A Dual-Enzyme Amplification Loop for the Sensitive Biosensing of Endopeptidases

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

View graph of relations

Author(s)

  • Xiaomin Li
  • Chunxi Liu
  • Michael H. W. Lam
  • Yun Wah Lam

Related Research Unit(s)

Detail(s)

Original languageEnglish
Pages (from-to)25592-25600
Journal / PublicationACS Omega
Volume8
Issue number28
Online published7 Jul 2023
Publication statusPublished - 18 Jul 2023

Link(s)

Abstract

A rapid and sensitive approach for the detection of endopeptidases via a new analyte-triggered mutual emancipation of linker-immobilized enzymes (AMELIE) mechanism has been developed and demonstrated using a matrix metallopeptidase, a collagenase, as the model endopeptidase analyte. AMELIE involves an autocatalytic loop created by a pair of selected enzymes immobilized on solid substrates via linkers with specific sites that can be proteolyzed by one another. These bound enzymes are spatially separated so that they cannot act upon their corresponding substrates until the introduction of the target endopeptidase analyte that can also cleave one of the linkers. This triggers the self-sustained loop of enzymatic activities to emancipate all the immobilized enzymes. In this proof of concept, signal transduction was achieved by a colorimetric horseradish peroxidase–tetramethylbenzidine (HRP-TMB-H2O2) reaction with HRP that are also being immobilized by one of the linkers. The pair of immobilized enzymes were collagenase and alginate lyase, and they were immobilized by an alginate linker and a short peptide chain containing the amino acid sequence of Leu-Gly-Pro-Ala for collagenase. A detection limit of 2.5 pg collagenase mL–1 with a wide linear range up to 4 orders of magnitude was achieved. The AMELIE biosensor can detect extracellular collagenase in the supernatant of various bacteria cultures, with a sensitivity as low as 103 cfu mL–1 of E. coli. AMELIE can readily be adapted to provide the sensitive detection of other endopeptidases. © 2023 The Authors. Published by American Chemical Society

Research Area(s)

  • SIGNAL AMPLIFICATION, MATRIX METALLOPROTEINASES, ELECTROCHEMICAL PEPTIDE, PROTEASE, IMMUNOSENSOR, CLEAVAGE, BACTERIA, ASSAY

Citation Format(s)

A Dual-Enzyme Amplification Loop for the Sensitive Biosensing of Endopeptidases. / Zhou, Chuanwen; Li, Xiaomin; Tang, Sze Wing et al.
In: ACS Omega, Vol. 8, No. 28, 18.07.2023, p. 25592-25600.

Research output: Journal Publications and Reviews (RGC: 21, 22, 62)21_Publication in refereed journalpeer-review

Download Statistics

No data available