Microbial sensor for penicillins using a recombinant strain of Escherichia coli.

E. coli harboring the multicopy plasmid pBR327, which codes for beta-lactamase synthesis, was immobilized on acetylcellulose membranes. These were placed in the vicinity of a flat pH electrode, thus constituting a penicillin biosensor based on the detection of changes in pH using a reference pH electrode. A response time of 8 min was obtained with at least 2 mg of cells cm-2 of membrane. A linear detection range between 5 and 30 mM of penicillin was achieved. Buffering capacity decreased the sensitivity, but to a lower extent as when compared with probes using purified enzyme. The sensor was completely stable for at least 13 days and proved to be useful for penicillin estimation in complex media such as fermentation broths and milk.

[1]  Isao Karube,et al.  Immobilized whole cell-based flow-type sensor for cephalosporins , 1979 .

[2]  L. C. Clark,et al.  ELECTRODE SYSTEMS FOR CONTINUOUS MONITORING IN CARDIOVASCULAR SURGERY , 1962 .

[3]  M. Flanagan,et al.  Thin‐film antimony–antimony‐oxide enzyme electrode for penicillin determination , 1986, Biotechnology and bioengineering.

[4]  J. Koryta.  Theory and applications of ion-selective electrodes: Part 6 , 1986 .

[5]  A. Turner,et al.  Analytical applications of immobilised proteins and cells , 1988 .

[6]  A. Mukherji,et al.  A penicillin selective enzyme electrode. , 1973, Analytical chemistry.

[7]  S. Enfors,et al.  Design and response characteristics of an enzyme electrode for measurement of penicillin in fermentation broth , 1979 .

[8]  A. Schleifer,et al.  Improved penicillin selective enzyme electrode. , 1974, Analytical chemistry.

[9]  I. Schneider.  Determination of penicillin V in fermentation samples by flow injection analysis , 1985 .

[10]  J. Rusling,et al.  Immobilized enzyme-based flowing-stream analyzer for measurement of penicillin in fermentation broths. , 1976, Analytical Chemistry.

[11]  G G Guilbault,et al.  Enzyme‐Based Electrodes as Analytical Tools , 1987, Biotechnology and applied biochemistry.

[12]  P. Vadgama,et al.  Microbial enzymes in biosensors. , 1986, The Journal of applied bacteriology.

[13]  Ashok Mulchandani,et al.  Developments and applications of biosensors , 1988 .

[14]  I. Karube,et al.  MICROBIAL ELECTRODES FOR FERMENTATION PROCESSES , 1979 .

[15]  C. O'callaghan,et al.  [5] β-Lactamase assays , 1975 .

[16]  F. Bolivar,et al.  Construction and characterization of new cloning vehicles. V. Mobilization and coding properties of pBR322 and several deletion derivatives including pBR327 and pBR328. , 1981, Gene.

[17]  R Tor,et al.  New enzyme membrane for enzyme electrodes. , 1986, Analytical chemistry.

[18]  D R Walt,et al.  Polymer immobilized enzyme optrodes for the detection of penicillin. , 1987, Analytical chemistry.

[19]  J. Janata,et al.  pH-based enzyme potentiometric sensors. Part 3. Penicillin-sensitive field effect transistor. , 1985, Analytical chemistry.

[20]  B. Danielsson,et al.  Enzyme Thermistor Analysis of Penicillin in Standard Solutions and in Fermentation Broth , 1981, Applied and environmental microbiology.

[21]  H. Mottola,et al.  Flow injection determination of penicillins using immobilized penicillinase in a single bead string reactor. , 1985, Analytical chemistry.

[22]  K. Mosbach,et al.  Determination of glucose, urea and penicillin using enzyme--pH-electrodes. , 1973, Biochimica et biophysica acta.

[23]  E. Hall,et al.  The developing biosensor arena , 1986 .

[24]  G G Guilbault,et al.  Enzyme electrode probes , 1971, Pure and applied chemistry. Chimie pure et appliquee.

[25]  O. H. Lowry,et al.  Protein measurement with the Folin phenol reagent. , 1951, The Journal of biological chemistry.

[26]  J. Anzai,et al.  Preparation of coated-wire potentiometric enzyme sensors. , 1986, Chemical & pharmaceutical bulletin.

[27]  Klaus Mosbach,et al.  An enzyme electrode for measurement of penicillin in fermentation broth: A step toward the application of enzyme electrodes in fermentation control , 1978 .

[28]  F. Bolivar,et al.  Construction and characterization of new cloning vehicles. IV. Deletion derivatives of pBR322 and pBR325. , 1980, Gene.

[29]  D. Margineanu,et al.  Bioselective electrodes with immobilized bacteria , 1985 .