The enzyme participates in the aerobic (late cobalt insertion) adenosylcobalamin biosynthesis pathway. The enzyme from the bacterium Pseudomonas denitrificans is a fusion protein with two active sites; one catalyses the methylation at C-15 followed by decarboxylation of the C-12 acetate side chain, while the other catalyses the methylation at C-5. The corresponding activities in the anaerobic adenosylcobalamin biosynthesis pathway are catalysed by EC 2.1.1.196, cobalt-precorrin-6B (C15)-methyltransferase [decarboxylating], and EC 2.1.1.289, cobalt-precorrin-7 (C5)-methyltransferase, respectively.
The expected taxonomic range for this enzyme is: Bacteria, Archaea
The enzyme participates in the aerobic (late cobalt insertion) adenosylcobalamin biosynthesis pathway. The enzyme from the bacterium Pseudomonas denitrificans is a fusion protein with two active sites; one catalyses the methylation at C-15 followed by decarboxylation of the C-12 acetate side chain, while the other catalyses the methylation at C-5. The corresponding activities in the anaerobic adenosylcobalamin biosynthesis pathway are catalysed by EC 2.1.1.196, cobalt-precorrin-6B (C15)-methyltransferase [decarboxylating], and EC 2.1.1.289, cobalt-precorrin-7 (C5)-methyltransferase, respectively.
Substrates: methylation and decarboxylation are catalyzed by 2 different enzyme encoded by gene cbiE, which is responsible for the 2 methylations at C5 and C15, and by gene cbiT, which is responsible for the decarboxylation, the genes are fused together, overview Products: -
Substrates: methylation and decarboxylation are catalyzed by 2 different enzymes in Propionibacterium freudenreichii, encoded by gene cbiE, organized in a orf/single gene with cbiG and cbiH, which is responsible for the methylations at C5 and C15, and by gene cbiT, which is responsible for the decarboxylation, steps of the anaerobic cobalamin biosynthesis, pathway overview Products: -
Substrates: methylation and decarboxylation are catalyzed by 2 different enzymes in Propionibacterium freudenreichii, encoded by gene cbiE, organized in a orf/single gene with cbiG and cbiH, which is responsible for the methylations at C5 and C15, and by gene cbiT, which is responsible for the decarboxylation, overview Products: -
Substrates: the organism performs the anaerobic cobalamin biosynthesis variant, the gene cbiE of Propionibacterium freudenreichii is organized in a single ORF with genes cbiH and cbiG, overview Products: -
Substrates: methylation and decarboxylation are catalyzed by 2 different enzymes in Propionibacterium freudenreichii, encoded by gene cbiE, organized in a orf/single gene with cbiG and cbiH, which is responsible for the methylations at C5 and C15, and by gene cbiT, which is responsible for the decarboxylation, steps of the anaerobic cobalamin biosynthesis, pathway overview Products: -
Substrates: the organism performs the anaerobic cobalamin biosynthesis variant, the gene cbiE of Propionibacterium freudenreichii is organized in a single ORF with genes cbiH and cbiG, overview Products: -
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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
purified recombinant apo-enzyme CbiT or in complex with S-adenosyl-L-methionine, hanging drop vapour diffusion method, 25°C, for the apo-enzyme: 5-10 mg/ml protein in 10% w/v PEG 8000, 100 mM MgCl2, and 5 mM Tric-HCl, pH 7.5, 0.002 ml equilibrated against 1 ml of well solution containing 16-22% PEG 8000, 0.2 M MgCl2, 10 mM Tris-HCl, pH 7.5, crystals appear after 1 day, for the binary complex: 1-2 mg/ml protein in 0.5 mM S-adenosyl-L-methionine, 0.1-0.2% PEG 8000, 50 mM MgCl2, 2.5 mM Tris-HCl, pH 7.5, 0.004 ml equilibrated against 1 ml of well solution containing 0.2-0.4% PEG 8000, 0.2 M MgCl2, and 5 mM Tris-HCl, pH 7.5, cryoprotection by soaking in 25% PEG 8000, 0.2 M MgCl2, and glycerol up to 25% w/v, freezing in liquid propane, X-ray diffraction structure determination and analysis at 1.9 A resolution, modeling
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CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene MT0146 or cbiT, fused to gene cbiE, resulting in cobL, DNA and amino acid sequence determination and analysis, promotor analysis, expression in Escherichia coli strain B834
genes cbiEGH and cbiT, the gene cbiE of Propionibacterium freudenreichii is organized in a single ORF with genes cbiH and cbiG, DNA and amino acid sequence determination and analysis, genetic organization, co-overexpression of enzymes and reconstruction of the anaerobic cobalamin biosynthetic pathway in Escherichia coli in a construct comprising several genes, overview
Genetic and sequence analysis of an 8.7-kilobase Pseudomonas denitrificans fragment carrying eight genes involved in transformation of precorrin-2 to cobyrinic acid
Biosynthesis of vitamin B12 in Pseudomonas denitrificans: the biosynthetic sequence from precorrin-6y to precorrin-8x is catalyzed by the cobL gene product
Isolation and characterization of 14 additional genes specifying the anaerobic biosynthesis of cobalamin (vitamin B12) in Propionibacterium freudenreichii (P. Shermanii)