The present study assessed the metabolites and bioactivities of Micromeria fruticosa plant parts extracted with various solvents (ethanol, n-hexane, and water) through the steeping extraction method. Thereafter, the extracts were analyzed using GC-MS. Moreover, the extracts were tested for their antioxidant, antimicrobial, and antitumor activities. The quali-quantitative analysis of Micromeria fruticosa crude extracts revealed the occurrence of 27 secondary metabolites. Some major bioactives identified were menthone, oleamide, pulegone, and menthol. Numerous antioxidant minerals, viz., Fe, Zn, and Mn, were present. The water extract of leaves exhibited the highest DPPH scavenging activity (89.73%), followed by the water extract of flowers (80.07%) at 100 μg/mL. The stems’ water extract demonstrated greater antimicrobial activity against all the bacteria species tested. The ethanolic leaf and aqueous stem extracts exhibited strong activity against C. albicans and E. coli. Flowers’ aqueo... [more]

Non-thermal technologies allow for the nutritional and sensory properties of foods to be preserved, something that consumers demand. Combining their use with antimicrobial peptides (AMPs) provides potential methods for food preservation that could have advantages over the use of chemical preservatives and thermal technologies. The aim of this review was to discuss the advances in the application of non-thermal technologies in combination with AMPs as a method for microbial inactivation. Published papers reporting studies on the combined use of power ultrasound (US), pulsed electrical fields (PEF), and high hydrostatic pressure (HHP) with AMPs were reviewed. All three technologies show a possibility of being combined with AMPs, generally demonstrating higher efficiency than the application of US, PEF, HHP, and AMPs separately. The most studied AMP used in combination with the three technologies was nisin, probably due to the fact that it is already officially regulated. However, the com... [more]

Carob syrup is one of the most important carob products, which can have applications in pastry and confectionery, as a fruit preservative, but also in the pharmaceutical field because of the antimicrobial activity due to its polyphenol content. Carob syrup is traditionally made through a very time-consuming process, involving solid−liquid extraction in boiling water and concentration at a high temperature (>100 °C), which potentially causes the degradation of the active compounds (i.e., procyanidins or flavonol glycosides). Therefore, in this work, an alternative and less drastic method based on ultrasound technology was proposed to produce carob syrup. Processing conditions (i.e., time, temperature, and liquid−solid ratio) influencing the extraction of total soluble solids (TSS) and total phenolic compounds (TPC) were optimized using a central composite design coupled to response surface methodology. Reliable mathematical models allowed us to predict the highest TSS (24 ± 2 °Brix) and... [more]

The integration of insect-derived extracts in feed and food products has become a field of growing interest in recent years. In this review, we collect different studies carried out on edible insects’ transformation processes and focus on the various treatment operations, extraction technologies, and solvents used in different processing steps. We include an overview of current insights into the different steps of the transformation process: insect reception, killing methods, pretreatments, storage, delipidation, protein extraction, as well as chitin and chitosan extraction. Finally, we reflect on the most important future challenges of this sector.

Coronaviruses have caused devastation in both human and animal populations, affecting both health and the economy. Amidst the emergence and re-emergence of coronaviruses, humans need to surmount the health and economic threat of coronaviruses through science and evidence-based approaches. One of these approaches is through biotechnology, particularly the heterologous production of biopharmaceutical proteins. This review article briefly describes the genome, general virion morphology, and key structural proteins of different coronaviruses affecting animals and humans. In addition, this review paper also presents the different systems in recombinant protein technology such as bacteria, yeasts, plants, mammalian cells, and insect/insect cells systems used to express key structural proteins in the development of countermeasures such as diagnostics, prophylaxis, and therapeutics in the challenging era of coronaviruses.

The large amounts of organic waste thrown into the garbage without any productivity, and the increase in the demand for electrical energy worldwide, has led to the search for new eco-friendly ways of generating electricity. Because of this, microbial fuel cells have begun to be used as a technology to generate bioelectricity. The main objective of this research was to generate bioelectricity through banana waste using a low-cost laboratory-scale method, achieving the generation of maximum currents and voltages of 3.71667 ± 0.05304 mA and 1.01 ± 0.017 V, with an optimal pH of 4.023 ± 0.064 and a maximum electrical conductivity of the substrate of 182.333 ± 3.51 µS/cm. The FTIR spectra of the initial and final substrate show a decrease in the peaks belonging to phenolic compounds, alkanes, and alkenes, mainly. The maximum power density was 5736.112 ± 12.62 mW/cm2 at a current density of 6.501 A/cm2 with a peak voltage of 1006.95 mV. The molecular analysis of the biofilm formed on the ano... [more]

Newcastle disease is a viral infection causing serious economic losses to the global poultry industry. The V protein of Newcastle disease virus (NDV) is a pathogenicity determinant having various functions such as the suppression of apoptosis and replication of the NDV. This study was designed to assess the resistance potential of plant flavonoids against the V protein of Newcastle disease virus. Sequence analysis was performed using EXPASY and ProtParam tools. To build the three-dimensional structure of V protein, a homology-modeling method was used. Plant flavonoids with formerly reported therapeutic benefits were collected from different databases to build a library for virtual screening. Docking analysis was performed using the modeled structure of V protein on MOE software. Interaction analysis was also performed by MOE to explain the results of docking. Sequence analysis and physicochemical properties showed that V protein is negatively charged, acidic in nature, and relatively u... [more]

A “foldamer” is an artificial oligomeric molecule with a regular secondary or tertiary structure consisting of various building blocks. A “stapled peptide” is a peptide with stabilized secondary structures, in particular, helical structures by intramolecular covalent side-chain cross-linking. Helical foldamers and stapled peptides are potential drug candidates that can target protein-protein interactions because they enable multipoint molecular recognition, which is difficult to achieve with low-molecular-weight compounds. This mini-review describes a variety of peptide-based foldamers and stapled peptides with a view to their applications in drug discovery, including our recent progress.

The local gas holdup details and behaviors in the cylindrical split airlift column by using an unconventional gamma-ray densitometry (GRD) measurement in non-invasive manner technique was investigated for the first time in this work for such kind of airlift column. With different gas velocities, 1, 2, and 3 cm/s, at three various axial planes (different levels) in z = 3, 60, and 110 cm were studied for local distribution in radial gas holdup profiles. The distribution in gas−liquid phases (air-water system) in the entire split reactor column, in the rising and descending sides, including their behavior in the upper and lower zones of the split plate, were investigated as well. The results of this study showed that approximately all reactor zones had exemplary gas−liquid phases and that there was a large magnitude over both the dividing ring and the top sections. The results further indicated that the distribution of which flow variable in the implementation of the cylindrical split rea... [more]

Paclitaxel (PX), plant alkaloid, is a chemotherapeutic agent intended for treating a wide variety of cancers. The objective of the present study was to formulate and evaluate the anticancer activity of PX loaded into a nanocarrier, mainly PEGylated nanoemulsion (NE) fabricated with myrrh essential oil. Myrrh essential oil has been estimated previously to show respectable anticancer activity. Surface modification of the formulation with PEG-DSPE would help in avoiding phagocytosis and prolong the residence time in blood circulation. Various NE formulations were developed after operating (22) factorial design, characterized for their particle size, in vitro release, and hemolytic activity. The optimized formula was selected and compared to its naked counterpart in respect to several characterizations. Quantitative amount of protein absorbed on the formulation surfaces and in vitro release with and without serum incubation were evaluated. Ultimately, MTT assay was conducted to distinguish... [more]

In this study, the evolution of the yeast microflora present on the berry surface, during the ripening of Barbera grapes, was monitored. Sampling was performed in three vineyards located in the “Nizza” Barbera d’Asti DOC zone and different methodologies have been employed. A culture-dependent method based on the identification of strains grown on solid media by ARDRA (Amplified Ribosomal DNA Restriction Analysis) and the D1-D2 domain of ribosomal 26S DNA capillary sequencing was coupled to NGS (Next Generation Sequencing) targeting ITS (Internal Transcribed Sequence) amplicons with the Illumina MiSeq platform. By using culture-dependent techniques, the most frequently detected species was the yeast-like fungus Aureobasidium pullulans, which was dominant in the culturable fraction. Among yeasts, the presence of oligotrophic basidiomycetes such as Cryptococcus spp., Rhodotorula graminis and Sporidiobolus pararoseus was observed at the beginning of ripening. Afterward, upon approaching th... [more]

Classical swine fever virus (CSFV) and porcine circovirus type 2 (PCV2) have caused severe diseases in swine populations worldwide. Here, a polycistronic baculovirus vector was developed to express a bivalent vaccine, consisting of the CSFV-E2 and PCV2-Cap protein, and an immunomodulator protein derived from the Flammulina velutipes, FVE-FIP, as well as the selection marker, green fluorescent protein. The simultaneous expression of the CSFV-E2 and PCV2-Cap protein was mediated by the 2A-like sequence derived from the Perina nuda virus (PnV), while the expression of the FVE-FIP was driven by the internal ribosome entry site (IRES) element derived from the Rhophalosipum padi virus (RhPV). The Western blot analysis result suggested that the CSFV-E2, PCV2-Cap, and FVE-FIP protein were successfully co-expressed by the infected Spodoptera frugiperda IPBL-Sf21 (Sf21) cell line. The extracted cell lysate containing all three recombinant proteins was administered to Balb/C mice with or without... [more]

This work explored the technical feasibility of using crab shell (CS) as a promising, low-cost biosorbent to individually and simultaneously remove Zn2+ and Ni2+ from aqueous solutions. It was found that in both monometallic and bimetallic systems, Zn2+ and Ni2+ biosorption by CS was strongly dependent on the solution pH, with the optimum biosorption occurring at a pH of 6.0 for both heavy metals. The obtained isotherms for Zn2+ and Ni2+ biosorption onto CS in monometallic and bimetallic systems demonstrated that CS has a higher affinity for Zn2+ than for Ni2+. The experimental equilibrium data for the bimetallic system revealed that when one heavy metal is present in the system, there is a decrease in the equilibrium biosorption capacity for the other heavy metal; therefore, the combined action of Zn2+ and Ni2+ was antagonistic. The Sips and Redlich−Peterson isotherm models best fitted the equilibrium biosorption data for Zn2+ and Ni2+ in the monometallic systems, while the modified S... [more]

Cyclic GMP-AMP (cGAMP) has lately been extensively investigated in cancer immunotherapy due its activation of the innate immunity stimulation of interferon genes (STING) pathway within antigen presenting cells (APC) leading to an increase in tumor specific CD8+ T cells. As negatively charged dinucleotides are prone to enzymatic degradation before being taken up by APC, there is a need for an appropriate carrier. Therefore, polyethylenimine (PEI), a gold standard for oligonucleotide delivery, was selected. Molecular weight, type of PEI and N/P ratio between PEI/cGAMP were investigated in terms of toxicity, efficacy and physicochemical properties of the nanocomplexes (NCs) such as size, zeta potential and shape. Due to lack of nano-medicine regulations and the need for a case-by case assessment, here we examine these parameters by several orthogonal methods, such as dynamic light scattering (DLS), nanoparticle tracking analysis (NTA) and online asymmetric flow field flow fractionation (A... [more]

The main objective is to effectively denature the autolysis enzymes of C. frondosa on the premise of avoiding the quality deterioration caused by overheating. The effects of the different thermal treatments (blanching at 40−80 °C for 45 min, boiling and steaming at 100 °C for 15−120 min) on the cooking yield, moisture content, protein degradation, texture, and enzyme inactivation were studied, and the inner relationship was investigated by multivariate analysis. The autolysis enzymes of C. frondosa were thermally stable and cannot be denatured completely by blanching. Boiling and steaming could efficiently inactivate the enzymes but overheating for 60−120 min reduced the cooking yield and texture quality. Boiling at 100 °C for 45 min was suitable for pre-treatment, with cooking yield of 70.3% and protein content of 78.5%. Steaming at 100 °C for at least 30 min was preferable for long-term storage and instant food, in which the relative activity was only 3.2% with better palatability.

Edible oil extraction is a large and well-developed sector based on solvent assisted extraction using volatile organic compounds such as hexane. The extraction of oil from oilseeds generates large volumes of oilseed by-products rich in proteins, fibres, minerals and secondary metabolites that can be valued. This work reviews the current status and the bio-macro-composition of oilseeds, namely soybean, rapeseed, sunflower and flaxseed, and the refining process, comprising the extraction of oil, the valorisation and separation of valuable secondary metabolites such as phenolic compounds, and the removal of anti-nutritional factors such as glucosinolates, while retaining the protein in the oilseed meal. It also provides an overview of alternative solvents and some of the unconventional processes used as a replacement to the conventional extraction of edible oil, as well as the solvents used for the extraction of secondary metabolites and anti-nutritional factors. These biologically active... [more]

The present study evaluated the effect of multiple variables (drying time, drying temperature, biomass/solvent ratio, glass beads/biomass ratio, extraction time, and extraction speed) in the solubilization of three different phycobiliproteins (C-PC, APC, and PE) from a thermotolerant Oscillatoria sp. The strain was grown in BG11 media (28 °C, light: dark cycle of 12:12 h at 100 µmol·m−2·s−1, 20 days) and the experiments were conducted according to a two-level randomized factorial design with six center points (38 runs). Results show that biomass/solvent ratio, glass beads/biomass ratio, and extraction time, are the most significant variables in the extraction of all three proteins, whereas the glass beads/biomass ratio and extraction time significantly affect their purity. The optimized conditions allow a statistical increase in the concentration of C-PC, APC, and PE extracted from the biomass; however, the purity was lower in comparison with the expected value. The latter occurs due t... [more]

The lipophilic yeast Malassezia furfur, is a member of the cutaneous commensal microbiota and is associated with several chronic diseases such as dandruff, pityriasis versicolor, folliculitis, and seborrheic dermatitis, that are often difficult to treat with current therapies. The development of alternatively effective antifungal therapies is therefore of paramount importance. In this study, we investigated the treatment effect of citral on M. furfur. The minimal inhibitory concentration of citral for M. furfur was 200 μg/mL, and the minimal fungicidal concentration was 300 μg/mL. Citral significantly increased the proportion of yeast cells to mycelial forms 2.6-fold. Phosphatidylserine externalization, DNA fragmentation, and metacaspase activation supported a citral-induced apoptosis in M. furfur. Moreover, citral at sub-minimum inhibitory concentrations reduced the invasion of M. furfur in HaCaT keratinocytes. Finally, we demonstrated that citral inhibited IL-6 and TLR-2 expression a... [more]

Cancer is a leading cause of death worldwide, and most of the currently available drugs for cancer treatment have limited potential. Natural products and their relatives continue to represent a very high percentage of the drugs used for cancer treatment. Curcumin is one of several natural drugs that has recently attracted much attention due to its putative cancer-preventive and anticancer properties. As well, Nitric Oxide (NO) holds a great potential for NO-based treatments for a wide variety of diseases. Here, for the first time, we tested the anti-cancer activities of an NO−Curcumin hybrid, hypothesizing that by joining the effects of curcumin and NO in one compound, the hybrid compound would be more potent than curcumin alone in treating colon cancer. To compare the anti-cancer activities of curcumin and NO−curcumin, we treated different colon cancer cell lines with either curcumin or NO−curcumin and tested their effects on cell proliferation and death. Our results show that NO−curc... [more]

In this study, several covalent methods of immobilization based on acrylic supports, Schiff bases and epoxides have been applied to a commercial cocktail with a high β-glucosidase activity secreted by Aspergillus fumigatus. This cocktail was preliminary compared to a commercial secretome of Aspergillus niger, which was also subjected to the aforementioned immobilization methods. Due to its higher activity, the cocktail from A. fumigatus immobilized on ReliZyme™ HA403 activated with glutaraldehyde was employed for pNPG and cellobiose hydrolysis in diverse operational conditions and at diverse enzyme loadings, showing a very high activity at high enzyme load. A kinetic model based on the Michaelis−Menten hypothesis, in which double inhibition occurs due to glucose, has been selected upon fitting it to all experimentally retrieved data with the lowest-activity immobilized enzyme. This model was compared to the one previously established for the free form of the enzyme, observing that cell... [more]

The high-osmotic-pressure environment of honey is not suitable for the survival of microorganisms, except for osmotic-tolerant fungal and bacterial spores. In this study, shotgun metagenomic sequencing technology was used to identify yeast species present in honey samples. As a result, Zygosaccharomyces spp. yeast, including Zygosaccharomyces rouxii, Z. mellis and Z. siamensis, were isolated. The intracellular trehalose and glycerin concentrations of yeast, as well as the antioxidant-related CAT, SOD and POD enzyme activities, increased under a high-glucose environment (60%, w/v). To learn more about the osmotic resistance of Z. mellis, iTRAQ-based proteomic technology was used to investigate the related molecular mechanism at the protein level, yielding 522 differentially expressed proteins, of which 303 (58.05%) were upregulated and 219 (41.95%) were downregulated. The iTRAQ data showed that the proteins involved in the pathway of the cell membrane and cell-wall synthesis, as well as... [more]

Actinomycetes are natural architects of numerous secondary metabolites including antibiotics. With increased multidrug-resistant (MDR) pathogens, antibiotics that can combat such pathogens are urgently required to improve the health care system globally. The characterization of actinomycetes available in Nepal is still very much untouched which is the reason why this paper showcases the characterization of actinomycetes from Nepal based on their morphology, 16S rRNA gene sequencing, and metabolic profiling. Additionally, antimicrobial assays and liquid chromatography-high resolution mass spectrometry (LC-HRMS) of ethyl acetate extracts were performed. In this study, we employed a computational-based dereplication strategy for annotating molecules which is also time-efficient. Molecular annotation was performed through the GNPS server, the SIRIUS platform, and the available databases to predict the secondary metabolites. The sequencing of the 16S rRNA gene revealed that the isolates BN6... [more]

Edible marine species are valuable sources of bioactive peptides. This study investigated the ACE-inhibitory activity of protein hydrolysates from the red algae Gracilaria tenuistipitata. Fifteen groups of protein hydrolysates were prepared by a two-step enzymatic hydrolysis of G. tenuistipitata: initial hydrolysis with several glycolytic enzymes, followed by three separate proteolytic reactions (Alcalase, Neutrase and Flavourzyme) for 2−10 h. Results showed that the hydrolysate GTN4H had the highest ACE-inhibitory activity in vitro. Furthermore, oral administration of GTN4H significantly reduced systolic blood pressure in spontaneously hypertensive rats. Fraction A derived from GTN4H displayed the highest ACE-inhibitory activity among fractions. Further purification of fraction A by RP-HPLC obtained a purified peptide (MW: 1776 Da) with 17 amino acids and 95.4% ACE-inhibitory activity.

A plant cell-based recombinant glucocerebrosidase was approved by the FDA in 2012 for the treatment of human inherited Gaucher disease, indicating that plant suspension cells have advantages in biosafety and a low production cost as a commercial pharmaceutical recombinant protein expression system. A low allergenic rice suspension cell-based recombinant protein expression system controlled by the αAmy3/RAmy3D promoter has been shown to result in relatively high protein yields in plant cell-based systems. Although several recombinant proteins have been produced in rice suspension cell-based systems, yields must be improved to compete with the current commercial protein expression systems. Different strategies were performed and showed successful improvements in recombinant protein yields in this rice system. The review updates and highlights strategies for potential improvements of the αAmy3-based rice suspension cell-based system.

Bioactive peptides with high potency against numerous human disorders have been regarded as a promising therapy in disease control. These peptides could be released from various dietary protein sources through hydrolysis processing using physical conditions, chemical agents, microbial fermentation, or enzymatic digestions. Considering the diversity of the original proteins and the complexity of the multiple structural peptides that existed in the hydrolysis mixture, the screening of bioactive peptides will be a challenge task. Well-organized and well-designed methods are necessarily required to enhance the efficiency of studying the potential peptides. This article, hence, provides an overview of bioactive peptides with an emphasis on the current strategy used for screening and characterization methods. Moreover, the understanding of the biological activities of peptides, mechanism inhibitions, and the interaction of the complex of peptide−enzyme is commonly evaluated using specific in... [more]