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Researches on the biocompatibility of dental biomaterials has become a hotspot. As a new subject, metabolomics has unique advantages in research of material compatibility. This paper aims to utilize 1H-NMR method to analyze and evaluate the biocompatibility of 3 common dental biomaterials (Titanium, Hydroxyapatite-calcium phosphate bioceramics and Poly-methyl-methacrylate). We utilize 1H-NMR method to analyze extracellular small molecular metabolites of MC3T3-E1 osteoblasts cultivated on 3 dental biomaterials above. Metabolite path is visualized using metPA network software to identify different characteristic metabolites, with the same software biocompatibility of these materials is also evaluated. The metabolic profiles of the 3 experimental groups are clearly distinguished from the control group. 25 characteristic metabolites are identified that could distinguish the metabolic profiles from experimental groups, many of which are mapped to pathway processes. Amino acid metabolism, glucose metabolism, and Lipid metabolism are correlated with the biocompatibility of these materials. Conclusions: 1H-NMR method is a method to analyze and evaluate the biocompatibility of dental biomaterials.

Aqueous two-phase systems (ATPS) have been widely used in the laboratory separation and purification of antibiotics. However, its large-scale application is limited due to the difficulty in the recovery of the phase-forming polymers. In this study, we aimed to develop a thermo-pH responsive recyclable ATPS using thermo-responsive polymer EO30PO70 and pH-responsive polymer PADB5.22 for lincomycin partition. This ATPS can rapidly form two-phase interface in less than 5 min and can sustainably be reused. The phase separation mechanism was studied by low-field nuclear magnetic resonance (LF-NMR) and surface tension. The effects of polymer concentration, lincomycin content, pH, temperature and salts type and concentrations on lincomycin partition were investigated. The 25 % (w/v) EO30PO70/4.5 % (w/v) PADB5.22 ATPS showed good partitioning at room temperature for lincomycin fermentation broth in the presence of 80 mM LiCl at pH 9.1, with a partition coefficient of 5.94 and extraction recovery of 94.68 %. In addition, the recycling of the two polymers showed a high efficiency, with 95.51 % and 96.18 % recoveries of EO30PO70 and PADB5.22 after four cycles, respectively. This recyclable ATPS has a promising industrial application in the separation and purification of antibiotics.

This study aimed to investigate the effect of repeated batch fermentation of cocultured Saccharomyces bayanus Y4 and Torulaspora delbrueckii Y7 on the quality of greengage wine. The acidity, as well as citric acid and L-malic acid content, decreased significantly with the batches. The citric acid content of the single culture in each batch was 58.59, 26.27, and 8.45 g/L, respectively. Additionally, the ester content increased with the batches. The respective values were 1627.93, 2249.46, and 2491.17 μg/L for the prophase of coculture. However, volatile content in the fresh wine of Y4 yeast in each batch was high. Ketone and terpene contents in the first batch of coculture significantly increased from 31.64 to 95.60 μg/L and 58.58–95.13 μg/L, respectively. Furthermore, a significant difference was observed between the microbial communities fermented using single culture and coculture. The change in fungal community diversity for coculture pattern was higher than that in the mash-inoculated single strain, which decreased the abundance of dominant fungi. Pichia, Stropharia, Suillus, Malassezia, Trichoderma, Faecalibacterium, and Butyricicoccus were associated with esters, terpenes, phenylethyl alcohol, and β-ionone. Our study provides a new avenue to improve the flavour and quality of wine brewed using a characteristic fruit.

Chemotherapy of hybrid-drugs has become one of the widely used therapies for the prevention of cancers with promising clinical results. This study aimed to create compostable polymeric nanoparticles (PPNPs) for the codelivery of capsaicin (CAPS) and biotin (BT) and to examine the anticancer activity of the drug delivery mechanism (CAPS/BT@PPNPs) against in vitro and in vivo gastric cancer cells. Because of the insufficiency of double antitumor drugs BT and CAPS compressed in polymer-based bio-degradable nanoparticles, the encapsulation of BT and CAPS antitumor drugs co-loaded with polyethylene glycol and polylactidecoglycolide nanoparticles (PPNPs) is effective. Transmission electron microscopy (TEM) was utilized to investigate the structure of BT@PPNPs, CAPS@PPNPs, and BT/CAPS@PPNPs, as well as their shape and size. Human gastric carcinoma cell lines, such as SGC-791 and NCI-N87, were also induced to apoptosis in vitro by BT/CAPS@PPNPs. Morphologies and cell death were observed using bio-chemical staining methods, AO/EB (acridine orange/ethidium bromide) and Hoechst. Flow cytometry in dual staining was also used to confirm the mechanistic investigations of apoptosis. Overall, this dual drug delivery strategy suggests that BT/CAPS@PPNPs could be used as a new method to increase the efficacy of gastric therapeutics.

Renal cell carcinoma (RCC) is a widespread type of urological tumor that derives from the highly heterogeneous epithelium of the kidney tissue. For the past decade, the treatment of kidney cancer cells has changed clinical care for RCC. Herein, we present a very easy and cost-effective method that incorporates tumor-specific targeting supramolecular nanoassembly, and therapeutically to overcome the different challenges raised by the distribution of the pharmaceutical potential anticancer drug Cisplatin (CIS-PT). On covalent conjugations of hydrophobic linoleic acid by carboxylic group, the CIS-PT prodrugs were skilled in impulsively nanoassembly into extremely steady nanoparticles size (∼100 nm). Electron microscopic techniques have verified the newly synthesized morphology of CIS-PT-NPs. The anticancer properties of CIS-PT and CIS-PT-NPs against Caki-1 and A-498 (renal carcinoma) cancer cell lines have been evaluated after successful synthesis. Other research, such as dual staining acridine orange/ethidium bromide (AO–EB), Hoechst 33,344 and flow cytometry study on the apoptosis mechanisms, have shown that proliferation in renal cancer cells is associated with apoptosis. Further the In vivo toxicity results displays the CIS-PT-NPs remarkably alleviated the toxicity of the potential anticancer drug CIS-PT In vivo while conserving the Pharmaceutical activity. Compared to CIS-PT, CIS-PT-NPs demonstrate excellent In vitro and In vivo property, this study clarified the CIS-PT-NPs as a healthy and positive RCC care chemotherapeutics technique and deserve further clinical evaluations.

The present paper reports the synthesis of an eco-friendly and low-cost chitosan modified hybrid Azadirachta indica leaves powder-Kaolinite clay (Chit/AILP-Kao) nanocomposite and further explored as a potential adsorbent to remove hazardous congo red (CR) and methylene blue (MB) dyes from aqueous solution. The optimum pH for the sequestration of CR and MB dye were found to be 6 and 7 respectively. Among the evaluated isotherms, equilibrium data was found to be the best fitted by Langmuir model on the basis of higher value of coefficient R2 (0.998). The maximum monolayer adsorption capacity (qm) for CR and MB dyes were found to be 104.6, 99.01 mgg−1 respectively at optimum pH and temperature 30 °C. To determine the adsorption process kinetic model was best obeyed by pseudo second order on the basis of higher value of coefficient R2 (0.998). In the thermodynamic parameters, negative values of ΔG° and positive values ΔHo confirms the spontaneous and endothermic nature of adsorption process. The values of maximum specific surface area for CR dye was found to be 210.7 m2 g-1 while for MB dye it was 223.7 m2 g-1. The regeneration study confirmed that the present nanocomposite can be regenerated upto fifth cycle successfully.

Pesticide poisoning is a menace in Indian subcontinent and is one of main reason behind the reported accidental, homicide or suicidal cases in the society. Conventional pesticide detection methods such as Chromatographic, Mass spectrometry, or High-performance liquid chromatography (HPLC) have several constraints including low sensitivity, costly, and more time consumption. These challenges can be overcome with emerging nanotechnology based electrochemical biosensing platform based on acetylcholinesterase (AChE) enzyme inhibition. These biosensors provide advantages to scientific fraternity of forensic laboratories to being fast in operation, portable, cheap, highly sensitive and user-friendly. In the presented work, an electrochemical biosensor for detection of Organophosphorus (OP) pesticides based upon AChE-inhibition is developed that operates in pico-molar range. The titanium oxide and molybdenum disulfide nanomaterials were synthesized and electrodeposited on a screen printed electrode. Further, AChE was immobilized on the modified electrode surface by crosslinking using glutaraldehyde and chitosan. The developed nano-interface was examined by different electrochemical measurements at every fabrication stage. The developed biosensor is expected to show higher stability and good reproducibility for OP pesticide detection. It was successfully validated for low OP pesticide concentration identification in forensic visceral samples and the detection limit of the sensor was calculated as 50 pM.

We examine the relationship between wetland (WL) and dryland (DL) Mangifera indica L with dietary-induced metabolic homeostasis. The typical Indian mango fruit Mangifera indica L has shown promise in the prevention of diarrhea and its complications. However, there is little information available regarding the characterization of organic compounds, metabolites, small molecules, and phytochemicals derived from Mangifera indica L. (Mango). In this study, we performed a global target profiling investigation of phytochemicals and inhibitors in Mangifera indica L.using proton nuclear magnetic resonance (1H NMR) signals. The glutathione (GSH), trimethylamine-N-oxcide (TMAO), and trimethylamine (TMO) are associated with disruptions of oxidative stress metabolisms. Metabolic differences of WL and DL mango were investigated with network analysis for revealing their metabolic discriminations. During plant life, fertilization, and biotic and abiotic stress reflected peculiar metabolomic profiles. The methodology is highly efficient, complex deconvolution, spectral baseline fitting that has transported to pattern recognition analysis. Fifty metabolites were identified and characterized which includes xanthones, phenolic acids, fatty acids, flavonoids, and amino acids. The current study is the first report of metabolites reglation by mango fruits. In conclusion, from Indian traditional mango fruits, we provide a perspective of metabolomic impacts and associated metabolic pathways with high-throughput analytical analysis.

Despite the serious damage of a wide range of agriculture and forestry by plant-parasitic nematodes, the use of many chemical pesticides has been declining in recent years due to concerns about their toxicity, tolerance, and environmental persistence issues. Therefore, there is an urgent need to develop efficient and environmentally safe alternatives to chemical pesticides. To this end, microbes have attracted great attention as sources for novel nematicidal agents, largely owing to their genetic diversity and metabolic capacity. However, their uses have often been limited due to their low productivity of natural products. Metabolic engineering of microbial cells, which entails rewiring of metabolic flux toward the target product, has emerged as a powerful method that can overcome the limitations of low yield. Here we review the recent metabolic engineering approaches for the high-production of nematicidal agents, including heterologous expression, metabolic flux control, and engineering of regulatory elements, transporters, and genes involved in post-modification.

Amylases have been widely applied for starch saccharification in the food, paper, pharmaceutical, and textile industries. In the present study, α-amylase DL-3-4−1 from B. amyloliquefaciens was successfully expressed in B. subtilis. The enzymatic activity exhibited high stability in a pH range from 4.0 to 11.0 and moderate thermostability. Furthermore, excellent stability was observed when incubated with metal ions, detergents, inhibitors, and organic solvents. Resistance against ethanol, urea, and guanidine−HCl was also observed. More importantly, a strategy to enhance the extracellular expression of α-amylase DL-3-4−1 was developed using 173 Sec-type signal peptides (SPs) as a library based on a fast and sequence-independent method. Fourteen SPs were identified with significantly increased yields of DL-3-4−1 via a high-throughput assay. Specifically, the enzymatic activity of the α-amylase-producing strain containing the signal peptide SP5D10 (YomL) was 3.74-fold higher than that of the strain with the wild type signal peptide (87.30 U/OD). In brief, the maximum specific activity of an α-amylase-producing strain containing the YomL signal peptide was 326.45 U/OD. This is the first report of an α-amylase that exhibits ethanol, urea, and guanidine−HCl tolerance. Hence, this α-amylase represents a promising candidate for future use in industry.

Understanding the potential of ionic liquids in biocatalysis is both useful and difficult. The unique properties of ionic liquids, such as their tunability, allow for the medium to be adjusted to the reaction, thereby supporting both substrate solubility and enzyme activity. Thus, the effect of 9 different ionic liquids with different structures (in terms of cation, anion, and the attached substituents) on the activity of β-galactosidase, an enzyme widely used in food industry processes, was investigated. According to the findings of this study, three of the nine tested ionic liquids promoted an increase in β-galactosidase activity, while two of them ([bmpyr][Cl] and [bmpyr][BF4]) promoted an inactivation (with EC50 of 20.4 and 10.7 mmol L−1, respectively), most likely due to the presence of the cation [bmpyr]. Some correlations between the structure of ionic liquids and the activity of β-galactosidase were established. To carry out these studies, a microfluidic automatic system was used, allowing for greater control of the reaction conditions as well as a reduction in reagent consumption and waste generation. It demonstrated the system's potential as a simple, versatile, robust, and fast analytical tool for the automation of enzymatic assays in ILs medium.

Biological pretreatment of lignocellulose by white-rot fungi is an environmentally friendly alternative to chemical and physical approaches to enhance enzymatic saccharification. However, inefficient lignin degradation and substantial cellulose consumption during fungal pretreatment can cause low sugar yields. In this study, the combined action of CuSO4 and MnSO4 effectively improved the degree and selectivity of delignification during solid-state fungal pretreatment of poplar wood by Phanerochaete chrysosporium. Compared to pretreatment without this supplementation, a 1.9 fold higher lignin degradation and 2.4 times higher delignification selectivity value were obtained due to the increased and prolonged manganese peroxidase activity. Enzymatic saccharification of supplemented pretreated wood resulted in a 2.9 times higher glucose yield compared to the non-supplemented system. This study has demonstrated that the combined application of Mn2+ and Cu2+ can significantly improve the fungal pretreatment process and that the beneficial effect of Cu2+ on delignification is not restricted to laccase-producing fungi.

Extracellular inulinase and invertase production by Aspergillus brasiliensis was evaluated in solid-state fermentation (SSF) and submerged fermentation (SmF) with a high sucrose concentration. The parameters estimated from online CO2 analysis revealed a lower Lag phase in SSF compared to SmF; the specific CO2 production rate (μCO2) decreased in both systems as the sucrose concentration increased. The highest maximum CO2 production rate (10.45 ± 0.72 mg/h mL) and total CO2 production (138.53 ± 6.48 mg/mL) were obtained in SSF with a sucrose concentration of 210 g/L. Inulinase and invertase production (1.13 ± 0.02 and 5.84 ± 0.12 U/mL, respectively) in SmF was independent of the initial sucrose concentration; meanwhile, both activities increased in SSF as the sucrose concentration increased, reaching maximum values of 5.88 ± 0.78 and 41.69 ± 3.60 U/mL, respectively, at the highest sucrose concentration (210 g/L). The zymographic analysis showed one band of inulinase and two bands of invertases involved in sucrose hydrolysis.

GLOBOCAN estimates in 2020 on the basis of occurrence, mortality, and development for 36 distinct types of tumor cells worldwide. Chemotherapy is the most preferred treatment for carcinoma, and also for hormonal, radiation, autoimmune, and genetic disorders. We propose a straightforward and cost-effective way to overcome the myriad barriers to the therapeutic potential of irinotecan (SN-38), including the rebuilding of medicines, targeting of supramolecular nanoassemblies, and therapeutic treatments. The drug SN-38 was capable by covalent combination of lipophilic linoleic acid by the end of carboxyl group action to provide an impulsive nanoassembly to extremely stable nanomaterials (NMs). The morphology of the recently synthesized SN-38-NM was tested by electron microscopy. The anticarcinoma properties of both HeLa and CaSki (cervical carcinoma lines) of SN-38 and SN-38-NMs were evaluated after successful synthesis. Further research on mechanistic cell death pathways showed that proliferation of cervical carcinoma cell lines was correlated with cells subjected to apoptosis, including AO/EB staining, nuclear staining, and flow cytometric studies. SN-38-NMs have higher biocompatibility in conjunction with free SN-38. This report describes SN-38-NMs as a positive and secure treatment device for cervical carcinoma, which requires further clinical assessment.

The etiological agent of melioidosis, Burkholderia pseudomallei, utilises a type III secretion system cluster 3 (T3SS3) to deliver proteins termed type III effectors (T3SEs) into the host cytoplasm in order to establish an intracellular lifecycle in phagocytic and non-phagocytic cells, thus playing an important role in pathogenesis. To gain insight into possible functional roles for BapA, a putative T3SE with unknown function, in the intracellular lifecycle of B.pseudomallei, bapA gene knockout mutant was constructed. The effect of the knockout on virulence to the otherwise isogenic parental strain, K96243, was studied by cellular infection assays and Caenorhabditis elegans killing assay. The attachment and subsequent entry into A549 cells was significantly (P < 0.05) attenuated in the Delta bapA compared to K96243. However, intracellular replication was not affected. Furthermore, the cell-to-cell spreading capacity of Delta bapA was impaired although the mutant exhibited no evident defect in its actin tail formation. Additionally, phagocytosis and intracellular replication rates of Delta bapA in U937 macrophage cells were significantly reduced relative to K96243 without phagosomal escape being affected. Based on these observations, we conclude that the BapA T3SE could play an important role in B.pseudomallei intracellular lifecycle, especially, in the early stages of attachment and entry into the host cell.

The investigation on the binding interaction mechanism of florfenicol with DNA in vitro was reported via multi-spectroscopy measurements and molecular docking approach in the present work. The static fluorescence quenching process accompanied with non-radiation energy transfer between florfenicol and DNA was confirmed by Stern-Volmer analysis, lifetime measurement and fluorescence resonance energy transfer study. Florfenicol was the A-T rich minor groove binding agent of DNA. Various experiments including site marker competitive experiments, viscosity measurements, helix melting studies, ionic strength measurements, thermal denaturation experiments, circular dichroism (CD) spectra measurements and UV–vis spectra characteristics were carried out to disclose the interaction mode. In addition, molecular docking stimulated appropriate binding orientation to further prove above experimental results. The binding constants were 6.92 × 103, 6.21 × 103 and 5.61 × 103 M−1 at 291, 301 and 311 K respectively and the binding distance r0 was 5.30 nm. Thermodynamic parameters, such as ΔHӨ =−7.90 kJ mol−1 and ΔSӨ = 46.39 J mol−1 K−1, showed that electrostatic attraction was the key force in the binding.

Neopterin (Np), a catabolic product of guanosine triphosphate (GTP), is synthesized by human macrophages and is an important indicator of cellular immune system activation. Np is associated with the activated cellular immune response and therefore this molecule is a potential biomarker for the diagnosis and follow-up of a wide variety of physiological conditions including cardiovascular and neurodegenerative diseases, autoimmune disorders and viral infections. Within the scope of this study, it was aimed to synthesize neopterin-imprinted cryogel columns (Np-MIPs), which can selectively recognize Np in human body fluids. Np-MIPs were synthesized via the cryo-polymerization in presence of Np as the template molecule and characterized by swelling test, polymerization yield calculations, Brunauer-Emmett-Teller (BET) measurements and Scanning Electron Microscopy (SEM). The surface area was measured as 22 m2/g. Adsorption and desorption of Np from aqueous solutions were investigated, and selectivity studies were performed against pterine and glucose molecules. Maximum Np adsorption on Np-MIP was found to be 249.2 μg/g and Np-MIPs can adsorb Np 1.09 and 3.84 times selective than glucose and pterin, respectively. It was demonstrated that Np-MIP columns can selectively adsorb Np from serum and urine, with the adsorption capacities of 36 μg/g and 38.2 μg/g Np-MIP, respectively.

In the present study, maltotetraose was prepared by enzymatic hydrolysis of maltodextrin. The hydrolysates were further separated and purified using a simulated moving bed after essential pretreatments. For the first time, this method offers a continuous preparation process to produce high-purity maltotetraose. In addition, the purity and stability of the prepared maltotetraose product were analysed. The maltotetraose product showed a purity of 81.32 %, which is the highest reported purity derived from the continuous process. Moreover, the maltotetraose product was extremely stable during storage and thermal processing and exhibited unique and excellent antibacterial activity against Erwinia sp., which cause soft rot in fruits and plants. The results indicate that the maltotetraose product shows great potential to serve as a functional sugar source with high safety and no pollution.

Fungal liquefaction of coal is a promising technology to convert low rank coal into value-added products such as humic acids (HAs) that are extensively used as fertilizer. Use of Pakistani Thar lignite for humic acid extraction can promote agriculture industry in Pakistan. In this study, fifty-five fungal strains were isolated from Thar lignite and screened for their lignite solubilization efficiency. Isolates M13 and MI exhibited highest capacity for lignite solubilization and HA was extracted with 67.4 ± 4.3 percent (w/w) yield. Elemental analysis showed increased nitrogen (0.72–1.39 percent) and oxygen (41.7–48 percent), and decreased carbon (51.8–46.1 percent) and sulphur (1.18–0.73 percent) contents in bio-extracted HA (bHA) compared to chemically extracted HA (cHA). UV/Vis spectroscopy showed increased E4/E6 value from 5.6 ± 0.39 (cHA) to 6.28 ± 0.35 (bHA) indicating reduced molecular mass and aromaticity and increased bioactivity of molecule. FTIR analysis also confirmed these results. HPLC analysis of bHA and cHA showed typical choromatogram of humic acids. Enzyme activity staining indicated the presence of fungal cellulase and xylanase on SDS-PAGE gel. The degradation of lignite macromolecules by lignin degrading fungi is a complex process dependent on alkaline substances, chelators, surfactants and extracellular fungal enzymes.

The current study aims to synthesize and investigate the effects of Ferulic acid-copper(II) (FA-Cu) and Ferulic acid-Zinc(II) (FA-Zn) complexes on bone formation. The initial biocompatibility test and ALP activity assay showed that the FA-Cu and FA-Zn complexes have reached the optimum concentrations of 40 μM. An in-vitro, cellular level evaluation showed that these complexes promote the ALP activity and calcium deposition in osteoblasts. At the molecular level analysis, these complexes increased the osteoblast specific markers, Runx2 and type 1 collagen mRNAs and secretory protein levels of osteocalcin (OC) and osteonectin (ON). Also, the expression of the co-activator of osteoblast differentiation, TAZ increased and the effect of these complexes decreased the co-suppressor HDAC4. We tested the bone forming effect of these complexes in (1) zebrafish embryo bone development, (2) osteoblast distribution in scale, and (3) tail fracture models. FA-Cu and FA-Zn complex exposures have enhanced bone mineralization in zebrafish embryos' development, increased osteoblast distribution in adult zebrafish scales, and reinforced bone healing by increasing ossification at the injury site. In summary, our findings have shown that FA stimulates bone formation, and the complexes of FA-Cu and FA-Zn have further improved it.

Several pharmacological and biological activities of phenolic compounds have been reported in the literature. Some phenolic compounds have antibacterial properties, either alone or in combination with currently used antibiotics. With an increasing number of cases of bacterial infections caused by resistant bacteria, the search for more effective treatments has become critical. The goal of this study was to assess pyrogallol's ability to reverse bacterial resistance by inhibiting efflux pumps, as well as to validate its mechanism of interaction using computational dynamics. To determine the Minimum Inhibitory Concentration (MIC) of pyrogallol, broth microdilution tests were performed. The MIC reduction of tetracycline and ethidium bromide confirmed efflux pump inhibition. Pyrogallol showed direct activity against Staphylococcus aureus; however, the synergistic effect was observed only with the tested antibiotic, reducing its concentration from 128 μg/mL to 45.25 μg/mL, and this effect was not associated with efflux pump inhibition. According to molecular dynamics, this compound interacts with the surface of the bacterial membrane. However, more research is needed to identify potential mechanisms of interaction with existing antibiotics.

Food waste is a global issue costing roughly US$ 680 billion annually in industrialized countries. Food spoilage is a major contribution to food waste but artificial food preservatives are often not well accepted by the public. The use of natural alternatives to control spoilage is therefore of interest to the food industry. Lactiplantibacillus plantarum B21, originally isolated from a Vietnamese fermented meat product, produces an antimicrobial bacteriocin peptide which is pH and temperature stable and capable of inhibiting spoilage and pathogenic bacterial strains, including Listeria. In this study the sources of nitrogen and Tween are demonstrated to be essential for growth and bacteriocin production in L. plantarum B21. Conversely, carbohydrate source was not critical for bacteriocin production but was essential for growth. The bacteriocin was found to interact with cell membranes of target microorganisms resulting in cell leakage and subsequent cell death. This study provides insight into the growth conditions needed for the potential development of L. plantarum B21 as a natural biopreservative for food products.

β-1,3-oligoglucoside is widely used as a functional food additive and a physiologically active compound. To efficiently produce β-1,3-oligoglucosides through curdlan hydrolysis, the endo-β-1,3-glucanase gene from Trichoderma harzianum was expressed in Pichia pastoris KM71 for the first time in order to construct a high-yield endo-β-1,3-glucanase strain. Optimal conditions for cell culture and endo-β-1,3-glucanase induction were subsequently determined in shake flask experiments. Recombinant endo-β-1,3-glucanase activity was then evaluated in a 7 L bioreactor, reaching a maximum of 198.57 U/mL after of 118 h fermentation. The recombinant endo-β-1,3-glucanase optimally functioned at pH 5.5 and 50 °C in addition to being stable over a pH range of 4.0–6.0 and a temperature range of 30 °C – 50 °C. In addition, the expressed glucanase could efficiently hydrolyze scleroglucan and schizophyllan to produce branched oligoglucosides exhibiting different degrees of polymerization (DP 3–10) and structures. The high enzyme activity and specificity towards polysaccharides with β-1,3-linked glucose residues as primary chains renders recombinant endo-β-1,3-glucanase an ideal candidate enzyme for the industrial production of oligoglucosides.

Application of plant-mediated Microbial fuel cell (PMFC) for bioelectricity production with food crop yield has not yet been appropriately investigated. This study aims to reveal the ability of two essential food plants: Amaranthus viridis (P1) and Triticum aestivum (P2), for sustainable bioenergy production and food harvest for 180 days consecutively. The study showed that the highest power density obtained from P2-PMFC was higher than the P1-PMFC, and the organic-rich nutrient called bioslurry in PMFCs shows twofold intensification in the electrochemical reaction. The main role of bioslurry (brown gold) as a magical organic-rich nutrient increases the plant growth and the activity of electrogenic bacteria in PMFCs. Thus the current density of P1 (114.52 ± 20.05 mA/m2) and P2 (185.23 ± 15.10 mA/m2) without bioslurry was increased with a maximum current density of 185.23 ± 15.10 mA/m2 (P1) and 291.23 ± 7.50 mA/m2 (P2) with bioslurry. The high content screening technique and scanning electron micrograph confirm the biofilm formation on the rhizosphere-anode region of both the plants. Culturable rhizosphere-anode region bacterial species were isolated in P1 and P2 PMFCs, and their electrogenic activity was confirmed using the single-chambered Microbial fuel cells (MFCs) through polarization test. The constructed phylogenetic tree for the 31 isolates concludes the diversity of all bacterial species, of which the phylum Firmicutes dominates in both PMFCs. The individual polarization curve efficiency of the isolates ranges from 7.62 to 194.45 mW/m2, and among the 31 isolates, Hydrogenophaga sp. JBP18, Pseudomonas aeruginosa JBP1, Ralstonia sp. JBP24, Ochrobactrum intermedium strain JBP22 and Bacillus safensis JBP6 were selected as potential electrogenic bacteria. Accordingly, this study might lead to the development of MFC technology for sustainable electricity production using food crop plants and determining more rhizosphere-anode electrogenic bacterial species in the future.

The rhamnogalacturonan acetylesterase gene (named pp1113), encoding a novel cold-adapted rhamnogalacturonan acetylesterase, was cloned from Paenibacillus polymyxa. pp1113 was functionally expressed in Escherichia coli by inducing expression at 20 °C for 16 h. The pH and temperature optimums for the recombinant enzyme were 8.5 and 30 °C, respectively. As an SGNH-type acetylesterase, pp1113 demonstrated high activity against acetyl ester substrates and could maintain approximately 60 % activity at 0 °C.The specific activity of purified pp1113 was 65.95 U/mg (C2). The Km and Vmax values of purified pp1113 were 3.89 mM and 81.07 μmol/min, respectively. We obtained an R232L variant with improved thermal stability and catalytic activity through site-directed mutagenesis. T1/2 of R232L at 50 °C increased from less than 30 min to 6 h when compared to wild-type pp1113, and T1/2 extended to 8 h after immobilization. The R232L mutant improved the conversion rate of 0.1 M cinnamyl alcohol at 40 °C to 94 % in the reaction of synthesis of cinnamyl acetate. After purification, the R232L mutant was immobilized (pH 8.0 phosphate buffer 0.5 M, 20 h at 20 °C, protein loading reached 17 mg/g). This was the first report of the rhamnogalacturonan acetylesterase gene obtained from Paenibacillus polymyxa with detailed enzymatic properties.

2,3-Butanediol (2,3-BDO) is a chemical platform used in several applications, such as cosmetics, medicines, polymers, and fuels. Its levo-isomer is used as an antifreeze agent of industrial interest. Glycerol has been extensively investigated to produce 2,3-BDO by different bacteria. However, few studies involving Paenibacillus polymyxa and glycerol have been conducted so far. In this study, biodiesel-derived glycerol was used for the 2,3-BDO production by P. polymyxa PM 3605. About 14.5 g/L of 2,3-BDO was obtained from 50 g/L of crude glycerol in shaken flasks at 37 °C and 200 rpm. By supplementing the glycerol-containing medium with 10 g/L of sugarcane molasses, the final 2,3-BDO production was enhanced to up to 19 g/L. This was the highest 2,3-BDO titer produced by the P. polymyxa strain from crude glycerol as the main carbon source, and sugarcane molasses as a secondary substrate, until now. The selectivity of levo-2,3-BDO was high, and the production of meso-2,3-BDO and acetoin were not detected. As 2,3-BDO metabolism is dependent on strict oxygen control to ensure a microaerobic condition, the production found here has potential for improvement, indicating that the glycerol use by P. polymyxa PM 3605 is promising for the levo-2,3-BDO synthesis.

Thermophilic anaerobic bacteria have received increasing interest as a promising bioprocessing platform for bioethanol production due to their broad substrate range and higher operating temperature. However, ethanol concentration is still lower than the commercial requirements on account of the high by-product formation and low ethanol tolerance. Our previous transcriptome analysis showed the gene expression level of pyruvate formate lyase-activating protein (PflA) was significantly down-regulated when Thermoanaerobacterium aotearoense SCUT27/Δldh was stimulated by various concentrations of ethanol. Based on this, we constructed the mutant strain T. aotearoense SCUT27/Δldh/ΔpflA, which not only possesses higher ethanol tolerance and production but also has enhanced ethanol/acetate ratio that is important for economical ethanol production. Compared to T. aotearoense SCUT27/Δldh, the mutant strain could resist to 5% (v/v) exogenous ethanol and produce 49 % more ethanol (38.82 g/L vs. 26.05 g/L) in 5-L bioreactor. In addition, the final concentration of ethanol reached 45.01 g/L by immobilized cell fermentation, which is the highest value obtained in T. aotearoense SCUT27. Simultaneously, the ethanol/acetate ratio of the mutant strain achieved 6.82, which was 2.16-fold higher than that of the parental strain.

Limited information is available on polysaccharides in Brassica rapa L. (BRP) cultivated at the plateau with an altitude over 3000 m. In the present study, an efficient ultrasonic-assisted extraction (UAE) procedure for polysaccharides from Brassica rapa L. was investigated and optimized by response surface methodology (RSM). Based on the optimal extraction parameters, gradient ethanol precipitation was used as the purification procedure of BRPs, obtaining four fractions named as BRP-20, BRP-40, BRP-60, and BRP-80, respectively. The four fractions presented different physicochemical properties and antioxidant activities. According to the chemical composition detection and monosaccharide compositions analysis, the uronic acids contents of BRP-40 and BRP-60 were higher than the other two fractions. The results of monosaccharide composition suggested that these four fractions were all heteropolysaccharides, consisted of rhamnose, xylose, mannose, glucose, galactose, galacturonic acid, and glucuronic acid. Furthermore, BRP-80 gave the best performance in radical scavenging tests, which could attribute to its lowest molecular weight. What’s more, animal experiment suggested that BRP-80 had anti-fatigue and antioxidant activity in vivo. Overall, polysaccharides from Brassica rapa L. might be developed as a novel source of natural antioxidant and be a potential candidate of anti-fatigue dietary supplements.

Chinese quince is one kind of crop that has edibility and historical medical applications. However, the high amount of lignin makes processing Chinese quince fruit difficult; on the other hand, this lignin represents a valuable resource if a way of using it in industrial applications could be found. In this study, the structural features and antioxidant activities of Brauns native lignin (BNL) isolated from Chinese quince fruit after oven drying (60 °C, 75 °C, and 90 °C), sun drying, freeze-drying, and subcritical dimethyl ether drying were assessed. The BNL samples had low molecular weights (Mw, 2604–5511 g/mol), high carbohydrate content (6.10–9.95 %), and high phenolic OH content (1.76–2.29 mmol/g). NMR spectra showed that the BNL is of the SGH-type with the relative content of p-hydroxyphenyl units ranging from 13 to 21 %. Oven drying (90 °C) reduced the carbohydrate content and increased the phenolic hydroxyl content. Freeze-drying reduced the relative content of β-O-4′ linkages and increased the H-unit content. In terms of antioxidant activities, all samples possessed high DPPH free radical scavenging capacity (IC50, 0.12−0.22 mg/mL) and strong reducing power. These results indicate that BNL extracted from Chinese quince fruit is a promising source of natural antioxidant and biomass material.

Microbial fuel cell (MFC) technology is a promising solution for both organic and inorganic effluent treatment, as it is capable of handling a variety of complex contaminants with simultaneous energy and resource recovery. Unlike conventional treatment technologies, MFCs can achieve removal of organic matter, persistent organic compounds, heavy metals and nutrients from categorically different waste effluents, while recovering energy and valuable substances. This makes the technology economically attractive, and viable for a wide range of industries for their waste treatment applications. However, in order to implement this technology as an on-site treatment unit, limitations pertaining to costs, scale-up and performance have to be addressed, with due emphasis on developing strategies that can be easily applied in industrial settings. This review summarizes the recent progress in the field of waste treatment and environmental remediation using MFCs, and examines these operational challenges. Cost-effective materials that have been implemented on the field and at large scales have also been highlighted. In addition, the review highlights the fact that there is a need to optimize the technology depending on energy or resource recovery. The MFC-based technology utilizes the synergy of bioremediation and bioelectricity production from wastes, offering a sustainable approach to industrial waste management.

A novel microaerobically inducible pUC18-derived plasmid was produced in Escherichia coli chromosomally expressing Vitreoscilla hemoglobin. The plasmid contains a copy of the positive replication control molecule rnaII under the control of the Pvgb promoter. The plasmid was unstable in the engineered strain. Sequencing showed that the plasmid was fragmented during preculture development, probably due to interactions between the host and vector genetic elements. Therefore, the rnaII copy under the control of the microaerobic promoters Pvgb, Pyfid, or Pyfidm was inserted in the minimal plasmid pUC57mini. Such plasmids were successfully produced in the engineered strain. The maximum plasmid DNA yield from biomass was 16.6 ± 0.1 mg g−1 in microaerobic cultures of the engineered strain using the Pyifd promoter. This was 14-fold greater than that using the original pUC57mini and the wild-type strain. Our results show the importance of considering host-vector interactions during the biodesign process and the potential of microaerobic induction for intensifying pDNA manufacturing.

The present study is focused on developing microfibers based on the European fish eel skin gelatin (ESG) using the electrospinning process and evaluating its ability to encapsulate European fish eel oil (EO). Based on the scanning electron microscopy images, electrospinning of 15 % (w/v) gelatin solution in ethanol/water 40 % (v/v) was efficient to produce eco-friendly microfibers. Regarding electrospinning parameters, the increase of the voltage from 10 to 17 kV and the flow rate of feed solution from 0.04 to 0.2 mL/h improved the electrospinnability of ESG solution (15 %, w/v). Furthermore, ESG-based microfibers, loaded with EO, were prepared. The effects of the EO/ESG ratios (1/2 and 1:4, w/w) and two emulsification methods, homogenization (H) and homogenization followed by ultrasonication (HS) treatment, on the electrospun microfibers formation were investigated. The success of EO encapsulation was confirmed by FTIR spectroscopy analysis. The data also revealed that the highest microencapsulation efficiency encapsulation (89.79 %) was noted with EO-loaded microfibers prepared at EO/ESG ratio of 1:4 (w/w) by combining the two emulsification processes (HS). These results suggested that EO-loaded microfibers may be promising to be used as active encapsulating materials in food and nutraceutical fields., This study was funded by the Ministry of Higher Education and Scientific Research, Tunisia. Authors would also like to thank the Central Support Service for Experimental Research (SCSIE) of the University of Valencia for the electronic microscopy service.

Silver nanoparticles (AgNPs) have several applications in nanomedicine where the use of bacterial proteins for synthesis and capping of nanoparticles remain unstudied. Therefore, an extracellular protein of Enterobacter cloacae (SMP1) with ∼15.6 kDa size was partially purified and utilized for the synthesis of AgNPs. Synthesized AgNPs were detected by UV–vis spectrophotometer and characterized by dynamic light scattering (DLS), atomic force microscopy (AFM), cryo-scanning transmission electron microscopy (Cryo-STEM), and high-resolution transmission electron microscopy (HR-TEM). These AgNPs were monodispersed, spherical/quasi-spherical with zeta potential (ZP) of -26.9 mV with an average size of 58.75 nm measured by DLS. The encapsulation of AgNPs with protein was confirmed by electron elemental mapping and the potential mechanism of the synthesis of protein encapsulated AgNPs is outlined. These AgNPs showed broad spectrum bactericidal activity to clinical antibiotic-resistant bacteria. This study revealed that the extracellular protein of bacteria play a vital role in the synthesis of protein-encapsulated bactericidal AgNPs.

Two novel extracellular keratinases were produced by Actinomadura keratinilytica strain Cpt20. Both enzymes were purified to homogeneity using heat-treatment (60 °C for 30 min) and ammonium sulfate salt fractionation (40 %–70 %), followed by anion-exchange chromatography with fast protein liquid chromatography (FPLC) system. The purified keratinases, designated as KERA-71 and KERB-19, are monomeric and named according to their molecular masses of 71 kDa and 19 kDa, respectively, as estimated via sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), zymography, and high-performance liquid chromatography (HPLC). N-terminal residues of both enzymes exhibited high identity with other Actinomadura keratinases. Their hydrolytic activities were significantly inhibited by phenylmethylsulfonyl fluoride (PMSF) and di-iodopropyl fluorophosphates (DFP), classifying them in the serine proteases family. While KERA-71 was ideally active at 50 °C and pH 8, KERB-19 illustrated optimum activity at 40 °C and pH 7. The thermo-activity and thermo-stability of both enzymes were improved with 10 mM Ca2+. Interestingly, the KERA-71 displayed broader substrate specificity, higher catalytic efficiency (kcat/Km), and a high degree of hydrolysis (DH) than actinobacterial keratinases, including KERB-19, KERDZ, KERAK-29, Actinase E, and KERAB. Interestingly, both enzymes exhibited effective keratinase activities with high potential, illustrating their possibility to be used in the process of keratin-containing wastes valorization and leather industry.

Combinations of two potent chemotherapeutic drugs are often used to treat early stage lung cancer. To enhance the efficiency of the treatment, a combinational co-drug delivery system of gemcitabine (GEM) and 5-fluorouracil (5FU) is used owing to its great antitumor ability, as GEM and 5FU improve the anticancer ability of lung cells by stabilizing the tumor microenvironment. Nevertheless, encapsulation of GEM and 5FU potential anticancer drugs by co-loaded poly(lactic-co-glycolic acid) (PLGA)- Polyethylene glycol (PEG) nanoparticles (NPs) is not effective because of the incompatibility of the dual drug GEM and 5FU polymeric biodegradable system. Transmission electron microscopy was used to examine the morphology of GEM NPs, 5FU NPs, and GEM-5FU NPs, along with their nanoparticle morphology and size. GEM-5FU NPs also induced apoptosis in vitro in human lung cells such as A549 and HEL-299. Morphological changes and cell deaths were the result of different biochemical staining techniques such as acridine orange-ethidium bromide (AO-EB) and Hoechst staining. In addition, the mechanistic investigation of cell death was confirmed by Annexin V-Fluorescein isothiocyanate (FITC) using flow cytometry. Taken together, this dual co-drug delivery method revealed that GEM-5FU NPs might have a remarkable potential to enhance the efficacy of lung cancer therapy.

In this study, the effects of melatonin were investigated on the exosome release from bovine cumulus cells after 7 days. Cells were randomly allocated into three groups as follows; Control, 0.1 nM, and 0.1 mM Met groups. The physicochemical properties of exosomes such as size and zeta potential were measured by the dynamic light scattering. Both scanning electron microscope (SEM) and transmission electron microscope (TEM) were used to characterize exosome morphology. The effect of melatonin on cell viability was studied using an MTT assay. The expression of exosome markers like Alix, CD63, Rab27a, and Rab27b was monitored using real-time PCR analysis. We also investigated fatty acid composition by the gas chromatography method. TEM and SEM confirmed typical exosome's morphology. By increasing the concentration of melatonin, the viability of cumulus cells was significantly increased compared to the control group (p

Nelumbo nucifera (lotus) is a unique aquatic plant. Its leaves are covered with superhydrophobic wax layer which protects it from the environmental wearing. In this study, the extraction yield and the functional group profile of crude lotus leaf natural wax have been observed. The lotus leaf wax has been extracted by using many polar and non-polar solvents through different extraction methods. The superhydrophobic wax was successfully extracted from the non-polar solvents i.e. benzene, toluene, xylene, petroleum ether; whereas the polar solvents used (ethanol and ethyl acetate) did not extract the superhydrohobic wax. The results show the highest extraction yield was observed with xylene and the extract in toluene exhibited the best lotus leaf surface-like behavior. Conventional extraction method of maceration provides a slightly higher extraction yield compared to the Soxhlet extraction. For the extract in xylene, maceration provided an extraction yield of 4.37 % and Soxhlet extraction provided a yield of 3.54 %. The TLC performed on the extracts depicted the presence of both polar and non-polar compounds. The FTIR analysis confirmed the presence of alkane chains in the crude natural wax extract. If utilized properly, the lotus leaf natural wax can help mankind in providing cost-effective and affordable waterproofing solutions.

Cotton microdust (CMD) was pretreated by a two-stage alkali-acid pretreatment and subsequently subjected to high solids loading enzymatic hydrolysis (HSLEH) and fermentation. The efficiency of the HSLEH of pretreated CMD was enhanced by optimizing the pretreatment condition using central composite design (CCD) and further improvement of enzymatic hydrolysis was facilitated by the addition of surfactant polyethylene glycol (PEG). The predicted optimized pretreatment conditions for maximum digestibility of cellulose from the regression model were 0.8 % (w/v) H2SO4 and 163 °C. The pretreated CMD was efficiently hydrolyzed at high solids loading (35 %) to produce high glucose concentration. When PEG was added to the HSLEH mixture with a lower cellulase loading of 22 FPU/g glucan, the concentration of glucose produced was 134 g/L. The fermentation of glucose after the surfactant assisted HSLEH at 35 % solids loading produced 66 g/L of ethanol with 96 % metabolic yield. The observed levels of reduction of cellulase load and improvement of ethanol production are higher than the reported values for cotton gin wastes. Hence the observed results demonstrated the feasibility of producing high concentration of ethanol from CMD, which would be useful for further improvement and scale-up of the process.

A novel bioactive ganoderic acid, 3, 28-dihydroxy-lanosta-8, 24-dien-26-oic-acid (DHLDOA), was recently biosynthesized using a heterologous host Saccharomyces cerevisiae by our group, but its low fermentation titer was an obstacle to large-scale application. In this study, the positive effect of oxygen supply on DHLDOA production by the yeast fermentation in shake-flask was revealed. The highest cell mass with an OD600 value of 27.92 was obtained at initial KLa of 15.6 h−1, and the highest DHLDOA titer of 8.79 mg/L was obtained at initial KLa of 12.6 h−1, which was 1.64- and 1.77-fold that at 4.2 and 15.6 h−1, respectively. Slightly higher pyruvate accumulation, lower citrate level, and adequate precursor pools of DHLDOA anabolism were found to be associated with higher DHLDOA production at initial KLa of 12.6 h−1, in which relatively higher CYP5150L8 and CYP5139G1 activities were also observed. Taken together, the results indicated that oxygen supply caused the global changes of the yeast metabolism, including the DHLDOA biosynthesis. This work will be helpful to further improvement of DHLDOA production by the yeast fermentation.

A detailed characterisation of spent hen meat hydrolysate (SMH) powders was done in this study. Briefly, SMH was obtained by enzymatic hydrolysis of whole spent hen meat of Indian layer bird breed (BV-300) using Flavourzyme® at optimised temperature (54 °C) and pH 6.6 for 30 min. The spray-dried (SD-SMH) hydrolysate powder showed significantly (p

In this study, the treatability of real textile wastewater [dissolved organic carbon (DOC): 500 mg/L), color: 6383 Pt-Co, sulfate: 362 mg/L)] was investigated in sequential anaerobic moving bed bioreactor (AnMBBR) and aerobic membrane bioreactor (AeMBR). AnMBBR was filled with 40 % career material (Kaldnes K1). The effect of different hydraulic retention times (HRTs) (6−48 h) and powdered activated carbon (PAC) addition (1 g/L) was investigated for system optimization. Sequential AnMBBR- AeMBR was successfully operated for textile wastewater treatment corresponding to 92 %, and 90 % dissolved organic carbon (DOC) and color removals, respectively. The effluent quality, dropped under 45 Pt-Co and 18 mg/L effluent values, was relatively high and complianted with discharge standards. The Fourier transform infrared spectroscopy (FT-IR) analysis showed that primary amines are successfully removed in AeMBR effluent. Fouling in AeMBR was increased when HRT was reduced, however, improved by 90 % at PAC addition conditions. Except for soluble microbial products (SMP) and soluble extracellular polymeric substances (EPS), it was observed that silicon (Si) (13.16 %) has an important role in fouling which was confirmed by scanning electron microscopy (SEM) and Energy-Dispersive X-ray (EDX) analysis. Additionally, microbial community structures indicated that the membrane fouling can be caused by Alphaproteobacteria (80 %) which was the dominant culture in the biofilm layer at a maximum fouling rate.

In this research, 54 newly isolated strains of yeast from various microhabitats were screened in order to explore their potential for bioethanol production through bioconversion of date palm sap (DPS), as a renewable raw material. The highest ethanol concentration of 58.7 ± 0.23 g/L was achieved by the newly selected Saccharomyces cerevisiae strain X19G2 under anaerobic conditions. The effects of nitrogen supplementation, inoculum size and incubation temperature on bioethanol production from DPS by this strain were investigated. Bioethanol yield (0.43 ± 0.01 g/g consumed sugars) and ethanol productivity (1.49 ± 0.11 g/L/h) increased significantly when a mixture of ammonium sulfate and yeast extract (1:1) were used as nitrogen sources at 30 °C. Subsequently, batch and fed-batch (FB) fermentation were performed in a 2 L bioreactor. The highest ethanol yield was achieved in the FB experiment, reaching 0.48 g/g consumed sugars, which corresponded to over 90 % of the maximum theoretical yield. Based on these findings, the minimum bioethanol production potential using DPS was estimated to be approximately 11000 L per hectare of palm trees. This study indicated that DPS is a highly promising feedstock for bioethanol production by the yeast S. cerevisiae strain X19G2 with economic benefits for industrial scale production.

The most plentiful carbohydrate present on earth is in the form of cellulose which has a tremendous capacity to manufacture second or third-generation biofuels and other valuable metabolites. Cellulose is a significant part of banana pseudostem (BP) squander that causes numerous environmental issues. Hence, disposal of BP is nowadays a major obstacle for Indian farmers because of immense volume, delay in decay raise in a foul smell during debasement which attracts pests and insects to farm. The present study reports the identification of potential isolates, which efficiently debase BP. Out of many isolates, NAULH-4 showed the uppermost decomposition rate of cellulose, hemicellulose, and lignin at 45 °C and pH 8 up to 28 days incubation. Moreover, the taxonomic characterization using phenotypic microarray (PM) technology directs that the isolate NAULH-4 belongs to Bacillus with 0.71 SIM index in metabolic pathway network which is reinforced by 16 s rRNA analysis. Molecular data reveal that NAULH-4 contain laccase, xylanase, and cellulase gene. Overall, Bacillus wakoensis NAULH-4 can be applicable for promising microbe for the conversion of BP waste into value-added metabolites.

In this study, the extraction of rare metals from the spent automobile catalyst using Aspergillus niger (A. niger) was investigated. The effect of different factors including initial glucose, initial pH, inoculum percentage, and Solid to Liquid (S/L) ratio on the A.niger growth was done. Then in the condition of initial glucose of 100 g/L, initial pH of 5, inoculum percentage of 2.5 %, S/L ratio of 1 %(w/v), the extraction efficiency were 25.9 % of cerium (Ce), 43.0 % of aluminum (Al) for 240 h, and 23.9 % of lanthanum (La) for 48 h. After the bioleaching section, the spent residue was applied for platinum (Pt) and palladium (Pd) recovery using aqua regia, which results in a 100 % recovery of these metals. The shrinking core model results showed that both chemical reaction and diffusion on the surface controlled the overall reaction rate. Additionally, the characterization methods including SEM, FTIR, HPLC, and XRD before and after bioleaching were performed.

The sapN gene, encoding the extracellular subtilisin SAPN, a serine alkaline protease from Melghiribacillus thermohalophilus Nari2AT, was isolated, sequenced, and heterologously expressed in Escherichia coli BL21(DE3)pLysS using pUT57 and pTrc99A vectors and in E. coli BL21-AI™ using the Gateway™ pDEST™ 17 vector. Conversely, three cassettes encoding pre-pro-subtilisin (rSAPN/SP-Pro-M), pro-subtilisin (rSAPN/Pro-M), and the mature-subtilisin (rSAPN/M) were hyperexpressed in Pichia pastoris SMD1168 and X33 using the pPICZαC vector. rSAPNs were purified, characterized, and compared to wild-type SAPN. The deduced amino acid sequences exhibited high similarity with subtilisins from Bacillus strains. The highest sequence identity (96 %) was observed with the Bacillus licheniformis MP1 protease, with a 10-residue difference. Compared to SAPN and untagged rSAPNs, (His)6-tagged enzymes showed the highest activity and stability at alkaline pH and high temperature, the highest hydrolysis degree on crab and shrimp by-products, and the best catalytic efficiency. It was found that His6-rSAPN/SP-Pro-Ms expressed in P. pastoris strains was more active than those produced in E. coli. To initiate structure-function relationships, a 3D-model of the Pro-SAPN was built based on the available structures of common subtilisins. These data constitute a pivotal first step toward the creation of new efficient rSAPNs with enhanced catalytic properties and high potential for biotechnological and industrial uses.

The thermostable GH78 family α-L-rhamnosidase DthRha from Dictyoglomus thermophilum DSM3960 was found to have the ability to remove the α-L-rhamnose moiety that was directly linked to the aglycone by an α-1-rhamnosidic bond at the C-3 position of epimedium flavonoids. It exhibited high specific activity on p-nitrophenyl-α-L-rhamnopyranoside (pNPR) (281.06 U/mg), followed by epimedin C (30.42 U/mg), icariin (9.74 U/mg), baohuoside I (1.02 U/mg) and 2’-O-rhamonosylicariside II (3.74 U/mg). In addition, the thermostable glucosidase Dth3 from Dictyoglomus thermophilum DSM3960, which exhibits β-xylosidase activity and β-glucosidase activity, was purified. The main enzymatic properties of the two thermostable glycosidases were compared, and these glycosidases were successfully utilized in the preparation of pharmacologically active icaritin from the total flavonoid extract of epimedium. Finally, 5 g/L of the total flavonoid extract of epimedium was completely transformed into 207.95 mg/L icaritin at pH 5.5 and 80 °C for a total of 4 h. This is the first report on the efficient biotransformation of the total flavonoid extract of epimedium into icaritin by two thermostable enzymes with high productivity. The cost of icaritin preparation will be substantially reduced by converting all the major ingredients in the total flavonoid extract of epimedium.

Skin pigmentation depends not only on melanosomes produced by melanocytes but also on the migration of melanosomes to adjacent keratinocytes and their distribution on the skin surface. The inhibitory effects of cardanol diene (CADI), a major phenolic compound extracted from cashew nut shell liquid (CNSL), on melanin transfer in B16F10-HaCAT co-culture cells and a crude extracted compound of cashew nut shell oil (CNCE) on melanin production in zebrafish were investigated. β-arbutin was used as a positive control. CADI inhibited the expression levels of the melanin phagocytosis-associated protein PAR-2 and filopodia-associated protein NMDAR1, and the effect was better than that of β-arbutin at the same concentration. To further study the inhibitory mechanism of CADI in melanin transfer, fluorescence microspheres were used to simulate melanin and co-culture with keratinocytes. The results indicated that CADI inhibited melanin transfer through suppressing keratinocyte phagocytosis, dendrite formation and filopodia three channels. We also found that CNCE inhibited melanin production in zebrafish and showed significant inhibitory effects on TYR, DCT and α-MSH. Thus, the natural components extracted from CNSL effectively inhibited skin pigmentation, providing insight into the application of new anti-melanin drugs in cosmetics and medicine.

In this contribution, urease was efficiently immobilized onto the new MOF called NEQC-340 for utilizing toward safe blood cleaning and water treatment. Upon this immobilization, the biochemical properties, shelf-life, and reusability of the enzyme were enhanced. The NEQC-340 was synthesized using tris(p-aminophenyl)amine as the amino-linker. The NEQC-340 was characterized by FT-IR, EDX, XRD, SEM, and DLS analysis and then used for urease immobilization (NEQC-340@Urease). The immobilization efficiency was calculated as 90.0 % along with retaining about 87.7 % of enzyme initial activity at optimal conditions. The best pH for both urease and NEQC-340@Urease was found to be 7.0 while the optimum temperature was estimated at about 37.0 °C and 45.0 °C for the free urease and NEQC-340@Urease, respectively. The value of Km was calculated as 22.65 mM and 33.28 mM and the Vmax was found to be 5.26 μmol mg−1 min−1 and 4.41 μmol mg−1 min−1 for the free urease and NEQC-340@Urease, respectively. Moreover, 81.0 % of NEQC-340@Urease initial activity was saved after 15 days of storage, also, it was retained 55.0 % of its activity after 10 reuse. Since NEQC-340@Urease derivatives showed high blood compatibility, they were utilized for both safe blood cleaning (efficiency, 87.0 %) and water bio-treatment (efficiency, 91.0 %) via urea biodegradation.

In this study, an acidogenic compartment was attached to a dual-chambered MFC. Rice mill wastewater was acidified in the acidogenic chamber to enhance the biodegradation of complex organics before feeding to MFC. The system was operated under HRT of 12 h (A-MFC12) and 15 h (A-MFC15). The performance of the acidogenic chamber-MFC unit was compared with a conventional dual-chambered MFC (C-MFC15). The maximum total volatile fatty acid (VFA) accumulation was found to be mg 991.21 mg COD/L for A-MFC12, and 560 mg COD/L for A-MFC15, which corresponds to the acidification yield of 78.35 ± 1.19 % and 55.94 ± 1.36 %, respectively. The maximum volumetric power density achieved in A-MFC12 (4.08 ± 0.08 W/m3) was higher than that of A-MFC15 (2.34 ± 0.07 W/m3), and C-MFC15 (2.11 ± 0.04 W/m3). The findings suggest that integrating acidogenic pre-treatment induced a positive effect on bioelectricity generation and substrate removal efficiency of the MFC.

The co-metabolic biodegradation of p-xylene (as a non-growth substrate model) by Pseudomonas putida (well-known aromatic compounds degrading microorganism) in the presence of Benzene (B), Toluene (T), and Ethylbenzene (E) was studied. Michaelis-Menten/Monod kinetics coupled with co-metabolic degradation expression can be applied for systems containing one growth and one non-growth substrate; thus, the GMDH Group Method of Data Handling (GMDH) was proposed in this study to predict the co-metabolic transformation of non-growth substrates in the presence of more than one growth substrate. Thus, the GMDH Group Method of Data Handling (GMDH) was proposed for the first time to quantify p-xylene degradation (with