Strain Brevibacterium Research Articles

Some enzymatic properties of cholesterol oxidase produced by Brevibacterium sp

In this study we determined some properties of the cholesterol oxidase from a Brevibacterium strain isolated from buffalo's milk and identified the cholesterol degradation products by the bacterial cell. A small fraction of the enzyme synthesized by cells cultured in liquid medium for 7days was released into the medium whereas a larger fraction remained bound to the cell membrane. The extraction of this fraction was efficiently accomplished in 1 mM phosphate buffer, pH 7.0, containing 0.7% Triton X-100. The enzyme stability under freezing and at 45oC was improved by addition of 20% glycerol. The optimum temperature and pH for the enzyme activity were 53°C and 7.5, respectively. The only steroidal product from cholesterol oxidation by the microbial cell and by the crude extract of the membrane-bound enzyme was 4-colesten-3-one. Chromatographic analysis showed that minor no steroidal compounds as well as 4-colesten-3-one found in the reaction media arose during fermentation process and were extracted together with the enzyme in the buffer solution. Cholesterol oxidation by the membrane-bound enzyme was a first order reaction type.

Intracellular changes in ions and organic solutes in halotolerant brevibacterium sp. Strain JCM 6894 after exposure to hyperosmotic shock

In the present study we aimed to observe the intracellular responses when there was a hyperosmotic shock with a large shift in ionic strength in nutrient-rich and nutrient-poor external environments in order to clarify the availability of substrates. To do this, we used the halotolerant organism Brevibacterium sp. strain JCM 6894, which is able to grow in the presence of a wide range of salt concentrations. Hyperosmotic shock was induced by transferring cells in the late exponential phase of growth in a complex medium containing 0.5 M NaCl into either old or fresh culture medium containing 2 M NaCl. Changes in the growth rate, in the pH of the medium, and in the internal cation or organic solute concentrations in the cytosol after an upshock were analyzed as a function of incubation time. The cells exhibited very different responses to upshocks in fresh culture medium and in old culture medium; in fresh culture medium, growth was stimulated and the medium became more acidic, whereas the old culture medium repressed growth and the medium became more alkaline. The intracellular free Na+ concentrations remained low (80 nmol mg of protein-1) after an upshock in fresh culture medium, although they quickly increased twofold in the old culture medium. In contrast, K+ ions immediately accumulated in the cells in fresh culture medium, whereas K+ ions were taken up quite slowly in old culture medium. Furthermore, the cells placed in fresh culture medium transiently accumulated alanine and glutamine in response to the upshock, but the cells placed in old culture medium did not. Growth of the Brevibacterium strain at higher levels of salinity was supported by ectoine synthesis but was not observed after the shift to high-osmolarity conditions in the old culture. In the fresh culture, however, ectoine was vigorously synthesized in cells for more than 5 h after the upshock; the concentration of ectoine in cells was more than 3,500 nmol mg of protein-1 at 10 h, which corresponded to a ninefold increase compared to the concentration before the shock. These findings are consistent with the results of an analysis of the extracellular medium composition before and after the upshock.

Development of 16S rRNA Oligonucleotide Probes for Brevibacterium, Micrococcus/Arthrobacter and Microbacterium/Aureobacterium Used in Dairy Starter Cultures

Summary Oligonucleotide probes for bacteria of the cheese surface were developed. The complete 16S rRNA nucleotide sequences available from databanks of representatives of the genera Brevibacterium, Microbacterium/Aureobacterium and Micrococcus/Arthrobacter were aligned. Oligonucleotide probes were defined for the identification of these genera by selecting a common sequence of their 16S rRNA genes which are different from related genera like Corynebacterium, Propionibacterium or Bifidobacterium . 82 strains were tested of which 70 were of the high DNA G+C branch of bacterial evolution. It was shown by colony-hybridization experiments, that the selected oligonucleotide probes targeted to the 16S rDNA region hybridized only with nucleic acids from strains of Brevibacterium, Microbacterium/Aureobacterium and Micrococcus/Arthrobacter , respectively. For confirmation, a part of the 16S rDNA was amplified in vitro using the polymerase chain reaction technique with two specific oligonucleotides for each of these bacterial groups. The tested probes provide a highly efficient tool to investigate and identify these genera within the coryneform group of bacteria. They were especially developed for analyzing the composition of the microflora of the cheese surface. The use of them may contribute substantially to the isolation and study of new strains of the these genera from natural habitats like cheese.

Electro-optical properties of microbial cells as affected by acrylamide metabolism

The electro-optical (EO) properties of a microbial cell suspension of a Brevibacterium strain were studied during the process of metabolism of acrylamide and certain other amides. Upon optimisation of cell growth conditions, followed by EO measurements, we found that acrylamide produced a specific effect on the cell orientational spectra (OS) over the frequency range 10 to 502 kHz. The other amides used also affected the EO properties of the cells.

5591577 Mobile genetic element originated from brevibacterium strain : Tsuchiya Makoto; Moriya Mika; Miwa Kiyoshi Kawasaki, Japan Assigned to Ajinomoto Co Inc

5591577 Mobile genetic element originated from brevibacterium strain : Tsuchiya Makoto; Moriya Mika; Miwa Kiyoshi Kawasaki, Japan Assigned to Ajinomoto Co Inc

Acrylamide and acrylic acid determination using respiratory activity of microbial cells

The possibility of acrylamide and acrylic acid determination using respiratory activity of microbial cells of the strain Brevibacterium sp. has been investigated. The conditions to obtain specific respiratory activity (SRA) of the cells for both substrates have been examined. The substrate concentration dependences of SRA have been obtained; they exhibited linearity in the range of 0.01–0.1 g/l acrylic acid and 0.01–0.075 g/l acrylamide. The selectivity of the cell respiratory activity for various organic compounds and storage conditions of the active biomass have been studied. Acrylamide quantification in the real waste waters has been made.

6′-Mercaptoguanosine-resistance is related with purF gene encoding 5′-phosphoribosyl-1′-pyrophosphate amidotransferase in inosine-5′-monophosphate overproducing Brevibacterium ammoniagenes

From the gene library constructed with the chromosomal DNA of 6′-mercaptoguanosine (MGS)-resistant strain Brevibacterium ammoniagenes IPR-1, a DNA fragment which conferred MGS-resistance to the wild-type strain B. ammoniagenes ATCC6872 was cloned. The purF gene encoding 5′-phosphoribosyl-1′-pyrophosphate amidotransferase was identified from this fragment and its nucleotide sequence was determined. Wild type purF gene was also cloned by polymerase chain reaction using chromosomal DNA of ATCC6872 as the template and its sequence was determined. Two nucleotides, 583 ‘A’ and 1065 ‘A’, of MGS-resistant purF gene had been changed from 583 ‘G’ and 1065 ‘G’ by mutagenesis, respectively. Both changes at position 583 and 1065 were proved to be responsible for MGS-resistance by site-directed mutagenesis.

Selective determination of acrylonitrile, acrylamide and acrylic acid in waste waters using microbial cells

The possibility of using the respiratory activity of microbial cells for the separate determination of acrylonitrile, acrylamide or acrylic acid in a mixture of these compounds has been demonstrated. The experiments were conducted using the two bacterial strains Brevibacterium sp. and Pseudomonas pseudoalcaligenes , possessing different enzyme systems of acrylonitrile and acrylamide metabolism. The conditions to obtain high respiratory activity of Brevibacterium sp. cells for acrylamide and/or for acrylic acid and of P. pseudoalcaligenes cells for acrylonitrile have been optimized. For the quantitation of the mentioned compounds, the concentration dependences of the cell respiratory activity were established. They were linear in the concentration diapason 0.14–1.4 mM for acrylic acid, 0.14–1.0 mM for acrylamide and 0.02–0.10 mM for acrylonitrile. The selectivity of the cell respiratory activity for various organic amides, nitriles and acids has been studied.

Molecular cloning of a novel gene, dtsR, which rescues the detergent sensitivity of a mutant derived from Brevibacterium lactofermentum.

Several strains of Corynebacterium and Brevibacterium are known for their ability to secrete large amounts of amino acids, especially L-glutamate. We focused on the mechanism of L-glutamate secretion triggered by a detergent, namely polyoxyethylenesorbitan monopalmitate (PESP). A mutant strain, AJ11060, derived from Brevibacterium lactofermentum ATCC 13869 indicates the sensitivity to PESP. A multicopy suppresser gene that compliments the sensitivity of AJ11060 to the detergent was derived from a gene library of B. lactofermentum AJ12036. A 2855-bp DNA fragment was cloned and sequenced. An open reading frame was found that coded for the rescuer gene of the sensitivity to PESP of AJ11060 and was designated dtsR. The expression of the dtsR gene in B. lactofermentum was confirmed by using anti-DtsR antibody. The deduced DtsR protein indicated significant homology with some biotin enzymes such as the beta chain of propionyl-CoA carboxylase from rat (48.3%) and human (48.7%), or a 12S chain of methylmalonyl-CoA carboxyltransferase from Propionibacterium freudenreichii (43.1%).

Inhibition ofPhytophthora vignaeand Stem and Root Rot of Cowpea by Soil Bacteria

ABSTRACT Bacteria isolated from Sri Lankan cowpea fields significantly reduced in vitro mycelial growth and sporangial production of Phytophthora vignae by producing agar-diffusable and volatile inhibitors. Bacterial volatile inhibitors were evolved when the bacteria were grown on tryptic soy agar, nutrient agar, or King's B medium, although relatively less on the latter two media. The pH of agar exposed to the volatiles rapidly increased, suggesting that ammonia was involved in the inhibition. Volatile inhibitors were produced by the bacteria in soil amended with tryptic soy broth or cowpea seed extract, but not in unamended soil. Oospore—induced disease by P. vignae was inhibited by Brevibacterium strain DF-3101 in soil. Soil or seed treatment with three Sri Lankan bacteria, Brevibacterium linens (DF-3101); Bacillus thuringiensis (DF-7107), and Bacillus pumilus (DF-1481) in greenhouse studies suppressed the disease on cowpea caused by P. vignae. None of the bacteria tested functioned as plant growth-pro...

Transposon mutagenesis of coryneform bacteria.

The Corynebacterium glutamicum insertion sequence IS31831 was used to construct two artificial transposons: Tn31831 and miniTn31831. The transposition vectors were based on a gram-negative replication origin and do not replicate in coryneform bacteria. Strain Brevibacterium flavum MJ233C was mutagenized by miniTn31831 at an efficiency of 4.3 x 10(4) mutants per microgram DNA. Transposon insertions occurred at different locations on the chromosome and produced a variety of mutants. Auxotrophs could be recovered at a frequency of approximately 0.2%. Transposition of IS31831 derivatives led not only to simple insertion, but also to cointegrate formation (5%). No multiple insertions were observed. Chromosomal loci of B. flavum corresponding to auxotrophic and pigmentation mutants could be rescued in Escherichia coli, demonstrating that these transposable elements are useful genetic tools for studying the biology of coryneform bacteria.

Optimization of culture conditions of Brevibacterium sp. for the production of amidase and adipamidase

The culture conditions required for the maximum production of the wide spectrum amidase and the adipamidase were determined for two strains of Brevibacterium sp. applying two mathematical techniques: the Hadamard matrix and the analyses of response surface. A culture medium was optimized and the role of Zn 2+ on the activity of adipamidase was demonstrated. The enzyme was shown to be not subject to catabolic repression by glucose or ammonium sulfat and it was not inducible. The influence of physical and chemical factors (pH, temperature) on cell growth and enzyme activity was also studied.

Purification and Characterization of an Adipamidase from a Mutant Strain of Brevibacterium sp. Involved in Dinitrile Degradation.

The adipamidase of a mutant strain of Brevibacterium sp. involved in the degradation of adiponitrile into adipic acid was purified and characterized. The molecular wight of the native enzyme was estimated at 120,000 and that of the sub-units at 58,000, suggesting that this is a dimer structure. This enzyme has a large activity spectrum; all the amides tested were found to be substrates of the adipamidase. An acyltransferase activity was also detected for short chain substrates (acetamide, acetic acid), as well as long chain substrates (oleic acid).

Screening of lysine-producing strains of brevibacterium with a rotating liquid culture system using a large number of micro-holes for individual cell cultivation

With the technique of rotating liquid culture system in 7000 bottomless micro-holes, mutant strains having urease activity higher than that of the original strain were screened out in a medium with urea as a sole nitrogen source. A strain having a lysine yield of 12.5% and urease activity 20% higher than those of the original strain was isolated. The technique is suitable for screening strains where visual methods are available to estimate the amount of metabolite.

N-terminal amino acid sequence of mutant strain Brevibacterium sp. adipamidase.

The adipamidase of a mutant strain Brevibacterium sp. R312 involved in the degradation of adiponitrile to adipic acid was purified. Its N-terminal amino acid sequence was shown to be identical to Brevibacterium sp. R312 enantio selective amidase and Rhodococcus sp. N-774 amidase.

Production of volatile compounds in liquid cultures by six strains of coryneform bacteria

The aromatic profiles of four strains of Brevibacterium linens, one strain of Brevibacterium sp. and one strain of Microbacterium sp. were determined with some pure cultures of these microorganisms in standard trypcase soy liquid medium, which enabled four of these six strains to produce flavour compounds of ripened cheese. Thirty-two flavour compounds were identified by gas chromatography and mass spectrometry. The identified flavour compounds included the following: fatty acids, alcohols, methylketones, sulphur compounds, aromatic compounds and a pyrazine. Some important differences were found among the six strains studied. The four strains of B. linens had similar flavour profiles. Their typical flavour was probably due to dimethyltrisulphide. The two other strains did not appear to produce this compound. Three strains produced significant amounts of the floral aromas phenylethanol and phenylpropanone.

Formation and reversion of protoplasts in mutant strains ofBrevibacterium sp. M27

In mutant strains ofBrevibacterium sp. M27 requiring amino acids and resistant to antibiotics four methods were verified and a method inducing a high formation of protoplasts and their reversion was developed. Formation of protoplasts is 60–99% and their reversion 17–76%. In induced mutants a correlation between protoplast formation and growth rate could be demonstrated.

Characterization of oGA1, An Inducible Phage Particle from Brevibacterium Flavum

Eighteen related strains of Arthrobacter, Brevibacterium and Corynebacterium were used as indicator strains in an attempt to isolate corynephages from a large number of soils and from waste-water samples. Although no phages capable of producing plaques were isolated, one of the indicator strains used, Brevibacterium flavum ATCC 14067, was lysogenic for the inducible phage phi GA1. This phage was not observed to form plaques on any of the strains tested. phi GA1 is of B1-morphotype with a linear double-stranded DNA genome of 48.1 kb with cohesive ends; a restriction map is presented.

Microbial proteinases. The amidase from a brevibacterium strain: Study and applications. Advances in biochemical engineering/biotechnology. Vol. 36 Enzyme studies. Berlin: Akademie‐Verlag, 1988. 115 S., M 108,–, ISBN 3‐05‐500568‐6

Acta BiotechnologicaVolume 10, Issue 1 p. 48-48 Book Review Microbial proteinases. The amidase from a brevibacterium strain: Study and applications. Advances in biochemical engineering/biotechnology. Vol. 36 Enzyme studies. Berlin: Akademie-Verlag, 1988. 115 S., M 108,–, ISBN 3-05-500568-6 A. Steudel, A. SteudelSearch for more papers by this author A. Steudel, A. SteudelSearch for more papers by this author First published: 1990 https://doi.org/10.1002/abio.370100113AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article. Volume10, Issue11990Pages 48-48 RelatedInformation

Occurrence and Chemistry of Cell Wall Teichoic Acids in the Genus Brevibacterium

The cell walls of Brevibacterium casei NCDO 2048, B. epidermidis NCDO 2286 and 14 B. linens isolates from cheese were shown to contain teichoic acids as the dominant non-peptidoglycan polymers. The variation in teichoic acid composition was similar to that described earlier for B. linens strains and for B. iodinum. In B. casei NCDO 2048 a glycerol teichoic acid was found, whereas in B. epidermidis NCDO 2286 glycerol teichoic acid existed together with mannitol teichoic acid. The occurrence of teichoic acids in the cell walls of all Brevibacterium strains investigated constitutes a biochemical characteristic of the genus and is of chemotaxonomic relevance. The co-existence of glycerol and mannitol within the cell walls of the B. linens strains AC480 and AC577 was investigated in detail. The polyols belonged to separate polymers: a glycerol teichoic acid and a mannitol teichoic acid.