Bioelectrocatalytic sensor for triglycerides in human skin sebum based on enzymatic cascade reaction of lipase, glycerol kinase and glycerophosphate oxidase

J Biotechnol. 2014 Apr 10:175:7-14. doi: 10.1016/j.jbiotec.2014.01.036. Epub 2014 Feb 12.

Abstract

We report the development of an electrochemical biosensor for the quantification of triglycerides in human skin sebum, based on a multienzyme cascade reaction. The presence of excessive triglycerides in human sebum is one of the leading causes of various skin ailments. However, to the best of our knowledge, no bioelectrocatalytic approach for the quantification of sebum triglycerides has been made. In order to develop triglyceride biosensor, we fabricated a multienzyme-associated electrode incorporating lipase, glycerol kinase, and glycerophosphate oxidase. Enzymes were deposited by electrostatic force and further stabilized via crosslinking between enzymes and polymer matrices. The enzyme-modified biosensing electrode maintained its bioelectrocatalytic activity for five days. An additional constraint was the limited solubility of sebum triglycerides in aqueous electrolytes, impeding the analysis. To address this issue, triglyceride samples were prepared in the form of micelles, enabling efficient sample preparation for biosensor signaling. Calibration tests revealed that the designed assay had a detection range of 15-200mg/dL of micellar triglyceride, which covered the required determination range. The developed biosensing approach was successfully used to determine triglyceride concentrations in real sebum samples of unknown triglyceride content.

Keywords: Bioelectrocatalytic; Enzyme cascade; Skin sebum triglyceride; Triglyceride biosensor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biocatalysis
  • Biosensing Techniques / methods*
  • Calibration
  • Enzymes / metabolism
  • Glycerol Kinase / metabolism
  • Glycerolphosphate Dehydrogenase / metabolism
  • Humans
  • Lipase / metabolism
  • Micelles
  • Sebum / chemistry*
  • Sebum / metabolism
  • Triglycerides / analysis*

Substances

  • Enzymes
  • Micelles
  • Triglycerides
  • Glycerolphosphate Dehydrogenase
  • Glycerol Kinase
  • Lipase