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Table 2 Research on the biosynthesis of vitamin B12 and its intermediates in vivo and in vitro in E. coli

From: Microbial production of vitamin B12: a review and future perspectives

Strains Products Strategies References
In vivo
 E. coli Cobyric acid Expression of the cob I operon of B. megaterium and cbiP of S. typhimurium [77]
 E. coli Cobyrinic acid a,c-diamide Co-expression of the cobA gene from P. freudenreichii and the cbiAP, cbiCDETFGHJ, and cbiJKL genes from S. typhimurium [16]
 E. coli HBA Co-expression of cobA, cobI, cobJ, cobF, cobM, cobK, cobL, and cobH from R. capsulatus SB1003, and cobG from B. melitensis 16M or P. denitrificans [70]
 E. coli HBA Co-expression of cobA, cobI, cobJ, cobF, cobM, cobK, cobL, cobE, cobH and cobZ from R. capsulatus [35]
 E. coli Vitamin B12 Expression of the operon pduBAF-pocR-cbiABCDETFGHJKLMNQOP-cobUST from S. typhimurium [18]
 E. coli Vitamin B12 Co-expression of cobWNEMcbtAB, tonBcobOBRDCQchlID and cobGHIJLFK from P. denitrificans [83]
In vitro
 E. coli HBA E. coli native genes hemB, hemC, hemD, and cobA, cobI, cobG, cobJ, cobF, cobM, cobK, cobL, and cobH genes from P. denitrificans were heterologously expressed in E. coli. HBA was produced when ALA was added to the media and the cells were incubated aerobically in 200 ml Tris–HCl buffer, pH 8.0, containing SAM, NADH, and NADPH for 15 h at 30 °C with a mixture of twelve enzymes: HemB, HemC, HemD, crude enzyme extract of CobA, CobI, CobG, CobJ, CobM, CobF, CobK, CobL, and CobH [78]
 E. coli Precorrin3b and precorrin-4 The cobG and cobJ genes from P. denitrificans were heterologously expressed in E. coli. Aerobic incubation of precorrin-3A with the CobG enzyme from P. denitrificans alone yielded precorrin3B. When CobJ from P. denitrificans and S-adenosyl-l-methionine were included in the reaction, the product precorrin-4 was formed [18]
 E. coli Cob(II)yrinic acid a,c-diamide The cobN, cobS, and cobT from B. melitensis were expressed and purified from E. coli. In the presence of ATP, Co2+, and hydrogenobyrinic acid a,c diamide, these enzymes together produced cobyrinic acid [81]
 E. coli Cob(I)yrinic acid a,c-diamide CobR from B. melitensis was expressed and purified from E. coli. This enzmye was confirmed to have co(II)rrin reductase activity [82]