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Enzyme immobilization

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https://www.readbyqxmd.com/read/28820997/nh2fe3o4-sio2-supported-peroxidase-catalyzed-h2o2-for-degradation-of-endocrine-disrupter-from-aqueous-solution-roles-of-active-radicals-and-noms
#1
Jing Ai, Weijun Zhang, Guiying Liao, Hua Xia, Dongsheng Wang
In this work, magnetic Fe3O4 was utilized to immobilize horseradish peroxidase (IM-HRP) in order to improve its stability and reusability by crosslinking method process with glutaraldehyde. The physicochemical properties of NH2Fe3O4@SiO2 and IM-HRP were characterized by powder X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), Thermo-gravimetric Analysis (TGA) and Transmission electron microscopy (TEM). The thermal stability of immobilized-HRP was considerably improved in comparison with free counterpart...
August 13, 2017: Chemosphere
https://www.readbyqxmd.com/read/28820469/a-third-generation-glucose-biosensor-based-on-cellobiose-dehydrogenase-immobilized-on-a-glassy-carbon-electrode-decorated-with-electrodeposited-gold-nanoparticles-characterization-and-application-in-human-saliva
#2
Paolo Bollella, Lo Gorton, Roland Ludwig, Riccarda Antiochia
Efficient direct electron transfer (DET) between a cellobiose dehydrogenase mutant from Corynascus thermophilus (CtCDH C291Y) and a novel glassy carbon (GC)-modified electrode, obtained by direct electrodeposition of gold nanoparticles (AuNPs) was realized. The electrode was further modified with a mixed self-assembled monolayer of 4-aminothiophenol (4-APh) and 4-mercaptobenzoic acid (4-MBA), by using glutaraldehyde (GA) as cross-linking agent. The CtCDH C291Y/GA/4-APh,4-MBA/AuNPs/GC platform showed an apparent heterogeneous electron transfer rate constant (ks) of 19...
August 18, 2017: Sensors
https://www.readbyqxmd.com/read/28815641/advances-in-capillary-electrophoretically-mediated-microanalysis-for-on-line-enzymatic-and-derivatization-reactions
#3
Shengyun Huang, Prasanta Paul, Pranov Ramana, Erwin Adams, Patrick Augustijns, Ann Van Schepdael
This review summarizes recent developments, applications and innovations of capillary electrophoretically mediated microanalysis methods. As a follow up of an earlier review, it covers the literature from early 2015 - early 2017. This article is divided into three parts. In the first part, different types of mixing procedures and applications of enzyme mediated microanalysis are discussed; the second part summarizes immobilized enzyme reactors (IMERs), while the third part deals with recent advances in on-line derivatization reactions...
August 17, 2017: Electrophoresis
https://www.readbyqxmd.com/read/28815227/microparticle-parking-and-isolation-for-highly-sensitive-microrna-detection
#4
Jae Jung Kim, Lynna Chen, Patrick S Doyle
Isolating small objects, such as particles, cells, and molecules, in individual aqueous droplets is useful for chemical and biological assays. We have developed a simple microfluidic platform to immobilize (park) microparticles at defined locations, and isolate particles in monodisperse droplets surrounded by immiscible oil. While conventional methods can only achieve stochastic encapsulation of objects within larger droplets, our in situ method ensures that a single particle is entrapped in a similar-sized droplet, with ∼95% yield for parking and isolation...
August 17, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28813159/enhancing-enzyme-activity-and-immobilization-in-nanostructured-inorganic-enzyme-complexes
#5
Xuye Lang, Lingling Zhu, Yingning Gao, Ian Wheeldon
Understanding the chemical and physical interactions at the interface of protein surfaces and inorganic crystals has important implications in the advancement of immobilized enzyme catalysis. Recently, enzyme-inorganic hybrid complexes have been demonstrated as effective materials for enzyme-immobilization. The precipitation of phosphate nanocrystals in the presence of enzymes creates enzyme-Cu3(PO4)2-3H2O particles with high surface-to-volume ratios, enhanced activity, and increased stability. Here, we begin to develop a mechanistic understanding of enzyme loading in such complexes...
August 16, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/28809474/a-nano-in-nano-approach-for-enzyme-immobilization-based-on-block-copolymers
#6
Finizia Auriemma, Claudio De Rosa, Anna Malafronte, Rocco Di Girolamo, Chiara Santillo, Yuri Gerelli, Giovanna Fragneto, Robert David Barker, Vincenzo Pavone, Ornella Maglio, Angela Lombardi
We have set-up a facile approach for fabrication of supports with tailored nanoporosity for immobilization of enzymes. To this aim block copolymers self-assembly has been used to prepare nanostructured thin films with well-defined architecture containing pores of tailorable size delimited by walls with tailorable degree of hydrophilicity. In particular, we have employed a mixture of polystyrene-block-poly(L-lactide) (PS-PLLA) and polystyrene-block-poly(ethylene oxide) (PS-PEO) diblock copolymers to generate thin films with a lamellar morphology consisting of PS lamellar do-mains alternating with mixed PEO/PLLA blocks lamellar domains...
August 15, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28803973/improved-biotransformation-of-arsenic-by-arsenite-oxidase-chitosan-nanoparticle-conjugates
#7
Neha Pandey, Renu Bhatt
Recent developments in the potential use of nanoparticles as carriers of enzyme have attracted great attention. In the present study, arsenite oxidase (AOase) enzyme capable of transforming the more toxic arsenite [As(III)] to the less toxic arsenate [As(V)] was extracted from an arsenic resistant bacterium (Exiguobacterium sp. As-9) and partially purified. Chitosan nanoparticles were prepared on the basis of ionic gelation of chitosan with tripolyphosphate (TPP) anions. The purified AOase was immobilized efficiently by physical adsorption on to chitosan nanoparticles and were characterized for particle size, morphology, zeta potential, AOase loading efficiency and in vitro transformation assay...
August 10, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28803843/high-throughput-screening-of-hybridoma-supernatants-using-multiplexed-fluorescent-cell-barcoding-on-live-cells
#8
Mei Lu, Brian M Chan, Peter W Schow, Wesley S Chang, Chadwick T King
With current available assay formats using either immobilized protein (ELISA, enzyme-linked immunosorbent assay) or immunostaining of fixed cells for primary monoclonal antibody (mAb) screening, researchers often fail to identify and characterize antibodies that recognize the native conformation of cell-surface antigens. Therefore, screening using live cells has become an integral and important step contributing to the successful identification of therapeutic antibody candidates. Thus the need for developing high-throughput screening (HTS) technologies using live cells has become a major priority for therapeutic mAb discovery and development...
August 10, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28801712/design-and-characterization-of-alcalase-chitosan-conjugates-as-potential-biocatalysts
#9
Milena G Žuža, Nikola Z Milašinović, Marko M Jonović, Jelena R Jovanović, Melina T Kalagasidis Krušić, Branko M Bugarski, Zorica D Knežević-Jugović
In this study, alcalase (protease from Bacillus licheniformis) immobilization by adsorption, enzyme crosslinking and covalent enzyme binding to activated chitosan microbeads were examined. The biocatalysts highest activity was obtained by covalent immobilization of alcalase onto a solid support. The alcalase covalent immobilization onto different types of chitosan beads obtained by inverse emulsion technique and electrostatic extrusion was studied. Parameters examined under different conditions were beads diameter, enzyme loading, enzyme capacity yield, and biocatalyst activity...
August 11, 2017: Bioprocess and Biosystems Engineering
https://www.readbyqxmd.com/read/28794310/structured-mono-and-diacylglycerols-with-a-high-content-of-medium-chain-fatty-acids
#10
Alaina A Esperón-Rojas, R Baeza-Jiménez, Cynthia Cano-Sarmiento, Hugo S García
In the present work, direct enzyme-catalysed esterification of medium chain fatty acids (MCFA) from three different sources (Medium chain triacylglycerols, MCT; saponified MCT and a mixture of free MCFA) was evaluated to obtain structured mono- and diacylglycerols. The esterifications were carried out mixing the different sources of MCFA with glycerol at two weight ratios (1:1 and 4:1, w/w), using three immobilized lipases: Novozym 435, Lipozyme RM IM and Lipozyme TL IM; different reaction times (t = 0, 15, 30, 60, 120 min); enzyme loadings (5, 10 or 15% with respect to the total weight of substrates)...
August 8, 2017: Journal of Oleo Science
https://www.readbyqxmd.com/read/28792903/hydrocalumite-thin-films-for-polyphenol-biosensor-elaboration
#11
A Soussou, I Gammoudi, A Kalboussi, C Grauby-Heywang, T Cohen-Bouhacina, Z M Baccar
Hybrid thin films based on Hydrocalumite (Ca2AlCl layered double hydroxide LDH) and tyrosinase enzyme have been used for the elaboration of a high sensitive amperometric biosensor detecting polyphenols extracted from green tea. Structural properties of LDH nanomaterials were characterized by X-ray powder diffraction and Infra-Red spectroscopy, confirming its crystalline phase and chemical composition. Ca2AlCl-LDHs-thin films were deposited by spin-coating, and studied by Atomic Force Microscopy (AFM) to obtain information about the surface morphology of this host matrix before and after enzyme's immobilization...
August 7, 2017: IEEE Transactions on Nanobioscience
https://www.readbyqxmd.com/read/28792205/solid-phase-chemical-modification-for-sialic-acid-linkage-analysis-application-to-glycoproteins-of-host-cells-used-in-influenza-virus-propagation
#12
Shuang Yang, Ewa Jankowska, Martina Kosikova, Hang Xie, John F Cipollo
Differentiation between the sialyl linkages is often critical to understanding biological consequence. Here we present a facile method for determining these linkages in glycans. Analysis of sialic acids is challenging due to their labile nature during sample preparation and ionization. Derivatization is often required via chemical reaction. Amidation derivatizes all sialic acids regardless of linkage, while esterification enables differentiation between α2,3-linked and α2,6-linked sialic acids. Reactions have been primarily performed on free glycans in solution but have been recently adapted to solid-phase providing unique advantages such as simplified sample preparation, improved yield, and high throughput applications...
August 9, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28791512/purification-of-angiotensin-i-converting-enzyme-ace-inhibitory-peptides-from-casein-hydrolysate-by-imac-ni-2
#13
Shanguang Wu, Xuezhen Feng, Yuan Lu, Yuting Lu, Saisai Liu, Yuhong Tian
Casein proteins were hydrolyzed by papain to identify inhibitory peptides of angiotensin I-converting enzyme (ACE). The hydrolysate was fractionized by immobilized metal affinity chromatography (IMAC-Ni(2+)). The fraction with high ACE inhibitory activity was enriched and further chromatographed on a reverse-phase column to yield four fractions. Among the fractions, the L4 fraction exhibited the highest ACE inhibitory activity and was identified by sequence analysis as Trp-Tyr-Leu-His-Tyr-Ala (WYLHYA), with IC50 value of 16...
August 8, 2017: Amino Acids
https://www.readbyqxmd.com/read/28789962/expression-and-characterization-of-an-enhanced-recombinant-heparinase-i-with-chitin-binding-domain
#14
Shuqin Xu, Meiling Qiu, Xuanyue Zhang, Jinghua Chen
Heparinase I (Hep I) can efficiently depolymerize heparin and heparin sulfate to oligosaccharides or unsaturated disaccharides, which resulted in loss of physiological function such as blood coagulation. In order to realize the immobilization of Hep I on chitin carriers, we cloned Hep I with the chitin binding domain (ChBD) as a chitin-affinity tag, and the Small Ubiquitin-like MOdifier (SUMO) linker as a solvation enhancer in different fusion sequence. DNA and protein gels suggested that 4 kinds of recombinants were successfully constructed and expressed in Escherichia coli (E...
August 5, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28783899/development-of-a-phage-chemiluminescent-enzyme-immunoassay-with-high-sensitivity-for-the-determination-of-imidaclothiz-in-agricultural-and-environmental-samples
#15
Yuan Ding, Xiude Hua, Nana Sun, Jiachuan Yang, Jiaqi Deng, Haiyan Shi, Minghua Wang
In this study, we isolated six phage-displayed peptides by biopanning phage-displayed peptide libraries on an immobilized anti-imidaclothiz monoclonal antibody. After analyzing the relative sensitivity of the individual phage-displayed peptides, we subsequently developed and optimized both a phage enzyme immunoassay (P-ELISA) and a phage chemiluminescent enzyme immunoassay (P-CLEIA) to improve the sensitivity and linear range of imidaclothiz assays. The P-CLEIA (50% inhibition concentration (IC50) of 0.86ngmL(-1), linear range of 0...
July 31, 2017: Science of the Total Environment
https://www.readbyqxmd.com/read/28783510/in-situ-immobilization-of-cd-by-organic-amendments-and-their-effect-on-antioxidant-enzyme-defense-mechanism-in-mung-bean-vigna-radiata-l-seedlings
#16
Pia Muhammad Adnan Ramzani, Mark S Coyne, Shazia Anjum, Waqas-Ud-Din Khan, Muhammad Iqbal
In situ immobilization of Cd is desirable due to the damaging effects of ex situ remediation techniques on soil. In this greenhouse study, the role of biochar (BC), chitosan (CH), and green waste (GW) was studied for in-situ Cd immobilization and alleviating Cd toxicity in mung bean seedlings. Amendments were applied at rates of 0.5% and 1% (w/w). The minimum mean value of Cd, in root, shoot, and soil (DTPA-Cd) (12.2, 4.7, and 0.7 mg kg(-1), respectively), occurred in the Cd + 1% CH treatment compared to all Cd amended treatments...
July 27, 2017: Plant Physiology and Biochemistry: PPB
https://www.readbyqxmd.com/read/28783436/a-multi-channel-open-tubular-enzyme-reactor-on-line-coupled-with-mass-spectrometry-for-detecting-ricin
#17
Ole Kristian Brandtzæg, Bent Tore Røen, Siri Enger, Elsa Lundanes, Steven Ray Wilson
For counter-terrorism purposes, a selective nano liquid chromatography-mass spectrometry (nanoLC-MS) platform was developed for detecting the highly lethal protein ricin from castor bean extract. Manual sample preparation steps were omitted by implementing a trypsin/Lys C enzyme-immobilized multi-channel reactor (MCR) consisting of 126 channels (8 µm inner diameter in all channels) that performed on-line digestion of proteins (5 minute reaction time, instead of 4-16 hours in previous in-solution methods). Reduction and alkylation steps were not required...
August 7, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28782724/zwitterionic-sulfobetaine-polymer-immobilized-surface-by-simple-tyrosinase-mediated-grafting-for-enhanced-antifouling-property
#18
Ho Joon Kwon, Yunki Lee, Le Thi Phuong, Gyeung Mi Seon, Eunsuk Kim, Jong Chul Park, Hyunjin Yoon, Ki Dong Park
Introducing antifouling property to biomaterial surfaces has been considered an effective method for preventing the failure of implanted devices. In order to achieve this, the immobilization of zwitterions on biomaterial surfaces has been proven to be an excellent way of improving anti-adhesive potency. In this study, poly(sulfobetaine-co-tyramine), a tyramine-conjugated sulfobetaine polymer, was synthesized and simply grafted onto the surface of polyurethane via a tyrosinase-mediated reaction. Surface characterization by water contact angle measurements, X-ray photoelectron spectroscopy and atomic force microscopy demonstrated that the zwitterionic polymer was successfully introduced onto the surface of polyurethane and remained stable for 7days...
August 4, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28782534/variation-in-adrenal-and-thyroid-hormones-with-life-history-stage-in-juvenile-northern-elephant-seals-mirounga-angustirostris
#19
J A Jelincic, M S Tift, D S Houser, D E Crocker
The classical approach to quantifying the impact of stressors on wildlife is through characterization of hormones associated with the generalized stress response. However, interpretation of hormone data can be difficult due to the range of natural variation within a species and potential confounds of individual and life-history variables. Blood adrenal and thyroid hormones were measured in 144 chemically immobilized yearling northern elephant seals (Mirounga angustirostris) to characterize variation between sexes and across semiannual haul-outs...
August 3, 2017: General and Comparative Endocrinology
https://www.readbyqxmd.com/read/28782250/meropenem-induced-immune-thrombocytopenia-and-the-diagnostic-process-of-laboratory-testing
#20
Rong Huang, Guang-Qing Cai, Jun-Hua Zhang, Feng-Xia Liu, Jin-Qi Ma, Hong Liu, Xin-Min Nie, Rong Gui
BACKGROUND: Drug-induced immune thrombocytopenia (DITP) is a serious, life-threatening clinical syndrome, the diagnosis of which is consistently difficult. In this report, we present a case of DITP caused by meropenem that was confirmed by laboratory tests. CASE REPORT: A 59-year-old male patient developed severe thrombocytopenia 8 days after the administration of meropenem and cefoperazone-sulbactam. After other causes were ruled out, DITP was suspected. Drug-induced platelet (PLT) antibodies were detected by enzyme immunoassay, flow cytometry, and monoclonal antibody immobilization of PLT antigens (MAIPA)...
August 7, 2017: Transfusion
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