Microbial electrode and microbial sensor

• US 6,113,762 A
• Filed: 08/22/1997
• Issued: 09/05/2000
• Est. Priority Date: 02/22/1995
• Status: Expired due to Fees

• Current Assignee: Akebono Brake Industry Company Limited (Akebono Brake Corporation), Akebono Research and Development Centre Limited, Isao Karube
• Original Assignee: Akebono Brake Industry Company Limited (Akebono Brake Corporation), Akebono Research and Development Centre Limited, Isao Karube
• Inventors: Karube, Isao, Yoshida, Nobuyuki, Yano, Kiyoko, Morita, Takashi
• Primary Examiner(s): Tung, T.
• Assistant Examiner(s): NOGUEROLA, ALEXANDER STEPHAN
• Application Number:
  US08/913,473
• Time in Patent Office: 1,110 Days
• Field of Search: 204/403, 205/777.5, 435/817
• US Class Current: 204/403.06
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/004 mediator-assisted

Abstract

The microbial electrode is constructed having a substrate(3) made of an insulating material, an electric conductor(2) which is fixed on the substrate and a membrane containing microorganism cells(1) which is fixed on the conductor. The microbial sensor(20) is constructed having the microbial electrode above described and a conductor(11) to act as a counter electrode fixed on the insulating material of the surface of the substrate of the electrode which is opposite to the surface where the membrane containing microorganism cells is fixed.

Claims

1. A microbial sensor for measuring a chemical or biological component present in a solution, comprising:

(a) a microbial electrode comprising;

an electric conductor;

a microorganism cell layer fixed on and electronically contacted with the electric conductor to detect transfer of electrons generated by the microorganism cells when metabolizing the chemical or biological component; and

an insulating layer which insulates the electric conductor from the solution when in use;

(b) a counter electrode which is in contact with the solution when used;

(c) a support having two faces insulated from each other, wherein the electric conductor of the microbial electrode is fixed on one of the faces to sandwich said electric conductor between said microorganism cell layer and said support, and the counter electrode is fixed on the other face of said support;

(d) a generator of electric potential which generates an electric potential between the counter electrode and the microbial electrode to facilitate transfer of electrons therebetween generated by the microorganism cells; and

(e) a detector for measuring the electric current passing between the counter electrode and the microbial electrode;

whereby the chemical or biological component is measured based on a predetermined correlation between the measured electric current and the amount of electrons transferred by the microorganism cells when metabolizing the chemical or biological component.

2. The microbial sensor of claim 1, wherein said microorganism cells are procaryotic cells selected from the group consisting of;

E. coli, Bacillus, Gluconobacter, Pseudomonas, and Actinomycetes.

3. The microbial sensor of claim 1, wherein said microorganisms are included into a gel membrane, photo-setting resin membrane, or a polyacrylamide membrane, or a polymer membrane.

4. The microbial sensor of claim 3, wherein said gel membrane is selected from the group consisting of;

an alginate gel membrane, a carrageenan gel membrane, an agarose gel membrane, a cudlan gel membrane and an achitosan gel membrane.

5. The microbial sensor of claim 3, wherein said photo-setting resin membrane is a photo crosslinkable polyvinyl alcohol membrane.

6. The microbial sensor of claim 1, wherein said electric conductor is platinum, gold, silver, graphite, or carbon.

7. The microbial sensor of claim 1 further comprising a mediator for facilitating the transfer of electrons generating from the metabolism of the various chemical or biological components to the electrode.

8. The microbial sensor of claim 7, wherein said mediator is selected from the group consisting of pigments, biological oxidation/reduction materials, and metal EDTA compounds.

9. The microbial sensor of claim 8, wherein the pigments are selected from the group consisting of 1-methoxy-5-methylphenaxinium methylsulfonate, 2,6-dichloroindophenol, 9-dimethylaminobenzo-a-phenazoxonium chloride, methylene blue, indigotrisulfonic acid, phenosafranin, thionine, new methylene blue, 2,6-dichlorophenol, indophenol, azule B, N,N,N'"'"',N'"'"'-tetramethyl-p-phenylenediamine dihyrochloride, resorufine, safianine, sodium anthraquinone β

-sulfonate, and indigo carmine.

10. The microbial sensor of claim 8, wherein said biological oxidation/reduction materials are selected from the group consisting of riboflavin, L-ascorbic acid, flavin adenine dinucleotide, flavin mononucleotide, nicotine adenine dinucleotide, lumichrome, ubiquinone, hyroquinone, 2,6-dichlorobenzoquinone, 2-methylbenzoquinone, 2,5-dihyroxybenzoquinone, 2-hydroxy-1,4-naphtoquinone, glutathione, peroxidase, cytochrome C and ferredoxin.

11. The microbial sensor of claim 8, wherein said metal EDTA compounds are selected from the group consisting of Fe-EDTA, MN-EDTA, Zn-EDTA.

12. The microbial sensor of claim 7 wherein said mediator has a concentration of 40 nM to 100 mM.

13. The microbial sensor of claim 12 wherein said mediator has a concentration of 10 μ

M to 50 mM.

14. The microbial sensor of claim 1 further comprising a reference electrode used to set the potential of the microbial electrode.

15. The microbial sensor of claim 1 further comprising additional microbial electrodes allowing for the synchronous measurement of a plurality of chemical or biological components.

16. The microbial sensor according to claim 1, which is incorporated in a sample liquid collecting unit comprising:

a sample liquid-sucking inlet;

a liquid accumulation portion for storing sample liquid sucked through the sample liquid-sucking inlet; and

an aspiration pump portion for generating an aspirating power, wherein the microbial sensor is disposed in the sample liquid accumulation portion.