Wild-cultivated medicinal mushroom Ganoderma lucidum was morphologically identified and sequenced using phylogenetic software. In submerged-liquid fermentation (SLF), biomass, exopolysaccharide (EPS) and intracellular polysaccharide (IPS) production of the identified G.lucidum was optimised based on initial pH, starting glucose concentration and agitation rate parameters using response surface methodology (RSM). Molecularly, the G. lucidum strain QRS 5120 generated 637 base pairs, which was commensurate with related Ganoderma species. In RSM, by applying central composite design (CCD), a polynomial model was fitted to the experimental data and was found to be significant in all parameters investigated. The strongest effect (p < 0.0001) was observed for initial pH for biomass, EPS and IPS production, while agitation showed a significant value (p < 0.005) for biomass. By applying the optimized conditions, the model was validated and generated 5.12 g/L of biomass (initial pH 4.01, 32.09 g/L of glucose and 102 rpm), 2.49 g/L EPS (initial pH 4, 24.25 g/L of glucose and 110 rpm) and 1.52 g/L of IPS (and initial pH 4, 40.43 g/L of glucose, 103 rpm) in 500 mL shake flask fermentation. The optimized parameters can be upscaled for efficient biomass, EPS and IPS production using G. lucidum.
In this study, a selected γ-aminobutyric acid (GABA)-rich Malaysian strain Aspergillus oryzae NSK was collected from soy sauce koji. The strain was used to explore the effect of using renewable native sugar syrup, sugarcane, nipa, and molasses as fermentable substrates for developing a novel functional GABA soy sauce. We evaluated the strain using the chosen native sugars for 7 days using shake flask fermentation at 30 °C. The results showed optimum GABA concentration was achieved using cane molasses as the fermentable substrate (354.08 mg/L), followed by sugarcane syrup (320.7 mg/L) and nipa syrup (232.07 mg/L). Cane molasses was subsequently utilized as a substrate to determine the most suitable concentration for A. oryzae NSK to enhance GABA production and was determined as 50% g/L of glucose standard cane molasses. Our findings indicate that cane molasses can be used as a GABA-rich ingredient to develop a new starter culture for A. oryzae NSK soy sauce production.
In submerged-liquid fermentation, seven key parameters were assessed using one-factor-at-a-time to obtain the highest GABA yield using an industrial soy sauce koji Aspergillus oryzae strain NSK (AOSNSK). AOSNSK generated maximum GABA at 30 °C (194 mg/L) and initial pH 5 (231 mg/L), thus was able to utilize sucrose (327 mg/L of GABA) for carbon source. Sucrose at 100 g/L, improved GABA production at 646 mg/L. Single nitrogen sources failed to improve GABA production, however a combination of yeast extract (YE) and glutamic acid (GA) improved GABA at 646.78 mg/L. Carbon-to-nitrogen ratio (C8:N3) produced the highest cell (24.01 g/L) and GABA at a minimal time of 216 h. The key parameters of 30 °C, initial pH 5, 100 g/L of sucrose, combination YE and GA, and C8:N3 generated the highest GABA (3278.31 mg/L) in a koji fermentation. AOSNSK promisingly showed for the development of a new GABA-rich soy sauce.
Four types of mycelial extracts were derived from the airlift liquid fermentation (ALF) of Pleurotus flabellatus, namely exopolysaccharide (EX), endopolysaccharide (EN), hot water (WE), and hot alkali (AE) extracts. Such extracts were screened for their active components and biological potential. EN proved to be most effective in inhibition of lipid peroxidation (EC50 = 1.71 ± 0.02 mg/mL) and in Cupric ion reducing antioxidant capacity (CUPRAC) assay (EC50 = 2.91 ± 0.01 mg TE/g). AE exhibited most pronounced ability to chelate ferrous ions (EC50 = 4.96 ± 0.08 mg/mL) and to scavenge ABTS radicals (EC50 = 3.36 ± 0.03 mg TE/g). β-glucans and total phenols contributed most to the chelating ability and quenching of ABTS radicals. Inhibition of lipid peroxidation correlated best with total glucans, total proteins, and β-glucans. Total proteins contributed most to CUPRAC antioxidant capacity. Antifungal effect was determined against Candida albicans ATCC 10231 (MIC: 0.019-0.625 mg/mL; MFC: 0.039-2.5 mg/mL), and towards C. albicans clinical isolate (MIC and MFC: 10.0-20.0 mg/mL). Comparison of cytotoxicity against colorectal carcinoma HCT 116 cells (IC50: 1.8 ± 0.3-24.6 ± 4.2 mg/mL) and normal lung MRC-5 fibroblasts (IC50: 17.0 ± 4.2-42.1 ± 6.1 mg/mL) showed that EN, and especially AE possess selective anticancer activity (SI values 3.41 and 9.44, respectively). Slight genotoxicity was observed only for AE and EX, indicating the low risk concerning this feature. Notable antioxidative and anticandidal activities, selective cytotoxicity against colorectal carcinoma cells, and absence/low genotoxicity pointed out that ALF-cultivated P. flabellatus mycelium could be considered as a valuable source of bioactive substances.
Natural mycelial biomass (MB) and exopolysaccharides (EPS) of Malaysian tiger milk mushroom Lignosus rhinocerus are considered high-end components due to their high commercial potential value in drug discovery. This study aims to evaluate the toxicity of the mushroom extracts' generated in a bioreactor using the zebrafish embryo toxicity (ZFET) model assay as a new therapy for treating asthma. Both MB and EPS extracts, at concentrations 0.16-10 mg/mL, were tested for ZFET and early development effects on Zebrafish Embryos (ZE) during 24-120 h post-fertilisation (HPF). Findings revealed that MB was deemed safe with an LC50 of 0.77 mg/mL; the EPS were non-toxic (LC50 of 0.41 mg/mL). Neither MB nor EPS delayed hatching nor teratogenic defects in the treated ZE at a 2.5 mg/mL dose. There were no significant changes in the ZE heart rate after treatments with MB (130 beats/min) and EPS (140 beats/min), compared to that of normal ZE (120-180 beats/min). Mixing both natural compounds MB and EPS did not affect toxicity using ZFET testing; thus, intimating their safe future use as therapeutic interventions. This represents the first study to have used the ZFET assay on MB and EPS extracts of L. rhinocerus for future health applications.
Underutilized mycelium of Ganoderma lucidum BCCM 31549 has been a significant source of a glucan sulfate (GS) possessing therapeutic activities. GS have been evaluated for their antifungaldemelanizing properties and nitrite oxide production from stimulated RAW264.7 macrophages. GS exhibited antifungal activity against Aspergillus niger A60 with a minimum inhibitory concentration of 60 mg/mL and a minimum fungicidal concentration of 100 mg/mL. At 60mg/mL (sublethal) and 30mg/mL (subinhibitory) doses of GS, the mycelium of A. niger A60 was successfully demelanized with a conidiophore head and black pigment reduction. Additionally, GS successfully stimulated RAW264.7 macrophage cells at a concentration of 500 μg/mL to produce 0.45 μM of nitric oxide. The GS-stimulated RAW264.7 macrophages were morphologically similar to those treated with lipopolysaccharide. The results highlight a novel bifunctional property of mycelial GS from G. lucidum.
Natural mycelial exopolysaccharide (EPS) and endopolysaccharide (ENS) extracted from bioreactor-cultivated European Ganoderma applanatum mushrooms are of potential high commercial value for both food and adjacent biopharmaceutical industries. In order to evaluate their potential toxicity for aquaculture application, both EPS (0.01-10 mg/mL) and ENS (0.01-10 mg/mL) extracts were tested for Zebrafish Embryo Toxicity (ZFET); early development effects on Zebrafish Embryos (ZE) were also analyzed between 24 and 120 h post-fertilization (HPF). Both EPS and ENS are considered non-toxic with LC50 of 1.41 mg/mL and 0.87 mg/mL respectively. Both EPS and ENS did not delay hatching and teratogenic defect towards ZE with <1.0 mg/mL, respectively. No significant changes in the ZE heart rate were detected following treatment with the two compounds tested (EPS: 0.01-10 mg/mL: 176.44 ± 0.77 beats/min and ENS: 0.01-10 mg/mL: 148.44 ± 17.75 beats/min) compared to normal ZE (120-180 beats/min). These initial findings support future pre-clinical trials in adult fish models with view to safely using EPS and ENS as potential feed supplements for supplements for development of the aquaculture industry.
The fluctuation of domestic wastewater characteristic inhibits the current conventional microbial-based treatment. The bioremediation fungi has received attention and reported to be an effective alternative to treat industrial wastewater. Similar efficient performance is envisaged for domestic wastewater whereby assessed performance of fungi for varying carbon-to-nitrogen ratios in domestic wastewater is crucial. Thus, the performance of pre-grown wild-Serbian Ganoderma lucidum mycelial pellets (GLMPs) was evaluated on four different synthetic domestic wastewaters under different conditions of initial pH (pH 4, 5, and 7) and chemical oxygen demand (COD) to nitrogen (COD/N) ratio of 3.6:1, 7.1:1, 14.2:1, and 17.8:1 (C3.6N1, C7.1N1, C14.2N1, and C17.8N1). The COD/N ratios with a constant concentration of ammonia-nitrogen (NH3-N) were chosen on the basis of the urban domestic wastewater characteristics sampled at the inlet basin of a sewage treatment plant (STP). The parameters of pH, COD, and NH3-N were measured periodically during the experiment. The wild-Serbian GLMPs efficiently removed the pollutants from the synthetic sewage. The COD/N ratio of C17.8N1 wastewater had the best COD and NH3-N removal, as compared to the lower COD/N ratio, and the shortest treatment time was obtained in an acidic environment at pH 4. The highest percentage for COD and NH3-N removal achieved was 96.0% and 93.2%, respectively. The results proved that the mycelium of GLMP has high potential in treating domestic wastewater, particularly at high organic content as a naturally sustainable bioremediation system.
The growing demand for high value aromatic herb Polygonum minus-based products have increased in recent years, for its antioxidant, anticancer, antimicrobial, and anti-inflammatory potentials. Although few reports have indicated the chemical profiles and antioxidative effects of Polygonum minus, no study has been conducted to assess the benefits of micro-environmental manipulation (different shading levels) on the growth, leaf gas exchange and secondary metabolites in Polygonum minus. Therefore, two shading levels (50%:T2 and 70%:T3) and one absolute control (0%:T1) were studied under eight weeks and 16 weeks of exposures on Polygonum minus after two weeks. It was found that P. minus under T2 obtained the highest photosynthesis rate (14.892 µmol CO2 m-2 s-1), followed by T3 = T1. The increase in photosynthesis rate was contributed by the enhancement of the leaf pigments content (chlorophyll a and chlorophyll b). This was shown by the positive significant correlations observed between photosynthesis rate with chlorophyll a (r2 = 0.536; p ≤ 0.05) and chlorophyll b (r2 = 0.540; p ≤ 0.05). As the shading levels and time interval increased, the production of total anthocyanin content (TAC) and antioxidant properties of Ferric Reducing Antioxidant Power (FRAP) and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) also increased. The total phenolic content (TPC) and total flavonoid content (TFC) were also significantly enhanced under T2 and T3. The current study suggested that P.minus induce the production of more leaf pigments and secondary metabolites as their special adaptation mechanism under low light condition. Although the biomass was affected under low light, the purpose of conducting the study to boost the bioactive properties in Polygonum minus has been fulfilled by 50% shading under 16 weeks' exposure.
The pellet morphology and diameter range (DR) of Ganoderma lucidum were observed in a repeated-batch fermentation (RBF) for the trio total production of biomass, exopolysaccharide (EPS) and endopolysaccharide (ENS). Two factors were involved in RBF; broth replacement ratio (BRR: 60%, 75% and 90%) and broth replacement time point (BRTP: log, transition and stationary phase) in days. In RBF, 34.31 g/L of biomass favoured small-compact pellets with DR of 20.67 µm< d < 24.00 µm (75% BRR, day 11 of BRTP). EPS production of 4.34 g/L was prone to ovoid-starburst pellets with DR of 34.33 µm< d <35.67 µm (75% BRR, day 13 of BRTP). Meanwhile, the highest 2.43 g/L of ENS production favoured large-hollow pellets with DR of 34.00 µm< d < 38.67 µm (90% BRR, day 13 of BRTP). In addition, RBF successfully shortened the biomass-EPS-ENS fermentation period (31, 33 and 35 days) from batch to 5 days, in seven consecutive cycles of RBF. In a FTIR detection, β-glucan (BG) from EPS and ENS extracts were associated with β-glycosidic linkages (2925 cm-1, 1635 cm-1, 1077 cm-1, 920 cm-1 and 800 cm-1 wavelengths) with similar 1H NMR spectral behaviour (4.58, 3.87 and 3.81 ppm). Meanwhile, 4 mg/L of BG gave negative cytotoxic effects on normal gingival cell line (hGF) but induced antiproliferation (IC50 = 0.23 mg/mL) against cancerous oral Asian cellosaurus cell line (ORL-48). Together, this study proved that G. lucidum mycelial pellets could withstand seven cycles of long fermentation condition and possessed anti-oral cancer beta-glucan, which suits large-scale natural drug fermentation.
In a commercial oyster mushroom farm, from 300 g of the total harvest, only the cap and stem of the fruiting body parts are harvested (200 g) while the unused lower section called fruiting-body-base (FBB) is discarded (50 g). A new antioxidative FBB flour (FBBF) conversion to mixed-ratio chicken patty was recently developed which converts 16.67% of FBB into an edible flour. At the initial stage, pretreatments of FBBF were optimized at particle size (106 µm) and citric acid concentration (0.5 g/100 mL) to improve flour antioxidant responses. Such pretreatments boosted total phenolic content (2.31 ± 0.53 mg GAE/g) and DPPH (51.53 ± 1.51%) of pretreated FBBF. Mixed-ratio chicken patty containing FBBF (10%, 20%, 30%) significantly (P
The rapid expansion of industrialization and continuous population growth have caused a steady increase in energy consumption. Despite using renewable energy, such as bioethanol, to replace fossil fuels had been strongly promoted, however the outcomes were underwhelming, resulting in excessive greenhouse gases (GHG) emissions. Microalgal biochar, as a carbon-rich material produced from the pyrolysis of biomass, provides a promising solution for achieving net zero emission. By utilizing microalgal biochar, these GHG emissions can be captured and stored efficiently. It also enhances soil fertility, improves water retention, and conduct bioremediation in agriculture and environmental remediation field. Moreover, incorporating microalgal biochar into a zero-waste biorefinery could boost the employ of biomass feedstocks effectively to produce valuable bioproducts while minimizing waste. This contributes to sustainability and aligns with the concepts of a circular bioeconomy. In addition, some challenges like commercialization and standardization will be addressed in the future.
In the world of fast fashion, textile industries are blooming rapidly to meet the consumer's demands. However, vast amounts of wastewater have been constantly produced, and it is becoming a serious environmental problem in the waterways. Although the technology for treating textile wastewater has been well reported and established, more sustainable efforts have taken the attention nowadays. Through the use of living Malaysian Ganoderma lucidum mycelial pellets (GL) and activated dolomite (AD) in the treatment system, the study explores the synergy between biosorption and physisorption as alternative treatment for textile wastewater. In the current work, mixture of GL premixed with AD (50:50; v/v) is used to treat industrial textile wastewater. The morphology, adsorption characteristics, and antibacterial activity of the adsorbents were studied. The mixture of adsorbents is capable of removing colours by 77.8% and reducing chemical oxygen demand (COD) by 75% within 48 h contact. Furthermore, the kinetic and adsorption had been studied and follow the pseudo-first-order kinetic model while both adsorption of Langmuir and Freundlich model was deduced from the treatment. In addition, antimicrobial activities from the treatment potentially reduced 10 × 101 CFU/mL after 48 h. The synergistic treatment by Ganoderma lucidum mycelial pellets and activated dolomite has immense potential in future wastewater treatment technology to obtain cleaner water.
The fungi-based technology, wild-Serbian Ganoderma lucidum (WSGL) as myco-alternative to existing conventional microbial-based wastewater treatment is introduced in this study as a potential alternative treatment. The mycoremediation is highly persistent for its capability to oxidatively breakdown pollutant substrates and widely researched for its medicinal properties. Utilizing the nonhazardous properties and high degradation performance of WSGL, this research aims to optimize mycoremediation treatment design for chemical oxygen demand (COD) and ammonia nitrogen (AN) removal in domestic wastewater based on proposed Model 1 (temperature and treatment time) and Model 2 (volume of pellet and treatment time) via response surface methodology (RSM). Combined process variables were temperature (0C) (Model 1) and the volume of mycelial pellets (%) (Model 2) against treatment time (hour). Response variables for these two sets of central composite design (CCD) were the removal efficiencies of COD (%) and AN (%). The regression line fitted well with the data with R2 values of 0.9840 (Model 1-COD), 0.9477 (Model 1-AN), 0.9988 (Model 2-COD), and 0.9990 (Model 2-AN). The lack of fit test gives the highest value of sum of squares equal to 9494.91 (Model 1-COD), 9701.68 (Model 1-AN), 23786.55 (Model 2-COD), and 13357.02 (Model 2-AN), with probability F values less than 0.05 showing significant models. The optimized temperature for Model 1 was at 25 °C within 24 h of treatment time with 95.1% COD and 96.3% AN removals. The optimized condition (temperature) in Model 1 was further studied in Model 2. The optimized volume of pellet for Model 2 was 0.25% in 24-h treatment time with 76.0% COD and 78.4% AN removals. Overall, the ascended sequence of high volume of pellet considered in Model 2 will slow down the degradation process. The best fit volume of pellet with maximum degradation of COD and AN is equivalent to 0.1% at 25 °C in 24 h. The high performance achieved demonstrates that the mycoremediation of G. lucidum is highly potential as part of the wastewater treatment system in treating domestic wastewater of high organic loadings.
The processing of edible insects as an alternative source of nutrition may be a key driver in the development of a sustainable food and feed system. This review will study two industrial types of insects-mealworms and locusts-and summarize evidence related to the impact of processing on their micro- and macronutritional characteristics. The focus will be on their potential use as food for human consumption as opposed to animal feed. Literature has indicated that these two insects have the potential to provide protein and fat qualities comparable to or better than traditional mammalian sources. For example, mealworms-the larval form of the yellow mealworm beetlepossess a higher fat content, while adult locusts are rich in fibers, especially chitin. However, due to the different matrix and nutrient compositions, the processing of mealworms or locusts at a commercial scale needs to be tailored to minimize nutritional loss and maximize cost efficiency. The stages of preprocessing, cooking, drying, and extraction are the most critical control points for nutritional preservation. Thermal cooking applications such as microwave technology have demonstrated promising results, but the generation of heat may contribute to a certain nutritional loss. In an industrial context, drying using freeze dry is the preferred choice due to its uniformity, but it can be costly while increasing lipid peroxidation. During the extraction of nutrients, the use of green emerging technologies such as high hydrostatic pressure, pulsed electric field, and ultrasound may provide an alternative method to enhance nutrient preservation.
The versatility of a well-known fibrous crop, Hibiscus cannabinus (kenaf) is still relatively new to many. Kenaf's potential applications, which can be extended even into critical industries such as pharmaceutical and food industries, have always been overshadowed by its traditionally grown fiber. Therefore, this study aimed to venture into the biotechnological approach in reaping the benefits of kenaf through plant cell suspension culture to maximize the production of kenaf callus biomass (KCB) and exopolysaccharide (EPS), which is deemed to be more sustainable. A growth curve was established which indicates that cultivating kenaf callus in suspension culture for 22 days gives the highest KCB (9.09 ± 1.2 g/L) and EPS (1.1 ± 0.02 g/L). Using response surface methodology (RSM), it was found that sucrose concentration, agitation speed, and naphthalene acetic acid (NAA) concentration can affect the production of KCB and EPS significantly (p
Fungal biomass is the future's feedstock. Non-septate Ascomycetes and septate Basidiomycetes, famously known as mushrooms, are sources of fungal biomass. Fungal biomass, which on averagely comprises about 34% protein and 45% carbohydrate, can be cultivated in bioreactors to produce affordable, safe, nontoxic, and consistent biomass quality. Fungal-based technologies are seen as attractive, safer alternatives, either substituting or complementing the existing standard technology. Water and wastewater treatment, food and feed, green technology, innovative designs in buildings, enzyme technology, potential health benefits, and wealth production are the key sectors that successfully reported high-efficiency performances of fungal applications. This paper reviews the latest technical know-how, methods, and performance of fungal adaptation in those sectors. Excellent performance was reported indicating high potential for fungi utilization, particularly in the sectors, yet to be utilized and improved on the existing fungal-based applications. The expansion of fungal biomass in the industrial-scale application for the sustainability of earth and human well-being is in line with the United Nations' Sustainable Development Goals.
Numerous generations have been affected by hunger, which still affects hundreds of millions of people worldwide. The hunger crisis is worsening although many efforts have been made to minimize it. Besides that, food waste is one of the critical problems faced by most countries worldwide. It has disrupted the food chain system due to inefficient waste management, while negatively impacting the environment. The majority of the waste is from the food production process, resulting in a net zero production for food manufacturers while also harnessing its potential. Most food production wastes are high in nutritional and functional values, yet most of them end up as low-cost animal feed and plant fertilizers. This review identified key emerging wastes from the production line of mushroom, peanut, and soybean (MPS). These wastes (MPS) provide a new source for food conversion due to their high nutritional content, which contributes to a circular economy in the post-pandemic era and ensures food security. In order to achieve carbon neutrality and effective waste management for the production of alternative foods, biotechnological processes such as digestive, fermentative, and enzymatic conversions are essential. The article provides a narrative action on the critical potential application and challenges of MPS as future foods in the battle against hunger.