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Enzyme immobilization

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https://www.readbyqxmd.com/read/28331958/nucleic-acid-sensing-with-enzyme-dna-binding-protein-conjugates-cascade-and-simple-dna-nanostructures
#1
Gülsen Betül Aktas, Vasso Skouridou, Lluis Masip
A versatile and universal DNA sensing platform is presented based on enzyme-DNA binding protein tags conjugates and simple DNA nanostructures. Two enzyme conjugates were thus prepared, with horseradish peroxidase linked to the dimeric single-chain bacteriophage Cro repressor protein (HRP-scCro) and glucose oxidase linked to the dimeric headpiece domain of Escherichia coli LacI repressor protein (GOx-dHP), and used in conjunction with a hybrid ssDNA-dsDNA detection probe. This probe served as a simple DNA nanostructure allowing first for target recognition through its target-complementary single-stranded DNA (ssDNA) part and then for signal generation after conjugate binding on the double-stranded DNA (dsDNA) containing the specific binding sites for the dHP and scCro DNA binding proteins...
March 22, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/28330374/recent-developments-in-the-use-of-tyrosinase-and-laccase-in-environmental-applications
#2
Sidy Ba, Vaidyanathan Vinoth Kumar
Our current global environmental challenges include the reduction of harmful chemicals and their derivatives. Bioremediation has been a key strategy to control the massive presence of chemicals in the environment. Enzymes including the phenoloxidases, laccases and tyrosinases, are increasingly being investigated as "green products" in the removal of many chemical contaminants in waters and soils. Both phenoloxidases are widespread in nature and attractive biocatalysts due to their ability to use readily available molecular oxygen as sole cofactor for their catalytic elimination of a large number of chemicals...
March 22, 2017: Critical Reviews in Biotechnology
https://www.readbyqxmd.com/read/28330301/evaluation-of-p-cresol-degradation-with-polyphenol-oxidase-ppo-immobilized-in-various-matrices
#3
Vijayalakshmi A Edalli, Sikandar I Mulla, Syed Ali Musstjab Akber Shah Eqani, Gurumurthy D Mahadevan, Rohit Sharma, Yogesh Shouche, Chandrappa M Kamanavalli
p-Cresol is an environmental pollutant due to its vast use, toxicity and persistence, nevertheless, its degradation in an enzyme is unclear. In this study, we used Pleurotus sp. isolate VLECK02 polyphenol oxidase (PPO) for the determination of p-cresol degradation. On the basis of UV, FT-IR and chromatographic (HPLC and GC-MS) analysis, 4-methylcatechol was identified as the main metabolite of p-cresol catabolism. In addition, batch and semi-continuous degradation of p-cresol (10 and 20 mM) were studied and compared by free and immobilized PPO in different matrices like sodium alginate (SA), sodium alginate-polyvinyl alcohol (SA-PVA) and sodium alginate-polyvinyl alcohol-silver nanoparticles (SA-PVA-AgNPs)...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28330279/immobilization-of%C3%A2-bacillus-amyloliquefaciens-sp1-and-its-alkaline-protease-in-various-matrices-for-effective-hydrolysis-of-casein
#4
Shiwani Guleria, Abhishek Walia, Anjali Chauhan, C K Shirkot
An extracellular alkaline protease producing B. amyloliquefaciens SP1 was isolated from apple rhizosphere having multifarious plant growth-promoting activities. B. amyloliquefaciens SP1 protease was immobilized using various concentrations of calcium alginate, agar and polyacrylamide to determine the optimum concentration for formation of the beads. Enzyme activity before immobilization (at 60 °C, pH 8.0 for 5 min) was 3580 µg/ml/min. The results of immobilization with various matrices revealed that 3 % calcium alginate (2829...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28330256/immobilized-lipase-catalyzing-glucose-stearate-synthesis-and-their-surfactant-properties-analysis
#5
A Maria Sebatini, Manisha Jain, P Radha, S Kiruthika, Krishnamurthi Tamilarasan
Sugar fatty acid esters are practical importance and have a variety of applications that include surfactants and as an emulsifying agent. In this study, we report glucose stearate synthesis using lipase-Fe3O4 nanoparticles catalyst. The influence of various reaction factors, such as silica gel concentration, molar ratio of sugar/acid, reaction temperature and speed of agitation on esterification by immobilized enzyme was analyzed. The glucose stearate esterification degree of 87.2 % was obtained under the optimized condition: 1:2 molar ratio of glucose/stearic acid, 2 % (w/v) of silica gel at 120 rpm and 40 °C...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28330233/improvement-of-shelf-life-of-soymilk-using-immobilized-protease-of-oerskovia-xanthineolytica-ncim-2839
#6
A K Sahoo, V S Gaikwad, R C Ranveer, P B Dandge, S R Waghmare
Protease enzyme has lot of commercial applications, so the cost-effective production of protease using sunflower oil seed waste was carried out from Oerskovia xanthineolyitca NCIM 2839. The maximum protease production was after 24 h of incubation with 2.5 % oil seed waste concentration. O. xanthineolytica was found to produce two proteases-P1 and P2. The proteases were purified using 60 % cold acetone precipitation and DEAE-cellulose ion exchange chromatography. SDS-PAGE revealed molecular weight of P1 and P2 was 36 and 24 kDa, respectively...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28330200/use-of-ca-alginate-immobilized-pseudomonas-aeruginosa-for-repeated-batch-and-continuous-degradation-of-endosulfan
#7
Vijayalakshmi Pradeep, Usha Malavalli Subbaiah
The current investigation is taken up with the aim of studying repeated batch and continuous degradation of Endosulfan, using Ca-alginate immobilized cells of Pseudomonas aeruginosa isolated from an agricultural soil. The work involves the study of genes and enzymes involved in the degradation of the pesticide and was carried out with an objective of reducing the toxicity of Endosulfan by degrading it to less toxic metabolites. The long-term stability of Endosulfan degradation was studied during its repeated batch degradation, carried out over a period of 35 days...
December 2016: 3 Biotech
https://www.readbyqxmd.com/read/28330097/immobilization-of-halophilic-aspergillus-awamori-em66-exochitinase-on-grafted-k-carrageenan-alginate-beads
#8
Mona A Esawy, Ghada E A Awad, Walaa A Abdel Wahab, Magdy M M Elnashar, Ahmed El-Diwany, Saadia M H Easa, Fawkia M El-Beih
A novel extreme halophilic exochitinase enzyme was produced by honey isolate Aspergillus awamori EM66. The enzyme was immobilized successfully on k-carrageenan-alginate gel carrier (CA) with 93 % immobilization yield. The immobilization process significantly improved the enzyme specific activity 2.6-fold compared to the free form. The significant factors influencing the immobilization process such as enzyme protein concentration and loading time were studied. Distinguishable characteristics of optimum pH and temperature, stability at different temperatures and NaCl tolerance for free and immobilized enzyme were studied...
June 2016: 3 Biotech
https://www.readbyqxmd.com/read/28330084/covalent-immobilization-of-microbial-naringinase-using-novel-thermally-stable-biopolymer-for-hydrolysis-of-naringin
#9
Ghada E A Awad, Abeer A Abd El Aty, Abeer N Shehata, Mohamed E Hassan, Magdy M Elnashar
Naringinase induced from the fermented broth of marine-derived fungus Aspergillus niger was immobilized into grafted gel beads, to obtain biocatalytically active beads. The support for enzyme immobilization was characterized by ART-FTIR and TGA techniques. TGA revealed a significant improvement in the grafted gel's thermal stability from 200 to 300 °C. Optimization of the enzyme loading capacity increased gradually by 28-fold from 32 U/g gel to 899 U/g gel beads, retaining 99 % of the enzyme immobilization efficiency and 88 % of the immobilization yield...
June 2016: 3 Biotech
https://www.readbyqxmd.com/read/28330083/enzymatic-production-of-glucosamine-and-chitooligosaccharides-using-newly-isolated-exo-%C3%AE-d-glucosaminidase-having-transglycosylation-activity
#10
Sujata Sinha, Subhash Chand, Pushplata Tripathi
Exochitosanase secreting fungus (A. fumigatus IIT-004) was isolated from fish waste using 1 % (w/v) chitosan as sole carbon source after multistage screening. Chitosan-dependent exochitosanase enzyme production (6 IU ml(-1)) in log phase of growth (chitosan utilization rate 0.11 g g(-1) cell h(-1)) was observed for Aspergillus fumigatus in chitosan minimal salt medium and there was no enzyme production in glucose medium. Enzyme production was found to be extracellular and subjected to purification by a number of steps like acetone fractionation as well as column chromatography...
June 2016: 3 Biotech
https://www.readbyqxmd.com/read/28328089/capacitive-dna-sensor-for-rapid-and-sensitive-detection-of-whole-genome-human-herpes-virus-1-dsdna-in-serum
#11
Cheng Cheng, Rania Oueslati, Jayne Wu, Jiangang Chen, Shigetoshi Eda
This work presents a rapid, highly sensitive, low-cost and specific capacitive DNA sensor for detection of whole genome human herpes virus-1 DNA. This sensor is capable of direct DNA detection with a response time of 30 seconds, and it can be used to test standard buffer or serum samples. The sensing approach for DNA detection is based on AC electrokinetics. By applying an inhomogeneous AC electric field on sensor electrodes, positive dielectrophoresis is induced to accelerate DNA hybridization. The same applied AC signal also directly measures the hybridization of target with the probe on the sensor surface...
March 22, 2017: Electrophoresis
https://www.readbyqxmd.com/read/28325868/inositol-phosphates-and-phosphoinositides-activate-insulin-degrading-enzyme-while-phosphoinositides-also-mediate-binding-to-endosomes
#12
Eun Suk Song, HyeIn Jang, Hou-Fu Guo, Maria A Juliano, Luiz Juliano, Andrew J Morris, Emilia Galperin, David W Rodgers, Louis B Hersh
Insulin-degrading enzyme (IDE) hydrolyzes bioactive peptides, including insulin, amylin, and the amyloid β peptides. Polyanions activate IDE toward some substrates, yet an endogenous polyanion activator has not yet been identified. Here we report that inositol phosphates (InsPs) and phosphatdidylinositol phosphates (PtdInsPs) serve as activators of IDE. InsPs and PtdInsPs interact with the polyanion-binding site located on an inner chamber wall of the enzyme. InsPs activate IDE by up to ∼95-fold, affecting primarily Vmax The extent of activation and binding affinity correlate with the number of phosphate groups on the inositol ring, with phosphate positional effects observed...
March 21, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/28325051/bioavailability-of-immobilized-epidermal-growth-factor-covalent-versus-noncovalent-grafting
#13
Nesrine Riahi, Frederic Murschel, Sophie Lerouge, Yves Durocher, Olivier Henry, Gregory De Crescenzo
In an effort to rationalize and optimize an antiapoptotic coating combining chondroitin sulfate (CS) and epidermal growth factor (EGF) for vascular applications, the authors here report the comparison of two grafting strategies aiming to display EGF in an oriented fashion on CS. For that purpose, the authors produced, purified, and characterized a chimeric protein corresponding to EGF that was N-terminally fused to a cysteine and a coil peptide. The chimera was covalently immobilized via its free thiol group or captured via coiled-coil interactions at the surface of a biosensor or on a chondroitin sulfate coating in multiwell plates, mimicking the coating that was previously developed by them for stent-graft surfaces...
March 21, 2017: Biointerphases
https://www.readbyqxmd.com/read/28323015/simultaneously-achieve-soluble-expression-and-biomimetic-immobilization-of-candida-antarctica-lipase-b-by-introducing-polyamine-tags
#14
Xiaoxue Zhou, Yu Han, Zheng Lv, Xuemei Tian, Han Li, Panpan Xie, Liangyu Zheng
Polyamine tags fused in Candida antarctica lipase B (CalB) can help achieve high soluble expression of CalB in E. coli and can directly mediate silicification, which leads to rapid formation of a CalB-silica particle complex through a one-step approach. After optimization experiments, the fused lipase CalB tagged with 6-histidine at the N terminal and 10-lysine at the C terminal (6His-CalB-10Lys) is effectively expressed with high solubility (0.1mg/mL) and specific activity (10.1U/mg), and easily cross-linked in silica particles with a high immobilization efficiency of 96...
March 16, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28322107/surface-modulation-of-polymeric-nanocarriers-enhances-the-stability-and-delivery-of-proteins-and-small-molecules
#15
Zohreh Amoozgar, Michael S Goldberg
AIM: We aimed to enhance the stability and therapeutic efficiency of protein-based therapeutic formulations. MATERIALS & METHODS: Proteins were immobilized on the surface of nanoparticles (NPs) to improve both protein stability and protein function, especially enzymatic activity. The modularity of the platform was demonstrated by coating proteins of varied molecular weights and functionalities on the surface of poly(lactic-co-glycolic acid)-based NPs. RESULTS: Coating proteins to the particle surface greatly enhanced the stability of the NPs, preventing particle aggregation and improving enzymatic potency, including in vivo...
March 21, 2017: Nanomedicine
https://www.readbyqxmd.com/read/28322047/thiol-ene-monolithic-pepsin-microreactor-with-a-3d-printed-interface-for-efficient-uplc-ms-peptide-mapping-analyses
#16
Alexander Jönsson, Rasmus R Svejdal, Nanna B Bøgelund, Tam T T N Nguyen, Henrik Flindt, Jörg P Kutter, Kasper D Rand, Josiane Penelope Lafleur
To improve the sample handling, and reduce cost and preparation time, of peptide mapping LC-MS workflows in protein analytical research, we here investigate the possibility of replacing conventional enzymatic digestion methods with a polymer microfluidic chip-based enzyme reactor. Off-stoichiometric thiol-ene is utilized as both bulk material and as a monolithic stationary phase for immobilization of the proteolytic enzyme pepsin. The digestion efficiency of the, thiol-ene based, immobilized enzym reactor (IMER) is compared to that of a conventional, agarose packed bed, pepsin IMER column commonly used in LC-MS-based protein analyses...
March 21, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28319863/binding-affinity-and-adhesion-force-of-organophosphate-hydrolase-enzyme-with-soil-particles-related-to-the-isoelectric-point-of-the-enzyme
#17
Shah Md Asraful Islam, Shabina Yeasmin, Md Saiful Islam, Md Shariful Islam
The binding affinity of organophosphate hydrolase enzyme (OphB) with soil particles in relation to the isoelectric point (pI) was studied. Immobilization of OphB with soil particles was observed by confocal microscopy, Fourier transform infrared spectroscopy (FT-IR), and Atomic force microscopy (AFM). The calculated pI of OphB enzyme was increased from 8.69 to 8.89, 9.04 and 9.16 by the single, double and triple mutant of OphB enzyme, respectively through the replacement of negatively charged aspartate with positively charged histidine...
March 16, 2017: Ecotoxicology and Environmental Safety
https://www.readbyqxmd.com/read/28319379/immobilization-of-%C3%AE-galactosidases-from-lactobacillus-on-chitin-using-chitin-binding-domain
#18
Mai Lan Mai Pham, Tatjana Leister, Hoang Anh Nguyen, Bien-Cuong Do, Tuan-Anh Pham, Dietmar Haltrich, Montarop Yamabhai, Thu-Ha Nguyen, Tien-Thanh Nguyen
Two β-galactosidases from Lactobacillus including a heterodimeric, LacLM type enzyme from L. reuteri L103 and a homodimeric LacZ type β-galactosidase from L. bulgaricus DSM 20081 were studied for immobilization on chitin using a carbohydrate-binding domain (chitin-binding domain, ChBD) from a chitinolytic enzyme. Three recombinant enzymes, namely LacLM-ChBD, ChBD-LacLM and LacZ-ChBD, were constructed and successfully expressed in L. plantarum WCFS1. Depending on the structure of the enzymes, either homodimeric or heterodimeric, as well as the positioning of the chitin-binding domain in relation to the catalytic domains, i...
March 20, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28319222/co-localization-of-proteins-with-defined-sequential-order-and-controlled-stoichiometric-ratio-on-magnetic-nanoparticles
#19
Yi Yue, Yang-Yang Lu, Mei Li, Zi-Jian Zhang, Tian-Wei Tan, Li-Hai Fan
Co-immobilization of enzymes used in cascade reactions is important for improving the overall catalytic efficiency. In this work, we employed scaffoldins as a bridge and succeeded in a highly-ordered co-localization of multiple proteins on magnetic nanoparticles with a loading capacity of ∼0.831 μmol g(-1) supports.
March 20, 2017: Nanoscale
https://www.readbyqxmd.com/read/28319173/affinity-purification-of-human-m-calpain-through-an-intrinsically-disordered-inhibitor-calpastatin
#20
Hung Huy Nguyen, Mihaly Varadi, Peter Tompa, Kris Pauwels
Calpains are calcium-activated proteases that have biomedical and biotechnological potential. Their activity is tightly regulated by their endogenous inhibitor, calpastatin that binds to the enzyme only in the presence of calcium. Conventional approaches to purify calpain comprise multiple chromatographic steps, and are labor-intensive, leading to low yields. Here we report a new purification procedure for the human m-calpain based on its reversible calcium-mediated interaction with the intrinsically disordered calpastatin...
2017: PloS One
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