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Iron is essential for almost all bacteria, and iron homeostasis is precisely controlled by the ferric uptake regulator (Fur). The Fur regulons have been well characterized in some model bacteria, yet little is known in the common opportunistic pathogen Proteus vulgaris . In this study, Fur regulon and iron-responsive genes in P. vulgaris were mainly defined by in silico and proteomic analyses. The results showed that about 250 potential Fur-regulated operons including 14 transcriptional factors were predicted, while 559 proteins exhibited differential expression in response to iron deficiency, not all being directly regulated by Fur, such as transcriptional factors lexA, recA, narL , and arcA . Collectively, these results demonstrated that Fur functioned as a global regulatory protein to repress or activate expression of a large repertoire of genes in P. vulgaris ; besides, not all the iron-responsive genes were directly regulated by Fur, whereas indirectly regulated through other mechanisms such as additional transcriptional regulatory proteins.

Bacterial non-coding RNAs (ncRNAs) play important regulatory roles in various physiological metabolic pathways. In this study, a novel ncRNA CsiR (ciprofloxacin stress-induced ncRNA) involved in the regulation of ciprofloxacin resistance in the foodborne multidrug-resistant Proteus vulgaris ( P. vulgaris ) strain P3M was identified. The survival rate of the CsiR-deficient strain was higher than that of the wild-type strain P3M under the ciprofloxacin treatment condition, indicating that CsiR played a negative regulatory role, and its target gene emrB was identified through further target prediction, quantitative real-time PCR (qRT-PCR), and microscale thermophoresis (MST). Further studies showed that the interaction between CsiR and emrB mRNA affected the stability of the latter at the post-transcriptional level to a large degree, and ultimately affected the ciprofloxacin resistance of P3M. Notably, the base-pairing sites between CsiR and emrB mRNAs were highly conserved in other sequenced P. vulgaris strains, suggesting that this regulatory mechanism may be ubiquitous in this species. To the best of our knowledge, this is the first identification of a novel ncRNA involved in the regulation of ciprofloxacin resistance in P. vulgaris species, which lays a solid foundation for comprehensively expounding the antibiotic resistance mechanism of P. vulgaris .

Proteus vulgaris is a rod-shaped Gram-negative bacterium known to be the member of Enterobacteriaceae that is able to cause disease in human being. Generally, non-protein-coding RNAs (npcRNAs) do not code for proteins, but they play a vital role in gene regulation at the RNA level including pathogenicity. The present study aims at elucidating homologous npcRNAs from other bacteria in Proteus vulgaris . A comparative genomic analysis was carried out to identify npcRNA homolog of other Enterobacteriaceae pathogens in Proteus vulgaris . A total of 231 npcRNAs previously reported in Salmonella typhi, Salmonella typhimurium and Escherichia coli were screened using BLASTn tool against Proteus vulgaris genome. Interestingly, 33 npcRNAs are homologs to Proteus vulgaris . Northern blot analysis of 6 out of 33 npcRNA candidates confirmed their expression and showed that most of them are differentially expressed during lag, exponential and stationary growth phases. This study is the first approach of identification and characterization of npcRNAs in P. vulgaris . Hence, this could be a pioneer study to further validate the regulatory functions of these npcRNAs to fill the gaps in understanding of the pathogenicity of P. vulgaris .

Background: A pentacyclic lupenane-type natural triterpenoid, betulin, has attracted attention in the field of medicinal chemistry since it exhibited a variety of biological activities, including anticancer activity., Objective: The aim of this present work was to obtain derivatives of betulin through bacterial biotransformation and investigate its anticancer activity against A549, HepG2 and 5RP7 cancer cell lines., Methods: Bacterial biotransformation studies were continued in an MBH broth medium for 7 days at 35oC. Anticancer activities of betulin against A549, HepG2 and 5RP7 cell lines were carried out using XTT assay, and their selectivity was determined using a healthy cell line of NIH/3T3. Cell proliferation ELISA, BRDU (colorimetric) assay was used for measuring proliferation in replicative cells in which DNA synthesis occurs. Flow cytometric analysis was used for measuring apoptotic cell percentages, caspase 3 activation and mitochondrial membrane potential., Results: Bacterial biotransformation studies with 7 bacteria of Staphylococcus aureus ATCC 6538, Proteus vulgaris NRRL B-123, Bacillus subtilis NRRL B-4378, Streptomyces griseolus NRRL B-1062, Escherichia coli ATCC 8739, Staphylococcus aureus ATCC 43300 and Bacillus velezensis NRRL B-14580 produced no metabolite. In in vitro anticancer activity studies, betulin was found to exert anticancer activity against A549, HepG2 and 5RP7 cell lines with IC50 values of 207.7, 125.0 and 28.3 μg/mL, whereas SI values were found to be 30, 50 and 223, respectively. Early and late apoptotic percentages of betulin were found as 9.6, 12.1 and 85.4% on A549, HepG2 and 5RP7, respectively, while caspase 3 positive cell percentages were 2.3, 28.7 and 13.3% for IC50 concentrations. In addition, betulin caused G1 cell cycle arrest (49.5%) on 5RP7 cell line., Conclusion: The results have been shown that betulin activities against A549 and HepG2 cell lines were nonselective and limited its cytotoxic activity against healthy cells, but it is possible to say that it exerted selective activity against 5RP7 cell (28.33±1.53 μg/mL). Betulin effects on apoptosis were found to be dosedependent, while its effect on caspase 3 activation, mitochondrial membrane potential, and cell cycle arrest on G0/G1 phase was not dependent on doses. Therefore, betulin could be a good candidate for the treatment of H-ras active cancer types., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

One of the most challenging problems when trying to recycle urine for different purposes is the removal of urea. In this project we studied an ureolysis system using the bacterium Proteus vulgaris for the transformation of urea to ammonia and its subsequent oxidation to nitrogen at a Pt working electrode. Our system was tested under different pH, microbial reaction times, and urea and bacteria concentrations. Our results indicate that a pH8 is optimal for the combined Proteus vulgaris urease activity and the ammonia oxidation reaction at a Pt electrode. The reaction time and concentration dependence on the ammonia oxidation reaction current densities was also studied. Results showed limited ammonia oxidation under high urea concentrations in ~2.5×10 9 cfu/mL Proteus vulgaris in synthetic urine., (Copyright © 2018. Published by Elsevier B.V.)

Lactobacillus species possesses surface exposed Mucin Binding Protein (MucBP) which plays a role in adhesion to gastrointestinal mucin. MucBP contains one or more mucin binding domain (MBD), the functionality of which has yet not been characterized thoroughly. Here, we have characterized a 93-amino acid MBD (MBD 93 ) of MucBP (LAF_0673) from Lactobacillus fermentum. Multiple sequence alignment of L. fermentum MBD 93 exhibited ∼60% sequence homology with MBDs from other Lactobacillus species. Further, we cloned, expressed and purified MBD 93 from Escherichia coli as N-terminal histidine-tagged protein (6X His-MBD 93 ). The purified MBD 93 was able to bind to mucin and showed strong affinity towards the terminally expressed mucin glycans viz. N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc), Galactose (Gal), and Sialic acid (N-acetylneuraminic acid; Neu5Ac). In silico experiments further confirmed the interaction between homology modeled MBD 93 to mucin glycans through hydrogen-bonding with its surface amino acid residues Ser 57 , Pro 58 , Ile 60 , Tyr 63 and Ala 65 . We also have demonstrated that MBD 93 was able to inhibit the adhesion of enteric pathogens, including E. coli, Salmonella Paratyphi A, Shigella sonnei and Proteus vulgaris to mucin. Our results suggested that L. fermentum MBD 93 is a functionally sufficient unit to act as an adhesin and to protect from invading enteric pathogens., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Colistin has become a last-line antibiotic for the treatment of multidrug-resistant bacterial infections; however, resistance to colistin has emerged in recent years. Some bacteria, such as Proteus and Serratia spp., are intrinsically resistant to colistin although the exact mechanism of resistance is unknown. Here we identified the molecular support for intrinsic colistin resistance in Proteus spp. by comparative genomic, transcriptomic and proteomic analyses of colistin-susceptible (CSUR P1868_S) and colistin-resistant (CSUR P1867_R) strains of an atypical Proteus vulgaris. A significant difference in outer membrane glycoside structures in both strains that was corroborated by MALDI-TOF/MS analysis was found, which showed an absence of 4-amino-4-deoxy-l-arabinose (L-Ara4N) in the outer membrane lipid A moiety of the susceptible strain. Comparative genomic analysis with other resistant strains of P. vulgaris available in a local database found a mutation in the arnBCADTEF operon of the susceptible strain. Transcriptomic analysis of genes belonging to the arnBCADTEF operon showed a significant decrease in mRNA expression level of these genes in the susceptible strain, supporting addition of L-Ara4N in the outer membrane lipid A moiety as an explanation for colistin resistance. Insertion of the arnD gene that was suggested to be altered in the susceptible strain by in silico analysis led to a 16-fold increase of colistin MIC in the susceptible strain, confirming its role in colistin resistance in this species. Here we show that constitutive activation of the arn operon and addition of L-Ara4N is the main molecular mechanism of colistin resistance in P. vulgaris., (Copyright © 2017 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.)

An organic solvent-stable lipase from Proteus vulgaris K80 was used to produce the omega-3 polyunsaturated fatty acid ethyl esters (ω-3 PUFA EEs). First, the lyophilized recombinant lipase K80 (LyoK80) was used to perform the transesterification reaction of menhaden oil and ethanol. LyoK80 produced the ω-3 PUFA EEs with a conversion yield of 82 % in the presence of 20 % water content via a three-step ethanol-feeding process; however, in a non-aqueous condition, LyoK80 produced only a slight amount of the ω-3 PUFA EEs. To enhance its reaction properties, the lipase K80 was immobilized on a hydrophobic bead to derive ImmK80; the biochemical properties and substrate specificity of ImmK80 are similar to those of LyoK80. ImmK80 was then used to produce ω-3 PUFA EEs in accordance with the same transesterification reaction. Unlike LyoK80, ImmK80 achieved a high ω-3 PUFA EE conversion yield of 86 % under a non-aqueous system via a one-step ethanol-feeding reaction. The ω-3 PUFA EEs were purified up to 92 % using a urea complexation method.

Phenylpyruvic acid is a deaminated form of phenylalanine and is used in various areas such as development of cheese and wine flavors, diagnosis of phenylketonuria, and to decrease excessive nitrogen accumulation in the manure of farm animals. However, reported phenylpyruvic acid fermentation studies in the literature have been usually performed at shake-flask scale with low production. In this study, phenylpyruvic acid production was evaluated in bench-top bioreactors by conducting fed-batch and continuous fermentation for the first time. As a result, maximum phenylpyruvic acid concentrations increased from 1350 mg/L (batch fermentation) to 2958 mg/L utilizing fed-batch fermentation. Furthermore, phenylpyruvic acid productivity was increased from 48 mg/L/hr (batch fermentation) to 104 and 259 mg/L/hr by conducting fed-batch and continuous fermentation, respectively. Overall, this study demonstrated that fed-batch and continuous fermentation significantly improved phenylpyruvic acid production in bench-scale bioreactor production.

This work aimed to develop a whole-cell biotransformation process for the production of α-ketoisocaproate from L-leucine. A recombinant Escherichia coli strain was constructed by expressing an L-amino acid deaminase from Proteus vulgaris. To enhance α-ketoisocaproate production, the reaction conditions were optimized as follows: whole-cell biocatalyst 0.8 g/L, leucine concentration 13.1 g/L, temperature 35 °C, pH 7.5, and reaction time 20 h. Under the above conditions, the α-ketoisocaproate titer reached 12.7 g/L with a leucine conversion rate of 97.8%. In addition, different leucine feeding strategies were examined to increase the α-ketoisocaproate titer. When 13.1 g/L leucine was added at 2-h intervals (from 0 to 22 h, 12 addition times), the α-ketoisocaproate titer reached 69.1 g/L, while the leucine conversion rate decreased to 50.3%. We have developed an effective process for the biotechnological production of α-ketoisocaproate that is more environmentally friendly than the traditional petrochemical synthesis approach.

Alpha keto acids are deaminated forms of amino acids that have received significant attention as feed and food additives in the agriculture and medical industries. To date, their production has been commonly performed at shake-flask scale with low product concentrations. In this study, production of phenylpyruvic acid (PPA), which is the alpha keto acid of phenylalanine was investigated. First, various microorganisms were screened to select the most efficient producer. Thereafter, growth parameters (temperature, pH, and aeration) were optimized in bench scale bioreactors to maximize both PPA and biomass concentration in bench scale bioreactors, using response surface methodology. Among the four different microorganisms evaluated, Proteus vulgaris was the most productive strain for PPA production. Optimum temperature, pH, and aeration conditions were determined as 34.5 °C, 5.12, and 0.5 vvm for PPA production, whereas 36.9 °C, pH 6.87, and 0.96 vvm for the biomass production. Under these optimum conditions, PPA concentration was enhanced to 1,054 mg/L, which was almost three times higher than shake-flask fermentation concentrations. Moreover, P. vulgaris biomass was produced at 3.25 g/L under optimum conditions. Overall, this study demonstrated that optimization of growth parameters improved PPA production in 1-L working volume bench-scale bioreactors compared to previous studies in the literature and was a first step to scale up the production to industrial production.

A marine strain of Proteus vulgaris capable of activating multiple acylated homoserine lactone (AHL)-based reporter cultures was isolated. The cognate signal molecule was characterized as octanoyl homoserine lactone (OHL) and its production was observed to be growth dependent, with maximum production (5.675 µg l(-1)) at 24 h growth. The strain exhibited swarming, but its motility was not affected upon addition of pure OHL or culture supernatant. Phytochemicals such as quercitin and berberine chloride inhibited OHL production and reduced swarming. FliA, the predominantly upregulated protein during swarming, was considered as a possible target for these inhibitors, and docking of the two most active and two least active inhibitors to this protein suggested preferential binding of the former set of compounds. Apart from adding new evidence to AHL production in Proteus vulgaris, active inhibitors shortlisted from this study could help in identifying lead compounds to act against this opportunistic pathogen of the respiratory and gastrointestinal tract., (© 2014 The Authors.)

Bacterial toxin-antitoxin (TA) systems regulate key cellular processes to promote cell survival during periods of stress. During steady-state cell growth, antitoxins typically interact with their cognate toxins to inhibit activity presumably by preventing substrate recognition. We solved two x-ray crystal structures of the Proteus vulgaris tetrameric HigB-(HigA)2-HigB TA complex and found that, unlike most other TA systems, the antitoxin HigA makes minimal interactions with toxin HigB. HigB adopts a RelE family tertiary fold containing a highly conserved concave surface where we predict its active site is located. HigA does not cover the solvent-exposed HigB active site, suggesting that, in general, toxin inhibition is not solely mediated by active site hindrance by its antitoxin. Each HigA monomer contains a helix-turn-helix motif that binds to its own DNA operator to repress transcription during normal cellular growth. This is distinct from antitoxins belonging to other superfamilies that typically only form DNA-binding motifs upon dimerization. We further show that disruption of the HigB-(HigA)2-HigB tetramer to a HigBA heterodimer ablates operator binding. Taken together, our biochemical and structural studies elucidate the novel molecular details of the HigBA TA system.

In this work we investigated relationship between molecular structure of alkali metal o-, m-, p-anisate molecules and their antimicrobial activity. For this purpose FT-IR spectra for lithium, sodium, potassium, rubidium and caesium anisates in solid state and solution were recorded, assigned and analysed. Microbial activity of studied compounds was tested against Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Proteus vulgaris. In order to evaluate the dependency between chemical structure and biological activity of alkali metal anisates the statistical analysis (multidimensional regression and principal component) was performed for selected wavenumbers from FT-IR spectra and parameters that describe microbial activity of anisates. The obtained statistical equations show the existence of correlation between molecular structure of anisates and their biological properties., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Selected ion flow tube-mass spectrometry has been used to measure the volatile compounds occurring in the headspace of urine samples inoculated with common urinary tract infection (UTI)-causing microbes Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, Klebsiella pneumoniae, Enterococcus faecalis, or Candida albicans. This technique has the potential to offer rapid and simple diagnosis of the causative agent of UTIs., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Expression of the tna operon of Escherichia coli and of Proteus vulgaris is induced by L-tryptophan. In E. coli, tryptophan action is dependent on the presence of several critical residues (underlined) in the newly synthesized TnaC leader peptide, WFNIDXXL/IXXXXP. These residues are conserved in TnaC of P. vulgaris and of other bacterial species. TnaC of P. vulgaris has one additional feature, distinguishing it from TnaC of E. coli; it contains two C-terminal lysine residues following the conserved proline residue. In the present study, we investigated L-tryptophan induction of the P. vulgaris tna operon, transferred on a plasmid into E. coli. Induction was shown to be L-tryptophan dependent; however, the range of induction was less than that observed for the E. coli tna operon. We compared the genetic organization of both operons and predicted similar folding patterns for their respective leader mRNA segments. However, additional analyses revealed that L-tryptophan action in the P. vulgaris tna operon involves inhibition of TnaC elongation, following addition of proline, rather than inhibition of leader peptide termination. Our findings also establish that the conserved residues in TnaC of P. vulgaris are essential for L-tryptophan induction, and for inhibition of peptide elongation. TnaC synthesis is thus an excellent model system for studies of regulation of both peptide termination and peptide elongation, and for studies of ribosome recognition of the features of a nascent peptide.

A developed consortium-GR, consisting of Proteus vulgaris NCIM-2027 (PV) and Micrococcus glutamicus NCIM-2168 (MG), completely decolorized an azo dye Scarlet R under static anoxic condition with an average decolorization rate of 16,666 microg h(-1); which is much faster than that of the pure cultures (PV, 3571 microg h(-1); MG, 2500 microg h(-1)). Consortium-GR gave best decolorization performance with nearly complete mineralization of Scarlet R (over 90% TOC and COD reduction) within 3h, much shorter relative to the individual strains. Induction in the riboflavin reductase and NADH-DCIP reductase was observed in the consortium, suggesting the involvement of these enzymes during the fast decolorization process. The FTIR and GC-MS analysis showed that 1,4-benzenediamine was formed during decolorization/degradation of Scarlet R by consortium-GR. Phytotoxicity studies revealed no toxicity of the biodegraded products of Scarlet R by consortium-GR. In addition, consortium-GR applied for mixture of industrial dyes showed 88% decolorization under static condition with significant reduction in TOC (62%) and COD (68%) within 72 h, suggesting potential application of this microbial consortium in bioremediation of dye-containing wastewater.

Seven sesterterpene sulfates (1-7) were isolated from the tropical sponge Dysidea sp. and their inhibitory activities against isocitrate lyase (ICL) from Candida albicans were evaluated. Among the isolated natural products compound 6 and 7 were found to be strong ICL inhibitors. The isolated compounds (1-7) also showed potent antibacterial effect against Bacillus subtilis and Proteus vulgaris, but did not display antifungal activity.

The copper-resistant (1318 microM CuSO(4).5H(2)O) strain KNP3 of Proteus vulgaris was isolated from soil near the Panki power plant, Kanpur, India, and was used to inoculate pigeon pea (Cajanus cajan var. UPS-120) seeds grown in soil for 60 days in the presence of 600 microM CuSO(4).5H(2)O. A study of siderophore production (126.34 +/- 0.52 microg ml(-1)) and its subsequent effects on plant growth promotion under in situ conditions was conducted. The parameters that were monitored included the plants' wet weight, dry weight, shoot length, chlorophyll content, and concentration of copper in plant roots and shoots. The results showed that the strain caused a significant (p < 0.05) increase in wet weight, dry weight, root length, shoot growth, and chlorophyll content (57.8%, 60%, 19.7%, 47.8%, and 36.3%, respectively) in the presence of copper. Furthermore, the strain reduced accumulation of Cu in the roots and shoots to 36.8% and 60.5%, respectively. Apart from this, copper concentration in the soil was measured on 0, 7, 15, 30, and 45 days consecutively and the results indicated that the bioinoculant KNP3 causes a significant decrease in Cu concentration in soil (55.6%), which was unlikely in the control (10.5%) treatment. The data suggested that the bacterial strain has the ability to protect plants against the inhibitory effects of copper besides reducing the copper load of the soil.

Twelve bacterial strains belonging to eight taxonomic groups: Brevibacterium linens, Microbacterium foliorum, Arthrobacter arilaitensis, Staphylococcus cohnii, Staphylococcus equorum, Brachybacterium sp., Proteus vulgaris and Psychrobacter sp., isolated from different surface-ripened French cheeses, were investigated for their abilities to generate volatile aroma compounds. Out of 104 volatile compounds, 54 volatile compounds (identified using dynamic headspace technique coupled with gas chromatography-mass spectrometry [GC-MS]) appeared to be produced by the different bacteria on a casamino acid medium. Four out of eight species used in this study: B. linens, M. foliorum, P. vulgaris and Psychrobacter sp. showed a high flavouring potential. Among these four bacterial species, P. vulgaris had the greatest capacity to produce not only the widest varieties but also the highest quantities of volatile compounds having low olfactive thresholds such as sulphur compounds. Branched aldehydes, alcohols and esters were produced in large amounts by P. vulgaris and Psychrobacter sp. showing their capacity to breakdown the branched amino acids. This investigation shows that some common but rarely mentioned bacteria present on the surface of ripened cheeses could play a major role in cheese flavour formation and could be used to produce cheese flavours.

O-Polysaccharides were obtained from the lipopolysaccharides of Proteus mirabilis CCUG 10704 (OE) and Proteus vulgaris TG 103 and studied by chemical analyses and one- and two-dimensional (1)H and (13)C nuclear magnetic resonance spectroscopy, including rotating-frame nuclear Overhauser effect spectroscopy, H-detected (1)H,(13)C heteronuclear single-quantum spectroscopy and (1)H,(31)P heteronuclear multiple-quantum spectroscopy experiments. The Proteus mirabilis OE polysaccharide was found to have a trisaccharide repeating unit with a lateral glycerol phosphate group. The Proteus vulgaris TG 103 produces a similar O-polysaccharide, which differs in incomplete substitution with glycerol phosphate (c. 50% of the stoichiometric amount) and the presence of an O-acetyl group at position 6 of the 2-acetamido-2-deoxygalactose (GalNAc) residue. These structures are unique among the known bacterial polysaccharide structures. Based on the structural and serological data of the lipopolysaccharides, it is proposed to classify both strains studied into a new Proteus serogroup, O54, as two subgroups, O54a,54b and O54a,54c. The serological relatedness of the Proteus O54 and some other Proteus lipopolysaccharides is discussed.

Biochemical oxygen demand (BOD) is an international regulatory environmental index for monitoring organic pollutants in wastewater and the current legislated standard test for BOD monitoring requires 5 days to complete (BOD5 test). We are developing a rapid microbial technique, MICREDOX, for measuring BOD by eliminating oxygen and, instead, quantifying an equivalent biochemical co-substrate demand, the co-substrate being a redox mediator. Elevated concentrations of Proteus vulgaris, either as free cells or immobilised in Lentikat disks, were incubated with an excess of redox mediator (potassium hexacyanoferrate(III)) and organic substrate for 1h at 37 degrees C without oxygen. The addition of substrate increased the catabolic activity of the microorganisms and the accumulation of reduced mediator, which was subsequently re-oxidised at a working electrode generating a current quantifiable by a coulometric transducer. The recorded currents were converted to their BOD5 equivalent with the only assumption being a fixed conversion of substrate and known stoichiometry. Measurements are reported both for the BOD5 calibration standard solution (150 mg l(-1) glucose, 150 mg l(-1) glutamic acid) and for filtered effluent sampled from a wastewater treatment plant. The inclusion of a highly soluble mediator in place of oxygen facilitated a high ferricyanide concentration in the incubation, which in turn permitted increased concentrations of microorganisms to be used. This substantially reduced the incubation time, from 5 days to 1h, for the biological oxidation of substrates equivalent to those observed using the standard BOD5 test. Stoichiometric conversion efficiencies for the oxidation of the standard substrate by P. vulgaris were typically 60% for free cells and 35-50% for immobilised cells.

Lactic and butyric acid production by bacterial flocs in a continuous culture obeyed different physiological constraints. The butyric acid rate of production was constant and independent of the growth rate [0.012 +/- 0.001 gBUT/(L.h)], whereas lactic fermentation occurred only beyond a critical growth rate (0.25 +/- 0.05 h(-1)) and was apparently associated with an abrupt drop in biomass. Principles of modeling used to describe a Crabtree effect in Saccharomyces cerevisiae were found to apply to lactic acid production by flocs. A rank of "physiological unit" (or "metabolic unit") can be attributed to the bacterial floc. From a practical point of view, the production of fermentation products by stable flocs, naturally resistant to contamination, opens the possibility of industrial production by continuous cultivation by using flocs-forming consortia.

The core region of the lipopolysaccharides of Proteus group OX bacteria, which are used as antigens in Weil-Felix test for serodiagnosis of rickettsiosis, were studied by chemical degradations in combination with ESI FTMS, including infrared multi-photon dissociation (IRMPD) MS/MS and capillary skimmer dissociation. Structural variants of the inner core region were found to be the same as in Proteus non-OX strains that have been studied earlier. The outer core region has essentially the same structure in Proteus vulgaris OX19 (serogroup O1) and OX2 (serogroup O2) and a different structure in Proteus mirabilis OXK (serogroup O3). A fragmentation due to the rupture of the linkage between GlcN or GalN and GalA was observed in IRMPD-MS/MS of core oligosaccharides and found to be useful for screening of Proteus strains to assign structures of the relatively conserved inner core region and to select for further studies strains with distinct structures of a more variable outer core region.

Aim: To study the microbial transformation of sinenxan A., Methods: Choose two strains of Fungi (Mucor spinosus AS 3.3450 and Cunninghamella echinulata AS 3.3400) and a strain of bacterium (Proteus vulgaris AS 1.1208) to transform the substrate., Results: Three products were obtained and identified as 10-deacetylsinenxan A1, 6 alpha-hydroxy-10-deacetylsinenxan A2 and 9 alpha-hydroxy-10-deacetylsinenxan A3 respectively., Conclusion: Sinenxan A is facile to be transformed by microorganisms, the 10-acetyl group of which is an active group.

Formation of infectious urinary calculi is the most common complication accompanying urinary tract infections by members of the genus Proteus. The major factor involved in stone formation is the urease produced by these bacteria, which causes local supersaturation and crystallization of magnesium and calcium phosphates as carbonate apatite [Ca(10)(PO(4))(6).CO(3)] and struvite (MgNH(4)PO(4).6H(2)O), respectively. This effect may also be enhanced by bacterial polysaccharides. Macromolecules of such kind contain negatively charged residues that are able to bind Ca(2+) and Mg(2+), leading to the accumulation of these ions around bacterial cells and acceleration of the crystallization process. The levels of Ca(2+) and Mg(2+) ions bound by whole Proteus cells were measured, as well as the chemical nature of isolated LPS polysaccharides, and the intensity of the in vitro crystallization process was compared in a synthetic urine. The results suggest that the sugar composition of Proteus LPS may either enhance or inhibit the crystallization of struvite and apatite, depending on its chemical structure and ability to bind cations. This points to the increased importance of endotoxin in urinary tract infections.

A study has been performed to examine the effect of temperature and ethanolic stresses on the coulombic efficiency of a microbial fuel cell. The conventional-type fuel cell containing Gram-negative bacteria, Proteus vulgaris, was investigated as a model system. From current output measurements, it was found that the coulombic yields were altered by environmental stresses such as temperature shock or ethanol treatment to the bacteria. While high-temperature or ethanolic shock led to a remarkable decrement in coulombic output, the low-temperature shock induced a slight increase in microbial fuel cell efficiency. These results indicate that the membrane fluidity is affected considerably by environmental stress, which in turn affects the electron transfer process through the bacterial cell membrane to and from the electrode. This interpretation was confirmed by the cyclic voltammetric study of a mediator on an electrode surface modified with the lipids extracted from the membrane of P. vulgaris under the given stress. Markedly different electrochemical behaviors were observed depending on the environmental stress. A reciprocal relationship between coulomb output and the ratio of saturation/unsaturation of fatty acids has been observed. This is the first report, to our knowledge, that the structural adaptation of membrane fatty acids in response to the environmental shock can regulate the coulombic efficiency of a microbial fuel cell.

Proteus bacilli play a particularly important role in urinary tract infections (UTI). Fimbriae and adherence ability and hemolysins production (HpmA, HlyA) are one of the factors of pathogenicity of these bacteria. In this paper we describe the invasion of HCV T-29 transitional bladder urothelial cells carcinoma strains of P. penneri, as well as P. vulgaris strains belonging to different serogroups. The cytotoxic effect was observed at 8 hour of incubation of the tested cells with P. vulgaris O21 and the same effect (complete lysis) at 6 hours by P. vulgaris O4 (this strain manifests maximal activity in the production of HlyA hemolysin). P. penneri strains, produce different types of fimbriae, expressed similar bacterial invasiveness. The hydrophobic properties of 25 P. vulgaris strains were also tested and only 3 strains occur to have hydrophobic cell surface.

We placed 43 isolates belonging to the Proteus vulgaris complex into proposed DNA groups (genomovars) using five previously recommended tests (tests for esculin hydrolysis, production of acid from salicin, L-rhamnose fermentation, and elaboration of DNase and lipase). On the basis of the results of these five tests, 49% of the isolates fell into DNA groups 5 and 6, 37% fell into DNA group 2, and the remaining 14% fell into DNA groups 3 and 4. Sequencing of the 16S rRNA genes of 11 members of DNA groups 5 and 6 indicated that 10 of these isolates (91%) could be unambiguously assigned to one of these two genomospecies. Overall expression of selected enzymatic and virulence-associated characteristics did not differ significantly among DNA groups.

Mediator-coupled microbial fuel cells containing Proteus vulgaris were constructed and the cell performance was tested. Fuel cell efficiency depended on the carbon source in the initial medium of the microorganism. Maltose and trehalose were not utilized substantially by P. vulgaris; however, their presence in the initial medium resulted in enhanced cell performance. In particular, galactose showed 63% coulombic efficiency in a biofuel cell after P. vulgaris was cultured in a trehalose-containing medium. This work demonstrates that optimum utilization of carbon sources by microorganisms, which leads to the maximization of fuel cell performance, is possible simply by adjusting initial carbon sources., (Copyright 2000 John Wiley & Sons, Inc.)

The saccharide constituents of lipopolysaccharides (LPS) of Proteus spp. vary with the strain and contain unique components about which little is known. The biological activities of LPS and lipid A from S- and R-forms of 10 Proteus strains were examined. LPS from all S-form Proteus strains was lethal to D-(+)-galactosamine (GalN)-loaded, LPS-responsive, C3H/HeN mice, but not to LPS-hypo-responsive C3H/HeJ mice. P. vulgaris 025 LPS evoked strong anaphylactoid reactions in N-acetylmuramyl-L-alanyl-D-isoglutamine (MDP)-primed C3H/HeJ mice. LPS from S- and R-form Proteus strains induced production of nitric oxide (NO) and tumour necrosis factor (TNF) by macrophages isolated from C3H/HeN but not C3H/HeJ mice. Lipid A from Proteus strains also induced NO and TNF production, although lipid A was less potent than LPS. The effects of LPS were mainly dependent on CD14; LPS-induced NO and TNF production in CD14+ J774.1 cells was significantly greater than in CD14-J7.DEF.3 cells. All LPS from Proteus strains, and especially from P. vulgaris 025, exhibited higher anti-complementary activity than LPS from Escherichia coli or Pseudomonas aeruginosa. Polymyxin B inactivated proteus LPS in a dose-dependent manner, but these LPS preparations were more resistant to polymyxin B than E. coli LPS. CAP18(109-135), a granulocyte-derived peptide, inhibited proteus LPS endotoxicity only when the LPS:CAP18(109-135) ratio was appropriate, which suggests that CAP18(109-135) acts through a different mechanism than polymyxin B. The results indicate that LPS from Proteus spp. are potently endotoxic, but that the toxicity is different from that of LPS from E. coli or Salmonella spp. and even varies among different Proteus strains. The variation in biological activities among proteus LPS may be due to unique components within the respective LPS.

Investigation of postmortem blood can reveal the presence of significant ethanol levels. However, in some instances it cannot easily be determined if the source of ethanol is from ingestion or from postmortem endogenous fermentation by contaminating microbes. Described here is a robust polymerase chain reaction (PCR)-based method for detecting the presence of common ethanol producing microbial contaminants in human blood. A set of DNA primers were designed for use in PCR to amplify and detect the genomic DNA from humans and three test microorganisms Escherichia coli, Proteus vulgaris, and Candida albicans. A rapid and reproducible protocol was developed for isolating genomic DNA from mixed human blood-microorganism samples that yields a suitable template for PCR. The organism-specific primer pairs can detect the presence of the target microorganisms in human blood at concentrations as low as 10 colony forming units/mL. The PCR products readily can be detected after agarose gel electrophoresis. This method provides an additional means of rapidly identifying microbial contaminants in postmortem blood samples.

Lipid-bilayer experiments were performed with the related hemolysins from Proteus vulgaris and Morganella morganii (HlyA). The addition of the toxins to the aqueous phase bathing lipid-bilayer membranes composed of different lipids resulted in the formation of transient ion-permeable channels. Membranes formed of pure lipids were rather inactive targets for the hemolysins as compared with lipid mixtures such as asolectin. The channels had several different substrates. The major open state had single-channel conductances of 500 pS in 0.15 M KCl at small transmembrane voltages. Experiments with different salts suggested that the hemolysin-induced channels of P. vulgaris and M. morganii were exclusively cation selective at neutral pH, caused by negative charges localized at the channel mouth. The mobility sequence of the cations within the channels was similar if not identical to their mobility sequence in the aqueous phase. The single-channel data were consistent with wide, water-filled channels with estimated minimal diameters of about 1 nm since the large organic cation Tris+ can permeate the channels without any detectable interaction with its interior. Pore-forming properties of these hemolysins were compared with those of HlyA of Escherichia coli. All these toxins share common features, oligomerize probably to form pores in lipid-bilayer membranes and form channels with similar properties which suggests that their structures are more or less identical.

An exocellular polysaccharide produced by a clinical isolate of Proteus vulgaris (ATCC 49990) was shown by composition, methylation, periodate oxidation, and nuclear magnetic resonance analyses to be composed of repeating trisaccharide units containing D-galactose, 2-acetamido-2-deoxy-D-glucose and pyruvic acid (2:1:1), and having the structure: [formula: see text] The homologus smooth lipopolysaccharide of the P. vulgaris strain was determined to have an O-polysaccharide component having the same structure as the above extracellular polysaccharide.

Various computer programs for large-scale bioprocess control and optimization have been developed as well as software for simple laboratory routine analysis. In comparison, software can hardly be found that works on laboratory scale and provides the control of complex flow injection analysis (FIA) systems, multisubstrate determination, data evaluation as well as minimal process control abilities. The sensors applied can be of different type (luminometric or other optical as well as electrochemical biosensors). The development of such a software may be very helpful for the transfer of FIA/biosensor systems from the state of development to industrial processes. Hence, each analysing system--even a well established biosensor--has to be individually adapted to the process, a task which is best done under laboratory conditions. Such a flexible, computer-controlled FIA system for research level based on the software FIACRE is presented. Five FIA/(bio)sensor system can be controlled simultaneously. Additionally, common temperature and pH recordings are possible. Determinations of substrate concentrations are performed by means of calibration curves which can be recorded at different times. This allows supervising the activities of the sensors during a cell cultivation and controlling the bioprocess, e.g. by adding substrate to a cell culture. The automated monitoring of the degradation of glucose and urea by two different optical sensing principles during a cell cultivation under the control of one microcomputer is presented for the first time. For this purpose, already well examined biosensors (a urease optode and a luminometric glucose sensor) were employed and their properties discussed under the aspect of working in real cultivation media. It will also be shown that substrates being of interest for bioprocess control can be detected by slight modifications of known reactions. For example, substrates of NADH-dependent enzymatic reactions can be detected by the luminol chemiluminescence system, and optodes can be employed for pH, penicillin and glucose determination.

Encrustations are the most frequent complications occurring with indwelling catheters and urine drainage systems. The conditions for bacterial infections, using synthetic urine and controlled contamination by Proteus vulgaris, were standardised by using an in vitro model. Crystal deposits on catheters were analysed by infra-red spectroscopy and scanning electron microscopy. The main components of deposits in all investigations were struvite (MgNH4PO4.6H2O) 60-70% and carbonate apatite (Ca10(PO4,CO3)6 (OH,CO3)2) 30-40%. Investigations as to the quality and quantity of encrustations confirmed the analysis. Irrigation treatment was carried out with physiological saline solution and citric acid solution (Suby G) to study and quantify the dissolution of crystal deposits. Regular irrigation with citric solution resulted in a 70% dissolution of encrustations and ensured free flow as ascertained by measuring flow rates.

The effect of subinhibitory concentrations of netilmicin, ceftriaxone, cefotaxime, aztreonam and piperacillin on the adherence of Proteus species to uroepithelial cells was examined. Bacterial adhesion to human uroepithelial cells, measured microscopically, was affected by all five antibiotics but to different extents. The most effective was netilmicin. There was a correlation between the decreased rate of bacterial attachment and morphological changes in the drug-exposed bacteria.

O-acetylated peptidoglycan was purified from Proteus mirabilis grown in the presence of specifically radiolabelled glucosamine derivatives, and the migration of the radiolabel was monitored. Mild-base hydrolysis of the isolated peptidoglycan (to release ester-linked acetate) from cells grown in the presence of 40 microM [acetyl-3H]N-acetyl-D-glucosamine resulted in the release of [3H]acetate, as detected by high-pressure liquid chromatography. The inclusion of either acetate, pyruvate, or acetyl phosphate, each at 1 mM final concentration, did not result in a diminution of mild-base-released [3H]acetate levels. No such release of [3H]acetate was observed with peptidoglycan isolated from either Escherichia coli incubated with the same radiolabel or P. mirabilis grown with [1,6-3H]N-acetyl-D-glucosamine or D-[1-14C]glucosamine. These observations support a hypothesis that O acetylation occurs by N----O acetyl transfer within the sacculus. A decrease in [3H]acetate release by mild-base hydrolysis was observed with the peptidoglycan of P. mirabilis cultures incubated in the presence of antagonists of peptidoglycan biosynthesis, penicillin G and D-cycloserine. The absence of free-amino sugars in the peptidoglycan of P. mirabilis but the detection of glucosamine in spent culture broths implies that N----O transacetylation is intimately associated with peptidoglycan turnover.

The relationship between growth condition and production of cell-bound hemolysin by Proteus mirabilis S1959 strain was investigated. Hemolytic activity was not dependent on type of medium, oxygen accessibility and was inhibited in presence of N-ethylmaleimide, different sera or trypsine. Cell-free hemolysin was released to the medium during stationary-death phases of growth of fluid Proteus mirabilis and vulgaris cultures.

Bacterial chemotactic peptides are low molecular weight peptides which stimulate a wide range of neutrophil functions following binding to specific leucocyte receptors. Formyl methionyl leucyl phenylalanine (FMLP) is the major chemotactic peptide in Escherichia coli culture supernatants. This paper reports the development and validation of a radio-immunoassay (RIA) for FMLP and its application to the analysis of formyl peptide production by enteric bacteria in vitro. The assay was moderately sensitive (10 nmol/L FMLP) and highly specific showing cross reactivity with F-met-leu-tyr, F-nle-leu-phe and F-met-met-met sequences (ID50 = 200, 100 and 250 nmol/L, respectively) but no significant cross reactivity with non-formylated or other formylated di- and tri-peptides (ID50 = 10(5) nmol/L. Culture supernatants from five species of enteric bacteria were filtered, concentrated and fractionated by reverse phase high performance liquid chromatography before RIA. All five organisms produced immunoreactive F-met peptides. A major peak of immunoreactivity co-chromatographing with authentic FMLP was found in all supernatants, but additional peaks representing more hydrophobic peptides were found in Streptococcus faecalis and Bacteroides fragilis cultures. In E. coli culture supernatants, concentration of immunoreactive FMLP increased in a linear fashion during 3 h of log phase growth reaching 31.2 nmol/L(s.e.m. = 10) with final bacterial concentrations of 3 +/- 0.73 x 10(8)/mL (n = 6). These findings extend earlier work showing production of bioactive formyl oligopeptides by different species of enteric bacteria and suggest that a RIA for FMLP will be a useful tool for investigating the production and metabolic fate of such peptides in man.

A strain of Enterobacter cloacae, isolated from soil, reduces racemic abdtriglycinatocobalt(III), triglycinatocobalt(III), stereoselectively under anaerobic conditions; Washed suspensions oc cells, grown anaerobically in the absence of triglycinatocobalt(III) initially reduce the L-enantiomer stereospecifically under argon yielding a labile Co(II) chelate. An approximately 5-fold increase in the rate of reduction of the complex occurs using cells disrupted by 2-phenylethanol; decomposition now proceeds with a slight preference for the D-enantiomermreduction of triglycinatocobalt(III) under argon by cell-free extracts, prepared by sonication, exhibits similar stereoselectivities to cells disrupted by 2-phenylethanolmhowever, extracts contain a factor capable of reducing D-triglycinatocobalt(III) with high stereoselectivity under molecular hydrogenmthe behaviour of Enterobacter cloacae is compared with that of Aerobacter aerogenes, Proteus vulgaris, Escherichia anaerogenes and Serratia marcescensmstereoselective effects are also observed in the bacterial reduction of other cobalt(III) complexes.

6 typical bacteria species of the human intestinal flora (E. coli, Klebsiella pneumoniae, Proteus vulgaris, Streptococcus faecalis, Bacteroides fragilis, Bifidobacterium sp.) were incubated in a liquid medium for 48 h with [15N2]-urea and [15N]-ammonium chloride. The rates of [15N]-incorporation were calculated. They depend reproducible on the species examined, on the kind of the offered NPN-substance and on the amount of NPN-substance in the medium. With [15N2]-urea the minimal rate of incorporation was 3.8% (E coli) and the maximal one 95.6% (Bifidobacterium sp.). With [15N]-ammonium chloride the corresponding figures were 31.0 (Proteus vulg.) and 98.0% (Bifidobacterium sp.). The findings are discussed with regard to a possible enteral detoxification in uremic patients by bacterial utilization and elimination of urea and ammonia.

Head-space gas-liquid chromatography and mass spectrometry were used to detect and identify products formed by Proteus vulgaris, P. mirabilis, P. morganii, and P. rettgeri from a defined medium supplemented with either phenylalanine, methionine, valine, leucine, histidine, lysine, ornithine, threonine, asparagine, aspartic acid, or tryptophan. In a detailed study of the products formed by 68 strains of Proteus spp. from L-methionine, the production of large amounts of both dimethyl disulfide and methyl mercaptan was found to be a characteristic of the genus. Both sulfur products appeared within a few hours of inoculation. Dimethyl disulfide was a more sensitive indicator of growth than the spectrometric determination of optical density. This suggests that it could be useful for the rapid, automated detection of any species of Proteus.

The biodegradation and utilization of the antiphytoviral substance 2,4-dioxohexahydro-1,3,5-triazine (DHT) by soil microorganisms was investigated. Mixed cultures of microorganisms deriving from different soils diminish in nutrient broth the content of DHT with increasing duration of culture. Microorganisms from an Egyptian garden soil fully degrade 10(-3) mol/1 DHT in a culture without additional aeration within 28 days. Also in deficient media the mixed microorganisms reduce the amount of DHT, reaching in nitrogen free nutrient solution even a degradation rate up to 12 mg DHT per liter and day. Pure cultures of Rhizobium leguminosarum, Proteus vulgaris, Saccharomyces cerevisiae and especially Agrobacterium radiobacter diminish the content of DHT in nitrogen free media, too. No such effect was detectable in cultures of four other species of soil bacteria. The DHT degradation by the microorganisms is connected with significant cell multiplication, e.g. A. radiobacter in shaking cultures with DHT as sole source of nitrogen shows a typical growth cycle with a lag-phase of 24 hours. The short persistence time of DHT in soils is concluded to be mainly due to biodegradation by microorganisms.

Three combined media ensuring the determination of 8 characteristics typical of bacteria belonging to the group Proteus-Providencia (tryptophane deamination, urease activity, production of hydrogen sulfite and indol, fermentation of mannitol, maltose, adonitol and inositol) are presented.

Dimethylsulphoxide (DMSO) and trimethylamine oxide (TMAO) sustained anaerobic growth of Proteus vulgaris with the non-fermentable substrate lactate. Cytoplasmic membrane vesicles energized by electron transfer from formate to DMSO displayed anaerobic uptake of serine, which was hindered by metabolic inhibitors known to destroy the proton motive force. This showed that DMSO reduction was coupled with a chemiosmotic mechanism of energy conversion; similar data for TMAO respiration have been presented previously. All biochemical tests applied indicated that the oxides were reduced by the same reductase system. The DMSO and TMAO reductase activities showed the same mobility on ion-exchange chromatography, and polyacrylamide disc gel electrophoresis (pH 8.9), gradient gel electrophoresis, and gel isoelectric focusing; mol. wt. and pI determined were 95,000 and 4.6, respectively. DMSO inhibited reduction of [14C]TMAO in vesicles. The reductase was inducible to a certain extent; both oxides being equally efficient as inducers. TMAO was reduced at a higher rate than DMSO, explaining faster growth of cells and increased uptake of serine in vesicles with TMAO as electron acceptor. Comparative studies with Escherichia coli also gave evidence for common TMAO and DMSO reductase systems.

The isolation from lake mud of a bacterium which can use dimethyl sulfoxide (DMSO) as an electron acceptor for growth is described. The isolate, called strain DL-1, was a small, gram negative, non-motile spiral. The sole product of DMSO reduction was dimethyl sulfide (DMS). Other electron acceptors used by the isolate included sulfite, thiosulfate, elemental sulfur, methionine sulfoxide, tetramethylene sulfoxide, nitrate, and oxygen (microaerophilically). Sulfate was not reduced and could not even be assimilated. Lactate or succinate could serve as electron donors, with acetate as the main product. Hydrogen could be used as an electron donor if acetate was present in the medium as a carbon source. The organism has a c-type cytochrome, and most likely uses electron transport phosphorylation during DMSO reduction. Cultures of Desulfovibrio sp., Escherichia coli, Pseudomonas aeruginosa, and Proteus vulgaris were tested for growth using DMSO as an electron acceptor, and only the Proteus strain grew. Both Proteus and strain DL-1 are versatile at coupling reductions with energy generation. There is a marked resemblance between strain DL-1 and the recently described sulfur-reducing spirillum of Wolfe and Pfennig.