Pilot scale production of high-content mycoprotein using Rhizopus microsporus var. oligosporus by submerged fermentation and agro-industrial by-products.

While mycoprotein has gained traction as a human food source, its potential as a nutrient for animals remains largely unexplored. The mycoprotein-producing Rhizopus microsporus var. oligosporus, a fungus traditionally used for human food in Indonesia, is promising. It could revolutionise animal nutrition once it is Generally Recognized as Safe (GRAS) and is a biosafety level 1 (BSL1) organism. To enhance sustainably, we propose using sugar cane molasses (SM) and corn steep liquor (CSL) as nutrient sources. Also, we investigated the growth of R. microsporus var. oligosporus in five 14 L external-loop airlift bioreactors using CSL as the sole nutrient source. After 96 h of fermentation, at 25 °C and 0.5 vvm, the mycelium produced had an average biomass yield of 38.34 g L-1, with 70.18 % (m v-1) crude protein (mycoprotein). This bioprocess, which is scalable and economically viable, produces high amounts of mycoprotein for animal feed using CSL, a cost-effective agro-industrial by-product, providing a practical solution to the growing demand for animal protein.

Light conditions affect the growth, chemical composition, antioxidant and antimicrobial activities of the white-rot fungus Lentinus crinitus mycelial biomass.

The mycelial biomass of basidiomycetes is a promising source of compounds and represents an alternative for industrial and biotechnological applications. Fungi use light as information and hold photoresponse mechanisms, in which sensors respond to light wavelengths and regulate various biological processes. Therefore, this study aimed to investigate the effects of blue, green, and red lights on the growth, chemical composition, and antioxidant and antimicrobial activity of Lentinus crinitus mycelial biomass. The chemical composition of the mycelial biomass was determined by chromatographic methods, antioxidant activity was analyzed by in vitro assays, and antimicrobial activity was investigated by the microdilution assay. The highest mycelial biomass yield was observed under blue-light cultivation. Many primordia arose under blue or green light, whereas the stroma was formed under red light. The presence of light altered the primary fungal metabolism, increasing the carbohydrate, tocopherol, fatty acid, and soluble sugar contents, mostly mannitol, and reducing the protein and organic acid concentrations. Cultivation under red light increased the phenol concentration. In contrast, cultivation under blue and green lights decreased phenol concentration. Benzoic and gallic acids were the main phenolic acids in the hydroalcoholic extracts, and the latter acids increased in all cultures under light, especially red light. Mycelial biomass cultivated under red light showed the highest antioxidant activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The ferric reducing antioxidant power (FRAP) method showed that all light wavelengths increased the antioxidant activity of mycelial biomass, with the highest value under red light. Moreover, the ß-carotene/linoleic acid co-oxidation (BCLA) assay demonstrated that the antioxidant activity was affected by light cultivation. Mycelial biomass grown under all conditions exhibited antibacterial and antifungal activities. Thus, mycelial biomass cultivation of L. crinitus under light conditions may be a promising strategy for controlling the mycelial chemical composition and biomass yield.

Mycelial biomass cultivation of Lentinus crinitus / Crescimento micelial de Lentinus crinitus

Lentinus crinitus is a medicinal basidiomycete, little studied regarding the basic cultivation conditions, which is used in bioremediation and consumed by native Indians from the Brazilian Amazon. Also, it produces a fungal secondary metabolite panepoxydone that has been described as an essential regulator of the inflammatory and immune response. This study aimed to evaluate basic conditions of temperature, pH, and nitrogen concentration and source in the cultivation of L. crinitus mycelial biomass. In order to evaluate fungal growth temperature, 2% malt extract agar (MEA) medium, pH 5.5, was utilized from 19 to 40 °C. For pH, MEA had pH adjusted from 2 to 11 and cultivated at 28 °C. Urea or soybean meal was added to MEA to obtain final concentration from 0.5 and 16 g/L of nitrogen, pH of 5.5, cultivated at 28 °C. The best temperature growth varies from 31 to 34 ºC and the optimal one is 32.7º C, and the best pH ranges from 4.5 to 6.5 and the optimal one is 6.1. Protein or non-protein nitrogen concentration is inversely proportional to the mycelial biomass growth. Nitrogen concentrations of 2.0 g/L soybean meal and urea inhibit mycelial biomass growth in 11% and 12%, respectively, but high concentrations of 16.0 g/L nitrogen inhibit the growth in 46% and 95%, respectively. The fungus is robust and grows under extreme conditions of temperature and pH, but smaller adaptation with increasing nitrogen concentrations in the cultivation medium, mainly non-protein nitrogen.

Lentinus crinitus é um basidiomiceto medicinal consumido por índios nativos da Amazônia brasileira. Este fungo tem sido estudado quanto ao potencial de biorremediação de metais, mas ainda carece de estudos sobre às condições básicas de crescimento. L. crinitus produz panepoxidona - um metabólito secundário fúngico - descrito como regulador da resposta inflamatória e imune em células animais. Este trabalho teve como objetivo avaliar as condições básicas de temperatura, pH e concentração e fonte de nitrogênio para o crescimento micelial de L. crinitus. O fungo foi crescido em meio agar extrato de malte a 2% (MEA), pH 5,5 e mantido entre 19 e 40 °C. Para a avaliação de pH o MEA teve o pH ajustado de 2 a 11 e o crescimento foi realizado a 28 °C. As fontes de nitrogênio estudadas foram a uréia e o farelo de soja adicionado ao MEA para obter entre 0,5 a 16 g/L de nitrogênio, pH de 5,5, cultivado a 28 ° C. A melhor faixa temperatura para o crescimento micelial foi de 31 a 34 ºC com ótimo a 32,7 º C; a melhor faixa de pH de 4,5 a 6,5 e com ótimo de 6,1. A concentração de nitrogênio proteico ou não proteico é inversamente proporcional ao crescimento do fungo. Concentrações de nitrogênio de 2,0 g/L reduzem o crescimento da biomassa micelial em 11% e 12%, respectivamente e meios com nitrogênio de 16,0 g/L reduzem o crescimento em 46% e 95%, respectivamente. O fungo é robusto e cresce sob condições extremas de temperatura e pH, mas menor adaptação em meios com alta concentração de nitrogênio, principalmente não proteico.

Pulp wash: a new source for production of ligninolytic enzymes and biomass and its toxicological evaluation after biological treatment.

Pulp wash was used as substrate for the activity of ligninolytic enzymes of the fungus Pleurotus sajor-caju. Activity of laccase (Lac) and manganese peroxidase (MnP) as well as fungal biomass occurred under four conditions: different pulp wash concentrations, pH variation at the optimal pulp wash concentration, different glucose concentrations, and different concentrations of ammonium nitrate. The best enzyme activity and biomass production were obtained with in natura pulp wash and pH corrected to 5.0 (4884 IU/L Lac; 82 IU/L MnP; 25 g/100 mL biomass). However, the addition of glucose and ammonium nitrate to the pulp wash was not necessary for increasing the enzyme activity and biomass production. Efficient removal of pulp wash chemical oxygen demand (99.66%) and biochemical oxygen demand (83.27%) occurred after the mycoremediation with P. sajor-caju in the optimized conditions. Lactuca sativa L. seeds germination bioassay showed a four-fold reduction in the residue toxicity (EC50 28.72%) after the treatment with the fungus. Our findings are consistent with the notion that pulp wash is an excellent substrate for inducing the activity of ligninolytic enzymes and producing fungal biomass, and that the biological treatment is efficient to reduce effluent toxicity.

Production of mycelial biomass by the Amazonian edible mushroom Pleurotus albidus

ABSTRACT Edible mushroom species are considered as an adequate source of food in a healthy diet due to high content of protein, fiber, vitamins, and a variety of minerals. The representatives of Pleurotus genus are characterized by distinct gastronomic, nutritional, and medicinal properties among the edible mushrooms commercialized worldwide. In the present study, the growth of mycelial biomass of Pleurotus albidus cultivated in submerged fermentation was evaluated. Saccharose, fructose, and maltose were the three main carbon sources for mycelial biomass formation with corresponding yields of 7.28 g L−1, 7.07 g L−1, and 6.99 g L−1. Inorganic nitrogen sources did not stimulate growth and the optimal yield was significantly higher with yeast extract (7.98 g L−1). The factorial design used to evaluate the influence of saccharose and yeast extract concentration, agitation speed, and initial pH indicated that all variables significantly influenced the production of biomass, especially the concentration of saccharose. The greater amount of saccharose resulted in the production of significantly more biomass. The highest mycelial biomass production (9.81 g L−1) was reached in the medium formulated with 30.0 g L−1 saccharose, 2.5 g L−1 yeast extract, pH 7.0, and a speed of agitation at 180 rpm. Furthermore, P. albidus manifested different aspects of morphology and physiology under the growth conditions employed. Media composition affected mycelial biomass production indicating that the diversification of carbon sources promoted its improvement and can be used as food or supplement.

Production of mycelial biomass by the Amazonian edible mushroom Pleurotus albidus

Edible mushroom species are considered as an adequate source of food in a healthy diet due to high content of protein, fiber, vitamins, and a variety of minerals. The representatives of Pleurotus genus are characterized by distinct gastronomic, nutritional, and medicinal properties among the edible mushrooms commercialized worldwide. In the present study, the growth of mycelial biomass of Pleurotus albidus cultivated in submerged fermentation was evaluated. Saccharose, fructose, and maltose were the three main carbon sources for mycelial biomass formation with corresponding yields of 7.28 g L1, 7.07 g L1, and 6.99 g L1. Inorganic nitrogen sources did not stimulate growth and the optimal yield was significantly higher with yeast extract (7.98 g L1). The factorial design used to evaluate the influence of saccharose and yeast extract concentration, agitation speed, and initial pH indicated that all variables significantly influenced the production of biomass, especially the concentration of saccharose. The greater amount of saccharose resulted in the production of significantly more biomass. The highest mycelial biomass production (9.81 g L1) was reached in the medium formulated with 30.0 g L1 saccharose, 2.5 g L1 yeast extract, pH 7.0, and a speed of agitation at 180 rpm. Furthermore, P. albidus manifested different aspects of morphology and physiology under the growth conditions employed. Media composition affected mycelial biomass production indicating that the diversification of carbon sources promoted its improvement and can be used as food or supplement.(AU)

Production of mycelial biomass by the Amazonian edible mushroom Pleurotus albidus.

Edible mushroom species are considered as an adequate source of food in a healthy diet due to high content of protein, fiber, vitamins, and a variety of minerals. The representatives of Pleurotus genus are characterized by distinct gastronomic, nutritional, and medicinal properties among the edible mushrooms commercialized worldwide. In the present study, the growth of mycelial biomass of Pleurotus albidus cultivated in submerged fermentation was evaluated. Saccharose, fructose, and maltose were the three main carbon sources for mycelial biomass formation with corresponding yields of 7.28gL(-1), 7.07gL(-1), and 6.99gL(-1). Inorganic nitrogen sources did not stimulate growth and the optimal yield was significantly higher with yeast extract (7.98gL(-1)). The factorial design used to evaluate the influence of saccharose and yeast extract concentration, agitation speed, and initial pH indicated that all variables significantly influenced the production of biomass, especially the concentration of saccharose. The greater amount of saccharose resulted in the production of significantly more biomass. The highest mycelial biomass production (9.81gL(-1)) was reached in the medium formulated with 30.0gL(-1) saccharose, 2.5gL(-1) yeast extract, pH 7.0, and a speed of agitation at 180rpm. Furthermore, P. albidus manifested different aspects of morphology and physiology under the growth conditions employed. Media composition affected mycelial biomass production indicating that the diversification of carbon sources promoted its improvement and can be used as food or supplement.

Microscopia de epifluorescência para a determinção de biomassa micelial em substrato de cultivo sólido particulado / Microscopía e epifluorescencia para la deteminación de biomasa micelial en sustrato de cultivo sólido particulado / Epifluorscence mmicroscopy for the deterination of mycelial biomass in particulated solid substrate cultivatoon

A determinação de biomassa micelial fúngica crescida em substratos de cultivo sólido particulado (SCSP) é ainda um desafio devido à dificuldade de separação do micélio e o substrato. O objetivo deste trabalho foi avaliar a técnica de microscopia de epifluorescência para determinação da biomassa micelial de Pleurotus ostreatus em SCSP. Para determinação da exatidão da metodologia P. ostreatus foi crescido em meio líquido de extrato de malte e; a biomassa micelial foi separada por centrifugação, liofilizada e moída. Concentrações conhecidas do pó do micélio foram misturadas ao SCSP, composto de bagaço de cana de açúcar e fibra de soja, previamente autoclavado. Em seguida, a biomassa micelial foi determinada por microscopia de epifluorescência. Para promover a variação da biomassa micelial a ser determinada por microscopia de epifluorescência, SCSP adicionado de diferentes concentrações de ferro foram utilizados para o crescimento do fungo. Concluiu-se que a técnica apresenta baixa precisão e exatidão, o que implica na necessidade de maiores estudos para aplicação desta técnica para a determinação de biomassa micelial crescida em SCSP.

Determination of fungal mycelial biomass grown on solid substrate cultivation (SSC) is still a challenge due to the difficulty in separating mycelium and substrate. The objective of this study was to evaluate the epifluorescence microscopy to determine the mycelial biomass of Pleurotus ostreatus grown in SSC. P. ostreatus was grown in malt extract liquid medium and mycelial biomass was separated by centrifugation. It was then lyophilized and milled. Mycelial powder was added at known concentrations to SSC composed of sugarcane bagasse and soy fiber, previously autoclaved. Mycelial biomass was determined by epifluorescence microscopy. In order to promote mycelial biomass variation for the determination by epifluorescence microscopy, SSC added at different iron concentrations was used for fungus growth. It was concluded that the technique has low precision and accuracy, which implies the need for further studies in order to apply this technique for the determination of mycelial biomass grown in SSC.

Microscopia de epifluorescência para a determinação de biomassa micelial em substrato de cultivo sólido particulado / Epifluorescense microscopy for the determination of mycelial biomass in particulated solid substrate cultivation / Microscopia de pifluorescencia para la determinación de biomasa micelial en sustrato de cultivo solido particulado

A determinação de biomassa micelial fúngica crescida em substratos de cultivo sólido particulado (SCSP) é ainda um desafio devido à dificuldade de separação do micélio e o substrato. O objetivo deste trabalho foi avaliar a técnica de microscopia de epifluorescência para determinação da biomassa micelial de Pleurotus ostreatus em SCSP. Para determinação da exatidão da metodologia P. ostreatus foi crescido em meio líquido de extrato de malte e; a biomassa micelial foi separada por centrifugação, liofilizada e moída. Concentrações conhecidas do pó do micélio foram misturadas ao SCSP, composto de bagaço de cana de açúcar e fibra de soja, previamente autoclavado. Em seguida, a biomassa micelial foi determinada por microscopia de epifluorescência. Para promover a variação da biomassa micelial a ser determinada por microscopia de epifluorescência, SCSP adicionado de diferentes concentrações de ferro foram utilizados para o crescimento do fungo. Concluiu- -se que a técnica apresenta baixa precisão e exatidão, o que implica na necessidade de maiores estudos para aplicação desta técnica para a determinação de biomassa micelial crescida em SCSP.(AU)

Determination of fungal mycelial biomass grown on solid substrate cultivation (SSC) is still a challenge due to the difficulty in separating mycelium and substrate. The objective of this study was to evaluate the epifluorescence microscopy to determine the mycelial biomass of Pleurotus ostreatus grown in SSC. P. ostreatus was grown in malt extract liquid medium and mycelial biomass was separated by centrifugation. It was then lyophilized and milled. Mycelial powder was added at known concentrations to SSC composed of sugarcane bagasse and soy fiber, previously autoclaved. Mycelial biomass was determined by epifluorescence microscopy. In order to promote mycelial biomass variation for the determination by epifluorescence microscopy, SSC added at different iron concentrations was used for fungus growth. It was concluded that the technique has low precision and accuracy, which implies the need for further studies in order to apply this technique for the determination of mycelial biomass grown in SSC.

La determinación de biomasa micelial fúngica crecida en sustratos de cultivo sólido particulado (SCSP) es todavía un reto debido a la dificultad de separación del micelio y el sustrato. El objetivo de este estudio fue evaluar la técnica de microscopía de epifluorescencia para determinación de la biomasa micelial de Pleurotus ostreatus en SCSP. Para determinación de exactitud de la metodología P. ostreatus se cultivó en medio líquido de extracto de malta y; la biomasa micelial separada por centrifugación, liofilizada y molida. Concentraciones conocidas del polvo del micelio fueron mezcladas al SCSP, compuesto de bagazo de caña de azúcar y fibra de soya, previamente tratado en autoclave. A continuación, la biomasa micelial a ser determinada por microscopía de epifluorescencia. Para promover la variación de la biomasa micelial a ser determinada por microscopía de epifluorescencia, SCSP añadida con diferentes concentraciones de hierro fueron utilizados para el crecimiento del hongo. Se concluye que la técnica presenta baja precisión y exactitud, lo que implica en la necesidad de realizar más estudios para aplicación de esta técnica para la determinación de biomasa micelial crecida en SCSP.(AU)

Microscopia de epifluorescência para a determinação de biomassa micelial em substrato de cultivo sólido particulado / Epifluorescense microscopy for the determination of mycelial biomass in particulated solid substrate cultivation / Microscopia de pifluorescencia para la determinación de biomasa micelial en sustrato de cultivo solido particulado

A determinação de biomassa micelial fúngica crescida em substratos de cultivo sólido particulado (SCSP) é ainda um desafio devido à dificuldade de separação do micélio e o substrato. O objetivo deste trabalho foi avaliar a técnica de microscopia de epifluorescência para determinação da biomassa micelial de Pleurotus ostreatus em SCSP. Para determinação da exatidão da metodologia P. ostreatus foi crescido em meio líquido de extrato de malte e; a biomassa micelial foi separada por centrifugação, liofilizada e moída. Concentrações conhecidas do pó do micélio foram misturadas ao SCSP, composto de bagaço de cana de açúcar e fibra de soja, previamente autoclavado. Em seguida, a biomassa micelial foi determinada por microscopia de epifluorescência. Para promover a variação da biomassa micelial a ser determinada por microscopia de epifluorescência, SCSP adicionado de diferentes concentrações de ferro foram utilizados para o crescimento do fungo. Concluiu- -se que a técnica apresenta baixa precisão e exatidão, o que implica na necessidade de maiores estudos para aplicação desta técnica para a determinação de biomassa micelial crescida em SCSP.

Determination of fungal mycelial biomass grown on solid substrate cultivation (SSC) is still a challenge due to the difficulty in separating mycelium and substrate. The objective of this study was to evaluate the epifluorescence microscopy to determine the mycelial biomass of Pleurotus ostreatus grown in SSC. P. ostreatus was grown in malt extract liquid medium and mycelial biomass was separated by centrifugation. It was then lyophilized and milled. Mycelial powder was added at known concentrations to SSC composed of sugarcane bagasse and soy fiber, previously autoclaved. Mycelial biomass was determined by epifluorescence microscopy. In order to promote mycelial biomass variation for the determination by epifluorescence microscopy, SSC added at different iron concentrations was used for fungus growth. It was concluded that the technique has low precision and accuracy, which implies the need for further studies in order to apply this technique for the determination of mycelial biomass grown in SSC.

La determinación de biomasa micelial fúngica crecida en sustratos de cultivo sólido particulado (SCSP) es todavía un reto debido a la dificultad de separación del micelio y el sustrato. El objetivo de este estudio fue evaluar la técnica de microscopía de epifluorescencia para determinación de la biomasa micelial de Pleurotus ostreatus en SCSP. Para determinación de exactitud de la metodología P. ostreatus se cultivó en medio líquido de extracto de malta y; la biomasa micelial separada por centrifugación, liofilizada y molida. Concentraciones conocidas del polvo del micelio fueron mezcladas al SCSP, compuesto de bagazo de caña de azúcar y fibra de soya, previamente tratado en autoclave. A continuación, la biomasa micelial a ser determinada por microscopía de epifluorescencia. Para promover la variación de la biomasa micelial a ser determinada por microscopía de epifluorescencia, SCSP añadida con diferentes concentraciones de hierro fueron utilizados para el crecimiento del hongo. Se concluye que la técnica presenta baja precisión y exactitud, lo que implica en la necesidad de realizar más estudios para aplicación de esta técnica para la determinación de biomasa micelial crecida en SCSP.