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V D Bokkenheuser

Publications and source records attributed to V D Bokkenheuser.

50 records · Page 3Linked to original sources
PubMed1979

Characterization of a C21 neutral steroid hormone transforming enzyme, 21-dehydroxylase, in crude cell extracts of Eubacterium lentum.

A strain of the obligate anaerobe, Eubacterium lentum, isolated from human feces, catalyzes the 21-dehydroxylation of 11-deoxycorticosterone to progesterone. A quantitative radiochromatographic assay was developed to measure 21-dehydroxylase activity in cell extracts. Maximum enzyme activity in cell extracts required both a reduced pyridine nucleotide and an oxidized flavin coenzyme. However, photochemically reduced flavin (FMNH2) could replace the requirement for NAD(P)H plus oxidized flavin. NAD(P)H : flavin (either FMN or FAD) oxidoreductase activity was detected spectrophotometrically in cell extracts assayed under anaerobic conditions. 21-Dehydroxylase was active from pH 5.4 to 8.5 with an apparent optimum between 6.4 and 6.8 using mixtures of NADH plus FMN as coenzymes. The substrate concentration at half-maximal reaction velocity was 8.0 microM and a specific acitivity of 5.8 nmol [3H]progesterone formed . h-1 . mg-1 protein was determined using [3th]deoxycorticosterone as substrate. Atabrine, rotenone, acriflavin, and 2,4-dinitrophenol (all at 1 mM) inhibited 21-dehydroxylase activity in cell extracts by 25, 24, 35 and 84%, respectively. These results suggest that 21-dehydrogenase may be coupled to a NAD(P)H : flavin oxidoreductase system in E. lentum.

Autoradiography↗
PubMed1979

Bacterial metabolism of corticoids with particular reference to the 21-dehydroxylation.

Clostridium paraputrificum, an obligate anaerobe recovered from human feces, reduces the alpha,beta-unsaturated carbonyl of deoxycorticosterone, corticosterone, cortisone, and cortisol. The same steroids are 21-dehydroxylated by Culture 116, recently isolated from human feces, and by a closely related organism, Eubacterium lentum. The 21-dehydroxylase has no effect on hydroxyl groups at carbon atoms 11 and 17.

Adrenal Cortex Hormones↗
PubMed1979

New markers for Eubacterium lentum.

Of 37 strains of Eubacterium lentum and phenotypically similar organisms, 26 (70%) synthesized a corticoid 21-dehydroxylase and/or a 3 alpha-hydroxysteroid dehydrogenase. It appeared that the corticoid 3 alpha-hydroxysteroid dehydrogenase was identical to the bile acid 3 alpha-hydroxysteroid dehydrogenase. Steroid-metabolizing enzymes were found both in E. lentum and in phenotypically similar organisms. E. lentum is characterized by nitrate reduction and enhanced growth in the presence of arginine. Many phenotypically similar organisms possess either one or the other of the two markers. In contrast, using the steroid-metabolizing enzymes as markers, a "steroid-active" and a "steroid-inactive" group were established with minimal overlapping of metabolic characteristics. Synthesis of the steroid enzymes was positively correlated with production of gas from H2O2 and formation of H2S. A simple method for the detection of corticoid 21-dehydroxylase and 3 alpha-hydroxysteroid dehydrogenase, one or both of which were present in 92% of the steroid-active group, is described.

3-Hydroxysteroid Dehydrogenases↗
PubMed1979

Detection of enteric campylobacteriosis in children.

Campylobacter fetus subsp. jejuni was recovered as the sole bacterial pathogen from 31% of 0- to 8-month-old children with acute gastroenteritis and from 5% of asymptomatic children (P less than 0.05). In children 8 to 24 months old, the respective recovery rates were 38 and 40%. With the exception of one case of simultaneous bacteremia, the clinical course of the symptomatic infection was benign and rarely lasted more than a week. The isolates were sensitive to most commonly used antibiotics. Several isolates shared antigenic determinants with C. fetus subsp. intestinalis. A bacteriophage specific for C. fetus subsp. jejuni lysed 73% of the strains. Several phage-resistant isolates carried a phage that lysed the stock strain of C. fetus subsp. jejuni (NADC 917).

Anti-Bacterial Agents↗
PubMed1978

Metabolism of biliary steroids by human fecal flora.

Biliary excreted steroid hormones are metabolized by the intestinal flora. The metabolic pathways of deoxycorticosterone and progesterone, including the bacterial species involved, have been established. The pathways for other biliary steroids are unknown. Information on the biological activities of the metabolites is lacking, but methods for their investigation are now available. Evidence suggests that the formation of the metabolites is influenced by dietary habits, and that, in some cases, it may be controlled by suppression of the responsible bacteria.

Bacteria↗
PubMed1977

Isolation and characterization of human fecal bacteria capable of 21-dehydroxylating corticoids.

It has been known for a decade that human intestinal flora include organisms capable of 21-dehydroxylating corticoids. Yet the identity of the organisms synthesizing 21-dehydroxylase has remained unknown. Using diluted human feces, we determined the prevalence of colonies of 21-dehydroxylating organisms on a variety of media. Isolation from the medium of colonies with the highest prevalence yielded an obligate anaerobe capable of 21-dehydroxylating deoxycorticosterone and tetrahydrodeoxycorticosterone. This transformation could be carried out in a prereduced medium by the microbial culture alone or in an aerobic medium reduced by growth of Escherichia coli. The culture shares many characteristics with Eubacterium lentum, the neotype strain of which elaborated both 21-dehydroxylase and 3alpha-hydroxysteroid dehydrogenase.

Anaerobiosis↗
PubMed1975

Metabolism of deoxycorticosterone by human fecal flora.

21-Dehydroxylation, a feature of metabolism of corticoids in humans, was observed in mixed cultures of fecal flora of normal individuals on a Western diet. The model substrate, 11-deoxycorticosterone (DOC), was metabolized to 3alpha-21-dihydroxy-5beta-pregnan-20-one (THDOC), 3alpha-hydroxy-5beta-pregnan20-one (pregnanolone), and to two unidentified structures, metabolites X and Y. DOC was not metabolized in all media supporting growth of fecal flora. Conversion required an initial pH between 6.0 and 8.0. 21-Dehydroxylation occurred within 4 days of incubation in media inoculated with 10-minus 1 to 10-minus 7 fecal suspensions. In higher dilutions, containing obligatory anaerobes only, DOC was converted to metabolite X and sometimes also to metabolite Y. The yield of pregnanolone was related to the promptness with which the specimen was processed, to the presence of cysteine in the medium, and to the concentrationof substrate (optimum, 16 to 64 mug of DOC per ml). The yield of THDOC was related to the delay in the processing of the specimen, the concentration of substrate (maximum at 256 mug/ml), and aeration of the culture. Pure cultures of aerobic organism of fecal origin either failed to metabolize DOC or converted it to metabolite Y. Pure cultures of fecal anaerobes converted DOC to metabolite X and sometimes also to metabolite Y. Neither THDOC nor pregnanolone was produced by pure cultures.

Aerobiosis↗
PubMedDate not supplied

The 21-acetylation of corticosteroids by Clostridium sporogenes.

A strain of Clostridium sporogenes, an anaerobic bacterium, isolated from sewage in New York City synthesizes two constitutive enzymes with action on steroid molecules: (i) an enzyme capable of selectively acetylating the 21-hydroxyl function of certain steroids and (ii) the corresponding esterase. Under our experimental conditions the enzymes have a strict structural requirement for 3-keto-4-ene and C-20-keto or 20 alpha-hydroxyl group and convert their respective substrates to a mixture of free and acetylated products.

Acetylation↗