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

Brazilian Journal of Microbiology

Helen Cristina Fávero Lisboa²  Carolina Rabal Biasetto²  João Batista De Medeiros²  Ângela Regina Araújo²  Dulce Helena Siqueira Silva²  Helder Lopes Teles¹  Henrique Celso Trevisan² 

[1] ,Universidade Estadual Paulista Julio de Mesquita Filho Instituto de Química Araraquara SP ,Brazil

关键词: esterase;    pH indicator;    enzyme activity;    endophytic fungi;   

DOI  :  10.1590/S1517-83822013005000067

来源: SciELO

【 摘 要 】

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 -carboxylesterases) 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 asapotential 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.

【 授权许可】

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