Endophytic fungi producing of esterases: evaluation in vitro of the enzymatic activity using pH indicator

Abstract

A sensitive and efficient colorimetric method was optimized for detection of esterase enzymes produced by endophytic fungi for development of High-Throughput Screening (HTS). The fungi were isolated and obtained previously from plant species of Cerrado and Atlantic Forest located in areas of environmental preservation in the State of Sao Paulo / Brazil, as part of the project “Chemical and biological prospecting endophytic fungi associated to plant species of Cerrado and Atlantic Forest”. The compounds ethyl butyrate, ethyl acetate and methyl propionate were used as standards esters which were hydrolyzed by extracellular enzyme from endophytic fungi (EC. 3.1.1.1 - carboxyl-esterases) for production of carboxylic acids. Thus, the reduction of the pH increases the protonated indicator concentration (bromothymol blue), changing the color of the reaction medium (from blue to yellow), that can be observed and measured by spectrophotometry at 616 nm. The methodology with acid-base indicator was performed on 13 microorganisms, aiming Periconia atropurpurea as a potential source of esterase for biotransformation of short chain esters. The results also evidenced that this methodology showed to be efficient, fast, cheap, having low consumption of reagents and easy development, and can be applied to screen carboxylic-ester hydrolases in a large number of microorganisms.

Bioprospecting and Structure of Fungal Endophyte Communities Found in the Brazilian Biomes, Pantanal, and Cerrado

“…Diaporthe was the dominant genus in both plants (Figure 3 ; Table S1 ). This genus has also been described as dominant in S. adstringens by Carvalho et al ( 2012 ) and is widely distributed in different plants of several biomes, such as the Pantanal (Hokama et al, 2017 ; Pietro-Souza et al, 2017 ), Mangrove forest (de Souza Sebastianes et al, 2013 ), Atlantic Rainforest (Correia et al, 2017 ), Cerrado and Caatinga (Ferreira et al, 2012 ; Lisboa et al, 2013 ). The ability of Diaporthe species to colonize different plants may result from the produced metabolites that bypass the plants defenses, or even by acting on host development and defense against plant pathogens (Nicoletti and Fiorentino, 2015 ; Santos et al, 2016 ; Tahir et al, 2017 ).…”

Gut microbial degradation of organophosphate insecticides-induces glucose intolerance via gluconeogenesis

“…Eighty microliters of suspension were used for esterase assay with ethyl butyrate as substrate as per Lisboa et al [49]. The formation of carboxylic acid due to hydrolysis of substrates mediated by esterase causes a reduction in the pH, which changes the color of the medium from blue to yellow.…”

Chemical, diversity and biotechnological potential of endophytic fungi isolated from Brazilian Cerrado plants

“…Owing to this biotechnological potential, these microorganisms have been the target of various chemical and biological studies. Lisboa et al (2013) evaluated a new method for the detection of esterase producing microorganisms via the in vitro detection of enzymatic activity using a pH indicator (bromothymol blue). This method evaluated the production of esterases using endophytic fungi isolated from Cerrado and Atlantic Forest plants.…”