Novel Pathway for Corrinoid Compounds Production in Lactobacillus

Abstract

Vitamin B12 or cobalamin is an essential metabolite for humans, which makes it an interesting compound for many research groups that focus in different producer-strains synthesis pathways. In this work, we report the influence of key intermediaries for cobalamin synthesis added to the culture medium in two Lactobacillus (L.) strains, L. reuteri CRL 1098 and L. coryniformis CRL 1001. Here, we report that addition of Co2+ and 5,6-dimethylbenzimidazole increased the corrinoid compounds production in both strains while addition of L-threonine increased only the corrinoid compounds production by CRL 1001 strain. Then, we purified and characterized by LC-MS the corrinoid compounds obtained. Physiological studies besides in silico analysis revealed that L. reuteri CRL 1098 and L. coryniformis CRL 1001 follow different pathways for the last steps of the corrinoid compounds synthesis.

Metabolism Characteristics of Lactic Acid Bacteria and the Expanding Applications in Food Industry

“…A strain of Latilactobacillus sakei UONUMA promotes the content of vitamin B2 (riboflavin), B3 (nicotinic acid and nicotinamide), and B6 (pyridoxine) in the traditional sweet Japanese beverage (koji amazake) ( Oguro et al, 2017 ). Limosilactobacillus reuteri CRL 1098 and Lactobacillus coryniformis CRL 1001 are well-known cobalamin producing strains ( Torres et al, 2018 ).…”

Microbial and Genetic Resources for Cobalamin (Vitamin B12) Biosynthesis: From Ecosystems to Industrial Biotechnology

“…The anaerobic pathway also requires five methyltransferases CysG, CbiK/X, and CbiLHF ( Figure 1 ). The cobalt chelation is catalyzed by the ATP-dependent heterotrimeric enzyme in the aerobic P. denitrificans consisting of the subnit CobN-magnesium chelatase (pfam02514), the subnit CobS-cobalamin 5′-phosphate synthase (TIGR01650), and the subunit CobT-cobalt chelatase (TIGR01651) or by the ATP-independent enzymes CbiK/X in the facultative anaerobes S. typhimurium / B. megaterium [ 31 , 34 , 35 ]. However, CobNST in the aerobic pathway are nongomologous to the enzymes with the same symbols in the anaerobic pathway due to their discovery history.…”

“…They only take up an extracellular cobinamide (Cbi I salvage pathway), which has to be converted it into the real intermediate, the last precursor AdoCbi-phosphate, assemble the nucleotide loop and attach the lower ligand ( Figure 1 ). Both de novo and salvage pathways are often themselves of lacking some genes or having alternative unknown genes and their uncertain organization in a chromosome [ 7 , 12 , 34 , 35 ]. The partial biosynthetic pathways predominantly lacking the first step, with the potential to salvage Cbi or tetrapyrrole precursors have been shown for Cbi-15.1%, ALA-0.3%, porphobilinogen (PBG)–0.6%, hydroxymethylbilane (HMB)–0.6%, UroIII–0.04%, precorrin-2–0.6%, where 78% salavgers of tetrapyrrole precursors are host-assosiated bacteria [ 7 ].…”

“…The de novo AdoCbl biosynthesis and Cbi I salvage pathways include the follow common genes: cobQ/cbiP encoding for the dimeric glutamine-hydrolyzing adenosylcobyric acid synthase CobQ/CbiP, with glutamine amidotransferase domain (5′-deoxy-5′-adenosylcobyrinic-acid- a,c -diamide:L-glutamine amido-ligase, EC 6.3.5.10) that catalyzes the four-step amidation sequence from cobyrinic acid a,c -diamide into cobyric acid; cobD/cbiB encoding for the AdoCbi-phosphate synthase CobD/CbiB (EC 6.3.1.10), attaching of an aminopropanol linker to the free carboxylic acid [ 20 , 29 ]. The aminopropanol is derived from threonine by the action of either PduX (in Salmonella -like bacteria) or nonorthologous BluE(F) (in Rhodobacterales) encoding for a kinase (EC 2.7.1.177), resulting in threonine phosphorylation, which is decarboxylated by the threonine-phosphate decarboxylase CobC (EC 4.1.1.81) for formation of ( R )-1-amino-2-propanol O-2-phosphate [ 7 , 35 , 42 ] ( Figure 1 ).…”

Media optimization studies and production of adenosylcobalamin (Vitamin B12) by environment friendly organism Rhizobium spp

“…The analysis suggests that the biosynthetic steps from co(II)byrinic acid a,c-diamide to adocobalamin are same in both the anaerobic and aerobic pathways. The yield of Vitamin B 12 from a genetically engineered, recombinant E. coli strain could be increased by more than ∼250-fold to 307.00 µg g −1 DCW by metabolic engineering and optimizing the favorable conditions required for fermentation [23].…”