[212] Some intermediates in the biosynthesis of vitamin B12

Renz, Paul

doi:10.1016/s0076-6879(71)18013-5

Cited by 53 publications

(26 citation statements)

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“…In addition prominent fragments (MH ϩ -CN) due to loss of the cyanide ligand occurred at m/z ϭ 1,318.8 for pseudovitamin B 12 and at m/z ϭ 1,332.6 for factor A. The FAB mass spectra confirmed the molecular formulae C 59 Very specific information on the structure of pseudovitamin B 12 and factor A in aqueous solutions was obtained from thorough NMR spectroscopic investigations. Complete assignment of all but three exchange labile hydroxyl and amino protons of the nucleotide moiety and of all carbons was obtained for the spectra of pseudovitamin B 12 and factor A.…”

Section: Guidedmentioning

confidence: 84%

“…Crystalline factor A was from the collection of the late W. Friedrich and presumably originated from sewage sludge (P. Renz, personal communication). Cobinamide was prepared by degradation of an aqueous solution of crystalline cyanocobalamin (Hoffmann-LaRoche, Basel, Switzerland) with cerous hydroxide by adaptation of the method described by Renz (59). Separation of cobinamide and ␣-D-ribazole formed in this reaction was achieved by preparative high-pressure liquid chromatography (HPLC) on an RP-18 column.…”

Section: Methodsmentioning

confidence: 99%

“…(13). In line with this, guided biosynthesis has also been employed for the preparation of nonnatural cobamides, such as naphthimidazolyl-and imidazolylcobamides (22,45,59,65), using cultures of Propionibacterium freudenreichii and P. shermanii.…”

mentioning

confidence: 99%

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Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B₁₂and Factor A

et al. 2000

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The corrinoids from the obligate anaerobe Clostridium cochlearium were extracted as a mixture of Co ␤ -cyano derivatives. From 50 g of frozen cells, approximately 2 mg (1.5 mol) of B 12 derivatives was obtained as a crystalline sample. Analysis of the corrinoid sample of C. cochlearium by a combination of high-pressure liquid chromatography and UV-Vis absorbance spectroscopy revealed the presence of three cyano corrinoids in a ratio of about 3:1:1. The spectroscopic data acquired for the sample indicated the main components to be pseudovitamin B 12 (Co ␤ -cyano-7؆-adeninylcobamide) (60%) and factor A (Co ␤ -cyano-7؆-[2-methyl]adeninylcobamide) (20%). Authentic pseudovitamin B 12 was prepared by guided biosynthesis from cobinamide and adenine. Both pseudovitamin B 12 and its homologue, factor A, were subjected to complete spectroscopic analysis by UV-Vis, circular dichroism, mass spectrometry, and by one-and two-dimensional 1 H, 13 C-, and 15 N nuclear magnetic resonance (NMR) spectroscopy. The third component was indicated by the mass spectra to be an isomer of factor A and is likely (according to NMR) to be 7؆-[N 6 -methyl]-adeninylcobamide, a previously unknown corrinoid. C. cochlearium thus biosynthesizes as its native "complete" B 12 cofactors the 7؆-adeninylcobamides and two homologous corrinoids, in which the nucleotide base is a methylated adenine.

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Section: Guidedmentioning

confidence: 84%

Section: Methodsmentioning

confidence: 99%

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Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B₁₂and Factor A

et al. 2000

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“…Preparation and incubation of the bacterial homogenate P. shermanii was grown under omission of a cobaltous salt as previously described [12]; 60 g frozen cell paste was pressed at -20°C in an X-press (Bachofer, Reutlingen, FRG) and suspended in 120 ml 0.29 M phosphate pH 6.5 (prepared by adding 1.93 ml 85% H3P04, 0.33 ml 5 M KOH and 0.1 ml 1 M MgC12 to 50 ml water, adjusting the pH to 6.5 with 50% NaOH, and adding water to a final volume of 100 ml). After addition of a small amount of deoxyribonuclease I, the suspension was stirred in an ice bath until it was homogeneous.…”

Section: Enzymatic Transformation Of Riboflavin Into 56-dimethylbenzmentioning

confidence: 99%

Biosynthesis of vitamin B12

Lingens

Schild

Vogler

et al. 1992

European Journal of Biochemistry

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The 5,6-dimethylbenzimidazole moiety of vitamin B12 is formed from riboflavin in aerobic and some aerotolerant bacteria. Thereby C1' of riboflavin is transformed into C2 of the vitamin B12 base. In the present publication a study on this transformation with riboflavin 'H-labeled in the 1'R or 1's position is described. This study was undertaken in order to find out if one of the two hydrogens at C1' is transferred to C2 of 5,6-dimethylbenzimidazole.The 2H-labeled riboflavin samples were synthesized starting from unlabeled or 1-'H-labeled ribose and 3,4-dimethylaniline yielding N-P-~-ribopyranosyl-3,4-dimethylaniline. The unlabeled riboside was reduced to N-~-ribityl-3,4-dimethylaniline with sodium cyanoborotrideuteride, the 'H-labeled riboside with sodium cyanoborohydride. The ribityl derivatives were transformed into N-~-ribityl-2-phenylazo-4,5-dimethylaniline, and condensed with barbituric acid to riboflavin.

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“…Factor Y was degraded as described in [3]. On degradation of factor V by the same method part of the nucleoside was transformed into a very basic product, presumably by substitution of the 2-methylsulfonyl group in the ammonia solution by an NH2 group catalyzed by cerous ion.…”

Section: Degradation Of Factors Y and V With Cerous Hydroxide To Cobimentioning

confidence: 99%

Two new vitamin B‐12 factors from sewage sludge containing 2‐methylsulfinyladenine or 2‐methylsulfonyladenine as base component

Renz

Blickle

Friedrich

1987

European Journal of Biochemistry

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Two hitherto unknown vitamin B-12 factors were isolated from sewage sludge. They were degraded with cerous hydroxide to cobinamide and the corresponding nucleoside. The nucleosides were further split with dilute hydrochloric acid to the bases and D-ribose. The structure of the two bases was found to be 2-methylsulfinyladenine and 2-methylsulfonyladenine. This was revealed by mass, infrared and nuclear magnetic resonance spectroscopy, and by comparison with the synthetic compounds.On addition of the synthetic bases to fermentations with Propionibacterium acidi-propionici the vitamin B-12 factors containing the corresponding base were formed. They were identical with the 2-methylsulfinyladenylcobamide and 2-methylsulfonyladenylcobamide originally isolated from sewage sludge.Several years ago two vitamin B-12 factors were isolated from sewage sludge. They were purified to homogeneity by preparative cellulose chromatography. Thus 2 g of one factor (named factor Y) and approximately 100mg of the other factor (named factor V) were obtained in crystalline form.Here we describe the elucidation of the structure of these compounds, and the biochemical preparation starting from the synthetic base components. Abbreviations. Factor Y, 2-methylsulfinyladenylcobamide; factor V, 2-methylsulfonyladenylcobamide; pseudovitamin B-12, adenylcobamide. For simplicity, we do not use the complete systematic name of cobamides, as for instance Coa-[a-(aden-9-yl)]-Cop-cyanocobamide for pseudovitamin B-12 (see [I]). In all natural purinylcobamides the a-glycosidic bond to the ribose is from N-7 of the purine base and bonding to the cobalt through N-9. Since the corrinoids mentioned in this paper were isolated in the presence of cyanide, their /3-ligand is always the cyano group. MATERIALS AND METHODS Preparative isolationtallized from aqueous acetone. The more slowly moving factor (factor Y) amounted to approximately 2 g, the faster migrating corrinoid (factor V) to approximately 100 mg. Degradation of factors Y and V with cerous hydroxide to cobinamide and the nucleosideFactor Y was degraded as described in [3]. On degradation of factor V by the same method part of the nucleoside was transformed into a very basic product, presumably by substitution of the 2-methylsulfonyl group in the ammonia solution by an NH2 group catalyzed by cerous ion. Therefore factor V was degraded with cerous hydroxide suspended in 0.1 M aqueous sodium cyanide essentially as described in [4].The nucleoside was separated from cobinamide and purified by chromatography on Amberlite XAD-2 (Serva, Heidelberg, 0.05 -0.1 mm; 42 x 2 cm). 6-ml fractions were collected. On elution with water the nucleoside left the column between fractions 60 and 120. Cobinamide was eluted with methanol/water (8/2, v/v). Acid hydrolysis of the nucleosides10 mg of nucleoside, dissolved in 2 ml 0.2 M HCI, was kept at 80°C for 10 min. The mixture was placed on a column (2 x 40 cm) of Amberlite XAD-2. On elution with water (6-ml fractions) ribose was found in fractions 9 -15, the purine base in...

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[212] Some intermediates in the biosynthesis of vitamin B12

Cited by 53 publications

References 11 publications

Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B₁₂and Factor A

Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B₁₂and Factor A

Biosynthesis of vitamin B12

Two new vitamin B‐12 factors from sewage sludge containing 2‐methylsulfinyladenine or 2‐methylsulfonyladenine as base component

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[212] Some intermediates in the biosynthesis of vitamin B12

Cited by 53 publications

References 11 publications

Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B12and Factor A

Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B12and Factor A

Biosynthesis of vitamin B12

Two new vitamin B‐12 factors from sewage sludge containing 2‐methylsulfinyladenine or 2‐methylsulfonyladenine as base component

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Product

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Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B₁₂and Factor A

Native Corrinoids fromClostridium cochleariumAre Adeninylcobamides: Spectroscopic Analysis and Identification of Pseudovitamin B₁₂and Factor A