A Convenient Fluorescence-Based Assay for the Detection of Sucrose Transport and the Introduction of a Sucrose Transporter from Potato into Clostridium Strains

Molecules. 2019 Sep 26;24(19):3495. doi: 10.3390/molecules24193495.

Abstract

A convenient and effective sucrose transport assay for Clostridium strains is needed. Traditional methods, such as 14C-sucrose isotope labelling, use radioactive materials and are not convenient for many laboratories. Here, a sucrose transporter from potato was introduced into Clostridium, and a fluorescence assay based on esculin was used for the analysis of sucrose transport in Clostridium strains. This showed that the heterologously expressed potato sucrose transporter is functional in Clostridium. Recombinant engineering of high-level sucrose transport would aid sucrose fermentation in Clostridium strains. The assay described herein provides an important technological platform for studying sucrose transporter function following heterologous expression in Clostridium.

Keywords: Clostridium; esculin; fluorescence assay; sucrose transporter.

MeSH terms

  • Biological Assay*
  • Biological Transport
  • Clostridium / genetics*
  • Clostridium / metabolism*
  • Fluorescence
  • Gene Expression*
  • Membrane Transport Proteins / genetics*
  • Membrane Transport Proteins / metabolism
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Solanum tuberosum / metabolism*
  • Sucrose / metabolism*

Substances

  • Membrane Transport Proteins
  • Plant Proteins
  • sucrose transport protein, plant
  • Sucrose