Development of a new chromogenic method for putrescine quantification using coupling reactions involving putrescine oxidase

Anal Biochem. 2020 Mar 15:593:113607. doi: 10.1016/j.ab.2020.113607. Epub 2020 Feb 5.

Abstract

Quantification of polyamines, including putrescine, is generally performed using high-performance liquid chromatography (HPLC) or gas chromatography. However, these methods are time-consuming because of sample derivatization and analytical reagent preparation. In this study, we developed a simple and high-throughput putrescine quantification method on a 96-well microtiter plate using putrescine oxidase from Rhodococcus erythropolis NCIMB 11540, peroxidase, 4-aminoantipyrine, and N-ethyl-N-(3-sulfopropyl)-3-methylaniline sodium salt. The developed method (named as PuO-POD-4AA-TOPS method) was applicable to bacterial culture supernatants. Furthermore, putrescine concentrations determined by the developed method roughly corresponded to the concentrations determined by HPLC.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ampyrone / chemistry
  • Chromogenic Compounds / chemistry
  • Colorimetry / methods
  • Oxidoreductases Acting on CH-NH Group Donors / chemistry
  • Proteus mirabilis / metabolism*
  • Putrescine / analysis*
  • Rhodococcus / enzymology

Substances

  • Chromogenic Compounds
  • Ampyrone
  • putrescine oxidase
  • Oxidoreductases Acting on CH-NH Group Donors
  • Putrescine

Supplementary concepts

  • Rhodococcus erythropolis