Urea, as an end product of protein metabolism and an abundant polar compound, significantly complicates the metabolomic analysis of urine by GC-MS. We developed a sample preparation method removing urea from urine samples prior the GC-MS analysis. The method based on urease immobilized on magnetic microparticles was compared with the others that are conventionally used (liquid-liquid extraction, free urease protocol), and samples without any treatment. To study the impact of sample preparation approaches on the quality of analytical data, we employed comprehensive metabolomic analysis (using both GC-MS and LC-MS/MS platforms) of standard material based on human urine. Multivariate statistical analysis has shown that immobilized urease treatment provides similar results to a free urease approach. However, significant alterations in the profiles of metabolites were observed in the samples without any treatment and after the extraction. Compared to other approaches that were tested, the immobilization of urease on microparticles reduces both the number of artifacts and the variability of the metabolites (average CV of extraction 19.7%, no treatment 11.4%, free urease 5.0%, and immobilized urease 2.5%). The method that was developed was applied in a GC-MS metabolomic experiment of glutaric aciduria type I, where both known diagnostically important biomarkers and unknowns, as the most discriminating compounds, were found.
- MeSH
- analýza hlavních komponent MeSH
- chromatografie kapalinová metody MeSH
- enzymy imobilizované moč MeSH
- glutaryl-CoA-dehydrogenasa nedostatek metabolismus MeSH
- lidé MeSH
- magnetické jevy * MeSH
- metabolické nemoci mozku metabolismus MeSH
- metabolom MeSH
- metabolomika metody MeSH
- metody pro přípravu analytických vzorků * MeSH
- močovina metabolismus MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí metody MeSH
- reprodukovatelnost výsledků MeSH
- studie proveditelnosti MeSH
- tandemová hmotnostní spektrometrie MeSH
- ureasa moč MeSH
- vrozené poruchy metabolismu aminokyselin metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Physiological aggregation of a recombinant enzyme into enzymatically active inclusion bodies could be an excellent strategy to obtain immobilized enzymes for industrial biotransformation processes. However, it is not convenient to recycle "gelatinous masses" of protein inclusion bodies from one reaction cycle to another, as high centrifugation forces are needed in large volumes. The magnetization of inclusion bodies is a smart solution for large-scale applications, enabling an easier separation process using a magnetic field. RESULTS: Magnetically modified inclusion bodies of UDP-glucose pyrophosphorylase were recycled 50 times, in comparison, inclusion bodies of the same enzyme were inactivated during ten reaction cycles if they were recycled by centrifugation. Inclusion bodies of sialic acid aldolase also showed good performance and operational stability after the magnetization procedure. CONCLUSIONS: It is demonstrated here that inclusion bodies can be easily magnetically modified by magnetic iron oxide particles prepared by microwave-assisted synthesis from ferrous sulphate. The magnetic particles stabilize the repetitive use of the inclusion bodies .
- MeSH
- biotransformace fyziologie MeSH
- buněčná inkluze metabolismus MeSH
- centrifugace metody MeSH
- Publikační typ
- časopisecké články MeSH
The annual food and agricultural waste production reaches enormous numbers. Therefore, an increasing need to valorize produced wastes arises. Waste materials originating from the food and agricultural industry can be considered as functional materials with interesting properties and broad application potential. Moreover, using an appropriate magnetic modification, smart materials exhibiting a rapid response to an external magnetic field can be obtained. Such materials can be easily and selectively separated from desired environments. Magnetically responsive waste derivatives of biological origins have already been prepared and used as efficient biosorbents for the isolation and removal of both biologically active compounds and organic and inorganic pollutants and radionuclides, as biocompatible carriers for the immobilization of diverse types of (bio)molecules, cells, nano- and microparticles, or (bio)catalysts. Potential bactericidal, algicidal, or anti-biofilm properties of magnetic waste composites have also been tested. Furthermore, low cost and availability of waste biomaterials in larger amounts predetermine their utilization in large-scale processes.
Smart materials of biological origin are attracting a lot of attention nowadays, especially as catalysts, carriers or adsorbents. Among them, magnetically modified biomaterials are especially important due to their response to external magnetic field. This report demonstrates that naturally occurring micrometer sized, high aspect ratio material (native and autoclaved Leptothrix sp. sheaths) efficiently bind synthetically prepared magnetite and maghemite nanoparticles and their aggregates. Magnetic modification of Leptothrix sheaths enables to prepare a promising material for advanced biotechnology and environmental technology applications. The prepared magnetically responsive sheaths were tested as inexpensive adsorbent for crystal violet removal from aqueous solutions. The observed maximum adsorption capacity was 243.1mg of dye per 1g of adsorbent.
- MeSH
- genciánová violeť chemie MeSH
- Leptothrix chemie MeSH
- magnetické nanočástice chemie MeSH
- Publikační typ
- časopisecké články MeSH
Bacterial cellulose (BC) produced by Komagataeibacter sucrofermentans was magnetically modified using perchloric acid stabilized magnetic fluid. Magnetic bacterial cellulose (MBC) was used as a carrier for the immobilization of affinity ligands, enzymes and cells. MBC with immobilized reactive copper phthalocyanine dye was an efficient adsorbent for crystal violet removal; the maximum adsorption capacity was 388mg/g. Kinetic and thermodynamic parameters were also determined. Model biocatalysts, namely bovine pancreas trypsin and Saccharomyces cerevisiae cells were immobilized on MBC using several strategies including adsorption with subsequent cross-linking with glutaraldehyde and covalent binding on previously activated MBC using sodium periodate or 1,4-butanediol diglycidyl ether. Immobilized yeast cells retained approximately 90% of their initial activity after 6 repeated cycles of sucrose solution hydrolysis. Trypsin covalently bound after MBC periodate activation was very stable during operational stability testing; it could be repeatedly used for ten cycles of low molecular weight substrate hydrolysis without loss of its initial activity.
- MeSH
- Acetobacteraceae chemie MeSH
- bakteriální polysacharidy chemie MeSH
- celulosa chemie MeSH
- enzymy imobilizované chemie MeSH
- genciánová violeť chemie MeSH
- imobilizované buňky cytologie MeSH
- indoly chemie MeSH
- magnetismus MeSH
- nosiče léků chemie MeSH
- organokovové sloučeniny chemie MeSH
- Saccharomyces cerevisiae cytologie MeSH
- skot MeSH
- trypsin chemie MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Cytokinins (CKs) are pivotal plant hormones that have crucial roles in plant growth and development. However, their isolation and quantification are usually challenging because of their extremely low levels in plant tissues (pmol g-1 fresh weight). We have developed a simple microscale magnetic immunoaffinity-based method for selective one-step isolation of CKs from very small amounts of plant tissue (less than 0.1 mg fresh weight). The capacity of the immunosorbent and the effect of the complex plant matrix on the yield of the rapid one-step purification were tested using a wide range of CK concentrations. The total recovery range of the new microscale isolation procedure was found to be 30-80% depending on individual CKs. Immunoaffinity extraction using group-specific monoclonal CK antibodies immobilized onto magnetic microparticles was combined with a highly sensitive ultrafast mass spectrometry-based method with a detection limit close to one attomole. This combined approach allowed metabolic profiling of a wide range of naturally occurring CKs (bases, ribosides and N9 -glucosides) in 1.0-mm sections of the Arabidopsis thaliana root meristematic zone. The magnetic immunoaffinity separation method was shown to be a simple and extremely fast procedure requiring minimal amounts of plant tissue.
The formation of bacterial biofilm on various surfaces has significant negative economic effects. The aim of this study was to find a simple procedure to decrease the Pseudomonas aeruginosa biofilm formation in a water environment by using different food waste biological materials as signal molecule adsorbents. The selected biomaterials did not reduce the cell growth but affected biofilm formation. Promising biomaterials were magnetically modified in order to simplify manipulation and facilitate their magnetic separation. The best biocomposite, magnetically modified spent grain, exhibited substantial adsorption of signal molecules and decreased the biofilm formation. These results suggest that selected food waste materials and their magnetically responsive derivatives could be applied to solve biofilm problems in water environment.
BACKGROUND: Barley straw, an agricultural by-product, can also serve as a low-cost and relatively efficient adsorbent of various harmful compounds. In this case, adsorption of four water-soluble dyes belonging to different dye classes (specifically Bismarck brown Y, representing the azo group; methylene blue, quinone-imine group; safranin O, safranin group; and crystal violet, triphenylmethane group) on native and citric acid-NaOH-modified barley straw, both in magnetic and non-magnetic versions, was studied. RESULTS: The adsorption was characterized using three adsorption models, namely Langmuir, Freundlich and Sips. To compare the maximum adsorption capacities (qmax), the Langmuir model was employed. The qmax values reached 86.5-124.3 mg of dye per g of native non-magnetic straw and 410.8-520.3 mg of dye per g of magnetic chemically modified straw. Performed characterization studies suggested that the substantial increase in qmax values after chemical modification could be caused by rougher surface of adsorbent (observed by scanning electron microscopy) and by the presence of higher amounts of carboxyl groups (detected by Fourier transform infrared spectroscopy). The adsorption processes followed the pseudo-second-order kinetic model and thermodynamic studies indicated spontaneous and endothermic adsorption. CONCLUSION: The chemical modification of barley straw led to a significant increase in maximum adsorption capacities for all tested dyes, while magnetic modification substantially facilitated the manipulation with adsorbent.
- MeSH
- adsorpce MeSH
- barvicí látky chemie MeSH
- biomasa MeSH
- chemické látky znečišťující vodu chemie MeSH
- ječmen (rod) chemie MeSH
- kinetika MeSH
- magnetismus * MeSH
- mikroskopie elektronová rastrovací MeSH
- průmyslový odpad MeSH
- stonky rostlin chemie MeSH
- termodynamika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The majority of algal cells can interact with a wide range of nano- and microparticles. Upon interaction the modified cells usually maintain their viability and the presence of foreign material on their surfaces or in protoplasm can provide additional functionalities. Magnetic modification and labeling of microalgal biomass ensures a wide spectrum of biotechnological, bioanalytical and environmental applications. Different aspects of microalgal cell magnetic modification are covered in the review, followed by successful applications of magnetic algae. Modified cells can be employed during their harvesting and removal, applied in toxicity microscreening devices and also as efficient adsorbents of different types of xenobiotics.
Large amounts of biochar are produced worldwide for potential agricultural applications. However, this material can also be used as an efficient biosorbent for xenobiotics removal. In this work, biochar was magnetically modified using microwave-synthesized magnetic iron oxide particles. This new type of a magnetically responsive biocomposite material can be easily separated by means of strong permanent magnets. Magnetic biochar has been used as an inexpensive magnetic adsorbent for the removal of water-soluble dyes. Five dyes (malachite green, methyl green, Bismarck brown Y, acridine orange and Nile blue A) were used to study the adsorption process. The dyes adsorption could be usually described with the Langmuir isotherm. The maximum adsorption capacities reached the value 137 mg of dye per g of dried magnetically modified biochar for Bismarck brown Y. The adsorption processes followed the pseudo-second-order kinetic model and the thermodynamic studies indicated spontaneous and endothermic adsorption. Extremely simple magnetic modification of biochar resulted in the formation of a new, promising adsorbent suggested for selected xenobiotics removal.
