Construction of Bioreactors for Obtaining Mycelium from Agaricus Brunnescens and Pleurotus Ostreatus Mushrooms Using the Honey Tek Technique

Mycelium cultivation plays an important role in mushroom production and in various biotechnological studies. Submerged fermentation has received much attention as a promising alternative for the efficient production of mushroom biomass and active metabolites. For the following research, bioreactors were constructed to obtain pure mycelium from Pleurotus ostreatus and Agaricus brunnescens mushrooms using the Honey technique. This method is based on using honey as the main nutrient for the growth of microorganisms on an industrial scale. In the beginning, spore impressions were extracted by the dry chamber method and inoculated into the honey broth. The bioreactors were dimensioned with a 2.5 L culture chamber, 2 L production, controlled temperature of 28∘C, pH = 4, and agitation of 120 revolutions per minute. Conidia/milliliter was quantified every three days with a Neubauer chamber, and the Kjeldahl method was used to identify protein as the main bioactive metabolite of the mycelium. Finally, the mycelium obtained was evaluated on three types of substrates comparing their colonization time. The use of bioreactors in mycelium cultivation is efficient as better mycelium quality, higher biomass yield, and more dispersion on substrates are obtained. It also has the benefits of much better oxygen mass transfer and culture homogeneity. With this technique, physical, chemical, and biological factors can be controlled to produce mushrooms in a much shorter time. Keywords: bioreactor, honey technic, mycelial biomass, Agaricus brunnescens, Pleurotus ostreatus. Resumen El cultivo de micelio juega un papel importante en la producción de hongos y en diversos estudios biotecnológicos, el usar la fermentación sumergida ha recibido mucha atención como alternativa prometedora para la producción eficiente de la biomasa de hongos y metabolitos activos. Para la siguiente investigación se construyeron biorreactores con el objetivo de obtener micelio puro de hongos Pleurotus ostreatus y Agaricus brunnescens usando la técnica Honey. El método Honey se basa en aprovechar la miel como nutriente principal para el crecimiento de microorganismos a escala industrial. En principio se extraen impresiones de esporas mediante el método de cámara seca y se inoculan en caldo honey. Los biorreactores se dimensionaron con una cámara de cultivo de 2,5 litros, producción de dos litros, temperatura controlada de 28 grados Celsius, pH igual a cuatro y agitación de 120 revoluciones por minuto. Se cuantificaron los conidios/mililitro cada tres días con una cámara de Neubauer y se usó el método Kjeldahl para identificar la proteína como principal metabolito bioactivo del micelio. Finalmente, el micelio obtenido fue evaluado sobre tres tipos de sustratos comparando su tiempo de colonización. El uso de biorreactores en el cultivo de micelio es eficiente debido a que se obtiene mejor calidad de micelio, mayor rendimiento de biomasa y más dispersión sobre sustratos. Así mismo, posee los beneficios de una transferencia de masa de oxígeno y una homogeneidad de cultivo mucho mejores. Con esta técnica se pueden controlar factores físicos, químicos y biológicos para producir setas en tiempos mucho más cortos. Palabras Clave: biorreactor, honey tek, biomasa micelial, Agaricus brunnescens, Pleurotus ostreatus

Construction of Bioreactors for Obtaining Mycelium from Agaricus Brunnescens and Pleurotus Ostreatus Mushrooms Using the Honey Tek Technique

Abstract Mycelium cultivation plays an important role in mushroom production and in various biotechnological studies. Submerged fermentation has received much attention as a promising alternative for the efficient production of mushroom biomass and active metabolites. For the following research, bioreactors were constructed to obtain pure mycelium from Pleurotus ostreatus and Agaricus brunnescens mushrooms using the Honey technique. This method is based on using honey as the main nutrient for the growth of microorganisms on an industrial scale. In the beginning, spore impressions were extracted by the dry chamber method and inoculated into the honey broth. The bioreactors were dimensioned with a 2.5 L culture chamber, 2 L production, controlled temperature of 28○C, pH = 4, and agitation of 120 revolutions per minute. Conidia/milliliter was quantified every three days with a Neubauer chamber, and the Kjeldahl method was used to identify protein as the main bioactive metabolite of the mycelium. Finally, the mycelium obtained was evaluated on three types of substrates comparing their colonization time. The use of bioreactors in mycelium cultivation is efficient as better mycelium quality, higher biomass yield, and more dispersion on substrates are obtained. It also has the benefits of much better oxygen mass transfer and culture homogeneity. With this technique, physical, chemical, and biological factors can be controlled to produce mushrooms in a much shorter time