Toxicity detection using lysosomal enzymes, glycoamylase and thioredoxin fused with fluorescent protein in Saccharomyces cerevisiae

J Biotechnol. 2015 Nov 20:214:141-6. doi: 10.1016/j.jbiotec.2015.09.028. Epub 2015 Sep 26.

Abstract

Saccharomyces cerevisiae is the simplest and a favorite eukaryotic system that contains lysosome and thus, is a suitable organism for monitoring some toxic effects in environmental pollution. In this study, S. cerevisiae was transformed with two recombinant plasmids. Sporulation-specific glycoamylase (SGA1), which was upregulated in response to arsenic, was fused with the blue fluorescent protein (BFP) for the construction of an oxidative stress-causing chemicals sensor. Additionally, thioredoxin (TRX2), a protein overexpressed exclusively under tetracycline's influence, fused with the cyan fluorescent protein (CFP) to create a detector for this kind of chemical. In summary, we developed two recombinant S. cerevisiae that facilitate the detection of both kinds of toxic chemicals, specifically visualized by different color indicators.

Keywords: Fluorescent proteins; Glycoamylase; Heavy metals; Lysosomal proteins; Pharmaceuticals; Thioredoxin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aspirin / toxicity
  • Biosensing Techniques / methods*
  • Daphnia / drug effects
  • Glucan 1,4-alpha-Glucosidase / chemistry
  • Glucan 1,4-alpha-Glucosidase / genetics
  • Glucan 1,4-alpha-Glucosidase / metabolism*
  • Green Fluorescent Proteins / chemistry
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Luminescent Proteins / chemistry
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism*
  • Lysosomes / enzymology
  • Metals, Heavy / toxicity
  • Pesticides / toxicity
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Saccharomyces cerevisiae / drug effects
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Tetracycline / toxicity
  • Thioredoxins / chemistry
  • Thioredoxins / genetics
  • Thioredoxins / metabolism*
  • Toxicity Tests / methods*

Substances

  • Cyan Fluorescent Protein
  • Luminescent Proteins
  • Metals, Heavy
  • Pesticides
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • blue fluorescent protein, Aequorea victoria
  • Green Fluorescent Proteins
  • Thioredoxins
  • Glucan 1,4-alpha-Glucosidase
  • SGA1 protein, S cerevisiae
  • Tetracycline
  • Aspirin