Yeast-driven whey biorefining to produce value-added aroma, flavor, and antioxidant compounds: technologies, challenges, and alternatives.

Whey is a liquid residue generated during the production of cheese and yogurt. It has a pH between 3.9 and 5.6, and a high chemical oxygen demand (COD), from 60 to 80 g/L. Whey contains lactose, proteins, and minerals. Globally, approximately 50% of the whey generated is untreated and is released directly into the environment, which represents an environmental risk. To overcome whey management problems, conventional thermo-physical valorization treatments have been explored, which are complex, costly and energy-intensive. As an alternative, whey fermentation processes employing bacteria, fungi and yeast are economical and promising methods. Among them, yeast fermentation creates value-added products such as antimicrobials, biofuels, aromas, flavors, and antioxidants with no need for previous conditioning of the whey, such as hydrolysis of the lactose, prior to whey biorefining. The biorefining concept applied to whey is discussed using chemical and biological transformation pathways, showing their pluses and minuses, such as technical drawbacks. The main challenges and solutions for the production of fusel alcohols, specifically for 2-phenylethanol, are also discussed in this review.

Immobilized laccase on polyimide aerogels for removal of carbamazepine.

Since it is known that conventional wastewater treatment plants cannot completely remove pharmaceutical compounds, such as carbamazepine, the need for their removal has intensified. The use of biocatalysts, such as enzyme is an environmentally friendly method for carbamazepine biodegradation. Nevertheless, enzyme immobilization is required to facilitate the recovery and reusability and avoid the loss of enzyme. In this work, laccase was immobilized on modified polyimide aerogels by means of covalent bonding. Results showed that the immobilized laccase on polyimide aerogels possesses significantly improved activity under acidic or basic pH range in comparison with the free enzyme. Furthermore, for all the temperature range the activity of the immobilized enzyme was higher compared to the free enzyme form. The storage stability improved by the immobilization on this support material. The reusability tests towards oxidation of 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonicacid) (ABTS) showed that the immobilized laccase maintained 22% of the initial activity after 7 cycles. Immobilized laccase on polyimide aerogels for carbamazepine (CBZ) degradation exhibited 76% and 74% removal in spiked water and secondary effluent, respectively. Furthermore, after 7 cycles the CBZ removal efficiency remained higher (50% and 65% for spiked water and secondary effluent, respectively).

Fabrication of nanobiocatalyst using encapsulated laccase onto chitosan-nanobiochar composite.

Laccase is one of the widely used enzymes for biotechnological processes. Immobilization of enzymes is a universally accepted approach to increase their reusability and stability. In this study, laccase enzyme from Trametes versicolor was encapsulated for the first time in a chitosan-nanobiochar matrix. The chitosan-tripolyphosphate gel formation technique was employed to produce homogeneous biocatalyst nanoparticles, with 35% effective binding efficiency and 3.5 Units/g apparent activity under the best configuration. The reusability of the encapsulated laccase was demonstrated towards the oxidation of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS) for several consecutive cycles, exhibiting 30% of the initial activity after 5 cycles. The encapsulated laccase showed a moderate increase in enzyme stability against pH and temperature variation compared to the free enzyme. Moreover, the storage stability of laccase at both 4 °C and 25 °C was increased after immobilization. Only 2% of laccase was leaked during a 5-day period from biocatalyst. Laccase in its free form showed no antibacterial activity against Gram positive and Gram-negative model microorganisms, while encapsulated laccase showed antibacterial activity towards Gram-positive ones. Thus, the encapsulation of the laccase is an efficient method to keep the enzyme active and stable for different applications.

Pinewood nanobiochar: A unique carrier for the immobilization of crude laccase by covalent bonding.

Nanotechnology-inspired biocatalytic systems attracted attention for many applications since nanosized supports for enzyme immobilization can improve efficiency-determining factors e.g. enhancing the surface area and loading capacity and reducing the mass transfer resistance. Among the nanomaterials, nanobiochar has unique features as a support for enzyme immobilization i.e. high surface to volume ratio, porous structure, and presence of functional groups on its surface. However, the performance of the immobilization is highly dependent on the immobilization conditions and the properties of the enzyme and the support material. In this research, crude laccase was covalently immobilized onto functionalized nanobiochar using a two-step method of diimide-activated amidation. The effect of different parameters was investigated. The optimal conditions were found to be 14 mg/mL of laccase concentration, 5 mg/mL of nanobiochar, 8.2 mM of cross-linker and 3 h of contact time. For investigating the pH, thermal, storage, and operational stability, the sample obtained from the optimized conditions was used. The results showed the higher stability of immobilized laccase against temperature and pH variation compared to free laccase. In addition, immobilized laccase maintained its catalytic performance up to seven cycles of utilization and showed more than 50% of initial activity after two months of room temperature storage.

Removal of pharmaceutical compounds in water and wastewater using fungal oxidoreductase enzymes.

Due to recalcitrance of some pharmaceutically active compounds (PhACs), conventional wastewater treatment is not able to remove them effectively. Therefore, their occurrence in surface water and potential environmental impact has raised serious global concern. Biological transformation of these contaminants using white-rot fungi (WRF) and their oxidoreductase enzymes has been proposed as a low cost and environmentally friendly solution for water treatment. The removal performance of PhACs by a fungal culture is dependent on several factors, such as fungal species, the secreted enzymes, molecular structure of target compounds, culture medium composition, etc. In recent 20 years, numerous researchers tried to elucidate the removal mechanisms and the effects of important operational parameters such as temperature and pH on the enzymatic treatment of PhACs. This review summarizes and analyzes the studies performed on PhACs removal from spiked pure water and real wastewaters using oxidoreductase enzymes and the data related to degradation efficiencies of the most studied compounds. The review also offers an insight into enzymes immobilization, fungal reactors, mediators, degradation mechanisms and transformation products (TPs) of PhACs. In brief, higher hydrophobicity and having electron-donating groups, such as amine and hydroxyl in molecular structure leads to more effective degradation of PhACs by fungal cultures. For recalcitrant compounds, using redox mediators, such as syringaldehyde increases the degradation efficiency, however they may cause toxicity in the effluent and deactivate the enzyme. Immobilization of enzymes on supports can enhance the performance of enzyme in terms of reusability and stability. However, the immobilization strategy should be carefully selected to reduce the cost and enable regeneration. Still, further studies are needed to elucidate the mechanisms involved in enzymatic degradation and the toxicity levels of TPs and also to optimize the whole treatment strategy to have economical and technical competitiveness.

Immobilized laccase on oxygen functionalized nanobiochars through mineral acids treatment for removal of carbamazepine.

Biocatalytic treatment with oxidoreductase enzymes, especially laccases are an environmentally benign method for biodegradation of pharmaceutical compounds, such as carbamazepine to less harmful compounds. However, enzymes are required to be immobilized on supports to be reusable and maintain their activity. Functionalization of support prior to immobilization of enzyme is highly important because of biomolecule-support interface on enzyme activity and stability. In this work, the effect of oxidation of nanobiochar, a carbonaceous material produced by biomass pyrolysis, using HCl, H2SO4, HNO3 and their mixtures on immobilization of laccase has been studied. Scanning electron microscopy indicated that the structure of nanobiochars remained intact after oxidation and Fourier transform infrared spectroscopy confirmed the formation of carboxylic groups because of acid treatment. Titration measurements showed that the sample treated with H2SO4/HNO3 (50:50, v/v) had the highest number of carboxylic groups (4.7mmol/g) and consequently the highest efficiency for laccase immobilization. Additionally, it was observed that the storage, pH and thermal stability of immobilized laccase on functionalized nanobiochar was improved compared to free laccase showing its potential for continuous applications. The reusability tests towards oxidation of 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) showed that the immobilized laccase preserved 70% of the initial activity after 3cycles. Finally, using immobilized laccase for degradation of carbamazepine exhibited 83% and 86% removal in spiked water and secondary effluent, respectively.

A review on the important aspects of lipase immobilization on nanomaterials.

Lipase is one of the most widely used enzymes and plays an important role in biotechnological and industrial processes including food, paper, and oleochemical industries, as well as in pharmaceutical applications. However, its aqueous solubility and instability make its application relatively difficult and expensive. The immobilization technique is often used to improve lipase performance, and the strategy has turned out to be a promising method. Immobilized lipase on nanomaterials (NMs) has shown superiority to the free lipase, such as improved thermal and pH stability, longer stable time, and the capacity of being reused. However, immobilization of lipase on NMs also sometimes causes activity loss and protein loading is relatively lowered under some conditions. The overall performance of immobilized lipase on NMs is influenced by mechanisms of immobilization, type of NMs being used, and physicochemical features of the used NMs (such as particle size, aggregation behavior, NM dimension, and type of coupling/modifying agents being used). Based on the specific features of lipase and NMs, this review discusses the recent developments, some mechanisms, and influence of NMs on lipase immobilization and their activity. Multiple application potential of the immobilized lipases has also been considered.

Development of adsorptive membranes by confinement of activated biochar into electrospun nanofibers.

Adsorptive membranes have many applications in removal of contaminants, such as heavy metals and organic contaminants from water. Recently, increasing concentrations of pharmaceutically active compounds, especially antibiotics, such as chlortetracycline in water and wastewater sources has raised concerns about their potentially adverse impacts on environment and human health. In this study, a series of polyacrylonitrile (PAN)/activated biochar nanofibrous membranes (NFMs) with different loadings of biochar (0-2%, w/w) were fabricated using electrospinning. The morphology and structure of fabricated membranes was investigated by scanning electron microscopy, Fourier transform infrared and thermogravimetric analysis. The results showed that at 1.5% of biochar loading, the surface area reached the maximum value of 12.4 m2/g and beyond this loading value, agglomeration of particles inhibited fine interaction with nanofibrous matrix. Also, the adsorption tests using chlortetracycline showed that, under environmentally relevant concentrations, the fabricated adsorptive NFMs had a potential for removal of these types of emerging contaminants from water and wastewaters.

Green and energy-efficient methods for the production of metallic nanoparticles.

In the last decade, researchers paid great attention to the concept of "Green Chemistry", which aims at development of efficient methods for the synthesis of nanoparticles (NPs) in terms of the least possible impact on human life and environment. Generally, several reagents including precursors, reducing agents, stabilizing agents and solvents are used for the production of NPs and in some cases, energy is needed to reach the optimum temperature for reduction. Therefore, to develop a green approach, researchers had the opportunity to investigate eco-friendly reagents and new energy transfer techniques. In order to substitute the harmful reagents with green ones, researchers worked on different types of saccharides, polyols, carboxylic acids, polyoxometalates and extracts of various plants that can play the role of reducers, stabilizers or solvents. Also, there are some reports on using ultraviolet (UV), gamma and microwave irradiation that are capable of reducing and provide uniform heating. According to the literature, it is possible to use green reagents and novel energy transfer techniques for production of NPs. However, these new synthesis routes should be optimized in terms of performance, cost, product quality (shape and size distribution) and scale-up capability. This paper presents a review on most of the employed green reagents and new energy transfer techniques for the production of metallic NPs.

A sulfuric-lactic acid process for efficient purification of fungal chitosan with intact molecular weight.

The most recent method of fungal chitosan purification, i.e., two steps of dilute sulfuric acid treatment, pretreatment of cell wall at room temperature for phosphate removal and extraction of chitosan from the phosphate free cell wall at high temperature, significantly reduces the chitosan molecular weight. This study was aimed at improvement of this method. In the pretreatment step, to choose the best conditions, cell wall of Rhizopus oryzae, containing 9% phosphate, 10% glucosamine, and 21% N-acetyl glucosamine, was treated with sulfuric, lactic, acetic, nitric, or hydrochloric acid, at room temperature. Sulfuric acid showed the best performance in phosphate removal (90%) and cell wall recovery (89%). To avoid depolymerisation of chitosan, hot sulfuric acid extraction was replaced with lactic acid treatment at room temperature, and a pure fungal chitosan was obtained (0.12 g/g cell wall). Similar pretreatment and extraction processes were conducted on pure shrimp chitosan and resulted in a chitosan recovery of higher than 87% while the reduction of chitosan viscosity was less than 15%. Therefore, the sulfuric-lactic acid method purified the fungal chitosan without significant molecular weight manipulation.