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.

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.

Delonix regia (common name: Flame tree) pods, an inexpensive lignocellulosic waste matrix, were successfully used to produce value-added bioethanol. Initially, the potentiality of D. regia pods as a lignocellulosic biomass was assessed by Fourier-transform infrared spectroscopy (FTIR), which revealed the presence of several functional groups belonging to cellulose, hemicellulose, and lignin, implying that D. regia pods could serve as an excellent lignocellulosic biomass. Response Surface Methodology (RSM) and Central Composite Design (CCD) were used to optimize pretreatment conditions of incubation time (10–70 min), H₂SO₄ concentration (0.5–3%), amount of substrate (0.02–0.22 g), and temperature (45–100 °C). Then, RSM-suggested 30 trials of pretreatment conditions experimented in the laboratory, and a trial using 0.16 g substrate, 3% H₂SO₄, 70 min incubation at 90 °C, yielded the highest amount of glucose (0.296 mg·mL⁻¹), and xylose (0.477 mg·mL⁻¹). Subsequently, the same trial conditions were chosen in the downstream process, and pretreated D. regia pods were subjected to enzymatic hydrolysis with 5 mL of indigenously produced cellulase enzyme (74 filter per unit [FPU]) at 50 °C for 72 h to augment the yield of fermentable sugars, yielding up to 55.57 mg·mL⁻¹ of glucose. Finally, the released sugars were fermented to ethanol by Saccharomyces cerevisiae, yielding a maximum of 7.771% ethanol after 72 h of incubation at 30 °C. Conclusively, this study entails the successful valorization of D. regia pods for bioethanol production.

The biotechnological process called solid-state fermentation (SSF) was applied for hyper production of protease by using a fungal strain called Aspergillus oryzae. From screening of 9 different local substrates (peanut shell, wheat bran, guava leaves, sugar cane bagasse, rice polish, wheat straw, corn straw, reed grass, and rice straw), peanut shells serve as the best substrates for protease production under optimized cultured conditions. The varying physiochemical parameters such as pH (2-9.5), temperature (30-52 °C), incubation time (1-10 days), inoculum size (1-8 mL), moisture level (20-125%), and substrate concentration (1-7 g) were optimized by response surface methodology (RSM). The highest activity of protease was recorded to be 1101.778 U/mL at 660 nm using peanut shell was optimum at pH 8, temperature 52 °C, incubation time 8 days, inoculum size 2 mL, moisture level 20%, and substrate concentration 2 g. The crude form of enzymes produced were further purified through ammonium sulfate precipitation, dialysis, and gel filtration chromatography. Then, purified enzymes were characterized at different pH, temperature, and incubation time. For characterization of purified protease, pH, temperature, and incubation time were 8, 52 °C, and 8 days for peanut shell and was done by one factor at a time method. Hence, isolated enzymes were alkaline in nature, i.e., alkaline proteases. Then, protease produced from peanut shells was applied to locally available detergents to increase their catalytic activity for strain removal. At last, the final results were interpreted in the form of 3D surface and contour plots using Microsoft Excel 2013 and Minitab 17 software. In conclusion, the utilization of A. oryzae and peanut shell as the substrate in the biotechnological process of SSF demonstrated successful hyper production of alkaline protease. The optimized conditions resulted in high enzyme activity and showcased the potential application of the isolated enzymes in improving the catalytic activity of locally available detergents.

Biofuels represent a sustainable alternative to fossil fuels, with non-food feedstock offering particular promise. This study evaluated Pongamia pinnata seeds as a dual feedstock for biodiesel and bioethanol production. Moisture content was determined at 8.0%, favorably lower than most lignocellulosic biomasses. FTIR analysis confirmed the presence of cellulose, hemicellulose, and lignin, indicating its suitability for biofuel production. Biodiesel was produced via alkaline transesterification, with the highest conversion (70%) obtained at a 5:1 alcohol-to-oil ratio. The de-oiled seed cake, rich in carbohydrates and low in lignin, was optimized for glucose release through Response Surface Methodology. Optimal conditions (100°C, 2.5 g substrate, 1.75% H ₂ SO ₄ , 60 min) yielded 429 mg g ⁻¹ glucose. Enzymatic hydrolysis further enhanced sugar release, with maximum glucose (118.21 mg g ⁻¹ ) achieved at 72 h with a 2.0 mL enzyme load. Subsequent fermentation with Saccharomyces cerevisiae produced 6.71% ethanol. These findings highlight the potential of P. pinnata as a feedstock for integrated biorefineries, supporting sustainable biofuel production.