title: 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.
authors: Instituto de Química, Universidade Estadual Paulista "Julio de Mesquita Filho", Araraquara, SP, Brazil. ; Instituto de Ciências Exatas e Naturais, Universidade Federal de Mato Grosso, Rondonópolis, MT, Brazil.
authors: Helen Cristina Fávero
authors: HC
authors: Lisboa
authors: Instituto de Química, Universidade Estadual Paulista "Julio de Mesquita Filho", Araraquara, SP, Brazil.
authors: Carolina Rabal
authors: CR
authors: Biasetto
authors: Instituto de Química, Universidade Estadual Paulista "Julio de Mesquita Filho", Araraquara, SP, Brazil.
authors: João Batista
authors: JB
authors: de Medeiros
authors: Instituto de Química, Universidade Estadual Paulista "Julio de Mesquita Filho", Araraquara, SP, Brazil.
authors: Angela Regina
authors: AR
authors: Âraújo
authors: Instituto de Química, Universidade Estadual Paulista "Julio de Mesquita Filho", Araraquara, SP, Brazil.
authors: Dulce Helena Siqueira
authors: DH
authors: Silva
authors: Instituto de Ciências Exatas e Naturais, Universidade Federal de Mato Grosso, Rondonópolis, MT, Brazil.
authors: Helder Lopes
authors: HL
authors: Teles
authors: Instituto de Química, Universidade Estadual Paulista "Julio de Mesquita Filho", Araraquara, SP, Brazil.
authors: Henrique Celso
authors: HC
authors: Trevisan
mesh_terms: D000085:Acetates / Q000378:metabolism; D001938:Brazil; D002087:Butyrates / Q000378:metabolism; D003124:Colorimetry / Q000379:methods*; D060026:Endophytes / Q000201:enzymology*; D004950:Esterases / Q000032:analysis*; D005658:Fungi / Q000201:enzymology* / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D010944:Plants / Q000382:microbiology; D011422:Propionates / Q000378:metabolism
chemical_list: D000085:Acetates; D002087:Butyrates; D007202:Indicators and Reagents; D011422:Propionates; C007650:ethyl acetate; D004950:Esterases; C045572:ethyl butyrate
keywords: endophytic fungi; enzyme activity; esterase; pH indicator