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An autoclavable glucose biosensor for microbial fermentation monitoring and control
Author(s) -
Phelps Michael R.,
Hobbs John B.,
Kilburn Douglas G.,
Turner Robin F. B.
Publication year - 1995
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260460604
Subject(s) - biosensor , glucose oxidase , fermentation , cellulose , chemistry , chromatography , bacterial cellulose , immobilized enzyme , matrix (chemical analysis) , biochemistry , enzyme
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 electode. Enzyme immobilization can be reversed by perfusing the cellulose matrix with a suitable eluting solution. An autocavable 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. © 1995 John Wiley & Sons, Inc.