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

ABSTRACT: 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.

SUBMITTER: Dudko D

PROVIDER: S-EPMC9925536 | biostudies-literature | 2023 Mar

REPOSITORIES: biostudies-literature

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

Dudko Darya D Holtmann Dirk D Buchhaupt Markus M

Antonie van Leeuwenhoek 20221117 3

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 microorgan ...[more]

PMID: 36385348

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Project description:BackgroundVitamin B12 is a crucial B-group vitamin, first isolated from the liver due to its role in combating pernicious anemia. It is distinguished by its unique and complex structure, which makes its chemical synthesis challenging and expensive. Consequently, vitamin B12 is alternatively obtained through microbial fermentations. Molasses, an affordable and safe agro-industrial waste, can be used as a carbon source for vitamin B12 production, offering a cost-effective alternative to expensive sugars in the production medium.ResultsA total of 87 yeast, actinomycete, and bacterial isolates were screened for vitamin B12 production, with 15 isolates showing high productivity. Bacillus isolates were selected for further analysis using MALDI-TOF and molecular identification. These isolates were identified as four strains of Bacillus subtilis (MZ08, JT10, BY11, and JT17), one strains of Bacillus sp. (CB09), and one strain of Peribacillus acanthi (MZ01). Genetic circuits associated with vitamin B12 production were demonstrated in a closely related strain of Peribacillus acanthi MZ01 strain. Three strains (MZ01, MZ08, and JT17) were selected for further evaluation of vitamin B12 productivity under different sugar types (glucose, sucrose, fructose, lactose, and galactose) and varying inoculum sizes. The inoculum size significantly impacted vitamin B12 production, with an increase from 5 to 10% enhancing yields. The ability of the strains to produce vitamin B12 varied depending on the type of sugar used. Peribacillus acanthi MZ01 strain showed the highest productivity and subsequently, selected for optimizing vitamin B12 production conditions using response surface methodology. Furthermore, the optimized conditions were then applied to molasses-based medium to achieve high vitamin B12 yields by MZ01 strain.ConclusionIn this study, Peribacillus acanthi was characterized for the first time as a vitamin B12 producer, demonstrating high productivity among various tested strains. The optimization of production conditions using response surface methodology, further enhanced vitamin B12 yields, showcasing the strain's efficiency in microbial fermentations. This research also highlights the potential of using molasses as a cost-effective alternative carbon source, significantly reducing production costs.

Dataset Information

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

Publications

Methylotrophic bacteria with cobalamin-dependent mutases in primary metabolism as potential strains for vitamin B<sub>12</sub> production.

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