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Biomedical subjects

Bo Mattiasson

Publications and source records attributed to Bo Mattiasson.

At least 37 records · Page 2Linked to original sources

Influence of agitation on the removal of nonylphenol by the white-rot fungi Trametes versicolor and Bjerkandera sp. BOL 13.

Bjerkandera sp. BOL 13 removed 95% of nonylphenol (at 9.7 mg nonylphenol L(-1) day(-1)) from aqueous medium after 5 days of incubation in agitated cultures. This removal rate decreased 2.5-fold in static cultures. By comparison, Trametes versicolor removed nonylphenol at 2.8 mg L(-1) day(-1) under conditions of static incubation, probably due to the action of laccase, but no growth was recorded in the agitated bottles.

Basidiomycota↗

Enhancement of sulphide production in anaerobic packed bed bench-scale biofilm reactors by sulphate reducing bacteria.

Two biofilm reactors, using pumice stone and Poraver as biofilm supports, were run, based on the optimization of sulphide production using a factorial design. The maximum H2S concentrations reached were 10 and 15 mM, respectively, both being appropriate for metal precipitation in effluents. The set-up of the pumice stone biofilm reactor is suitable for application in the mining area in the Bolivian Andean region, where this material is widely available. The use of specific primers for sulphate-reducing bacteria groups permits the identification of the sulphide-producing bacteria present in biofilms.

Bacteria, Anaerobic↗

Effects of polyols, saccharides, and glycoproteins on thermoprecipitation of phenylboronate-containing copolymers.

The copolymer of 3-(acrylamido)phenylboronic acid and N-isopropylacrylamide (82:18, Mn = 47000 g/mol) was prepared by free radical polymerization. The copolymer showed typical thermoprecipitation behavior in aqueous solutions; its phase transition temperature (TP) was 26.5 +/- 0.2 degrees C in 0.1 M glycine-NaOH buffer containing 0.1 M NaCl, pH 9.2. Due to specific complex formation of the pendant boronates with sugars, TP was strongly affected by the type of sugar and its concentration at pH 9.2. Fructose, lactulose, and glucose caused the largest increase in TP (up to 4 degrees C) at 0.56 mM concentration, attributed to the high binding affinity of the sugars to borate and phenylboronate. Among the sugars typical of nonreducing ends of oligosaccharides, N-acetylneuraminic acid had the strongest effect on TP (ca. 2 degrees C at 0.56 mM concentration and pH 9.2), while the effects of other sugars are well expressed at the higher concentrations (16 and 80 mM) and decreased in the order xylose approximately galactose >or= N-acetyllactosamine >or= mannose approximately fucose >> N-acetylglucosamine. The effect exerted on the phase transition by glycoproteins was the strongest with mucin from porcine stomach and decreased in the series mucin > horseradish peroxidase > human gamma-globulin at pH 9.2. As a first approximation, the weight percentage and/or the number of oligosaccharides in glycoproteins determined the character of their interaction with the pendant phenylboronates and, therefore, the effect on the copolymer phase transition.

Acrylamides↗

Nesterenkonia aethiopica sp. nov., an alkaliphilic, moderate halophile isolated from an Ethiopian soda lake.

Strain DSM 17733(T), isolated from the shore of Lake Abjata in Ethiopia, is a heterotrophic, alkaliphilic, moderately halophilic, Gram-positive, strictly aerobic, non-motile,non-endospore-forming bacterium. The organism grows optimally at 30-37 degrees C, pH 9 and 3 % (w/v) NaCl. Analysis of the cell wall showed the presence of murein of the type L-lys-gly-L-Glu, variation A4alpha. The G + C content of the genomic DNA was 69.0 mol%. Sequence analysis of 16S rRNA gene sequence of strain DSM 17733(T) placed the isolate in the genus Nesterenkonia. DNA-DNA hybridization of DSM 17733(T) with those organisms with the closest phylogenetic affiliation, i.e. Nesterenkonia halobia, Nesterenkonia lacusekhoensis and Nesterenkonia xinjiangensis, gave relatedness values of 48.5 %, 63.7 % (repetition, 57.2 %) and 35.7 % (repetition, 29.3 %), respectively. On the basis of both phenotypic and phylogenetic criteria and the low levels of DNA-DNA relatedness with the phylogenetically closest species N. xinjiangensis and N. halobia, it is proposed that the isolate be classified in a novel species, Nesterenkonia aethiopica sp. nov. The type strain is DSM 17733(T) (=CCUG 48939(T)).

Ethiopia↗

Clostridium lundense sp. nov., a novel anaerobic lipolytic bacterium isolated from bovine rumen.

A strictly anaerobic, mesophilic, endospore-forming, lipolytic bacterium, designated strain R1T, was isolated from bovine rumen fluid and characterized. Cells of this isolate were Gram-positive, non-motile rods that formed spherical terminal spores. The overall biochemical and physiological characteristics indicated that this strain should be placed in the genus Clostridium. The strain grew at temperatures between 25 and 47 degrees C (optimum, 37 degrees C), at pH between 5.0 and 8.5 (optimum pH 5.5-7.0) and in NaCl concentrations of 0-3% (w/v). The isolate was not able to utilize glucose or other carbohydrates as carbon sources. The DNA G + C content was 31.2 mol%. Sequence analysis of the 16S rRNA gene of R1T revealed that it has the closest match (98 % similarity) with Clostridium tetanomorphum DSM 4474T. The highest levels of DNA-DNA relatedness of the isolate were 61.9 and 54.3% with Clostridium pascui DSM 10365T and C. tetanomorphum DSM 4474T, respectively. Based on 16S rRNA gene sequence similarity, phylogenetic analysis, DNA G + C content, DNA-DNA hybridization data and distinct phenotypic characteristics, strain R1T (= DSM 17049T = CCUG 50446T) was classified in the genus Clostridium, as a member of a novel species, for which the name Clostridium lundense sp. nov. is proposed.

Animals↗

Cloning, sequence analysis, and expression of a gene encoding an endoxylanase from Bacillus halodurans S7.

The gene encoding an alkaline active xylanase of Bacillus halodurans S7, containing an open reading frame of 1188 nucleotides encoding 396 amino acids, was cloned and expressed in Escherchia coli. On the basis of sequence similarity, possible -10 and -35, ribosome binding, and transcription terminator regions were identified. Analysis of the deduced amino acid sequence revealed that the protein was a single domain enzyme belonging to family 10 and designated as xyn10A. The calculated molecular mass and isoelectric point (pI) of the mature peptide were 42.6 and 4.5 kDa, respectively. Xylanase activity expressed by the recombinant organism was detected in the cytoplasm, periplasm and the extracellular medium. In an 18-h old culture, about 39% of the xylanase was detected in the medium. The stability and activity profile of the recombinant xylanase was similar to the properties of the enzyme produced by the wild-type organism.

Amino Acid Sequence↗

Supermacroporous cryogel matrix for integrated protein isolation. Immobilized metal affinity chromatographic purification of urokinase from cell culture broth of a human kidney cell line.

A new type of supermacroporous, monolithic, cryogel affinity adsorbent was developed, allowing the specific capture of urokinase from conditioned media of human fibrosarcoma cell line HT1080. The affinity adsorbent was designed with the objective of using it as a capture column in an integrated perfusion/protein separation bioreactor setup. A comparative study between the utility of this novel cryogel based matrix and the conventional Sepharose based affinity matrix for the continuous capture of urokinase in an integrated bioreactor system was performed. Cu(II)-ion was coupled to epoxy activated polyacrylamide cryogel and Sepharose using iminodiacetic acid (IDA) as the chelating ligand. About 27-fold purification of urokinase from the conditioned culture media was achieved with Cu(II)-IDA-polyacrylamide cryogel column giving specific activity of about 814 Plough units (PU)/mg protein and enzyme yields of about 80%. High yields (95%) were obtained with Cu(II)-IDA-Sepharose column by virtue of its high binding capacity. However, the adsorbent showed lower selectivity as compared to cryogel matrix giving specific activity of 161 PU/mg protein and purification factor of 5.3. The high porosity, selectivity and reasonably good binding capacity of Cu(II)-IDA-polyacrylamide cryogel column make it a promising option for use as a protein capture column in integrated perfusion/separation processes. The urokinase peak pool from Cu(II)-IDA-polyacrylamide cryogel column could be further resolved into separate fractions for high and low molecular weight forms of urokinase by gel filtration chromatography on Sephacryl S-200. The selectivity of the cryogel based IMAC matrix for urokinase was found to be higher as compared to that of Cu(II)-IDA-Sepharose column.

Acrylamides↗

Sequential removal of heavy metals ions and organic pollutants using an algal-bacterial consortium.

The residual algal-bacterial biomass from photosynthetically supported, organic pollutant biodegradation processes, in enclosed photobioreactors, was tested for its ability to accumulate Cu(II), Ni(II), Cd(II), and Zn(II). Salicylate was chosen as a model contaminant. The algal-bacterial biomass combined the high adsorption capacity of microalgae with the low cost of the residual biomass, which makes it an attractive biosorbent for environmental applications. Cu(II) was preferentially taken-up from the medium when the metals were present both separately and in combination. There was no observed competition for adsorption sites, which suggested that Cu(II), Ni(II), Cd(II), and Zn(II) bind to different sites and that active Ni(II), Cd(II) and Zn(II) binding groups were present at very low concentrations. Therefore, special focus was given to Cu(II) biosorption. Cu(II) biosorption by the algal-bacterial biomass was characterized by an initial fast cell surface adsorption followed by a slower metabolically driven uptake. pH, Cu(II), and algal-bacterial concentration significantly affected the biosorption capacity for Cu(II). Maximum Cu(II) adsorption capacities of 8.5+/-0.4 mg g-1 were achieved at an initial Cu(II) concentration of 20 mg l-1 and at pH 5 for the tested algal-bacterial biomass. These are consistent with values reported for other microbial sorbents under similar conditions. The desorption of Cu(II) from saturated biomass was feasible by elution with a 0.0125 M HCl solution. Simultaneous Cu(II) and salicylate removal in a continuous stirred tank photobioreactor was not feasible due to the high toxicity of Cu(II) towards the microbial culture. The introduction of an adsorption column, packed with the algal-bacterial biomass, prior to the photobioreactor reduced Cu(II) concentration, thereby allowing the subsequent salicylate biodegradation in the photobioreactor.

Adsorption↗

Degradation of toxaphene by Bjerkandera sp. strain BOL13 using waste biomass as a cosubstrate.

The white-rot fungus Bjerkandera sp. strain BOL13 was capable of degrading toxaphene when supplied with wood chips, wheat husk or cane molasses as cosubstrates in batch culture experiments. Approximately 85% of toxaphene was removed when wheat husk was the main substrate. The production of lignin peroxidase was only stimulated when wheat husk was present in the liquid medium. Although xylanase was always detected, wheat husk supported the highest xylanase production. A negligible amount of beta-glucosidase and cellulase were found in the batch culture medium. To the best of our knowledge, this is the first reported case of toxaphene degradation by white-rot fungi.

Basidiomycota↗

Anion-exchange supermacroporous monolithic matrices with grafted polymer brushes of N,N-dimethylaminoethyl-methacrylate.

Graft polymerization using potassium diperiodatocuprate as initiator was found to be an effective and convenient method for grafting functional polymer of N,N-dimethylaminoethyl methacrylate (DMAEMA) onto superporous polyacrylamide gels, so-called cryogels (pAAm cryogels). It was possible to achieve grafting degrees up to 110% (w/w). The two-step graft polymerization i.e. first activation of the matrix followed by displacement of initiator solution with the monomer solution, decreased pronouncedly the soluble homopolymer formation. The efficiency of graft polymerization using a two-step technique increased up to 50% (w/w) at a monomer conversion of 10%, compared to 10% graft efficiency with 60-70% monomer conversion for one-step direct graft polymerization. The pAAm cryogels grafted in one-step and two-step procedures, respectively, behaved similarly when binding low-molecular weight ligand but showed very different behavior for sorption of a high-molecular-weight ligand, bovine serum albumin (BSA). The differences in behavior were rationalized assuming different structure of the graft polymer layers and tentacle-type BSA binding to the grafted polymer.

Anion Exchange Resins↗

Variations of wettability and protein adsorption on solid siliceous carriers grafted with poly(N-isopropylacrylamide).

Poly(N-isopropylacrylamide), a thermally responsive polymer, was end-grafted to mercaptopropyl derivatives of silica gel, plane glass sheets and glass capillary tubing by free radical polymerization of the monomer in 1,4-dioxane at 100 degrees C. The polymer monolayer attached to the glass carriers provided them with thermally controlled wettability registered by two independent methods: direct measurements of the water contact angle and capillary rise. The water contact angle changed from 54+/-3 degrees to 68+/-3 degrees in the temperature range from 20 to 50 degrees C. The polymer grafting to silica gel (pore diameter 100 A, particle size 5 microm) resulted in 15-30-fold reduction in protein adsorption on the carrier at 35 degrees C. Adsorption isotherms of myoglobin indicate completely different characters of the protein adsorption to silica gel and its polyNIPAM-grafted derivative. Cooling of the grafted carrier containing adsorbed myoglobin to 9 degrees C led to a partial release of the protein to the contacting solution, whereas heating of the system to 35 degrees C resulted in reversible binding of the protein. Adsorption of myoglobin on polyNIPAM-coated silica was ca. 2-fold higher at 35 than at 9 degrees C, most probably due to steric repulsion displayed by the swollen copolymer at the lower temperature.

Acrylamides↗

Immobilised peptide displaying phages as affinity ligands. Purification of lactoferrin from defatted milk.

An affinity purification procedure for the direct purification of lactoferrin from defatted (skimmed) milk has been developed. The procedure is based on using selected phage clones expressing a peptide with high binding affinity for lactoferrin which were covalently coupled to macroporous poly(dimethylacrylamide) monolithic column. Large pore size (10-100 microm) of macroporous poly(dimethylacrylamide) makes it possible to couple long (1 microm) phage particles as ligands without any risk of blocking the monolithic column. Bound lactoferrin was eluted using 1M NaCl with a purity of >95%. The technique presents a good alternative to conventional immunoaffinity chromatography for purification of a protein of interest from complex samples due to (i) the robustness of the system in terms of recovery and ligand leakage and (ii) economical aspect in terms of low ligand cost.

Acrylamides↗

Degradation of toxaphene in aged and freshly contaminated soil.

Degradation of toxaphene in soil from both newly contaminated (from Sweden) and aged spills (from Nicaragua) were studied. The newly contaminated soil contained approximately 11 mg kg(-1) toxaphene while the aged Nicaraguan soil contained approximately 100 mg kg(-1). Degradation was studied in anaerobic bioreactors, some of which were supplied with lactic acid and others with Triton X-114. In this study we found that the lower isomers Parlar 11, 12 were degraded while the concentration of isomer Parlar 15 increased. This supported an earlier evaluation which indicated that less chlorinated isomers are formed from more heavily isomers. Lactic acid when added to the soil, interfere with the degradation of toxaphene. Lactic acid was added; several isomers appeared to degrade rather slowly in newly contaminated Swedish soil. The Swedish soil, without any external carbon source, showed the slowest degradation rate of all the compounds studied. When Triton X-114 at 0.4 mM was added, the degradation rate of the compounds increased. This study illustrates that biodegradation of toxaphene is a complex process and several parameters have to be taken into consideration. Degradation of persistent pollutants in the environment using biotechnology is dependent on bioavailability, carbon sources and formation of metabolites.

Anaerobiosis↗

Amperometric biosensor for the detection of hydrogen peroxide using catalase modified electrodes in polyacrylamide.

A simple biosensor for the detection of hydrogen peroxide in organic solvents has been developed and coupled to a flow injection analysis (FIA) system. Catalase was entrapped in polyacrylamide gel and placed on the surface of platinum (working electrode) fixed in a Teflon holder with Ag-wire (auxiliary electrode), followed by addition of filter paper soaked in KCl. The entrapped catalase gel was held on the electrode using membranes. The effects of cellulose and polytetrafluroethylene (PTFE) membranes on the electrode response towards hydrogen peroxide have been studied. The modified electrode has been used to study the detection of hydrogen peroxide in solvents like water, dimethyl sulfoxide (DMSO), and 1,4-dioxane using amperometric techniques like cyclic voltammetry (CV) and FIA. The CV of modified catalase electrode showed a broad oxidation peak at -150 mV and a clear reduction peak at -212 mV in the presence of hydrogen peroxide. Comparison of CV with hydrogen peroxide in various solvents has been carried out. The electrode showed an irreversible kinetics with DMSO as the solvent. A flow cell has been designed in order to carry on FIA studies to obtain calibration plots for hydrogen peroxide with the modified electrode. The calibration plots in several solvents such as water, dimethyl sulfoxide, 1,4-dioxane have been obtained. The throughput of the enzyme electrode was 10 injections per hour. Due to the presence of membrane the response time of the electrode is concentration dependent.

Acrylic Resins↗

Screening of peptide affinity tags using immobilised metal affinity chromatography in 96-well plate format.

A method for high throughput screening of Green Fluorescent Proteins carrying metal binding tags in bacteria was developed. A random four amino acids tag-peptide library was successfully generated in E. coli. A 96-microtiter plate assembled with metal-iminodiacetic acid small cryogel columns was used for library screening. For the first time we were able to simultaneously screen a metal binding peptide tags library obtained from E. coli against different metal ions. From screening 25 different tags, three clones were able to bind to all metal ions studied (Ni2+, Zn2+, Co2+ and Cd2+). It was clearly demonstrated that the new construct could facilitate the screening of large peptide libraries.

Affinity Labels↗

Capture of bacterial endotoxins using a supermacroporous monolithic matrix with immobilized polyethyleneimine, lysozyme or polymyxin B.

Bacterial endotoxins (BEs) are integrated part of Escherichia coli, a microorganism widely used for the production of recombinant proteins. BEs should be eliminated in the course of down stream processing of target protein produced by these bacteria. Supermacroporous monolith (continuous bed) columns, so called cryogel columns, with immobilized polyethyleneimine (PEI), polymyxin B (PMB) and lysozyme were employed for BEs capture. Due to the large interconnected pores it was possible to use cryogel columns at flow rates as high as 10 ml/min. The columns packed with Sepharose CL-4B with immobilized PEI, PMB and lysozyme were impossible to use at these high flow rates due to the collapse of the bed. The dynamic capacities of the cryogel columns were nearly independent of the flow rate. In the presence of EDTA, BEs were quantitatively captured from mixtures with a model protein, bovine serum albumin (BSA) at pH 7.2 with practically no protein losses. At pH 3.6 BEs were captured directly from non-clarified E. coli cell lysate resulting in more than 10(4) times BEs clearance.

Chromatography, Affinity↗

Binding of adenosine to pendant phenylboronate groups of thermoresponsive copolymer: a quantitative study.

Binding of adenosine to the thermosensitive copolymer of N-isopropylacrylamide and 3-(acrylamido)aminophenylboronic acid (82:18, Mn = 47,000 g . mol(-1)) was studied by equilibrium dialysis at 22 degrees C and 37 degrees C, in a 0.1 M glycine buffer containing 0.1 M NaCl at pH 9.2. The copolymer exhibited a the phase transition temperature (T(p)) of 26.5 degrees C under the above conditions. At 22 degrees C the binding of adenosine to the water-soluble copolymer was well described by a Langmuir model, accounting for preferential ionisation of the boronate-nucleoside complexes and, therefore, restricted reactivity of the rest of boronates. At saturation, the copolymer contained 38% of its phenylboronic acid groups in the form of complexes, whereas the association constant was 1,400 M(-1). At 37 degrees C no binding of adenosine to thermally precipitated copolymer was found, presumably owing to interaction of the phenylboronates with hydrophobic segments of polyNIPAM. At high loading of the copolymer by the reversibly bound adenosine the T(p) steeply increases with increasing fraction of the phenylboronate-adenosine complexes in the chains. The increase of the T(p) observed above the saturating adenosine concentration (>1 x 10(-3) M, 22 degrees C) very probably testifies to competition of the nucleoside with hydrophobic polyNIPAM segments for binding to the pendant phenylboronates.

Adenosine↗

Biodegradation of nonylphenol in a continuous bioreactor at low temperatures and effects on the microbial population.

A packed-bed bioreactor inoculated with a mixed culture obtained from a contaminated site was used to continuously treat a saturated solution of nonylphenol. The reactor was operated at feeding rates of 13-112 ml h-1 and temperatures of 5.5, 10, and 15 degrees C. Optimal bioreactor performance was achieved at 10 degrees C and at a feeding rate of 84 ml h-1 (with a removal rate of 43 mg l-1 day-1 of nonylphenol). No endocrine activity was observed in the effluent of the bioreactor at any of the temperatures tested, and the only metabolic products found were branched carboxylic acids and alkanes (lacking an aromatic ring). The study of the microbial populations in the biofilm at the three temperatures tested using fluorescence in situ hybridization showed that all the bacterial species that could be identified belonged to the phylum Proteobacteria. The most abundant class identified at all three temperatures was beta-Proteobacteria. The proportions of bacteria that bound to the specific probes among the total population, identified with the bacterial probe EUB338MIX, were 60, 43, and 24% at 15, 10, and 5.5 degrees C, respectively.

Alkanes↗