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Analytical and Bioanalytical Chemistry

, Volume 391, Issue 4, pp 1255–1261 | Cite as

Optical enzyme sensor for determining l-lysine content using l-lysine oxidase from the rockfish Sebastes schlegeli

  • Hideaki Endo
  • Yoshitaka Hayashi
  • Yoichiro Kitani
  • Huifeng Ren
  • Tetsuhito Hayashi
  • Yuji Nagashima
Original Paper

Abstract

l-Lysine (l-Lys) in living bodies is critical for metabolism; therefore, determination of its levels in food is important. Most enzymatic methods for l-Lys analysis are performed using l-lysine oxidase (LyOx), but commercially manufactured LyOx is generally not highly selective for l-Lys among amino acids. We previously isolated LyOx as an antibacterial protein secreted from the skin of the rockfish Sebastes schlegeli. In the present study, we developed an optical enzyme sensor system for rapid and continuous determination of l-Lys using this LyOx. The system comprised an immobilized LyOx membrane, an optical oxygen probe, a flow system, and a personal computer. The amount of l-Lys was detected as a decrease in the oxygen concentration due to the LyOx reaction. The specificity of the sensor was examined against various amino acids. The sensor response was specific for l-Lys. Good reproducibility was obtained in 58 assays. The response of the sensor using commercially prepared LyOx was unstable compared with the response using LyOx isolated in our laboratory. Our sensor system could be used for 5 weeks without our having to change the enzyme membrane. The calibration curve for a standard l-Lys solution was linear from 0.1 to 3.0 mmol L−1. One assay could be completed within 2 min. The sensor was applied to determine the l-Lys content in food samples such as bonito cooking water and scallop hepatopancreas. The values obtained using the sensor and conventional high-performance liquid chromatography methods were well correlated.

Keywords

Biosensor Enzyme sensor Lysine Food Fish 

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Hideaki Endo
    • 1
  • Yoshitaka Hayashi
    • 1
  • Yoichiro Kitani
    • 2
  • Huifeng Ren
    • 1
  • Tetsuhito Hayashi
    • 1
  • Yuji Nagashima
    • 2
  1. 1.Department of Ocean SciencesTokyo University of Marine Science and TechnologyTokyoJapan
  2. 2.Department of Food Science and TechnologyTokyo University of Marine Science and TechnologyTokyoJapan

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