Document Detail

Development of a D-alanine sensor for the monitoring of a fermentation using the improved selectivity by the combination of D-amino acid oxidase and pyruvate oxidase.
MedLine Citation:
PMID:  14623466     Owner:  NLM     Status:  MEDLINE    
A D-alanine (D-Ala) sensor for the monitoring of a fermentation process was developed using flow injection analysis (FIA). The FIA system consisted of a D-amino acid oxidase (D-AAOx) reactor, a Pyruvate oxidase (PyOx) electrode and a contrast electrode in the flow cell, and through the oxidation of D-amino acids in the D-AAOx reactor, pyruvic acid was formed only from D-Ala. The pyruvic acid was further oxidized with PyOx via the D-AAOx reaction. The amount of oxygen consumed in the PyOx reaction was proportional to the amount of D-Ala. It was possible to continuously repeat the assay up to 60 times at pH 6.8 and a flow rate of 0.18-ml min(-1). A linear relationship was obtained in the range of 0.1-1 mM D-Ala with a correlation coefficient of 0.987 and the detection limit was 0.05 mM. The relative standard deviation (R.S.D.) was 4.9% (n=5) for 0.5 mM D-Ala. The D-Ala content in some fish sauces was also determined using the proposed sensor system. The results obtained indicated a linear relationship between the amounts of D-Ala determined by the proposed sensor system and the conventional method. From the results, even if the substrate specificity of the enzyme (D-AAOx) was low, it was evident that the concentration of the original material (D-Ala) could be determined specifically when the first reaction product was changed by the second reaction (PyOx).
Yohei Inaba; Kentaro Mizukami; Naoko Hamada-Sato; Takeshi Kobayashi; Chiaki Imada; Etsuo Watanabe
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article; Validation Studies    
Journal Detail:
Title:  Biosensors & bioelectronics     Volume:  19     ISSN:  0956-5663     ISO Abbreviation:  Biosens Bioelectron     Publication Date:  2003 Dec 
Date Detail:
Created Date:  2003-11-18     Completed Date:  2004-08-09     Revised Date:  2009-07-14    
Medline Journal Info:
Nlm Unique ID:  9001289     Medline TA:  Biosens Bioelectron     Country:  England    
Other Details:
Languages:  eng     Pagination:  423-31     Citation Subset:  IM    
Department of Food Science and Technology, Tokyo University of Fisheries, 5-7 Kounan 4, Minato-ku, Tokyo 108-8477, Japan.
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MeSH Terms
Alanine / analysis*,  chemistry*
Biosensing Techniques / instrumentation*,  methods
Cell Culture Techniques / instrumentation*,  methods
D-Amino-Acid Oxidase / chemistry*
Electrochemistry / instrumentation*,  methods
Equipment Design
Equipment Failure Analysis
Flow Injection Analysis / instrumentation*,  methods
Online Systems
Pyruvate Oxidase / chemistry*
Reproducibility of Results
Sensitivity and Specificity
Reg. No./Substance:
56-41-7/Alanine; EC Oxidase; EC Oxidase

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