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AN AUTOCLAVABLE GLUCOSE BIOSENSOR FOR MICROBIAL FERMENTATION MONITORING AND CONTROL

TitleAN AUTOCLAVABLE GLUCOSE BIOSENSOR FOR MICROBIAL FERMENTATION MONITORING AND CONTROL
Publication TypeJournal Article
Year of Publication1995
AuthorsPhelps, MR, Hobbs, JB, Kilburn, DG, Turner, RFB
JournalBiotechnology and Bioengineering
Volume46
Pagination514-524
Date PublishedJun
Type of ArticleArticle
ISBN Number0006-3592
Keywords(CBD), AMPEROMETRIC BIOSENSORS, BATCH, BIOSENSOR, CELLULOSE BINDING DOMAIN, CELLULOSE-BINDING DOMAIN, ELECTROCHEMICAL DETECTOR, ENZYME ELECTRODE, ESCHERICHIA-COLI, FERMENTATION MONITORING, FLOW-INJECTION ANALYSIS, GLUCOSE BIOSENSOR, GLUCOSE OXIDASE, OPTIMIZATION, OXIDASE ELECTRODE, OXYGEN
Abstract

The design, construction, and characterization of a prototype-regenerable glucose biosensor based on the reversible immobilization of glucose oxidase (GOx) using cellulose binding domain (CBD) technology is described. GOx, chemically linked to CBD, is immobilized by binding to a cellulose matrix on the sensor-indicating electrode. Enzyme immobilization can be reversed by perfusing the cellulose matrix with a suitable eluting solution. An autoclavable sensor membrane system is employed which is shown to be practical for use in real microbial fermentations. The prototype glucose biosensor was used without failure or deterioration during fed-batch fermentations of Escherichia coli reaching a maximum cell density of 85 g (dry weight)/L. Medium glucose concentration based on sensor output correlated closely with off-line glucose analysis and was controlled manually at 0.44 +/- 0.2 g/L for 2 h based on glucose sensor output. The sensor enzyme component could be eluted and replaced without interrupting the fermentation. To our knowledge, no other in situ biosensor has been used for such an extended period of time in such a high-cell-density fermentation. (C) 1995 John Wiley & Sons, Inc.

URL<Go to ISI>://A1995QY63000003