Search Results (25)

Biotechnological valorization of Flourensia cernua foliage was carried out using fungal solid-state fermentation; several outcomes of this bioprocess were identified which added value to the plant material. F. cernua leaves placed in aluminum trays were inoculated with Aspergillus niger; extracts of this plant were evaluated and the foliage was tested for in vitro digestibility. The solid bioprocess was carried out at 75% humidity for 120 h and after the fermentation, β-glucosidase activity; phenolics and in vitro digestibility were quantified and measured. Two high β-glucosidase production levels were detected at 42 and 84 h with 3192 and 4092 U/L, respectively. Several phenolics of industrial importance were detected with a HPLC-ESI-MS, such as glycosides of luteolin and apigenin. The other outcome was a substantial improvement in anaerobic digestibility. The unfermented sample registered a 30% in vitro degradability, whereas samples subjected to 84 h of fungal fermentation increased degradability by up to 51%. This bioprocess was designed to detect more than one product, which can contribute to an increase in the added value of F. cernua foliage. Full article

Solid-state fermentation (SSF) involves the growth of microorganisms on solid substrates, mimicking natural environments of many species. Due to sustainability concerns, transforming agro-industrial by-products into value-added products through SSF has been increasingly studied. Brewer’s spent grain (BSG), the main by-product of beer production, mostly consists of barley grain husks, making BSG a great support for microorganism cultivation. Although autoclaving remains the standard sterilization and pretreatment method of substrates, electric field technologies and their attendant ohmic heating (OH) have great potential as an alternative technology. In the present work, pretreatment of BSG by OH was explored in SSF with Aspergillus niger to produce commercially valuable enzymes. OH favored the solubilization of phenolic compounds, total protein, and reducing sugars significantly higher than autoclaving. SSF of treated BSG led to the production of lignocellulosic enzymes, with xylanases being the most active, reaching 540 U/g, a 1.5-fold increase in activity compared to autoclaved BSG. Protease activity was also improved 1.6-fold by OH, resulting in 49 U/g. Our findings suggest that OH treatment is an effective alternative to autoclaving and that its integration with SSF is a sustainable strategy to enhance by-product valorization through enzyme production with many industrial applications, according to circular economy guidelines. Full article

Creosote bush (Larrea tridentata), a shrub distributed across approximately 19 Mha of arid North American regions, has traditional applications in folk medicine due to the presence of bioactive molecules such as nordihydroguaiaretic acid (NDGA) and ellagic acid (EA). This study investigated the implementation of a solid-state fermentation (SSF) optimization process employing creosote bush leaves as substrate using Aspergillus niger GH1 to improve NDGA and EA extraction. This study was based on previous research by our group that identified key parameters for NDGA production in a related SSF system. Creosote bush is a recognized source of these bioactive compounds, which possess antioxidant and anti-inflammatory properties. Conventional extraction methods often exhibit limitations in efficiency and sustainability. The efficacy of A. niger GH1 in SSF has been previously established with diverse substrates. In this study, A. niger GH1 was employed in an SSF process utilizing creosote bush leaves as a substrate using a Box–Behnken experimental design. The accumulation of NDGA and EA, which were quantified by HPLC-MS, yielded values of 1.20 ± 0.32 mg g⁻¹ for EA and 7.39 ± 0.52 mg g⁻¹ for NDGA. In vitro antioxidant assays (DPPH and ABTS) demonstrated significant antioxidant activity, with inhibition percentages of 55.69% and 84.84%, respectively. These results indicate that A. niger GH1-mediated SSF using Creosote bush leaves is a viable and sustainable strategy for producing these valuable bioactive compounds. Full article

The by-products from three varieties of dates—Mozafati, Sayer, and Kabkab—were subjected to solid-state fermentation using Aspergillus niger alone or in co-culture with Lactiplantibacillus plantarum or Limosilactobacillus reuteri to enhance their phenolic and flavonoid content, along with antioxidant and antimicrobial activities. Solid-state fermentation, being environmentally friendly and cost-effective, is particularly suitable for agricultural residues. Significant increases (p < 0.05) in total polyphenol content (TPC), total flavonoid content (TFC), and antioxidant power were observed post-fermentation, especially under co-culture conditions. The highest TPC (12.98 ± 0.29 mg GA/g) and TFC (1.83 ± 0.07 mg QE/g) were recorded in the co-culture fermentation of by-products from the Mozafati and Sayer varieties, respectively. HPLC analysis revealed changes in polyphenol profiles post-fermentation, with reductions in gallic and ferulic acids and increases in caffeic acid, p-coumaric acid, rutin, quercetin, and kaempferol. FT-IR analysis confirmed significant alterations in polyphenolic functional groups. Enhanced antimicrobial activity was also observed, with inhibition zones ranging from 8.26 ± 0.06 mm for Kabkab to 17.73 ± 0.09 mm for Mozafati. These results suggest that co-culture solid-state fermentation is a promising strategy for valorizing date by-products, with potential applications in nutraceuticals and/or pharmaceutical products and as valuable additives in the food industry. Full article

Grape pomace seeds contain abundant phenolic compounds, which are also present in both soluble and insoluble forms, similar to many other plant matrices. To further increase the extractable soluble phenolics and their antioxidant activities, grape pomace seeds were fermented with different fungi. Results showed that solid-state fermentation (SSF) with Aspergillus niger, Monascus anka, and Eurotium cristatum at 28 °C and 65% humidity had a significantly positive impact on the release of soluble phenolics in grape pomace seeds. Specifically, SSF with M. anka increased the soluble phenolic contents by 6.42 times (calculated as total phenolic content) and 6.68 times (calculated as total flavonoid content), leading to an overall improvement of antioxidant activities, including DPPH (increased by 2.14 times) and ABTS (increased by 3.64 times) radical scavenging activity. Furthermore, substantial changes were observed in the composition and content of individual phenolic compounds in the soluble fraction, with significantly heightened levels of specific phenolics such as chlorogenic acid, syringic acid, ferulic acid, epicatechin gallate, and resveratrol. Notably, during M. anka SSF, positive correlations were identified between the soluble phenolic content and hydrolase activities. In particular, there is a strong positive correlation between glycosidase and soluble phenols (r = 0.900). The findings present an effective strategy for improving the soluble phenolic profiles and bioactivities of grape pomace seeds through fungal SSF, thereby facilitating the valorization of winemaking by-products. Full article

Solid by-products with lignocellulosic structures are considered appropriate substrates for solid-state fermentation (SSF) to produce enzymes with diverse industrial applications. In this work, brewer’s spent grain (BSG), rice husk (RH), and vine shoot trimmings (VSTs) were employed as substrates in SSF with Aspergillus niger CECT 2088 to produce cellulases, xylanases, and amylases. The addition of 2% (NH₄)₂SO₄ and 1% K₂HPO₄ to by-products had a positive effect on enzyme production. Substrate particle size influenced enzyme activity and the overall highest activities were achieved at the largest particle size (10 mm) of BSG and RH and a size of 4 mm for VSTs. Optimal substrate composition was predicted using a simplex centroid mixture design. The highest activities were obtained using 100% BSG for β-glucosidase (363 U/g) and endo-1,4-β-glucanase (189 U/g), 87% BSG and 13% RH for xylanase (627 U/g), and 72% BSG and 28% RH for amylase (263 U/g). Besides the optimal values found, mixtures of BSG with RH or VSTs proved to be alternative substrates to BSG alone. These findings demonstrate that SSF bioprocessing of BSG individually or in mixtures with RH and VSTs is an efficient and sustainable strategy to produce enzymes of significant industrial interest within the circular economy guidelines. Full article

Plant feedstuffs are the main ingredients of animal feed. Owing to food–feed competition, increasing the utilization efficiency of these feedstuffs is important for animal nutrition. This can be achieved via solid-state fermentation (SSF). SSF of a plant feedstuff mixture (PFM) (25% rapeseed meal, soybean meal, rice bran, and sunflower meal) by three fungi (Aspergillus ibericus MUM 03.29, Aspergillus niger CECT 2088, and Aspergillus niger CECT 2915) resulted in an increase in protein content by 5%, irrespective of fungi, a reduction in cellulose content by 9 to 11%, and of hemicellulose content by 21 to 34%, relative to unfermented PFM. Enzyme production was measured: the highest cellulase (123.7 U/g), xylanase (431.8 U/g), and beta-glucosidase (117.9 U/g) activity were achieved with A. niger CECT 2088. Principal component analysis showed a positive correlation between all fermented PFMs and enzyme production, protein content, digestibility, and fiber reduction. Bioprocessing of the PFM by SSF increased its nutritional value and digestibility, making it more appealing for animal feeds. Full article

Tea (Camellia sinensis) is an evergreen shrub that is recognized worldwide for its functional properties. The current global production of green tea is approximately 5.3 million tons per year. Green tea processing has severely affected the generation of agro-industrial waste. One strategy for reducing waste accumulation is the revalorization of agro-industrial wastes via solid-state fermentation (SSF). The aim of this study was to valorize green tea processing residues to produce tannase under SSF using an endemic strain from Western Ghats, Aspergillus niger TBG 28A. SSF was performed in Erlenmeyer flasks with spent green tea leaves inoculated with spores of A. niger TBG 28A. Bioprocess optimization was carried out by employing the Box–Benkhen experimental design, achieving a high enzymatic yield of 246.82 (U/g). The present study shows the complexity of the degradation of tannins and the different patterns of expression of fungal tannase obtained from A. niger TBG 28 A. The enzyme was further purified to obtain a fold purification of 16.35% and a molecular mass of 150 kDa. Producing tannase with a novel strain of A. niger TBG 28A is an interesting strategy to revalorize green tea waste. Full article

Lignocellulosic biomass is a promising feedstock for added value compound production in biotechnological processes such as solid-state fermentation (SSF). Although these solid materials can be directly used as substrates in fermentations in a solid state, a pretreatment is often required, especially if the microorganism selected is unable to produce lignocellulosic enzymes. In the present work, several pretreatment strategies were applied to a 50% (w/w) mixture of olive and sunflower cakes before SSF for lipase production by the oleaginous yeast Yarrowia lipolytica W29. Co-culture strategies with Y. lipolytica and Aspergillus niger did not improve lipase production by the oleaginous yeast. Biological pretreatment with a fungal enzymatic extract led to a significant increase in sugar availability in the substrate mixture after a short incubation period, improving yeast growth. Microwave and ultrasound were the physical pretreatments selected and microwave irradiation proved to be the best method, resulting in 44% and 17% increases in yeast growth and lipase production, respectively, compared to the untreated mixture. An improvement in lipase activity was also observed after ultrasonic treatment in semi-solid fermentations, leading to a 2-fold increase in this enzyme activity compared to the control. The utilization of pretreatments before SSF with Y. lipolytica can increase sugars availability and result in structural changes in the solid substrate, which can improve the bioprocesses’ productivity. Full article

Extracted sugar beet press pulp (SBPP) is a promising agricultural residue for saccharification and further bioconversion. Combining solid-state fermentation of SBPP with engineered Aspergillus niger for enzyme production followed by hydrolysis of additionally added SBPP in the same bioreactor was studied to produce a sugar solution (hydrolysate) in a one-pot process. The initial aerobic solid-state fermentations were carried out in duplicate on non-milled, wet SBPP (moisture content of 72% (w/v)) with an A. niger strain engineered for constitutive pectinase production for 96 h, and this resulted in polygalacturonase activities of up to 256 U mL⁻¹ in the wet media. Afterwards, water was added to the bioreactor, and the remaining solids were suspended by stirring to dissolve the hydrolytic enzymes. Metabolic activities of A. niger were inactivated by a N₂-atmosphere and by increasing the temperature to 50 °C. High solid loads of milled SBPP were added to the stirred-tank reactor with a delay of 24 h to enable sugar yield calculations based on the compositional analysis of the SBPP used. The resulting final sugar concentrations of the hydrolysate after 166 h were 17 g L⁻¹ d-glucose, 18.8 g L⁻¹ l-arabinose, and 12.5 g L⁻¹ d-galacturonic acid, corresponding to sugar yields of 98% d-glucose, 86% l-arabinose, and 50% d-galacturonic acid, respectively. Including the other sugars released during enzymatic hydrolysis in the one-pot process (d-xylose, d-mannose, d-galactose), a total sugar concentration of 54.8 g L⁻¹ was achieved in the hydrolysate. The one-pot process combining hydrolytic enzyme production in solid-state fermentation with high solid loads during enzymatic hydrolysis of the milled SBPP reduces hydrolytic process costs by replacing chemical pre-treatments, enabling the in situ production of SBPP-adapted hydrolytic enzymes, as well as avoiding intermediate enzyme extraction and preparation steps. Full article

This study aimed to explore the viability of converting potato peel waste into bioethanol using a custom-produced multi-enzyme preparation. Various pretreatment approaches were employed on the potato peels, including thermal, chemical, and thermo-chemical methods. These methods involved boiling for 30 and 60 min, steaming at different pressures and durations, and applying different concentrations of chemicals, including H₂SO₄, HNO₃, CH₃COOH, HCl, NaOH, Ca(OH)₂, KOH, NH₃, and H₂O₂, either individually or in combination with steam treatment. The pretreated potato peels were subsequently subjected to enzymatic hydrolysis using a crude multi-enzyme cocktail obtained from solid-state fermentation of wheat bran by a naturally occurring strain of Aspergillus niger P-19. This enzyme cocktail consisted of cellulases, hemicellulases, pectinase, and amylases. The most effective pretreatment combination involved the use of 3% H₂SO₄ followed by steam treatment under pressure, and enzymatic hydrolysis utilizing the crude multi-enzyme preparation. This combination resulted in the highest concentration of reducing sugars (141.04 ± 12.31 g/L), with a carbohydrate conversion rate of 98.49% when a substrate loading of 20% was used. As a result, an ethanol yield of 43.2 ± 3.82 g/L, representing 21.6% based on dry matter, was achieved. Furthermore, supplementing the medium with peptone, (NH₄)(H₂PO₄), and ZnSO₄ at a concentration of 0.1% w/v each, along with solid loadings of 22% and 24%, respectively, achieved yield improvements of 51.67 g/L and 54.75 g/L. However, the maximum productivity of 23.4% was observed with a 22% loading, compared to a yield of 22.8% with a 24% solid loading, based on dry matter. Full article

Background: Lipase is one of the most important enzymes produced from microbial fermentation. Agricultural wastes are a good source of enzyme production because they are cost-effective and production rates are also higher. Method: In this study, eight lignolitic substrates were screened for lipase production. Results: Out of these substrates, guava leaves showed maximum activity of 9.1 U/mL from Aspergillus niger by using the solid-state fermentation method. Various factors such as temperature, pH, incubation period, moisture content, inoculum size, and substrate size that influence the growth of fungi were optimized by response surface methodology (RSM), and then characterization was performed. When all physical and nutritional parameters were optimized by RSM, the maximum lipase activity obtained was 12.52 U/mL after 4 days of incubation, at pH 8, 40 °C temperature, 3 mL inoculum size, 20% moisture content, and 6 g substrate concentration. The enzyme was partially purified through 70% ammonium sulfate precipitation. After purification, it showed 34.291 U/mg enzyme activity, increasing the purification fold to 1.3. The enzyme was then further purified by dialysis, and the purification fold increased to 1.83 having enzyme activity of 48.03 U/mg. Furthermore, activity was increased to 132.72 U/mg after column chromatography. A purification fold of 5.07 was obtained after all purification steps. Full article

The present work describes the purification of an enzyme capable of degrading punicalagin. The enzyme was produced by Aspergillus niger GH1 by solid-state fermentation, and the enzyme production was induced by using ellagitannins as the sole carbon source. The purification steps included the concentration by lyophilization, desalting, anionic exchange, and gel filtration chromatography. The enzyme kinetic constants were calculated by using punicalagin, methyl gallate, and sugar beet arabinans. The molecular mass of the protein was estimated by SDS-PAGE. The identified bands were excised and digested using trypsin, and the peptides were submitted to HPLC-MS/MS analysis. The docking analysis was conducted, and a 3D model was created. The purification fold increases 75 times compared with the cell-free extract. The obtained Km values were 0.053 mM, 0.53% and 6.66 mM for punicalagin, sugar beet arabinans and methyl gallate, respectively. The optimal pH and temperature for the reaction were 5 and 40 °C, respectively. The SDS-PAGE and native PAGE analysis revealed the presence of two bands identified as α-l-arabinofuranosidase. Both enzymes were capable of degrading punicalagin and releasing ellagic acid. Full article

The large family of enzymes, known as polyphenols oxidases, includes laccase. Due to the inclusion of a copper atom in their catalytic core, laccases are frequently referred to as multi-copper oxidases. Laccases are versatile enzymes that can catalyze the oxidation of a wide range of phenolic and non-phenolic substances. In the current study, a local strain of Aspergillus niger was used for solid-state fermentation to produce fungal laccase, as well as purify and optimize laccase. The enzyme profile, which was acquired using guaiacol to measure enzyme activity, showed that after five days of incubation, wheat straw provided the highest level of laccase activity, or 2.551 U/mL. A technique called response surface methodology (RSM) was used to examine the effects of various conditions on the production of enzymes. The RSM results demonstrated that after five days of incubation, the enzyme activity was at its highest at 45 °C, pH 5.5, and 30% moisture level, inoculated with 2 mL mycelium. Through ammonium sulphate precipitation and dialysis, the enzyme was purified. Additionally, column chromatography was used to further purify laccase. The next step was enzyme characterization to evaluate how temperature and pH affected enzyme activity. At 45 °C and pH 5.5, the isolated enzyme produced its highest level of activity. The findings of the current study showed that A. niger is capable of producing laccase in an economical and environmentally friendly way. Due to its unique oxidative and catalytic features, this enzyme has received a lot of attention recently. Full article

The ability of Aspergillus niger strains to support citric acid production using solid-state fermentation of agricultural processing coproducts was examined in this review. Citric acid has been shown to have a number of commercial applications in the food and beverage industries. The A. niger strains capable of elevated citric acid production are known to contain genetic mutations that stimulate overproduction of the organic acid likely involving citric acid cycle reactions. The agricultural processing coproducts previously examined for their ability to support citric acid production by A. niger solid-state fermentation include fruit processing wastes, sugarcane bagasse, starch vegetable processing wastes and cereal grain processing coproducts. A comparison of citric acid production by A. niger strains using solid-state fermentation demonstrated that certain agricultural processing coproducts were more effective in supporting a high level of acid synthesis. In particular, fruit processing wastes, such as apple pomace, banana peels, grape pomace and orange peels, supported high levels of citric acid by the fungal strains following solid-state fermentation. On the other hand, processing coproducts of cereal grains, such as brans and ethanol processing coproducts, supported low levels of citric acid production by the A. niger strains using solid-state fermentation. It appeared that the cereal processing coproducts provided less available sugar content to support citric acid production by the fungal strains. It was concluded that the level of citric acid produced by the A. niger strains during solid-state fermentation was dependent on the sugar content of the agricultural processing coproduct utilized. Full article