Enzyme microgels in packed-bed bioreactors with downstream amperometric detection using microfabricated interdigitated microsensor electrode arrays
| Autor: | Sean Brahim, Dyer Narinesingh, Anthony Guiseppi-Elie, Norman F. Sheppard |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Bioengineering
Biosensing Techniques Applied Microbiology and Biotechnology Sampling Studies Glucose Oxidase Bioreactors Enzyme Stability Sodium Glutamate Electrochemistry Bioreactor Glucose oxidase L-glutamate oxidase Electrodes Packed bed Detection limit Chromatography Dose-Response Relationship Drug biology Chemistry technology industry and agriculture Reproducibility of Results Hydrogen-Ion Concentration Enzymes Immobilized Streptomyces Amperometry Glucose Flow Injection Analysis Self-healing hydrogels biology.protein Aspergillus niger Gels Biosensor Biotechnology |
| Zdroj: | Biotechnology and Bioengineering. 75:475-484 |
| ISSN: | 1097-0290 0006-3592 |
| DOI: | 10.1002/bit.10069 |
| Popis: | In this article, we describe the use of pH- responsive hydrogels as matrices for the immobilization of two enzymes, glucose oxidase (GOx) and glutamate oxidase (GlutOx). Spherical hydrogel beads were prepared by inverse suspension polymerization and the enzymes were immobilized by either physical entrapment or covalent immobilization within or on the hydrogel surface. Packed-bed bioreactors were prepared containing the bioactive hydrogels and these incorporated into flow injection (FI) systems for the quantitation of glucose and monosodium glutamate (MSG) respectively. The FI amperometric detector comprised a microfabricated interdigitated array within a thin-layer flow cell. For the FI manifold incorporating immobilized GOx, glucose response curves were found to be linear over the concentration range 1.8–280 mg dL−1 (0.1–15.5 mM) with a detection limit of 1.4 mg dL−1 (0.08 mM). Up to 20 samples can be manually analyzed per hour, with the hydrogel-GOx bioreactor exhibiting good within-day (0.19%) precision. The optimized FI manifold for MSG quantitation yielded a linear response range of up to 135 mg dL−1 (8 mM) with a detection limit of 3.38 mg dL−1 (0.2 mM) and a throughput of 30 samples h−1. Analysis of commercially produced soup samples gave a within-day precision of 3.6%. Bioreactors containing these two physically entrapped enzymes retained > 60% of their initial activities after a storage period of up to 1 year. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 475–484, 2001. |
| Databáze: | OpenAIRE |
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