Application of liquid-based colorimetric method for high throughput screening of bioplastic-degrading strains using esterase assay

Anal Biochem. 2024 Jan 15:685:115390. doi: 10.1016/j.ab.2023.115390. Epub 2023 Nov 10.

Abstract

To alleviate environmental problems caused by using conventional plastics, bioplastics have garnered significant interest as alternatives to petroleum-based plastics. Despite possessing better degradability traits compared to traditional plastics, the degradation of bioplastics still demands a longer duration than initially anticipated. This necessitates the utilization of degradation strains or enzymes to enhance degradation efficiency, ensuring timely degradation. In this study, a novel screening method to identify bioplastic degraders faster was suggested to circumvent the time-consuming and laborious characteristics of solid-based plate assays. This liquid-based colorimetric method confirmed the extracellular esterase activity with p-nitrophenyl esters. It eliminated the needs to prepare plastic emulsion plates at the initial screening system, shortening the time for the overall screening process and providing more quantitative data. p-nitrophenyl hexanoate (C6) was considered the best substrate among the various p-nitrophenyl esters as substrates. The screening was performed in liquid-based 96-well plates, resulting in the discovery of a novel strain, Bacillus sp. SH09, with a similarity of 97.4% with Bacillus licheniformis. Furthermore, clear zone assays, degradation investigations, scanning electron microscopy, and gel permeation chromatography were conducted to characterize the biodegradation capabilities of the new strain, the liquid-based approach offered a swift and less labor-intensive option during the initial stages.

Keywords: Biodegradation; Bioplastic; Screening; esterase; p-nitrophenyl ester.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biopolymers
  • Colorimetry
  • Esterases* / chemistry
  • High-Throughput Screening Assays
  • Plastics* / chemistry

Substances

  • Plastics
  • Esterases
  • Biopolymers