Enzyme microgels in packed-bed bioreactors with downstream amperometric detection using microfabricated interdigitated microsensor electrode arrays

Anthony Guiseppi-Elie, Norman F. Sheppard, Sean Brahim, Dyer Narinesingh

Research output: Contribution to journalArticle

55 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)475-484
Number of pages10
JournalBiotechnology and Bioengineering
Volume75
Issue number4
DOIs
StatePublished - Nov 20 2001

Keywords

  • Amperometry
  • Glucose
  • Glucose oxidase
  • Glutamate oxidase
  • Hydrogels
  • Interdigitated microsensor electrode arrays
  • Monosodium glutamate

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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