The effects of trace elements, cations, and environmental conditions on protocatechuate 3,4-dioxygenase activity

Andréa Scaramal Da Silva,Flávio Anastácio De Oliveira Camargo,Fátima Menezes Bento,Robson Andreazza,Rodrigo Josemar Seminoti Jacques

doi:10.1590/s0103-90162013000200002

Andréa Scaramal Da Silva, Flávio Anastácio De Oliveira Camargo + Show 3 more

Open Access

https://doi.org/10.1590/s0103-90162013000200002

Copy DOI

Abstract

Phenanthracene is a highly toxic organic compound capable of contaminating water and soils, and biodegradation is an important tool for remediating polluted environments. This study aimed to evaluate the effects of trace elements, cations, and environmental conditions on the activity of the protocatechol 3,4-dioxygenase (P3,4O) enzyme produced by the isolate Leifsonia sp. in cell-free and immobilized extracts. The isolate was grown in Luria Bertani broth medium (LB) amended with 250 mg L-1 of phenanthrene. Various levels of pH (4.0-9.0), temperature (5-80 °C), time (0-90 min), trace elements (Cu2+, Hg2+ and Fe3+), and cations (Mg2+, Mn2+, K+ and NH4+) were tested to determine which conditions optimized enzyme activity. In general, the immobilized extract exhibited higher enzyme activity than the cell-free extract in the presence of trace elements and cations. Adding iron yielded the highest relative activity for both cell-free and immobilized extracts, with values of 16 and 99 %, respectively. Copper also increased enzyme activity for both cell-free and immobilized extracts, with values of 8 and 44 %, respectively. Enzyme activity in the phosphate buffer was high across a wide range of pH, reaching 80 % in the pH range between 6.5 and 8.0. The optimum temperatures for enzyme activity differed for cell-free and immobilized extracts, with maximum enzyme activity observed at 35 ºC for the cell-free extract and at 55 ºC for the immobilized extract. The cell-free extract of the P3,4O enzyme exhibited high activity only during the first 3 min of incubation, when it showed 50 % relative activity, and dropped to 0 % after 60 min of incubation. By contrast, activity in the immobilized extract was maintained during 90 min of incubation. This isolate has important characteristics for phenanthrene biodegradation, producing high quantities of the P3,4O enzyme that forms part of the most important pathway for PAH biodegradation.

Highlights

Polycyclic aromatic hydrocarbons (PAH) are highly toxic compounds that pose risks to humans and animals
This study aimed to evaluate the effects of trace elements, cations, and environmental conditions on the activity of the protocatechol 3,4-dioxygenase (P3,4O) enzyme produced by the isolate Leifsonia sp. in cell-free and immobilized extracts
The optimum temperatures for enzyme activity differed for cell-free and immobilized extracts, with maximum enzyme activity observed at 35 oC for the cellfree extract and at 55 oC for the immobilized extract

Summary

Introduction

Polycyclic aromatic hydrocarbons (PAH) are highly toxic compounds that pose risks to humans and animals. Factors such as trace elements and environmental conditions interfere with microorganisms’ metabolisms for PAH biodegradation. Peripheral enzymes are responsible for recognizing and converting the PAHs in degradable molecules, and fission enzymes pass these molecules through the common pathways for energy generation and carbon routing in the microbial cells (Mishra et al, 2001). The meta pathway that originates the 2-hydroximuconic semialdehyde will be transformed in pyruvate and acetaldehyde In this pathway, catechol 2,3-dioxygenase belongs to the class of enzymes that cleave the aromatic ring to form the core intermediary catechol, and protocatechol 4,5dioxygenase to the class that cleave the aromatic ring to form the compound protocatechol (Caldwell, 2000). Enzyme activity and its applicability for biodegradation require more study

Objectives

Methods

Findings

Discussion

Conclusion