Studies on the Thermal Stability of Peroxidase from Leaf of Oil Palm (Elaeis guineensis)

Peroxidase was extracted from leaves of oil palm tree with 0.01M phosphate buffer pH 7.0. It was partially purified using 70% ammonium sulphate ((NH4)2SO4) precipitation. This resulted in peroxidase with activity of (26U/ml) and specific activity of 35.8U/mg. Effect of heat on the activity of peroxidase was studied at temperature of 323-363°K. After gel filtration on sephadex G100, the peroxidase activity increased to 27U/ml, with specific activity of 55U/mg .The overall purification fold was 4 with 51.9% enzyme recovery. The peroxidase partially purified from leaves of oil palm tree showed pH and temperature optima of 5.0 and 50°C respectively. High pH and temperature stabilities of pH 5.0 to pH 9.0 and 50°C to 70°C were obtained respectively. Also, the activation energy (Ea) of the reaction was - 21.616kj/mol. The free energy changes (ΔG) were 96008.64, 96315.59, 97901.63, 94132.33 and 97146.75kj/mol at 323,333,343,353 and 363°K respectively. It was observed that the D-values were decreasing with increasing temperature with a Z-value of 0.044. The enthalpy results suggest that the reaction was exothermic, non-spontaneous and reversible.

Development of a Hybrid Bioinorganic Nanobiocatalyst: Remarkable Impact of the Immobilization Conditions on Activity and Stability of β-galactosidase

Hybrid bioinorganic biocatalysts have received much attention due to their simple synthesis, high efficiency, and structural features that favor enzyme activity and stability. The present work introduces a biomineralization strategy for the formation of hybrid nanocrystals from β-galactosidase. The effects of the immobilization conditions were studied, identifying the important effect of metal ions and pH on the immobilization yield and the recovered activity. For a deeper understanding of the biomineralization process, an in silico study was carried out to identify the ion binding sites at the different conditions. The selected β-galactosidase nanocrystals showed high specific activity (35,000 IU/g biocatalyst) and remarkable thermal stability with a half-life 11 times higher than the soluble enzyme. The nanobiocatalyst was successfully tested for the synthesis of galacto-oligosaccharides, achieving an outstanding performance, showing no signs of diffusional limitations. Thus, a new, simple, biocompatible and inexpensive nanobiocatalyst was produced with high enzyme recovery (82%), exhibiting high specific activity and high stability, with promising industrial applications.

One-pot green synthesis of iron oxide nanoparticles from Bauhinia tomentosa: Characterization and application towards synthesis of 1, 3 diolein

The green synthesis of NPs through plant extracts can be a modest, one-pot alternative synthesis to the conventional physical or chemical method. The prime focus of this study is to produce MNPs by the reducing effect of Bauhinia tomentosa leaf extract, and it was immobilized in porcine pancreatic lipase (PPL). Synthesized NPs were characterized by field emission scanning electron microscopy (FESEM), X-ray diffraction (XRD) and Raman spectroscopy, UV–Vis Spectrometry, Thermogravimetry, and Differential Scanning Calorimeter (DSC), Zeta potential test, VSM, BET and Fourier Transform Infrared Spectroscopy (FTIR). The effect of process parameters was studied, about the efficiency of immobilization are enzyme stability, the extent of enzyme reusability, its separation from products, the activity of immobilized enzyme, recovery, and its loss. Finally, the immobilized lipase was used for the synthesis of 1,3-diolein using enzyme-mediated esterification of oleic acid and glycerol. Under optimized condition (reaction temp-55 $$^\circ $$ ∘ C; molar ratio-2.5:1; pH-7) diolein yield was achieved to be 94%. Therefore, this work was further used for the industrial production of 1,3-diacylglycerol since a perfect enzyme-catalyzed process was observed.

Immobilization of Thermoalkalophilic Esterase Onto Magnetic-Cornstarch Nanoparticle

The immobilization of the biocatalysts onto magnetic nanoparticles (MNPs) has been extensively applied since the external magnetic field facilitates the enzyme recovery from the reaction mixture. In the present study, glutaraldehyde-modified magnetite-cornstarch nanoparticles (MCNs) were successfully synthesized, elaborately characterized by ZetaSizer and surface-enhanced raman spectroscopy (SERS), and used for the immobilization of a thermoalkalophilic esterase from Geobacillus sp. The optimal immobilization conditions were obtained at 65oC, 2:3 molar ratios of Fe2+:Fe3+ and 1 g cornstarch resulted in approximately 90 nm magnetic particles in size. Also, immobilization yield and entrapment efficiency of the esterase were found as 74% and 82%, respectively. Scanning Electron Microscopy (SEM) micrographs showed that MCNs were uniform, spherical in shape, and well dispersed and esterase immobilized MCNs displayed similar morphology as free MCNs. The maximum activity of free and immobilized esterase was obtained at 80oC and pH 9. Immobilization onto glutaraldehyde-modified MCNs significantly enhanced the esterase thermostability. Additionally, the immobilized esterase kept its residual activity of 75% after three sequential cycles, suggesting that it has favorable operational stability.

Enzyme Recovery from Biological Wastewater Treatment

Enzymes are high value industrial bio-catalysts with extensive applications in a wide range of manufacturing and processing sectors, including the agricultural, food and household care industries. The catalytic efficiency of enzymes can be several orders higher compared to inorganic chemical catalysts under mild conditions. However, the nutrient medium necessary for biomass culture represents a significant cost to industrial enzyme production. Activated sludge (AS) is a waste product of biological wastewater treatment and consists of microbial biomass that degrades organic matter by producing substantial quantities of hydrolytic enzymes. Therefore, enzyme recovery from AS offers an alternative, potentially viable approach to industrial enzyme production. Enzyme extraction from disrupted AS flocs is technically feasible and has been demonstrated at experimental-scale. A critical review of disruption techniques identified sonication as potentially the most effective and suitable method for enzyme extraction, which can be scaled up and is a familiar technology to the water industry. The yields of different enzymes are influenced by wastewater treatment conditions, and particularly the composition, and can also be controlled by feeding sludge with specific target substrates. Nevertheless, hydrolytic enzymes can be effectively extracted directly from waste AS without specific modifications to standard wastewater treatment processes. Purification, concentration and stabilisation/immobilisation techniques can also greatly expand the industrial application and increase the economic value and marketability of enzyme products extracted from AS. Concentrated and purified AS enzymes could readily substitute inorganic and/or commercial bioenzyme catalysts in many industrial applications including, for example, leather processing, and in detergent and animal feed formulation. Enzyme extraction from AS therefore offers significant economic benefits to the Water Industry by recovering valuable resources from wastewater. They can also optimise important waste treatment processes, such as the anaerobic digestion (AD) of sewage sludge, increasing biogas and renewable energy production. The enzyme-extracted sludge exhibits improved treatment properties, such as increased settleability, dewaterability, and anaerobic digestibility for biogas production, assisting sludge management by wastewater treatment plants (WWTPs) and enabling the further utilisation of the residual sludge. Graphic Abstract

Valorization of Wheat Byproducts for the Co-Production of Packaging Material and Enzymes

Waste management systems are overloaded with huge streams of plastic, a large part of this being originated from packaging. Additionally, the production of wheat, one of the most cultivated crops in the world, generates low-value lignocellulosic materials, which are mostly discarded. In this study, the wheat lignocellulosic byproducts straw and bran were used for the co-production of enzymes and bio-based materials with possible application as packaging via the compression molding method. The mechanical properties of the films were studied based on the effects of the removal of lignin by alkali and biological pretreatment, the growth of filamentous fungi, the size of the particles, and the enzyme recovery. Generally, the straw films were stiffer than the bran ones, but the highest Young’s modulus was obtained for the biologically pretreated bran (1074 MPa). The addition of a step to recover the fungal cellulases produced during the cultivation had no statistical effect on the mechanical properties of the films. Moreover, alkali and biological pretreatments improved the anaerobic biodegradability of the straw films. Thus, the wheat bran and straw can be used for the co-production of enzymes, materials, and biogas, potentially changing how wheat and packaging wastes are managed.