Valorization of Bread Waste to a Fiber- and Protein-Rich Fungal Biomass

Filamentous fungi can be used for the valorization of food waste as a value-added product. The goal of this study was the valorization of bread waste through fungal cultivation and the production of value-added products. The fungal cultivation was verified for upscaling from shake flasks to a bench-scale bioreactor (4.5 L) and a pilot-scale bioreactor (26 L). The fungus showed the ability to grow without any additional enzymes or nutrients, and it was able to consume a bread concentration of 4.5% (w/v) over 48 h. The biomass concentration in the shake flasks was 4.1 g/L at a 2.5% bread concentration, which increased to 22.5 g/L at a 15% bread concentration. The biomass concentrations obtained after 48 h of cultivation using a 4.5% bread concentration were 7.2–8.3 and 8.0 g/L in 4.5 and 26 L bioreactors, respectively. Increasing the aeration rate in the 4.5 L bioreactor decreased the amount of ethanol produced and slightly reduced the protein content of the fungal biomass. The initial protein value in the bread was around 13%, while the protein content in the harvested fungal biomass ranged from 27% to 36%. The nutritional value of the biomass produced was evaluated by analyzing the amino acids and fatty acids. This study presents the valorization of bread waste through the production of a protein- and fatty-acid-rich fungal biomass that is simultaneously a source of microfibers.

Dioscorea alata as Alternative Culture Media for Fungal Cultivation and Biomass Production

Dioscorea alata (purple yam) is a tuber crop that contains plenty of nutrients. It is widely cultivated in Thailand, but it is underutilized. In this study, the suitability of purple yam to replace potato dextrose media for fungal growth was investigated. Mushrooms and molds were grown on purple yam dextrose agar (PYDA), whereas yeasts were cultured in purple yam dextrose broth (PYDB). Response surface methodology (RSM) with Box-Behnken design (BBD) was used to optimize the culture conditions for Saccharomyces cerevisiae biomass production. The growth profile of S. cerevisiae in PYDB under optimized culture conditions was also studied. All test mushrooms and molds recorded the highest colony diameter and mycelial dry weight on PYDA containing 40% purple yam. Similar to mushrooms and molds, yeasts in PYDB with 40% purple yam showed the highest number of cells. The growth of fungi on purple yam dextrose media was significantly higher than those on potato dextrose media under standard conditions. The optimal conditions from the RSM results for the biomass production of S. cerevisiae in PYDB were purple yam concentration of 49.61%, dextrose concentration of 4.87%, pH value of 5.74, and inoculum size of 7.00%. The biomass of S. cerevisiae in PYDB under the optimal conditions obtained from the results of the optimization by RSM was thirty times higher than S. cerevisiae biomass in potato dextrose broth under standard conditions. Our results suggest that purple yam could be an alternative to potato dextrose media for fungal cultivation.

Effect of pH and Temperature on the Growth and Laccases Production in the Cultivation of Pleurotus sajor-caju PS-2001 in Stirred-tank Bioreactor

Background: Laccases are multi-copper enzymes that oxidize phenolic/aromatic compounds and represent a promising alternative to environmental decontamination processes and biotechnological applications. Objective: The effects of pH and temperature on the growth and the production of laccases during the cultivation of Pleurotus sajor-caju PS-2001 in stirred-tank bioreactor were assessed. Methods: Assays were performed at fixed pH values from 4.5 to 7.5 (28°C) and at temperatures from 24 to 36°C (pH 6.5). Results: In pH testing, larger biomass concentration (4.5 g L-1) was reached at pH 5.5, whereas concentrations of 3.7, 3.1 and 1.7 g L-1 were measured at pH 4.5, 6.5 and 7.5, respectively. With ABTS as substrate, peaks of laccases activity of 50, 30 and 24 U mL-1, at pH 6.5, 5.5 and 7.5, respectively, were detected. Under different temperatures, higher mycelial concentrations (3.0 g L-1) were quantified at 66 hours at 28°C, while concentrations below 2.0 g L-1 were observed at 24, 32, and 36°C. Maximum laccases activities of 50, 42, 6 and 5 U mL-1 were obtained at 28, 32, 24, and 36°C, respectively. In all tests, the presence of other phenol oxidases – total peroxidase, manganese peroxidase, lignin peroxidase and veratryl alcohol oxidase – was observed. Conclusion: The results indicate that variations in pH and temperature during fungal cultivation strongly affect the enzymatic activity and growth kinetics of P. sajor-caju PS-2001 in a stirredtank bioreactor.

A Liquid Chromatographic Method for Rapid and Sensitive Analysis of Aflatoxins in Laboratory Fungal Cultures

Culture methods supplemented with high-performance liquid chromatography (HPLC) technique provide a rapid and simple tool for detecting levels of aflatoxins (AFs) produced by fungi. This study presents a robust method for simultaneous quantification of aflatoxin (AF) B1, B2, G1, and G2 levels in several fungal cultivation states: submerged shake culture, liquid slant culture, and solid-state culture. The recovery of the method was evaluated by spiking a mixture of AFs at several concentrations to the test medium. The applicability of the method was evaluated by using aflatoxigenic and non-aflatoxigenic Aspergilli. A HPLC coupled with the diode array (DAD) and fluorescence (FLD) detectors was used to determine the presence and amounts of AFs. Both detectors showed high sensitivity in detecting spiked AFs or AFs produced in situ by toxigenic fungi. Our methods showed 76%–88% recovery from medium spiked with 2.5, 10, 50, 100, and 500 ng/mL AFs. The limit of quantification (LOQ) for AFs were 2.5 to 5.0 ng/mL with DAD and 0.025 to 2.5 ng/mL with FLD. In this work, we described in detail a protocol, which can be considered the foremost and only verified method, to extract, detect, and quantify AFs employing both aflatoxigenic and non-toxigenic Aspergilli.

Mycoprotein: environmental impact and health aspects

The term mycoprotein refers to the protein-rich food made of filamentous fungal biomass that can be consumed as an alternative to meat. In this paper, the impact caused by the substitution of animal-origin meat in the human diet for mycoprotein on the health and the environment is reviewed. Presently, mycoprotein can be found in the supermarkets of developed countries in several forms (e.g. sausages and patties). Expansion to other markets depends on the reduction of the costs. Although scarce, the results of life cycle analyses of mycoprotein agree that this meat substitute causes an environmental impact similar to chicken and pork. In this context, the use of inexpensive agro-industrial residues as substrate for mycoprotein production has been investigated. This strategy is believed to reduce the costs involved in the fungal cultivation and lower the environmental impact of both the mycoprotein and the food industry. Moreover, several positive effects in health have been associated with the substitution of meat for mycoprotein, including improvements in blood cholesterol concentration and glycemic response. Mycoprotein has found a place in the market, but questions regarding the consumer’s experience on the sensory and health aspects are still being investigated.

Trade-offs in an ant–plant–fungus mutualism

Species engaged in multiple, simultaneous mutualisms are subject to trade-offs in their mutualistic investment if the traits involved in each interaction are overlapping, which can lead to conflicts and affect the longevity of these associations. We investigate this issue via a tripartite mutualism involving an ant plant, two competing ant species and a fungus the ants cultivate to build galleries under the stems of their host plant to capture insect prey. The use of the galleries represents an innovative prey capture strategy compared with the more typical strategy of foraging on leaves. However, because of a limited worker force in their colonies, the prey capture behaviour of the ants results in a trade-off between plant protection (i.e. the ants patrol the foliage and attack intruders including herbivores) and ambushing prey in the galleries, which has a cascading effect on the fitness of all of the partners. The quantification of partners' traits and effects showed that the two ant species differed in their mutualistic investment. Less investment in the galleries (i.e. in fungal cultivation) translated into more benefits for the plant in terms of less herbivory and higher growth rates and vice versa. However, the greater vegetative growth of the plants did not produce a positive fitness effect for the better mutualistic ant species in terms of colony size and production of sexuals nor was the mutualist compensated by the wider dispersal of its queens. As a consequence, although the better ant mutualist is the one that provides more benefits to its host plant, its lower host–plant exploitation does not give this ant species a competitive advantage. The local coexistence of the ant species is thus fleeting and should eventually lead to the exclusion of the less competitive species.