154-92-7Relevant articles and documents
Study the effect of trypsin enzyme activity on the screening of applying frontal affinity chromatography
Qian, JunQing,Zhao, ChangYan,Tong, Jun,Jiang, ShengLan,Zhang,Lu, ShiYong,Guo, Hui
, p. 740 - 751 (2019)
Frontal affinity chromatography (FAC) combined with enzyme has been widely used for drug screening. In this paper, the effect of target enzyme activity on screening of bioactive compounds was studied through applying FAC. Trypsin with different degree of inactivation were prepared as target enzyme by thermal denaturation. Their primary structure was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and use Fourier transform infrared (FTIR) and ultraviolet-visible (UV–vis) spectroscopy to detect group structure. Ultimately, it was found that the main structure of enzyme with decreased activity remained unchanged. The oxymatrine and matrine which can interact with trypsin were selected to study their binding to trypsin with different activities in FAC. The results showed that oxymatrine and matrine had a significant difference in the breakthrough volume among seven kinds of columns prepared by trypsins with different activities, at the different concentration. It indicated that trypsins with different activities in FAC could combine with oxymatrine and matrine. The binding constant (Kd) variation between oxymatrine, matrine and trypsin with different activities are 5.520 ± 0.038 and 3.577 ± 0.071, within error range, which indicated that the activity of target enzyme with primary unchanged structure has no effect on screening of bioactive components by FAC.
Synthesis of papain-polyacrylamide hydrogel microspheres and their catalytic application
Fei, Xu,Li, Yao,Ma, Yuan,Tian, Jing,Wang, Yi,Xu, Longquan
, p. 16696 - 16704 (2021)
Although immobilized enzyme technologies effectively improve the reusability and stability of an enzyme, the introduction of covalent bonds or/and charge interaction causes the change for the secondary structure of an enzyme, thereby reducing the catalytic activity. In this work, a mild and efficient strategy for enzyme immobilization is proposed. A size-controlled hydrogel microsphere was designed and synthesized as an immobilized enzyme carrier. Papain was immobilized on the surface and pores of hydrogel microspheres by hydrogen bonding interactions, which have no adverse effect on the secondary structure of an enzyme. The activity of the papain-polyacrylamide hydrogel microsphere was 106.41% of that of free papain. In addition, the immobilized papain maintained 59.60% of free papain activity after 10 cycles. The results of practical catalytic application proved that the obtained papain-polyacrylamide hydrogel microspheres were very suitable for catalytic hydrolysis of allergen protein in cow's milk without introducing any other hetero protein.
Biocatalytic synthesis, antimicrobial properties and toxicity studies of arginine derivative surfactants
Fait, M. Elisa,Garrote, Graciela L.,Clapés, Pere,Tanco, Sebastian,Lorenzo, Julia,Morcelle, Susana R.
, p. 1465 - 1477 (2015)
Abstract Two novel arginine-based cationic surfactants were synthesized using as biocatalyst papain, an endopeptidase from Carica papaya latex, adsorbed onto polyamide. The classical substrate N α-benzoyl-arginine ethyl ester hydrochloride for the determination of cysteine and serine proteases activity was used as the arginine donor, whereas decyl- and dodecylamine were used as nucleophiles for the condensation reaction. Yields higher than 90 and 80 % were achieved for the synthesis of N α-benzoyl-arginine decyl amide (Bz-Arg-NHC10) and N α-benzoyl-arginine dodecyl amide (Bz-Arg-NHC12), respectively. The purification process was developed in order to make it more sustainable, by using water and ethanol as the main separation solvents in a single cationic exchange chromatographic separation step. Bz-Arg-NHC10 and Bz-Arg-NHC12 proved antimicrobial activity against both Gram-positive and Gram-negative bacteria, revealing their potential use as effective disinfectants as they reduced 99 % the initial bacterial population after only 1 h of contact. The cytotoxic effect towards different cell types of both arginine derivatives was also measured. Bz-Arg-NHCn demonstrated lower haemolytic activity and were less eye-irritating than the commercial cationic surfactant cetrimide. A similar trend could also be observed when cytotoxicity was tested on hepatocytes and fibroblast cell lines: both arginine derivatives were less toxic than cetrimide. All these properties would make the two novel arginine compounds a promising alternative to commercial cationic surfactants, especially for their use as additives in topical formulations.
Enzyme kinetics in crowded solutions from isothermal titration calorimetry
Maximova, Ksenia,Wojtczak, Jakub,Trylska, Joanna
, p. 96 - 105 (2019)
Isothermal titration calorimetry (ITC) is a universal technique that directly measures the heat absorbed or released in a process. ITC is typically used to determine thermodynamic parameters of association of molecules without the need to label them. However, ITC is still rarely applied to study chemical reactions catalyzed by enzymes. In addition, these few studies of enzyme kinetic measurements that have been performed were in diluted solutions. Yet, to estimate realistic kinetic parameters, we have to account for the fact that enzymatic reactions in cells occur in a crowded environment because cells contain 200–400 g/L of macromolecular crowders such as proteins, ribosomes and lipids. Thus we expanded the ITC application for solutions mimicking the cellular environment by adding various macromolecular crowders. We investigated how these crowders affect the kinetics of trypsin-catalyzed reactions and determined the Michaelis-Menten parameters for hydrolysis of two trypsin substrates: Nα-benzoyl-L-arginine ethyl ester (BAEE) and Nα-benzoyl-DL-arginine β-naphthylamide (BANA). Since ITC enables investigations of complex and turbid solutions with label-free reagents, it seems a perfect technique for kinetic analyses in crowded solutions. ITC also offers the opportunity to control enzyme-crowder and substrate-crowder interactions.
Time-resolved dynamic nuclear polarization enhanced NMR spectroscopy
Bowen, Sean,Hilty, Christian
, p. 5235 - 5237 (2008)
(Chemical Presented) The sensitive touch: Sensitivity constraints in NMR spectroscopy typically call for long measurement times. Hyperpolarization can enhance the time resolution of NMR spectroscopy by removing the need for signal averaging. Reactions such as enzyme catalysis can be followed in real time by hyperpolarized NMR spectroscopy through reduction in the intensity of the substrate resonance as well as the appearance of product resonances (see picture).
Comparative Studies on the Interaction of Spermidine with Bovine Trypsin by Multispectroscopic and Docking Methods
Momeni, Lida,Shareghi, Behzad,Saboury, Ali Akbar,Farhadian, Sadegh
, p. 9632 - 9641 (2016)
The effect of spermidine on the kinetics, conformation, and dynamics of native trypsin was studied by steady-state thermal stability, intrinsic fluorescence, circular dichroism (CD), ultraviolet-visible (UV-vis) spectroscopy, and kinetic techniques, as well as molecular docking, at the temperatures of 298 and 308 K. The Stern-Volmer quenching constants (Ksv) for the trypsin-spermidine complex were obtained at two temperatures, revealing that spermidine quenched the intensity of trypsin through the static mode of the quenching mechanism. The corresponding thermodynamic parameters, Gibbs free-energy, enthalpy, and entropy changes, showed that the binding process was spontaneous. These values and the molecular docking technique revealed that the hydrogen bonding and van der Waals forces played a major role in stabilizing the complex. CD, absorption, and fluorescence results also indicated that spermidine binding had a partial effect on trypsin structure. Spermidine could also influence the activity of trypsin. Upon spermidine binding, the Vmax value of the enzyme was increased and the kcat/Km values were enhanced slightly. The Tm of the trypsin-spermidine complex was enhanced probably due to the higher H-bond formation and lower surface hydrophobicity after spermidine modification, as confirmed by UV-vis spectroscopy and fluorescence spectra. UV absorption and CD studies also indicated that the binding of spermidine to trypsin had induced microenvironmental changes around the enzyme, leading to changes in its secondary structure.
Synthetic approaches to a challenging and unusual structure—an amino-pyrrolidine guanine core
Rippel, Rafael,Pinheiro, Luís,Lopes, Mónica,Louren?o, Ana,Ferreira, Luísa M.,Branco, Paula S.
, (2020/02/25)
The synthesis of an unreported 2-aminopyrrolidine-1-carboxamidine unit is here described for the first time. This unusual and promising structure was attained through the oxidative decarboxylation of amino acids using the pair of reagents, silver(I)/peroxydisulfate (Ag(I)/S2O82?) followed by intermolecular (in the case of L-proline derivative) and intramolecular trapping (in the case of acyl L-arginine) by N-nucleophiles. The L-proline approach has a broader scope for the synthesis of 2-aminopyrrolidine-1-carboxamidine derivatives, whereas the intramolecular cyclization afforded by the L-acylarginines, when applied, results in higher yields. The former allowed the first synthesis of cernumidine, a natural alkaloid isolated in 2011 from Solanum cernuum Vell, as its racemic form.
Pyrazine-derived disulfide-reducing agent for chemical biology
Lukesh, John C.,Wallin, Kelly K.,Raines, Ronald T.
supporting information, p. 9591 - 9594 (2014/08/18)
For fifty years, dithiothreitol (DTT) has been the preferred reagent for the reduction of disulfide bonds in proteins and other biomolecules. Herein we report on the synthesis and characterization of 2,3-bis(mercaptomethyl)pyrazine (BMMP), a readily accessible disulfide-reducing agent with reactivity under biological conditions that is markedly superior to DTT and other known reagents. This journal is the Partner Organisations 2014.
Nanostructured layered double hydroxide aerogels with enhanced adsorption properties
Touati, Souad,Mansouri, Hela,Bengueddach, Abdelkader,De Roy, Andre,Forano, Claude,Prevot, Vanessa
supporting information; scheme or table, p. 7197 - 7199 (2012/08/13)
Aerogels of layered double hydroxides were prepared by a simple and eco-friendly method involving a quick coprecipitation followed by supercritical CO2 drying. Such aerogels display high surface areas and enhanced adsorption behavior.
Encapsulation of enzyme in large mesoporous material with small mesoporous windows
Malvi, Bharmana,Gupta, Sayam Sen
supporting information; experimental part, p. 7853 - 7855 (2012/09/05)
Trypsin has been encapsulated in the mesopores of a hierarchical mesoporous silica material synthesized via Cu(i) catalyzed azide-alkyne click reaction between azide functionalized large spherical SBA-15 particles and alkyne functionalized mesoporous sili
