Enzyme-integrated metal-organic framework platform for cascade detection of α-amylase

Int J Biol Macromol. 2024 May;268(Pt 1):131870. doi: 10.1016/j.ijbiomac.2024.131870. Epub 2024 Apr 24.

Abstract

As one of the most important industrial enzymes, α-amylase is widely used in food processing, such as starch sugar and fermentation, bringing high added value to industry of more than a trillion dollars. We developed a multi-enzyme system (Glu&Gox@Cu-MOF-74) prepared by embedding α-glucosidase (Glu) and glucose oxidase (Gox) into the biomimetic metal-organic framework Cu-MOF-74 using in situ encapsulation within 15 min at room temperature for efficient and sensitive detection of α-amylase activity. Benefitting from the remarkable peroxidase-mimicking property and rigid skeleton of Cu-MOF-74, the biocatalytic platform exhibited excellent cascade activity and tolerance in various extremely harsh environments compared to natural enzymes. On this basis, a cascade biocatalytic platform was constructed for the detection of α-amylase activity with wide linear range (5-100 U/L) and low limit of detection (1.45 U/L). The colorimetric cascade scheme is important for the sensitive and selective determination of α-amylase in complex fermentation samples, and the detection time is short (∼0.5 h). This work provides new ideas for the detection of α-amylase based on the cascade amplification method.

Keywords: Cascade catalysis; Enzyme immobilization; Metal-organic framework; α-Amylase detection.

MeSH terms

  • Biocatalysis
  • Biosensing Techniques / methods
  • Colorimetry / methods
  • Copper / analysis
  • Copper / chemistry
  • Enzymes, Immobilized / chemistry
  • Enzymes, Immobilized / metabolism
  • Glucose Oxidase* / chemistry
  • Glucose Oxidase* / metabolism
  • Limit of Detection
  • Metal-Organic Frameworks* / chemistry
  • alpha-Amylases* / analysis
  • alpha-Amylases* / chemistry
  • alpha-Amylases* / metabolism
  • alpha-Glucosidases / analysis
  • alpha-Glucosidases / metabolism