Method for determining the concentration of an L-amino acid in fermentation

• US 4,299,669 A
• Filed: 03/27/1979
• Issued: 11/10/1981
• Est. Priority Date: 03/28/1978
• Status: Expired due to Term

• Current Assignee: Aerpat AG
• Original Assignee: Ajinomoto Co., Inc.
• Inventors: Suzuki, Shuichi, Karube, Isao, Yasuda, Takeo, Obana, Haruo, Hikuma, Motohiko, Shirakawa, Tadashi
• Primary Examiner(s): Kaplan, G. L.
• Application Number:
  US06/024,330
• Time in Patent Office: 959 Days
• Field of Search: 204/1 T, 204/195 B, 204/195 P, 204/1 N, 204/1 P, 204/1 E, 435/817, 435/37, 435/25, 435/26, 435/28, 435/182, 435/288, 128/635
• US Class Current: 205/778
• CPC Class Codes: C12Q 1/001 Enzyme electrodes
• CPC Class Codes: C12Q 1/02 involving viable microorganisms
• CPC Class Codes: Y10S 435/817 Enzyme or microbe electrode

Abstract

Electrochemical method for determining the concentration of a carbon source and L-amino acid in a fermentation medium or cultured broth by contacting a microbial electrode consisting of an oxygen-sensitive electrode or a carbon dioxide gas-sensitive electrode combined with fixed microbial cells of a micro-organism capable of assimilating said carbon source or decarboxylating said L-amino acid with a sample solution and sensing electrically the rate of current decrease which is caused by the consumed oxygen in the solution in proportion to the concentration of carbon source; or the electric motive force which is caused by the liberation of carbon dioxide in the sample solution in proportion to the concentration of L-amino acid. Microbial electrodes and systems useful in carrying out the aforementioned method.

Claims

1. A method for determining the concentration of a L-amino acid in a culture medium or cultured broth solution which comprises:

contacting said culture anaerobically with a microbial electrode comprising a fixed or immobilized microorganism capable of anaerobically decarboxylating said L-amino acid and thereby liberating carbon dioxide into said culture, and a carbon dioxide gas sensitive electrode;

electrochemically measuring the liberation of carbon dioxide into said culture with said electrode and thereby correspondingly determining the concentration of said L-amino acid in said culture.

2. The method of claim 1 wherein said fixed or immobilized microorganisms are microbial cells.

3. The method of claim 1 wherein said microorganisms are immobilized by dispersion in a continuous solid matrix.

4. The method of claim 1 wherein said microorganism is selected from the group consisting of Escherichia Coli ATCC 8739, Citrobacter Freundii ATCC 10787, Clostridium welchii ATCC 13124, Rhodotorula glutinis IFO 0413, Escherichia coli ATCC 23226, Bacterium cadaveris ATCC 9760, Streptococcus faecalis ATCC 12984, Pseudononus saccharophia ATCC 16946, Bacillus subtilis ATCC 15037, and Lactobacillus casei ATCC 7469.

5. The method of claim 1 wherein said amino acid is L-glutamic acid or L-glutamine.