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PubMed2004

Development of a 16S rRNA primer for the detection of Brevibacterium spp.

AIM: To develop a PCR method for the rapid identification of the genus Brevibacterium. METHODS AND RESULTS: Genus-specific primers were designed by aligning and comparing the 16S sequence of all Brevibacterium spp. with closely related genera. The primer set was tested with all validly described Brevibacterium spp. and their closest neighbours. SIGNIFICANCE: Until today brevibacteria could only be identified with laborious and time-consuming phenotypic characterization. The primer from this study offers a rapid alternative to the detection of Brevibacterium spp. Brevibacteria have been isolated from food, blood, ear discharge, from a wound and from an intravascular catheter.

Blood↗
PubMed1994

Transcriptional analysis and regulatory signals of the hom-thrB cluster of Brevibacterium lactofermentum.

Two genes, hom (encoding homoserine dehydrogenase) and thrB (encoding homoserine kinase), of the threonine biosynthetic pathway are clustered in the chromosome of Brevibacterium lactofermentum in the order 5' hom-thrB 3', separated by only 10 bp. The Brevibacterium thrB gene is expressed in Escherichia coli, in Brevibacterium lactofermentum, and in Corynebacterium glutamicum and complements auxotrophs of all three organisms deficient in homoserine kinase, whereas the Brevibacterium hom gene did not complement two different E. coli auxotrophs lacking homoserine dehydrogenase. However, complementation was obtained when the homoserine dehydrogenase was expressed as a fusion protein in E. coli. Northern (RNA) analysis showed that the hom-thrB cluster is transcribed, giving two different transcripts of 2.5 and 1.1 kb. The 2.5-kb transcript corresponds to the entire cluster hom-thrB (i.e., they form a bicistronic operon), and the short transcript (1.1 kb) originates from the thrB gene. The promoter in front of hom and the hom-internal promoter in front of thrB were subcloned in promoter-probe vectors of E. coli and corynebacteria. The thrB promoter is efficiently recognized both in E. coli and corynebacteria, whereas the hom promoter is functional in corynebacteria but not in E. coli. The transcription start points of both promoters have been identified by primer extension and S1 mapping analysis. The thrB promoter was located in an 87-bp fragment that overlaps with the end of the hom gene. A functional transcriptional terminator located downstream from the cluster was subcloned in terminator-probe vectors.

Amino Acid Sequence↗
PubMed1994

Human infections caused by Brevibacterium casei, formerly CDC groups B-1 and B-3.

Forty-one clinical strains of CDC coryneform groups B-1 and B-3 were compared biochemically, by analysis of cell wall sugars, amino acids, and cellular fatty acids, and by DNA relatedness to the type strains of Brevibacterium casei, Brevibacterium epidermidis, and Brevibacterium linens. Twenty-two strains were shown to be B. casei, while five other strains formed a phenotypically inseparable genomospecies in the same genus. The remaining isolates were genetically heterogeneous, and most are probably members of the genus Brevibacterium. They were not further identified, but they were biochemically distinguishable from B. casei. Eleven of the clinical strains of B. casei were isolated from blood, and two each were isolated from cerebrospinal fluid and from pleural fluid. At least five isolates were from multiple blood or cerebrospinal fluid cultures. To our knowledge, these strains are the first described clinical isolates identified as B. casei, which was previously considered to be a nonpathogenic species.

Actinomycetales Infections↗
PubMed2005

Industrial importance of the genus Brevibacterium.

The genus Brevibacterium has long been difficult for taxonomists to classify due to its close morphological similarity to other genera. Since it was proposed in 1953, the genus has often been redefined. The genus is best known for its important role in the ripening of certain cheeses (B. linens) and for its supposed over-production of L: -amino acids. Other interesting industrial applications, including the production of ectoine, have recently been proposed. The general characteristics, the occurrence and the recent taxonomy of Brevibacterium are reviewed here. Furthermore, known and potential industrial applications for Brevibacterium species are briefly discussed.

Amino Acids↗
PubMed2006

Application of Streptomyces and Brevibacterium cholesterol oxidase for total serum cholesterol assay by the enzymatic kinetic method.

BACKGROUND: Using non-esterified cholesterol standard, Brevibacterium and Streptomyces are found as suitable sources of cholesterol oxidase for kinetic cholesterol assay. For clinical use, we investigated the suitability of these enzymes for cholesterol determination in human serum. METHODS: We compared the performance of reagents containing 2 enzymes for the kinetic determination of total serum cholesterol with the standardized endpoint method. RESULTS: Reagent containing Streptomyces enzyme was more sensitive than that of Brevibacterium, with linearity up to 20.7 and 2.6 mmol/l, respectively. The analytical reaction for Streptomyces showed a shorter lag phase (148 s) and a steeper slope (absorbance vs. time) than that of Brevibacterium (246 s). The assay using Streptomyces reagent was precise and accurate and compared favorably with the endpoint method (y=1.06x-0.15, r=0.996, bias=0.21 mmol/l). Hemoglobin as high as 7.5 g/l did not interfere while turbidity greater than 2+ (absorbance >0.778 at 670 nm) and bilirubin concentrations >171.0 micromol/l did interfere (in a negative interference). Reagent was stable up to at least 8 weeks. CONCLUSIONS: The Streptomyces cholesterol oxidase, with 3,4-dichlorophenol, proved a suitable source for serum total cholesterol determination by the kinetic method.

Artifacts↗
PubMed1999

Reclassification of Brevibacterium oxydans (Chatelain and Second 1966) as Microbacterium oxydans comb. nov.

Phylogenetic and chemotaxonomic analyses indicate that Brevibacterium oxydans is closely related to species of the genus Microbacterium, namely Microbacterium liquefaciens, Microbacterium luteolum and Microbacterium saperdae. DNA-DNA reassociation values of less than 60% between Brevibacterium oxydans and these three Microbacterium species support the distinctness of this misclassified Brevibacterium species, which is reclassified as Microbacterium oxydans comb. nov.

Actinomycetales↗
PubMed2001

Brevibacterium paucivorans sp. nov., from human clinical specimens.

Seven isolates from various human body sites displayed general chemotaxonomic and phenotypic characteristics of the genus Brevibacterium. This was corroborated by the 16S rRNA gene sequence analysis of strain CF62T, showing a sequence similarity of 99% to Brevibacterium mcbrellneri. However, DNA-DNA hybridization, a peculiar amino acid content of the cell wall and some phenotypic properties clearly suggested that these strains belong to a new species, for which the name Brevibacterium paucivorans sp. nov. is proposed. The type strain of B. paucivorans is CF62T (= DSM 13657T = LMG 19814T). The DNA G+C content of the type strain is 55.8 mol%.

Brevibacterium↗
PubMed2003

Brevibacterium lutescens sp. nov., from human and environmental samples.

Three strains of coryneform rods isolated from clinical samples and one of environmental origin exhibited phenotypic and chemotaxonomic properties characteristic of the genus Brevibacterium and their 16S rRNA gene sequences were closely related (98.5-99.0 %) to that of Brevibacterium otitidis. However, DNA-DNA hybridization of one strain (CF87(T)) showed only 59.6 % relatedness to the type strain of B. otitidis, DSM 10718(T), and 75-82 % relatedness to the three other strains. The four strains could be differentiated from B. otitidis by cellular fatty acid composition and some phenotypic characteristics. These findings suggest that the four strains belong to a novel species, for which the name Brevibacterium lutescens sp. nov. is proposed. The type strain of B. lutescens is CF87(T) (=DSM 15022(T)=CCUG 46604(T)).

Brevibacterium↗
PubMed2004

Brevibacterium celere sp. nov., isolated from degraded thallus of a brown alga.

Two whitish yellow, Gram-positive, non-motile, aerobic bacteria were isolated from enrichment culture during degradation of the thallus of the brown alga Fucus evanescens. The bacteria studied were chemo-organotrophic, mesophilic and grew well on nutrient media containing up to 15 % (w/v) NaCl. The DNA G+C content was 61 mol%. The two isolates exhibited a conspecific DNA-DNA relatedness value of 98 %, indicating that they belong to the same species. A comparative analysis of 16S rRNA gene sequences revealed that strain KMM 3637(T) formed a distinct phyletic lineage in the genus Brevibacterium (family Brevibacteriaceae, class Actinobacteria) and showed the highest sequence similarity (about 97 %) to Brevibacterium casei. DNA-DNA hybridization experiments demonstrated 45 % binding with the DNA of B. casei DSM 20657(T). Physiological and chemotaxonomic characteristics (meso-diaminopimelic acid in the peptidoglycan, major cellular fatty acids 15 : 0ai and 17 : 0ai) of the bacteria studied were consistent with the genomic and phylogenetic data. On the basis of the results of this study, a novel species, Brevibacterium celere sp. nov., is proposed. The type strain is KMM 3637(T) (=DSM 15453(T)=ATCC BAA-809(T)).

Aerobiosis↗
PubMed2002

Evaluation of methods for molecular typing and identification of members of the genus Brevibacterium and other related species.

The genus Brevibacterium includes pleomorphic Gram-positive bacteria with a high mol% G+C content. Species in the genus are difficult to identify by classical methods. The discriminatory power of DNA-based methods is assessed. Strains representing the four well established Brevibacterium species, and other related bacteria, were compared by amplified ribosomal DNA restriction analysis (ARDRA), repetitive-sequence-based PCR (rep-PCR) and ribotyping. Fingerprinting by rep-PCR and ribotyping provided complex genomic profiles with the highest discriminatory potential for molecular typing at the strain level, whereas ARDRA showed differentiation from the genus to the species levels. A high degree of heterogeneity within the genus Brevibacterium is apparent, thus indicating that the taxonomy of the genus should be further studied.

Bacterial Typing Techniques↗
PubMed1980

Enzymes involved in 3,5-diaminohexanoate degradation by Brevibacterium sp.

Cell-free extracts of Brevibacterium sp. L5 grown on DL-erythro-3,5-diaminohexanoate were found to contain a 3-keto-5-aminohexanoate cleavage enzyme that converts 3-keto-5-aminohexanoate and acetyl-coenzyme A (CokA) to 3-aminobutyryl-CoA and acetoacetate and a deaminase that coverts L-3-aminobutyryl-CoA to crotonyl-CoA. The cleavage enzyme has been purified extensively, and some of its properties have been determined for comparison with the 3-keto-6-acetamido-hexanoate cleavage enzyme of Pseudomonas sp. B4. The deaminase has been partially purified and characterized. Both the cleavage enzyme and the deaminase are induced by growth on 3,5-diaminohexanoate. The presence of these and other accessory enzymes in Brevibacterium sp. extracts accounts for the results of earlier tracer experiments which showed that C-1 and C-2 of 3-keto-5-aminohexanoate are converted mainly to acetoacetate and acetate, whereas C-3 to C-6 are converted mainly to 3-hydroxybutyrate or its coenzyme A thiolester. The enzymes observed in extracts of Brevibacterium sp. can account for the conversion of 3,5-diaminohexanoate to acetyl-CoA.

Amino Acids, Diamino↗
PubMed1984

Identification of Brevibacterium from clinical sources.

Coryneform bacteria of the genus Brevibacterium occur on the normal skin surface, but reports of human infection with this genus are lacking. A number of cultures of coryneform bacteria sent to the National Collection of Type Cultures for identification have been identified as Brevibacterium spp on the basis of their cell wall composition and ability to produce methane-thiol from L-methionine. We describe a rapid method for the detection of methane-thiol and confirmatory tests which differentiate Brevibacterium from morphologically similar genera.

Bacteriological Techniques↗
PubMed2005

Efficient utilization of ectoine by halophilic Brevibacterium species and Escherichia coli subjected to osmotic downshock.

Halophilic and non-halophilic bacteria subjected to osmotic downshock, from 0.7 M NaCl to deionized water, were examined for their survival, with the uptake and utilization of the cyclic amino acid ectoine, one of the representative compatible solutes, being taken into account. The uptake of ectoine added externally and survival of the cells were monitored as a function of incubation time in the presence and absence of NaCl. The halophilic Brevibacterium sp. JCM 6894 and B. epidermidis JCM 2593 actively accumulated ectoine regardless of the presence of NaCl, which led to cell survival. Brevibacterium casei JCM 2594 belonging to the same Brevibacterium species, however, revealed Na+-dependence of its uptake activity of ectoine. Non-halophilic Escherichia coli K-12 did not accumulate ectoine, and thereby this strain failed to survive irrespective of whether NaCl was present. The physiological meanings of the downshock procedure are discussed in connection with the uptake and the subsequent utilization of ectoine.

Amino Acids, Diamino↗
PubMed1995

The influence of the pH-value on the growth of Brevibacterium epidermidis in continuous culture.

Brevibacterium epidermidis is a major component of the bacterial flora of certain skin surface biotopes, characterized by a comparatively high pH-value. The presence of Brevibacterium epidermidis seems to be linked to the production of malodour. Skin surface pH has been found to be a major factor of bacterial growth on the skin. In order to find out if this might also apply to Brevibacterium epidermidis, this microorganism was grown in vitro in continuous culture using a chemostat. Specific growth rate and density of colony forming units were well correlated. While the organism grew readily from pH 5.5 to 8.5, this was not the case with a pH of 5.0. Thus pH-shifts induced by cosmetic procedures can only prevent unpleasant body odour due to abundant growth of bacteria if the pH-value is decreased to 5.0 or less.

Brevibacterium↗
PubMed1995

Lethal effect of carbonyl cyanide m-chlorophenylhydrazone on Escherichia coli and a halotolerant Brevibacterium species.

An attempt was made to examine quantitatively the survival of Escherichia coli and the halotolerant Brevibacterium species, as a function of the exposure time to carbonyl cyanide m-chlorophenylhydrazone (CCCP), a proton conductor. Growth rates, viability, and protein concentrations of E. coli grown in the absence of glucose were unaffected by the addition of 100 microM CCCP. In the presence of glucose the viability was reduced after 24 h incubation with CCCP. Such a high efficiency of CCCP lethality for E. coli cells in the presence of glucose was attributed to not only the growth phase but also the acidic pH of the culture due to metabolites from glucose, mainly lactic acid. The culture of E. coli appeared to be in a syncopic state hovering between life and death when it was exposed to CCCP in the presence of more than 30 mM glucose. In contrast, growth rates of Brevibacterium were reduced in proportion to the exposure time to CCCP. The lethal effect of CCCP to Brevibacterium was slightly enhanced by the addition of glucose into the culture.

Anti-Bacterial Agents↗
PubMed1963

OCCURRENCE OF NUCLEOTIDES IN CULTURE FLUIDS OF MICROORGANISMS. V. EXCRETION OF ADENOSINE CYCLIC 3',5'-PHOSPHATE BY BREVIBACTERIUM LIQUEFACIENS SP. N.

Okabayashi, Tadashi (Shionogi & Co., Ltd., Fukushima-ku, Osaka, Japan), Akihiro Yoshimoto, and Misao Ide. Occurrence of nucleotides in culture fluids of microorganisms. V. Excretion of adenosine cyclic 3',5'-phosphate by Brevibacterium liquefaciens sp. n. J. Bacteriol. 86:930-936. 1963.-Brevibacterium liquefaciens sp. n., when grown in a medium containing amino acids as the nitrogen source, excreted a considerable amount of an adenine ribonucleotide, which had not previously been noticed. The nucleotide was identified as adenosine cyclic 3',5'-phosphate by analysis, ultraviolet-absorption spectra, infrared-absorption spectra, paper chromatography, paper electrophoresis, and by comparison of behavior in hydrolysis by HCl, NaOH, and Ba(OH)(2); also, behavior in digestion with a crude enzyme preparation of adenosine cyclic 3',5'-phosphate phosphodiesterase was compared with that of an authentic sample. Preliminary examination of culture conditions revealed that, at least superficially, the substitution of dl-alanine for Casamino Acids (as nitrogen source) is one of the causes of the excretion of adenosine cyclic 3',5'-phosphate.

Adenine↗
PubMed1993

Expression of a synthetic DNA region containing a consensus promoter and two lac operators in Brevibacterium sp.

The cat reporter gene was used to assess expression of two promoters, previously strongly expressed in Escherichia coli, in Brevibacterium sp. R312 strain. The tac promoter (de Boer et al., 1983, Proc. Natl. Acad. Sci. USA 80, 21-25) was poorly expressed in Brevibacterium sp. In contrast, the AatII-SalI fragment of plasmid pYEJ001 (Pharmacia LKB Biotechnology, Sweden) containing two lac operators, a consensus sequence promoter and the cat structural gene clearly revealed chloramphenicol acetyltransferase activity and the presence of a 25,600-kDa protein, corresponding to the monomeric CAT protein, in cell extracts.

Base Sequence↗
PubMed1979

DNA-DNA homology studies among strains of Arthrobacter and Brevibacterium.

Sixteen named strains of Arthrobacter and two strains of Brevibacterium were investigated by nucleic acid hybridisation. The Arthrobacter strains show homology values ranging between 11 and 55% to the type strain A. globiformis DSM 20124 (ATCC 8010), indicating only a low to moderate relationship. Two strains of A. globiformis, DSM 20124 and DSM 20125, exhibit only poor relationship to one another (30%). Among all the Arthrobacter strains the homology data range between 10 to 70% demonstrating separate status of almost all species. Only A. polychromogenes DSM 20136 was found to be a subspecies of A. oxydans DSM 20119. The type strain of A. citreus, DSM 20133 shows a remarkable lack of homology to four other strains of A. citreus, deposited as ATCC 15170, ATCC 17775, ATCC 21040 and ATCC 21348 (11--13%) which themselves can be separated into two groups according to the homology data (24--31%). Each of the two strains of Brevibacterium share high genetic relatedness with one of these A. citreus groups (71 and 73%, respectively). According to the DNA-DNA homology data, most of the species of Arthrobacter can actually be ranged taxonomically as species.

Arthrobacter↗