Biotecnología

 

Transformation of trans-Anethole using the plant pathogenic fungus Colletotrichum acutatum as biocatalyst

 

Transformación de trans-Anetol usando el hogo fitopatogénico Colletotrichum acutatum como biocatalizador

 

R. Velasco-Bucheli¹, A. Mesa², J. Gil^{2, 3}, C. García², D. Durango²*

 

¹ Departamento de Bioquímica y Biología Molecular I, Facultad de Biología, Universidad Complutense de Madrid, Madrid, España.

² Universidad Nacional de Colombia, Sede Medellín. Facultad de Ciencias, Escuela de Química, Grupo de Química de los Productos Naturales y los Alimentos, Medellín, Colombia. * Corresponding author. E-mail: dldurango@unal.edu.co Tel. +57-4-4309888 Ext. 46367

³ Universidad Nacional de Colombia, Facultad de Ciencias Agrarias, Departamento de Ingeniería Agrícola y Alimentos, Medellín, Colombia.

 

Received April 15, 2015;
Accepted September 29, 2015.

 

Abstract

Microbial transformation of propenylbenzenes may offer a cleaner and cheaper alternative to natural production of flavors and fragrances. In the present study, the biotransformation of trans-anethole using cells of a Colombian strain of the fungus Colletotrichum acutatum was investigated. Initially, fungitoxicity of this compound against C. acutatum was evaluated; trans-anethole displayed a relatively weak toxiciiy against the microorganism (<70%, at 200 μg/mL and after 48 hours) and apparently a detoxification mechanism was present. Then, the microorganism was incubated with the substrate atroom conditions, using three different culture media (Czapek-Dox, Sabouraud and PDB). Results show that trans-anethole is mainly degraded through an epoxide-diol pathway (trans-anethole to anethole-epoxide, then to syn- and anti-anethole-diol, p-anisaldehyde, p-anisic acid and p-anisic alcohol). However, other minor metabolites [e.g. 3-(4-methoxyphenyl)-1-propanol, 1-(4-methoxyphenyl)-2-propanol, ethyl ester of anisic acid], possibly proceeding from other metabolic pathways were also found. Additionally, it was demonsfrated that: the concenfration of metabolic products is dependent on culture medium used, being anethole-diol the mapr product obtained in all media used. Interestingly, some of the compounds generated in the biotransformation have been utilized as flavors and fragrances. Based on the identified metabolites, a possible metabolic pathway of the biotransformation of trans-anethole by C. acutatum was proposed.

Keywords: fungitoxicity, biotransformation, C. acutatum, metabolic pathway, culture media.

 

Resumen

La transformación microbiana de propenilbencenos puede ofrecer una alternativa más limpia y económica para la producción natural de aromas y fragancias. En el presente estudio se investigó la biotransformación de trans-anetol usando células de una cepa colombiana del hongo Colletotrichum acutatum. Inicialmente, se evaluó la toxicidad de este compuesto contra C. acutatum; trans-anetol exhibió una toxicidad relativamente baja contra el microorganismo (<70%, a 200 μ/mL y 48 horas) y aparentemente se presentó un mecanismo de desintoxicación. Luego, el microorganismo se incubó con el sustrato a condiciones ambientales, usando tres medios de cultivo (Czapek-Dox, Sabouraud y PDB). Los resultados muestran que trans-anetol es degradado principalmente a través de una ruta epóxido-diol (trans-anetol a anetol-epóxido, luego a syn- y anti-anetol-diol, p-anisaldehido, ácido p-anísico y alcohol p-anísico). Sin embargo, también se encontraron otros metabolitos minoritarios [por ejemplo, 3-(4-metoxifenil)-1-propanol, 1-(4-metoxifenil)-2-propanol, éster etílico del ácido anísico], posiblemente procedentes de otras rutas metabólicas. Adicionalmente, se demostró que la concentración de los productos metabólicos es dependiente del medio de cultivo usado, siendo anetol-diol el producto mayoritario obtenido en todos los medios. Interesantemente, algunos compuestos generados en la biotransformación se han utilizado como aromas y fragancias. Basados en los metabolitos identificados, se propuso una posible ruta metabólica para la biotransformación de trans-anetol por C. acutatum.

Palabras clave: fungitoxicidad, biotransformación, C. acutatum, ruta metabólica, medios de cultivo.

 

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