Optimización de la extracción de proteínas asistida por ultrasonido desde la cianobacteria marina Oscillatoria sp. y evaluación de su potencial bioestimulante en la germinación de Chenopodium quinoa.
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Date
2026
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Universidad de Concepción
Abstract
Las cianobacterias constituyen microorganismos fotosintéticos versátiles con un alto potencial biotecnológico, y la cepa marina Oscillatoria sp. CCM-UdeC-334 representa un aislado de interés debido a su capacidad de acumular compuestos proteicos bajo condiciones controladas. Sin embargo, la recuperación eficiente de proteínas intracelulares se ve limitada por estructuras celulares complejas que actúan como barrera en los procesos de ruptura celular. Este trabajo tuvo por objetivo optimizar la extracción de proteínas a partir de biomasa de Oscillatoria sp. mediante disrupción celular asistida por ultrasonido, utilizando metodología de superficie de respuesta (RSM) con diseño compuesto central (CCD), y evaluar el efecto de los extractos obtenidos sobre la germinación y crecimiento temprano de Chenopodium quinoa Willd.
La cepa Oscillatoria sp. (CCM-UdeC-334) se cultivó en medio Walne modificado con sobreoferta de nitrógeno y fósforo. En fase estacionaria se alcanzaron 5.428 ± 995.71μg L⁻¹ de clorofila a, una biomasa seca de 0,955 ± 0,098 g L⁻¹ y un contenido proteico de 30,6 ± 4,5% de la biomasa seca (~292 mg proteínas L⁻¹ de cultivo). Posteriormente, se implementó un diseño compuesto central para evaluar la amplitud (20–50%), tiempo de sonicación (5–15 min) y tipo de solvente (agua destilada y NaOH 0,1 M). La condición óptima correspondió al sistema acuoso (20% amplitud, 5 min), con una eficiencia de extracción de 97,6%, superando la mejor condición alcalina (NaOH 0,1 M; 83,7%).
La selección de los mejores rendimientos de extracción acuosa y alcalina se evaluó mediante un bioensayo de germinación de C.quinoa, alcanzando un alto 100% para el solvente acuoso a las condiciones de operación 20% de amplitud a 5 min. A las 72 h, el tratamiento T2 H₂O presentó la mayor longitud radicular (2,896 ± 0,651 cm), superando significativamente al control (2,18 ± 0,53 cm), lo que evidencia un efecto bioestimulante sobre el crecimiento radicular de las plántulas.
Los resultados indican que la disrupción asistida por ultrasonido en sistema acuoso es una estrategia eficiente para la recuperación de proteínas desde Oscillatoria sp., y permite obtener extractos crudos con actividad biológica detectables y potencial bioestimulante sobre el crecimiento radicular de C.quinoa con potencial de aplicación en la formulación de bioestimulantes para la industria agrícola.
Cyanobacteria are versatile photosynthetic microorganisms with considerable biotechnological potential. The marine strain Oscillatoria sp. CCM-UdeC-334 is of particular interest because of its ability to accumulate proteinaceous compounds under controlled conditions. However, the efficient recovery of intracellular proteins is constrained by complex cellular structures that act as barriers to cell disruption. This study aimed to optimize protein extraction from Oscillatoria sp. biomass through ultrasound-assisted cell disruption using response surface methodology (RSM) with a central composite design (CCD), and to evaluate the effects of the resulting extracts on germination and early growth of Chenopodium quinoa Willd. Oscillatoria sp. CCM-UdeC-334 was cultivated in modified Walne medium supplemented with excess nitrogen and phosphorus. During the stationary phase, cultures reached 5,428 ± 995.71 μg L⁻¹ chlorophyll a, a dry biomass concentration of 0.955 ± 0.098 g L⁻¹, and a protein content of 30.6 ± 4.5% of dry biomass, equivalent to approximately 292 mg protein L⁻¹ of culture. A central composite design was then used to evaluate ultrasound amplitude (20–50%), sonication time (5–15 min), and solvent type (distilled water and 0.1 M NaOH). The optimal condition was obtained in the aqueous system at 20% amplitude and 5 min of sonication, achieving a protein extraction efficiency of 97.6%. This result exceeded the best alkaline condition, which reached 83.7% extraction efficiency using 0.1 M NaOH. The highest-yielding aqueous and alkaline extracts were evaluated using a C. quinoa germination bioassay. The aqueous extract obtained at 20% amplitude and 5 min achieved 100% germination. After 72 h, the T2 H₂O treatment produced the greatest root length (2.896 ± 0.651 cm), significantly exceeding the control treatment (2.18 ± 0.53 cm), indicating a biostimulant effect on seedling root growth. These findings indicate that ultrasound-assisted cell disruption in an aqueous system is an efficient strategy for protein recovery from Oscillatoria sp. biomass. In addition, the process yields crude extracts with detectable biological activity and potential use as biostimulant ingredients for agricultural applications.
Cyanobacteria are versatile photosynthetic microorganisms with considerable biotechnological potential. The marine strain Oscillatoria sp. CCM-UdeC-334 is of particular interest because of its ability to accumulate proteinaceous compounds under controlled conditions. However, the efficient recovery of intracellular proteins is constrained by complex cellular structures that act as barriers to cell disruption. This study aimed to optimize protein extraction from Oscillatoria sp. biomass through ultrasound-assisted cell disruption using response surface methodology (RSM) with a central composite design (CCD), and to evaluate the effects of the resulting extracts on germination and early growth of Chenopodium quinoa Willd. Oscillatoria sp. CCM-UdeC-334 was cultivated in modified Walne medium supplemented with excess nitrogen and phosphorus. During the stationary phase, cultures reached 5,428 ± 995.71 μg L⁻¹ chlorophyll a, a dry biomass concentration of 0.955 ± 0.098 g L⁻¹, and a protein content of 30.6 ± 4.5% of dry biomass, equivalent to approximately 292 mg protein L⁻¹ of culture. A central composite design was then used to evaluate ultrasound amplitude (20–50%), sonication time (5–15 min), and solvent type (distilled water and 0.1 M NaOH). The optimal condition was obtained in the aqueous system at 20% amplitude and 5 min of sonication, achieving a protein extraction efficiency of 97.6%. This result exceeded the best alkaline condition, which reached 83.7% extraction efficiency using 0.1 M NaOH. The highest-yielding aqueous and alkaline extracts were evaluated using a C. quinoa germination bioassay. The aqueous extract obtained at 20% amplitude and 5 min achieved 100% germination. After 72 h, the T2 H₂O treatment produced the greatest root length (2.896 ± 0.651 cm), significantly exceeding the control treatment (2.18 ± 0.53 cm), indicating a biostimulant effect on seedling root growth. These findings indicate that ultrasound-assisted cell disruption in an aqueous system is an efficient strategy for protein recovery from Oscillatoria sp. biomass. In addition, the process yields crude extracts with detectable biological activity and potential use as biostimulant ingredients for agricultural applications.
Description
Tesis presentada para optar al título de Ingeniero/a en Biotecnología Marina y Acuicultura.
Keywords
Cianobacterias, Biotecnología, Quinoa