless, the rising popularity of veganism leads to a growing demand for vitamin B12 and thereby interest in alternative strains which can be used as efficient vitamin B12 sources. In this work, we demonstrate that methylotrophic microorganisms which utilize the ethylmalonyl-CoA pathway containing B12-dependent enzymes are capable of active vitamin B12 production. Several bacteria with an essential function of the pathway were tested for vitamin B12 synthesis. Among the identified strains, Hyphomicrobium sp. DSM3646 demonstrated the highest vitamin B12 levels reaching up to 17.9 ± 5.05 µg per g dry cell weight. These relatively high vitamin B12 concentrations achieved in simple cultivation experiments were performed in a mineral methanol medium, which makes Hyphomicrobium sp. DSM3646 a new promising cobalamin-producing strain.

Keywords: Cobalamin; Ethylmalonyl-CoA pathway; LC–MS; Methylotroph; Pseudovitamin B12; Vitamin B12.

MeSH terms

  • Bacteria / metabolism
  • Intramolecular Transferases*
  • Propionibacterium freudenreichii* / metabolism
  • Vitamin B 12 / metabolism
  • Vitamins


  • Vitamin B 12
  • Intramolecular Transferases
  • Vitamins