Enzyme immobilization is a technology that enables (bio-)catalysts to be applied in continuous-flow systems. However, there is a plethora of immobilization methods available with individual advantages and disadvantages. Here, we assessed the influence of simple and readily available methods with respect to the performance of 2-deoxy-d-ribose-5-phosphate aldolase (DERA) in continuous-flow conditions. The investigated immobilization strategies cover the unspecific attachment to carriers via epoxides, affinity-based attachment via metal ion affinity, StrepTag™-StrepTactin™ interaction as well as the covalent affinity attachment of an enzyme to a matrix tethered by the HaloTag. The metal-ion-affinity-based approach outperformed the other methods in terms of immobilized activity and stability under applied conditions. As most enzymes examined today already have a HisTag for purification purposes, effective immobilization may be applied, as simple as a standard purification, if needed.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9570893 | PMC |
| http://dx.doi.org/10.3390/molecules27196483 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Tyramine-enzyme conjugate repeats for an interdigitated capacitance immunosensing array in the detection of neuroblastoma biomarker neuron-specific enolase.
Analyst
January 2025
Department of Pediatric Surgery, The First Affiliated Hospital of Fujian Medical University, 20 Cha-Zhong Road, Taijiang District, Fuzhou 350005, China.
Methods based on enzyme labelling strategies have been widely developed for capacitance immunoassays, but most suffer from low sensitivity and are unfavorable for routine use in the early stages of diagnostics. Herein, we designed a highly efficient capacitance immunosensing method for the low-abundance neuroblastoma biomarker neuron-specific enolase (NSE) using an interdigitated micro-comb electrode. Initially, monoclonal mouse anti-human NSE capture antibodies were immobilized on the interdigitated gold electrodes using bovine serum albumin.
View Article and Find Full Text PDFA miniaturized colorimetric-photoacoustic dual-mode device for point-of-care testing of ascorbic acid and ascorbate oxidase.
Anal Chim Acta
March 2025
The Radiology Department of Shanxi Provincial People's Hospital Affiliated to Shanxi Medical University, Taiyuan, 030001, China. Electronic address:
Background: Real-time and rapid detection of ingredients in food has important significance for food safety. However, traditional detection methods not only require bulky and costly instruments but also are often based on single-mode analysis, limiting their accuracy and applications in point-of-care testing. Herein, an integrated and miniaturized dual-mode device based on colorimetric and photoacoustic (PA) principles is developed, using Au@Ag nanoparticles (Au@AgNPs) as signal probe and ascorbic acid (AA) and ascorbate oxidase (AAO) as analytes.
View Article and Find Full Text PDFTrehalose decorated nanostructures stabilize combined cross-linked enzyme aggregates (Combi-CLEAs) of β-galactosidase and glucose isomerase.
Int J Biol Macromol
January 2025
Department of Food Science, College of Agriculture and Life Sciences, Cornell University, Ithaca, NY 14853, USA. Electronic address:
Combined cross-linked enzyme aggregates (Combi-CLEAs) of β-Galactosidase (β-Gal) and Glucose Isomerase (GI) allow the transformation of d-lactose to lactose-fructose syrup through one-pot cascade biocatalytic reactions. Despite its promise, the low thermostability of β-Gal and high-temperature demands for GI limits this application. Trehalose is a protein-stabilizing disaccharide which has been utilized in immobilized enzyme systems to enhance protein thermostability.
View Article and Find Full Text PDFMultifunctional quaternary ammonium-modified TEMPO-oxidized cellulose nanofibers and MIL-100 with encapsulated laccase for efficient removal of anionic arund cationic dyes in wastewater.
Int J Biol Macromol
January 2025
Department of Cell and Molecular Biology, Faculty of Life Science and Biotechnology, Shahid Beheshti University, P.O. Box 19839-69411, Tehran, Iran. Electronic address:
The increasing prevalence of micropollutants like cationic and anionic dyes in wastewater creates an influential environmental challenge, mainly due to their toxic effects and persistence. Current methods often lack the efficiency and versatility to cope with a wide variety of contaminants. This study explores the modification of TEMPO-oxidized cellulose nanofibers (TOCNF) using (3-chloro-2-hydroxypropyl) trimethylammonium chloride (CHPTAC) to enhance their cationic properties.
View Article and Find Full Text PDFCofactor-directed co-immobilization of dual-enzyme on functionalized montmorillonite with enhanced catalytic performance.
Int J Biol Macromol
January 2025
School of Environment and Resource, Key Laboratory of Solid Waste Treatment and Resource Recycle of Ministry of Education, Southwest University of Science and Technology, Mianyang, Sichuan 621010, China.
Recently, multi-enzyme cascade catalysis has attracted increasing attention due to the advantages of integrating multiple enzymes, few side reactions and high catalytic efficiency. Herein, a novel dual-enzyme cascade system (GOx-FMt-HRP) was developed through cofactor-directed orientational co-immobilization of glucose oxidase (GOx) and horseradish peroxidase (HRP) onto functional montmorillonite (FMt). The presented method realizes the reconstitution of cofactors and apo-enzymes (enzymes without cofactors), which enables enzymes to be immobilized in specific orientations on the support, thereby effectively reducing changes in their conformation.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!