Method and apparatus for continuously controlling an enzymatic reaction

• US 3,919,052 A
• Filed: 01/08/1974
• Issued: 11/11/1975
• Est. Priority Date: 01/10/1973
• Status: Expired due to Term

• Current Assignee: Daniele Trosset, Jean Marie Fresnel
• Original Assignee: Daniele Trosset, Jean Marie Fresnel
• Inventors: Fresnel, Jean Marie, Trosset, Daniele
• Primary Examiner(s): Monacell, A. Louis
• Assistant Examiner(s): Penland, R. B.
• Application Number:
  US05/431,608
• Time in Patent Office: 672 Days
• Field of Search: 195/68,63,103.5,105,115,116,117-118,DIG.11 204/195B,195T
• US Class Current: 435/173.2
• CPC Class Codes: A23C 19/0245 with immobilized enzymes or microorganisms
• CPC Class Codes: B01J 31/003 containing enzymes
• CPC Class Codes: C12M 21/18 Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C25B 15/00 Operating or servicing cells
• CPC Class Codes: C25B 3/00 Electrolytic production of organic compounds
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

The invention relates to a control method and apparatus for carrying out enzymatic reactions at a constant rate. A liquid reaction medium containing a substance to be transformed by the desired reaction is contacted with an enzymatic electrode comprising the corresponding enzyme fixed on a solid electronically conductive support. Control of the enzymatic reaction is achieved by applying a potential to the enzymatic electrode and controlling the value of this potential during the reaction so as to compensate for variations in the reaction conditions and the enzyme activity and thereby ensure a constant reaction rate.

Claims

1. AN ASSEMBLY FOR COLLECTING AD CULTURING A SAMPLE OF A REACTIONS WHEREIN A SUBSTRATE IS CONTINUOUSLY TRANSFORMED BY CONTACT WITH AT LEAST ONE ENZYME FIXED TO A SOLID CARRIER, SAID METHOD COMPRISING THE STEPS OF:

A. PROVIDING AN ENZYMATIC ELECTRODE WITH AT LEAST ONE ENZYME FIXED TO THE SURFACE OF A SOLID ELECTRONICALLY CONDUCTIVE ENZYME SUPPORT B. CONTACTING A LIQUID REACTION MEDIUM SUBSTRAE, CONTAINING SUBSTANCES TO BE TRANSFORMED, WITH SAID ENZYMATIC ELECTRODE, AND C. REGULATING THE CATALYTIC ACTIVITY OF SAID ENZYME BY APPLYING A POTENTIAL TO SAID ELECTRODE WHILE ADJUSTING THE VALUE OF SAID POTENTIAL IN SUCH A MANNER AS TO TRANSFORM SAID SUBSTRATE SUBSTANCES AT A SUBSTANTIALLY UNIFORM RATE.

2. A method as defined in claim 1 for the simultaneous control of several enzymatic reactions, wherein the reaction substrate medium, containing substances to be each transformed by at least one of said reactions, is contacted with the enzymes fixed on at least one enzymatic electrode, and wherein the potential of said electrode is continuously adjusted in a manner that each of said substances on contact with said enzymatic electrodes is transformed at a constant rate.

3. A method as defined in claim 1 wherein said enzymatic electrode is alternately brought to a first potential and to a second potential which respectively provide electro-chemical reactions;

said first potential producing a substance participating in the desired enzymatic reaction and said second potential promoting the catalysis of this enzymatic reaction, and wherein the value of said second potential is adjusted so that the enzymatic reaction is caused to proceed at a constant rate.

4. A method as defined in claim 3, for controlling an enzymatic oxidation reaction, wherein an aqueous reaction medium, containing the substance to be oxidized, is contacted with said enzymatic electrode, which comprises an oxidase fixed on the support of said electrode, wherein said electrode is alternately brought to a first potential and to a second potential which first potential causes electrolysis of the water, to thereby generate oxygen on said electrode, and the enzymatic oxidation of said substance through the oxygen thus generated, and wherein said second potential is controlled in such manner that the enzymatic oxidation is caused to proceed at a constant rate.

5. An apparatus for governing the reaction rate of an enzymatic reaction comprising:

a. at least one enzymatic electrode having at least one enzyme fixed to the surface of a solid, electronically conductive support connected to means to apply an adjustable electrical potential to said electrode;

b. a reaction chamber which contains said enzymatic electrode associated with a counter-electrode and is adapted to receive said liquid reaction medium for contact with said electrodes and to discharge the products of the enzymatic reaction; and

c. adjustable electrical supply and voltage control means connected to said enzymatic electrode and to said counter-electrode and adapted to apply, between said electrodes, a variable potential difference which is adjustable so as to regulate the activity of said enzyme on said medium.

6. An apparatus as defined in claim 5, further comprising a reference electrode which is arranged in the vicinity of said enzymatic electrode and is associated with said control means so as to provide a fixed potential value as a reference for controlling the potential of said enzymatic electrode.