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Enzymes Research

15:25 EST 21st November 2018 | BioPortfolio

UFM1-Activating Enzyme 5 (Uba5) Requires an Extension to Get the Job Done Right.

A novel strategy to screen inhibitors of multiple aminoglycoside-modifying enzymes with ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry.

Resistance to aminoglycoside antibiotics occurs primarily as a result of aminoglycoside-modification enzymes (AMEs) that modify the antibiotics. In this work, a novel strategy to combat the effects of antibiotic resistance was developed by screening multiple AMEs inhibitors with ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UHPLC-QTOF MS). The method scr...

Effect of immobilization on the activity of catalase carried by poly(HEMA-GMA) cryogels.

Hydrogen peroxide is converted by catalase to molecular oxygen and water to remove oxidative stress. In this study, catalase immobilization was performed using poly(2-hydroxyethyl methacrylate-glycidyl methacrylate) (poly(HEMA-GMA)) cryogels with different amounts of GMA. Catalase adsorption capacity of 298.7 ± 9.9 mg/g was achieved at the end of 9 h using the poly(HEMA-GMA)-250 cryogel. ...

Inulinase immobilized gold-magnetic nanoparticles as a magnetically recyclable biocatalyst for facial and efficient inulin biotransformation to high fructose syrup.

To date, the high cost of enzyme production, lack of enzyme reusability and operational stability are the main limitations of the enzyme's application in industry. In this work, inulinase was covalently immobilized on the surface of glutathione-coated gold magnetic nanoparticles (GSH-AuMNPs). The synthesized NPs were fully characterized. The effects of different restriction factors such as substra...

Optimization of cloned enzyme donor immunoassay cut-offs for drugs of abuse in whole blood of drivers involved in road accidents.

Immunoassay (IA) tests should be able to detect low concentrations of illegal drugs when used for the screening of drugs in drivers. False negatives should be avoided, and false positives should be reduced as far as possible. In this study, semi-quantitative results for blood samples containing illicit drugs (cannabinoids, cocaine, amphetamines/methamphetamines, opiates and methadone) obtained wit...

Maternal stress and placental function, a study using questionnaires and biomarkers at birth.

Prenatal stress affects the health of the pregnant woman and the fetus. Cortisol blood levels are elevated in pregnancy, and fetal exposure to cortisol is regulated by the placenta enzyme 11β-HSD2. A decrease in enzyme activity allows more maternal cortisol to pass through the placental barrier. Combining the fetal and maternal cortisol to cortisone ratio into the adjusted fetal cortisol exposure...

How do Enzymes Orient when Trapped on Metal-Organic Framework (MOF) Surfaces?

Enzyme immobilization in Metal-Organic Frameworks (MOFs) offers retained enzyme integrity and activity, enhanced stability, and reduced leaching. Trapping enzymes on MOF surfaces would allow for catalysis involving large substrates. In both cases, the catalytic efficiency and selectivity depend not only on enzyme integrity/concentration but also orientation. However, it has been a challenge to det...

RNA-Mediated Dimerization of the Human Deoxycytidine Deaminase APOBEC3H Influences Enzyme Activity and Interaction with Nucleic Acids.

Human APOBEC3H is a single-stranded (ss)DNA deoxycytidine deaminase that inhibits replication of retroelements and HIV-1 in CD4+ T cells. When aberrantly expressed in lung or breast tissue APOBEC3H can contribute to cancer mutagenesis. These different activities are carried out by different haplotypes of APOBEC3H. Here we studied APOBEC3H haplotype II, which is able to restrict HIV-1 replication a...

Structural Stability of Human Butyrylcholinesterase under High Hydrostatic Pressure.

Human butyrylcholinesterase is a nonspecific enzyme of clinical, pharmacological and toxicological significance. Although the enzyme is relatively stable, its activity is affected by numerous factors, including pressure. In this work, hydrostatic pressure dependence of the intrinsic tryptophan fluorescence in native and salted human butyrylcholinesterase was studied up to the maximum pressure at a...

Catalytic activity, structure and stability of proteinase K in the presence of biosynthesized CuO nanoparticles.

Here, CuO nanoparticles were synthesized using Sambucus nigra (elderberry) fruit extract. Further, the binding of proteinase K, as a model enzyme with green synthesized nanoparticles was investigated. The results demonstrated that the structural changes in enzyme were induced by the binding of nanoparticles. These changes were accompanied by the decrease in the Michaelis-Menten constant at 298 K...

Role of Musa paradisiaca ascorbate peroxidase in the transformation of methyl phenyl sulfide to its sulfoxide.

An ascorbate peroxidase from a new source Musa paradisiaca leaf juice has been purified to homogeneity using a simple procedure involving concentration by ultra filtration and anion exchange chromatography on diethyl amino ethyl [DEAE] cellulose column. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis [SDS-PAGE] analysis of the purified enzyme has shown a single protein band of molecular...

Relationship between anaphylaxis and use of beta-blockers and angiotensin-converting enzyme inhibitors: a systematic review and meta-analysis of observational studies.

Beta-blockers (BBs) and angiotensin-converting enzyme inhibitors (ACEIs) have been associated with an increased risk and severity of anaphylaxis. However, the evidence supporting these findings is contradictory.

Bioinspired Dual-enzyme Colloidosome Reactors for High-Performance Biphasic Catalysis.

In this paper, a novel method for the construction of colloidosome as a microreactor for dual-enzyme cascade biphasic reaction have been reported. A lipase-glucose oxidase enzyme pair is employed in this system. A water-soluble enzyme glucose oxidase (GOx) is compartmentalized inside the colloidosomes. A hydrophobic environment-favored enzyme, Candida Antarctica lipase B (CalB) is adsorbed on the ...

Investigating the Effect of Two-point Surface Attachment on Enzyme Stability and Activity.

Immobilization on solid supports provides an effective way to improve enzyme stability and simplify downstream processing for biotechnological applications, which has been widely used in research and in applications. However, surface immobilization may disrupt enzyme structure due to interactions between the enzyme and the supporting substrate, leading to a loss of the enzyme catalytic efficiency ...

The role of conformational fluctuations of protein towards methylation in DNA by cytosine-5-Methyltransferase.

Methylation of cytosine is the common epigenetic modification in genomes ranging from bacteria to mammals, and aberrant methylation leads to human diseases including cancer. Recognition of cognate DNA sequence by DNA methyltransferases and flipping of target base into the enzyme active site pocket are the key steps in DNA methylation. Using molecular dynamics simulations and enhanced sampling tech...

Immobilization of β-galactosidase by complexation: Effect of interaction on the properties of the enzyme.

In the present work, we aimed to explore the molecular binding between alginate and β-galactosidase, as well as the effect of this interaction on the activity retention, thermal stability, and kinetic properties of the enzyme. The impact of pH and enzyme/alginate ratio on physicochemical properties (turbidity, morphology, particle size distribution, ζ-potential, FTIR, and isothermal titration ca...

Lipase-Catalyzed Regioselective Synthesis of Dextrin Esters.

Four lipase enzymes were investigated as catalysts in the synthesis of regioselectively mono-substituted dextrin esters from dextrin and vinyl acetate. An immobilized lipase enzyme (Lipozyme TL IM) exhibited the highest activity. This enzyme showed regioselective substitution of the dextrin at the primary hydroxyl group (C6 position) under optimal conditions (60°C for 24 hours, using a 1:3 molar ...

Zebrafish acid ceramidase: Expression in Pichia pastoris GS115and biochemical characterization.

Acid ceramidase (N-acylsphingosine deacylase EC 3.5.1.23; AC) catalyzes the hydrolysis of ceramide into sphingosine (SPH) and free fatty acid. Zebrafish acid ceramidase (AC) has 60% homology with the human AC). Mutations in the human AC gene asah1 are known to cause Farber disease and spinal muscular atrophy with progressive myoclonic epilepsy. Zebrafish AC was overexpressed in Pichia pastoris by ...

Significant enhancement of direct electric communication across enzyme-electrode interface via nano-patterning of synthetic glucose dehydrogenase on spatially tunable gold nanoparticle (AuNP)-modified electrode.

In this study, the effect of inter-enzyme steric hindrance that occurs during enzyme immobilization on the electrode, on direct electrical communications of enzyme with electrode was investigated via nano-patterning of enzymes on the electrode. Here, the nano-patterning of enzymes was achieved through the combination of DET-capable enzyme that was produced via fusion of site-specific gold binding ...

BRENDA in 2019: a European ELIXIR core data resource.

The BRENDA enzyme database (www.brenda-enzymes.org), recently appointed ELIXIR Core Data Resource, is the main enzyme and enzyme-ligand information system. The core database provides a comprehensive overview on enzymes. A collection of 4.3 million data for ∼84 000 enzymes manually evaluated and extracted from ∼140 000 primary literature references is combined with information obtained by text ...

Imidazole-pyrazole hybrids: Synthesis, characterization and in-vitro bioevaluation against α-glucosidase enzyme with molecular docking studies.

Herein, substituted imidazole-pyrazole hybrids (2a-2n) were prepared via a multi component reaction employing pyrazole-4-carbaldehydes (1a-1d), ammonium acetate, benzil and arylamines as reactants. All the new compounds were characterized through their spectral and elemental analyses. Further these compounds were tested against α-glucosidase enzyme. The compounds 2k, 2l and 2n possessed good inhi...

Disulfide bridge formation to increase thermostability of DFPase enzyme: A computational study.

Organophosphate compounds bioremediation by use of organophosphorus degradation enzymes such as DFPase is a developing interest in industry and medicine. The most important problem with the bio-catalytic enzymes is their instability on high temperatures. This work carried out to find suitable locations for introducing disulfide bridges in DFPase enzyme. We employed some computational approaches to...

Metal-enzyme nanoaggregates eradicate both Gram positive and negative bacteria and their biofilms.

In order to palliate the appearance of antimicrobial resistance (AMR), the use of bactericidal agents acting differently than conventional antibiotics and the elimination of bacterial biofilm, are the two most promising strategies. Here, we integrated these two complementary strategies into new antimicrobial metal-enzyme nanoaggregates (NAs) of α-amylase and silver (αAgNAs) able to eliminate bac...

Stereo-preference of camphor for H-bonding with phenol, methanol and chloroform: A combined matrix isolation IR spectroscopic and quantum chemical investigation.

Camphor is known to be held in the substrate pocket of cytochrome P450cam enzyme via H-bond with a tyrosine residue of the enzyme in a unique orientation. This structural exclusivity results in regio- and stereo-specific hydroxylation of camphor by the enzyme. We have carried out a combined IR spectroscopic and quantum chemical investigation to shed light on the factors influencing the conformatio...

Stabilization of polydopamine modified silver nanoparticles bound trypsin: Insights on protein hydrolysis.

This study presents an effective method to enhance the proteolytic activity of trypsin by designing a nanobioconjugate of trypsin and polydopamine coated silver nanoparticles (Ag-PDA). Ag-PDA nanocomposite was synthesized and trypsin was covalently attached to this matrix. A multifold enhancement in enzyme activity was observed after conjugation, with an effectiveness factor of η = 5.62. Conj...

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