Revista Mexicana de Ingeniería Química ISSN: 1665-2738 [email protected] Universidad Autónoma Metropolitana Unidad Iztapalapa México Velasco-Bucheli, R.; Mesa, A.; Gil, J.; García, C.; Durango, D. TRANSFORMATION OF trans- ANETHOLE USING THE PLANT PATHOGENIC FUNGUS Colletotrichum acutatum AS BIOCATALYST Revista Mexicana de Ingeniería Química, vol. 14, núm. 3, 2015, pp. 653-666 Universidad Autónoma Metropolitana Unidad Iztapalapa Distrito Federal, México Available in: http://www.redalyc.org/articulo.oa?id=62043088008 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative Revista Mexicana de Ingeniería Química Vol. 14, No.CONTENIDO 3 (2015) 653-666 TRANSFORMATIONVolumen 8, OF númerotrans- 3, 2009ANETHOLE / Volume 8, number USING 3, 2009 THE PLANT PATHOGENIC FUNGUS Colletotrichum acutatum AS BIOCATALYST TRANSFORMACI213ON´ Derivation DE andtrans- applicationANETOL of the Stefan-Maxwell USANDO equations EL HONGO FITOPATOGENICO´ Colletotrichum acutatum COMO BIOCATALIZADOR (Desarrollo y aplicación de las ecuaciones de Stefan-Maxwell) R. Velasco-Bucheli1, A. Mesa2, J. Gil2;3, C. Garc´ıa2, D. Durango2* Stephen Whitaker 1Departamento de Bioqu´ımicay Biolog´ıaMolecular I, Facultad de Biolog´ıa,Universidad Complutense de Madrid, Madrid, Espa˜na 2Universidad Nacional deBiotecnología Colombia, / Biotechnology Sede Medell´ın.Facultad de Ciencias, Escuela de Qu´ımica,Grupo de Qu´ımicade los Productos Naturales y los Alimentos, Medell´ın,Colombia. 245 Modelado de la biodegradación en biorreactores de lodos de hidrocarburos totales del petróleo 3Facultad de Ciencias Agrarias, Departamento de Ingenier´ıaAgr´ıcolay Alimentos, Medell´ın,Colombia intemperizadosReceived en suelos April y 15,sedimentos 2015; Accepted September 29, 2015 (Biodegradation modeling of sludge bioreactors of total petroleum hydrocarbons weathering in soil Abstract and sediments) Microbial transformation ofS.A. propenylbenzenes Medina-Moreno, S. mayHuerta-Ochoa, offer a cleanerC.A. Lucho-Constantino, and cheaper L. alternative Aguilera-Vázquez, to natural A. Jiménez- production of flavors and fragrances. In the presentGonzález study, y theM. Gutiérrez-Rojas biotransformation of trans-anethole using cells of a Colombian strain of the fungus Colletotrichum acutatum259was Crecimiento, investigated. sobrevivencia Initially, y adaptación fungitoxicity de Bifidobacterium of this compoundinfantis a condiciones against ácidasC. acutatum was evaluated; trans-anethole displayed a relatively(Growth, survival weak and toxicity adaptation against of Bifidobacterium the microorganism infantis to acidic (< conditions)70%, at 200 µg/mL and after 48 hours) and apparently a detoxificationL. Mayorga-Reyes, mechanism wasP. Bustamante-Camilo, present. Then, theA. Gutiérrez-Nava, microorganism E. Barranco-Florido was incubated y with A. Azaola- the substrate at room conditions, using three differentEspinosa culture media (Czapek-Dox, Sabouraud and PDB). Results show that trans-anethole is mainly degraded through265 an Statistical epoxide-diol approach pathway to optimization (trans- ofanethole ethanol fermentation to anethole-epoxide, by Saccharomyces then tocerevisiaesyn- and in anti-the anethole-diol, p−anisaldehyde, p−anisic acid and p−anisic alcohol). However, other minor metabolites [e.g. 3-(4-methoxyphenyl)-1- presence of Valfor® zeolite NaA propanol, 1-(4-methoxyphenyl)-2-propanol, ethyl ester of anisic acid], possibly proceeding from other metabolic pathways were also found. Additionally, (Optimización it wasdemonstrated estadística de la fermentación that the concentration etanólica de Saccharomyces of metabolic cerevisiae products en presencia is dependent de on culture medium used, being anethole-diolzeolita Valfor® the major zeolite NaA) product obtained in all media used. Interestingly, some of the compounds generated in the biotransformationG. Inei-Shizukawa, have been H. A. utilizedVelasco-Bedrán, as flavors G. F. Gutiérrez-López and fragrances. and H. BasedHernández-Sánchez on the identified metabolites, a possible metabolic pathway of the biotransformation of trans-anethole by C. acutatum was proposed. Keywords: fungitoxicity,Ingeniería biotransformation, de procesos / ProcessC. acutatum engineering, metabolic pathway, culture media. Resumen 271 Localización de una planta industrial: Revisión crítica y adecuación de los criterios empleados en esta decisión La transformacion´ microbiana de propenilbencenos puede ofrecer una alternativa mas´ limpia y economica´ para la (Plant site selection: Critical review and adequation criteria used in this decision) produccion´ natural de aromas y fragancias. En el presente estudio se investigo´ la biotransformacion´ de trans-anetol J.R. Medina, R.L. Romero y G.A. Pérez usando celulas´ de una cepa colombiana del hongo Colletotrichum acutatum. Inicialmente, se evaluo´ la toxicidad de este compuesto contra C. acutatum; trans-anetol exhibio´ una toxicidad relativamente baja contra el microorganismo (<70%, a 200 µg/mL y 48 horas) y aparentemente se presento´ un mecanismo de desintoxicacion.´ Luego, el microorganismo se incubo´ 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 traves´ de una ruta epoxido-diol´ (trans-anetol a anetol- epoxido, luego a syn- y anti-anetol-diol, p−anisaldehido, acido´ p−an´ısico y alcohol p−an´ısico). Sin embargo, tambien´ se encontraron otros metabolitos minoritarios [por ejemplo, 3-(4-metoxifenil)-1-propanol, 1-(4-metoxifenil)-2-propanol, ester´ et´ılico del acido´ an´ısico], posiblemente procedentes de otras rutas metabolicas.´ Adicionalmente, se demostro´ que la concentracion´ de los productos metabolicos´ es dependiente del medio de cultivo usado, siendo anetol-diol el producto mayoritario obtenido en todos los medios. Interesantemente, algunos compuestos generados en la biotransformacion´ se han utilizado como aromas y fragancias. Basados en los metabolitos identificados, se propuso una posible ruta metabolica´ para la biotransformacion´ de trans-anetol por C. acutatum. Palabras clave: fungitoxicidad, biotransformacion,´ C. acutatum, ruta metabolica,´ medios de cultivo. * Corresponding author. E-mail: [email protected] Tel. +57-4-4309888 Ext. 46367 Publicado por la Academia Mexicana de Investigacion´ y Docencia en Ingenier´ıa Qu´ımica A.C. 653 Tablas y figuras FIGURES Velasco-Bucheli et al./ Revista MexicanaFigure deIngenier 1. Structure´ıaQu ´ımica Vol. of 14, No.trans 3 (2015)-anethole 653-666 (A) 1 Introduction Flavors and fragrances are widely used in food, beverage, cosmetic, home and personal care, and in chemical and pharmaceutical industries (Palmer´ın- Carreno˜ et al., 2014; Freire et al., 2005; Kashi et al., 2007). Nowadays, most of the flavor compounds Fig. 1. Structure of trans-anethole (A) on the world market are produced by chemical synthesis or by direct extraction from plants (Bicas et al., 2010). However, chemical synthesis results the toxicity of organic substrates and products, and the frequently in environmentally unfriendly production culture medium (Bertrand et al., 2013; Baeza-Jimenez´ processes and the undesirable formation of by- et al., 2014; Leon´ et al., 1998). products, thereby reducing process efficiency and In this sense, microbial transformation of propenylbenzenes has attracted considerable attention increasing downstream cost (Longo and Sanroman,´ 2006; Wohlgemuth, 2010). Besides, natural flavor becoming one of the most important topics (Hua production by extraction from plants has some et al., 2007; Overhage et al., 2006; Yamada et al., 2007a, 2007b; Zhang et al., 2006). Propenylbenzenes, problems. First, the concentration of the desired compound is commonly low, making the extraction, extracted from plant essential oils, are common isolation and formulation very expensive (Xu et al., aromatic compounds often employed as starting intermediates for the production of various flavors 2007). Second, the level of flavor compounds in plants is influenced by different factors including and fragrances. These compounds can be transformed weather, agronomic conditions, subspecies genotype, by whole microorganisms or some enzymes to give plant age and diseases (LongoFigure and Sanrom 2. Mycelialan,´ 2006; growthnatural flavors inhibition and fragrances of which C. are acutatum products by (A) Bicas et al., 2010). Additionally, some flavors of high values (Koeduka et al., 2006; Xu et al., 2007). For example, the phenylpropanoids ferulic and fragrances are only. 100 found in exotic species. Owing to the demand for flavors and fragances, and acid, eugenol, and isoeugenol have been used for the problems associated with organic synthesis and preparing natural vanillin through biotechnological100 µg/mL processes. physical extraction from plants, the industry is actively 125 µg/mL pursuing new compounds and80 technologies for the This study reports for the first time the ability of production of natural flavors and fragrances (Shimoni one strain of C. acutatum, which is a cosmopolitan150 µg/mL et al., 2000). filamentous phytopathogenic fungus that attack175 in µg/mL harvest and postharvest, to biotransform trans- In addition to the above, US (US Code, 1985) and anethole [trans-1-(4-methoxyphenyl)-1-propene, (A),200 µg/mL European (The Council of the European