The tiny big world of solid lipid nanoparticles and nanostructured lipid carriers: an updated review.

Nanotechnology is currently a field of endeavour that has reached a maturation phase beyond the initial hypotheses with an undercurrent challenge to optimise the safety, and scalability for production and clinical trials. Lipid-based nanoparticles (LNP), namely solid lipid nanoparticles (SLN) and nanostructured lipid (NLC), carriers are presently among the most attractive and fast-growing areas of research. SLN and NLC are safe, biocompatible nanotechnology-enabled platforms with ubiquitous applications. This review presents a modern vision that starts with a brief description of characteristics, preparation strategies, and composition ingredients, benefits, and limitations. Next, a discussion of applications and functionalization approaches for the delivery of therapeutics via different routes of delivery. Additionally, the review presents a concise perspective into limitations and future advances. A brief recap on the prospects of molecular dynamics simulations in better understanding NP bio-interface interactions is provided. Finally, the alliance between 3D printing and nanomaterials is presented here as well.

Visible light photocatalytic disintegration of waste activated sludge for enhancing biogas production.

Biogas production using waste activated sludge (WAS) is one of the most demanding technologies for sludge treatment and generating energy in sustainable manner. The present study deals with the photocatalytic pretreatment of WAS using ZnO-ZnS@polyaniline (ZnO-ZnS@PANI) nanocomposite as means for increasing its degradability for improved biogas production by anaerobic digestion (AD). Photocatalysis accelerated the hydrolysis of WAS and increased the sCOD by 6.7 folds after 6 h and transform tCOD into bioavailable sCOD. After the AD of WAS, a removal of organic matter (60.6%) and tCOD (69.3%) was achieved in photocatalytic pretreated sludge. The biogas production was 1.6 folds higher in photocatalytic sludge with accumulative biogas up to 1645.1 ml L-1vs after 45 days compared with the raw sludge (1022.4 ml L-1VS). Moreover, the photocatalysis decrease the onset of methanogenesis from 25 to 12 days while achieve the maximum conversion rate of reducing sugars into organic acids at that time. These results suggested that photocatalysis is an efficient pretreatment method and ZnO-ZnS@PANI can degrade sludge efficiently for enhance biogas production in anaerobic digestion process.

Quality and biochemical changes of 'Hindi-Besennara' mangoes during shelf life as affected by chitosan, gallic acid and chitosan gallate.

Quality and biochemical changes of 'Hindi-Besennara' mangoes in response to chitosan, gallic acid (GA) and chitosan gallate (CG) postharvest dipping were studied during 2 weeks of storage at 20 ± 2 °C and 60-70% RH. Both GA and CG lowered decay and weight loss during storage. Chitosan and GA at high level and CG at both level maintained higher membrane stability index of peel than control. Fruits treated only CG and GA at high level and chitosan at both levels retained higher acidity and vitamin C but lower pH and total soluble solids (TSS) than control. All treatments resulted with fruits with higher flesh firmness and lower TSS/acid ratio than untreated fruits. GA at both rates gave lower total phenols after 1 week of storage than control. Both levels of GA and low level of chitosan resulted with fruits with higher antioxidant capacity (lower IC50 values) after 1 week of storage than control. All treatments decreased α-amylase activity of fruit peel compared to control. CG and GA at high level and chitosan at low level increased peroxidase activity compared to control. It was concluded that CG and GA dipping delayed ripening and maintained quality of 'Hindi-Besennara' mangoes during 2 weeks of shelf life.

Postharvest chitosan, gallic acid and chitosan gallate treatments effects on shelf life quality, antioxidant compounds, free radical scavenging capacity and enzymes activities of 'Sukkari' bananas.

The effect of postharvest chitosan, gallic acid (GA) and chitosan gallate (CG) dipping treatments at different concentrations on quality parameters, antioxidant compounds, free radical scavenging capacity (FRSC) and enzymes activities of 'Sukkari' bananas were studied during storage (ripening) at 20 ± 2 °C and 60-70% RH for 13 days. Weight loss and peel color index (the change from green to yellow) increased while, membrane stability index of peel tissues, pulp firmness and acidity decreased during storage. CG and GA treatments slowed down the changes in these parameters compared to control. Total soluble solids (TSS) concentration increased during storage and was lower at CG than other treatments. TSS/acid ratio increased during storage and showed higher value after storage than initial. This ratio was lower at 1% chitosan, 0.075% GA and CG treatments than control. Both vitamin C and total flavonoids concentrations decreased during storage and were not affected by the applied treatments. Total phenols concentration decreased during storage and was higher at acetic acid and the high rate of chitosan, GA and CG treatments than control. FRSC (DPPH IC50 values) of fruit peel ranged from 2.54 to 4.19 µg phenolics concentration among the treatments. FRSC was not affected by the applied treatments but increased (lower IC50 value) during shelf life. The possible relations of these biochemical changes with the activities of the enzymes α-amylase, xylanase, polygalacturonase, peroxidase and polyphenoloxidase were discussed. It is concluded that postharvest CG and GA treatments delayed ripening and maintained better quality parameters of 'Sukkari' bananas during 13 days of shelf life than control.

Influence of solid state fermentation by Trichoderma spp. on solubility, phenolic content, antioxidant, and antimicrobial activities of commercial turmeric.

The influence of solid state fermentation (SSF) by Trichoderma spp. on the solubility, total phenolic content, antioxidant, and antibacterial activities of turmeric was determined and compared with unfermented turmeric. The solubility of turmeric was monitored by increase in its phenolic content. The total phenolic content of turmeric extracted by 80% methanol and water after SSF by six species of Trichoderma spp. increased significantly from 2.5 to 11.3-23.3 and from 0.5 to 13.5-20.4 GAE/g DW, respectively. The antioxidant activities of fermented turmeric were enhanced using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis (3-ethylbenzo-thiazoline-6-sulfonic acid) (ABTS), and ferric ion-reducing antioxidant power (FRAP) assays. The antibacterial activity of fermented turmeric against human-pathogenic bacteria Escherichia coli, Streptococcus agalactiae, Staphylococcus aureus, Entreococcus faecalis, Methicillin-Resistant S. aureus, Klebsiella pneumonia, and Pseudomonas aeruginosae showed a broad spectrum inhibitory effect. In conclusion, the results indicated the potentials of using fermented turmeric as natural antioxidant and antimicrobial material for food applications.

Saccharification and hydrolytic enzyme production of alkali pre-treated wheat bran by Trichoderma virens under solid state fermentation.

BACKGROUND: In continuation of our previously interest in the saccharification of agriculture wastes by Bacillus megatherium in solid state fermentation (SSF), we wish to report an investigation and comparative evaluation among Trichoderma sp. for the saccharification of four alkali-pretreated agricultural residues and production of hydrolytic enzymes, carboxymethyl cellulase (CMCase), filter paperase (FPase), pectinase (PGase) and xylanase (Xylase) in SSF. The optimization of the physiological conditions of production of hydrolytic enzymes and saccharification content from Trichoderma virens using alkali-pretreated wheat bran was the last goal. METHODS: The physico-chemical parameters of SSF include incubation time, incubation temperature, moisture content of the substrate, incubation pH, supplementation with carbon and nitrogen sources were optimized. RESULTS: Saccharification of different solid state fermentation sources wheat bran, date's seeds, grass and palm leaves, were tested for the production of fermentable sugar by Trichoderma sp. The maximum production of hydrolytic enzymes CMCase, FPase, PGase and Xylase and saccharification content were obtained on wheat bran. Time course, moisture content, optimum temperature, optimum pH, supplementation with carbon and nitrogen sources were optimized to achieve the maximum production of the hydrolytic enzymes, protein and total carbohydrate of T. virens using alkali pre-treated wheat bran. The maximum production of CMCase, FPase, PGase, Xylase, protein and carbohydrate content was recorded at 72 h of incubation, 50-70 % moisture, temperature 25-35 °C and pH 5. The influence of supplementary carbon and nitrogen sources was studied. While lactose and sucrose enhanced the activity of PGase from 79.2 to 582.9 and 632.6 U/g, starch inhibited all other enzymes. This was confirmed by maximum saccharification content. Among the nitrogen sources, yeast extract and urea enhanced the saccharification content and CMCase, PGase and Xylase. CONCLUSIONS: The results of this study indicated that alkali pre-treated wheat bran was a better substrate for saccharification and production of hydrolytic enzymes CMCase, FPase, PGase and xylase by T. virens compared to other alkali-pretreated agricultural residues tested.

Solid fermentation of wheat bran for hydrolytic enzymes production and saccharification content by a local isolate Bacillus megatherium.

BACK GROUND: For enzyme production, the costs of solid state fermentation (SSF) techniques were lower and the production higher than submerged cultures. A large number of fungal species was known to grow well on moist substrates, whereas many bacteria were unable to grow under this condition. Therefore, the aim of this study was to isolate a highly efficient strain of Bacillus sp utilizing wheat bran in SSF and optimizing the enzyme production and soluble carbohydrates. RESULTS: A local strain Bacillus megatherium was isolated from dung sheep. The maximum production of pectinase, xylanase and α-amylase, and saccharification content (total soluble carbohydrates and reducing sugars) were obtained by application of the B. megatherium in SSF using wheat bran as compared to grasses, palm leaves and date seeds. All enzymes and saccharification content exhibited their maximum production during 12-24 h, at the range of 40-80% moisture content of wheat bran, temperature 37-45°C and pH 5-8. An ascending repression of pectinase production was observed by carbon supplements of lactose, glucose, maltose, sucrose and starch, respectively. All carbon supplements improved the production of xylanase and α-amylase, except of lactose decreased α-amylase production. A little increase in the yield of total reducing sugars was detected for all carbon supplements. Among the nitrogen sources, yeast extract induced a significant repression to all enzyme productivity. Sodium nitrate, urea and ammonium chloride enhanced the production of xylanase, α-amylase and pectinase, respectively. Yeast extract, urea, ammonium sulphate and ammonium chloride enhanced the productivity of reducing sugars. CONCLUSIONS: The optimization of enzyme production and sccharification content by B. megatherium in SSF required only adjustment of incubation period and temperature, moisture content and initial pH. Wheat bran supplied enough nutrients without any need for addition of supplements of carbon and nitrogen sources.

Purification and characterization of α-Amylase from Miswak Salvadora persica.

BACKGROUND: The miswak (Salvadora persica) is a natural toothbrush. It is well known that very little information has been reported on enzymes in miswak as medicinal plant. Recently, we study peroxidase in miswak. In the present study, the main goal of this work is to purify and characterize α-amylase from miswak. The second goal is to study the storage stability of α-amylase in toothpaste. METHOD: The purification method included chromatography of miswak α-amylase on DEAE-Sepharose column and Sephacryl S-200 column. Molecular weight was determined by gel filtration and SDS-PAGE. RESULTS: Five α-amylases A1, A4a, A4b, A5a and A5b from miswak were purified and they had molecular weights of 14, 74, 16, 30 and 20 kDa, respectively. α-Amylases had optimum pH from 6 to 8. Affinity of the substrates toward all enzymes was studied. Miswak α-amylases A1, A4a, A4b, A5a and A5b had Km values for starch and glycogen of 3.7, 3.7, 7.1, 0.52, 4.3 mg/ml and 5.95, 5.9 4.16, 6.3, 6.49 mg/ml, respectively. The optimum temperature for five enzymes ranged 40°C- 60°C. Miswak α-amylases were stable up to 40°C- 60°C after incubation for 30 min. Ca+2 activated all the miswak α-amylases, while Ni2+, Co+2 and Zn+2 activated or inhibited some of these enzymes. The metal chelators, EDTA, sodium citrate and sodium oxalate had inhibitory effects on miswak α-amylases. PMSF, p-HMB, DTNB and 1,10 phenanthroline caused inhibitory effect on α-amylases. The analysis of hydrolytic products after starch hydrolysis by miswak α-amylases on paper chromatography revealed that glucose, maltose, maltotriose and oligosaccharide were the major products. Crude miswak α-amylase in the toothpaste retained 55% of its original activity after 10 months of storage at room temperature. CONCLUSIONS: From these findings, α-amylases from miswak can be considered as beneficial enzymes for pharmaceuticals. Therefore, we study the storage stability of the crude α-amylase of miswak, which contained the five α-amylases, in toothpaste. The enzyme in the toothpaste retained 55% of its original activity after 10 months of storage at room temperature.

Immobilization of Trichoderma harzianum α-amylase on treated wool: optimization and characterization.

α-Amylase from Trichoderma harzianum was covalently immobilized on activated wool by cyanuric chloride. Immobilized α-amylase exhibited 75% of its initial activity after 10 runs. The soluble and immobilized α-amylases exhibited maximum activity at pH values 6.0 and 6.5, respectively. The immobilized enzyme was more thermally stable than the soluble one. Various substrates were hydrolyzed by immobilized α-amylase with high efficiencies compared to those of soluble α-amylase. The inhibition of the immobilized α-amylase by metal ions was low as compared with soluble enzyme. On the basis of the results obtained, immobilized α-amylase could be employed in the saccharification of starch processing.

Improvement of phenolic profile and biological activities of wild mustard sprouts.

The current study aimed to assess the effect of the germination process of wild mustard seeds on the phenolic profile, antioxidant, antibacterial, and antidiabetic properties, and some relevant enzyme activities. The total phenolic and flavonoid contents increased 5- and 10-fold, respectively, and were maximized on 5-days sprouts. One new phenolic compound was identified on 5-days sprout extract using HPLC. The concentrations of the identified phenolic compounds increased 1.5-4.3 folds on 5-days sprouts compared with dry seeds. The total antioxidant activity multiplied 17- and 21-fold on 5-days sprouts using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) assays, respectively. The activity of carbohydrate-cleaving, phenolic-synthesizing and antioxidant enzymes also increased during germination. On 5-days sprouts, there was a substantial correlation between the highest ß-glucosidase and peroxidase activities with highest phenolic and flavonoid levels and maximum antioxidant activity. The phenolic extract of 5-days sprouts exhibited antimicrobial activities against Escherichia coli and Staphylococcus aureus and showed potent antidiabetic activity established by its inhibitory effect against α-amylase and α-glucosidase compared to dry seeds.

Chia gum-gelatin-based encapsulation of chia sprouts phenolic compounds enhanced storage stability, bioavailability, antioxidant, antidiabetic, and antibacterial properties.

Chia seeds are currently gaining popularity as functional and healthy foods. The developed chia 7-day sprout phenolic extract (CSP) is an abundant supply of highly concentrated antioxidant phenolic compounds with health-promoting and antibacterial properties. The easy destruction against different environmental changes and low bioavailability of these phenolic compounds are the main limitations of their applications/utilization. This study aims to microencapsulate the phenolic compounds of developed CSP for use as valuable functional food additives. Three microcapsules were prepared using coating materials, chia gum (CG), gelatin (G), and their mixture (CG/G) via the freeze-drying technique. The prepared CG-, CG/G-, and G-microcapsules demonstrated high encapsulation efficiency percentages of 97.0, 98.1, and 94.5%, respectively. They retained most of the CSP-phenolics (91.4-97.2%) and increased total antioxidant activity (108-127.1%). The prepared microcapsules released more CSP-phenolic compounds into the simulated intestinal stage (70-82%) than the gastric stage (15-24%), demonstrating that the coating materials enhance protection during the gastric stage. The produced microcapsules exhibited higher storage stability at 40 °C for 60 days than the non-capsulated CSP, indicating that the encapsulation provided enhanced stability. The prepared microcapsules microstructures showed uniform, smoother surfaces, and hidden micropores compared to their coating material microstructures. In addition, the connection between the functional groups of coating materials and CSP-phenolic compounds was demonstrated by FTIR analysis. The prepared CG-, CG/G-, and G-microcapsules can perfectly inhibit the α-amylase and α-glucosidase activities by 65, 68, 60 and 74, 78, and 70%, respectively, compared to CSP (54, and 66%). The three prepared microcapsules displayed better antibacterial with low MBC values (0.36-0.68 mg ml-1) compared to CSP (0.53-0.74 mg ml-1). The prepared CSP microcapsules can be incorporated into various food products to enhance their antioxidant, antidiabetic, and antibacterial properties.

Development of chia gum/alginate-polymer support for horseradish peroxidase immobilization and its application in phenolic removal.

Chia gum's molecular structure with distinctive properties as well as the alginate-based hydrogel's three-dimensionally cross-linked structure can provide a potent matrix for immobilization of enzyme. Herein, chia gum (CG)/alginate (A)-polymeric complex was synthesized and employed as a support material for the immobilization of horseradish peroxidase (HRP). HRP was successfully immobilized on the developed ACG-polymeric support, and the highest immobilization recovery (75%) was observed at 1.0% CG and 2% A, pH 7.0, and 50 units of the enzyme. The structure, morphology, and thermal properties of the prepared ACG-HRP were demonstrated using Fourier Transform Infrared (FTIR), Scanning Electron Microscope, and Thermogravimetric (TGA) analyses. ACG-HRP showed a good reusability (60%) over ten reuses. The immobilized ACG-HRP displayed an acidic pH optimum (6.0), a higher temperature optimum (50 °C), and improved thermal stability (30-50 °C) compared to the soluble HRP at pH 7.0, 40 °C and (30-40 °C), respectively. ACG-HRP has a lower affinity for hydrogen peroxide (H2O2) and guaiacol and a higher oxidizing affinity for a number of phenolic substrates. The ACG-HRP demonstrated greater resistance to heavy metals, isopropanol, urea, Triton X-100, and urea, as well as improved efficiency for eliminating phenol and p-chlorophenol. The developed ACG-polymeric support provided improved enzyme properties, allowed the reuse of the immobilized HRP in 10 cycles, and made it promising for several biotechnological applications.

Antioxidant capacity of chewing stick miswak Salvadora persica.

BACKGROUND: Chewing stick (miswak Salvadora persica L.) is an effective tool for oral hygiene. It possessed various biological properties including significant antibacterial and anti-fungal effects. In the present study, we evaluated the antioxidant compounds in miswak. METHOD: Miswak root was extracted with 80% methanol. Methanol extract as antioxidant was evaluated by using DPPH, ABTS and phosphomolybdenum complex assays and analysis by GC-MS. Peroxidase, catalase and polyphenoloxidase assays were performed for crude extract of miswak root. RESULTS: The methanol extract of miswak contained the highest amount of crude extract among the various solvent extracts. The methanol extract showed a concentration dependent scavenging of DPPH and ABTS radicals with IC50 values 4.8 and 1.6 µg crude extract, respectively. The total antioxidant activities, based on the reduction of molybdenum (VI) to molybdenum (V), increased with increasing crude extract content. The correlation coefficients (R2) between total crude extract and DPPH, ABTS scavenging activities and the formation of phosphomolybdenum complex were 0.97, 0.99 and 0.95, respectively. The GC-MS analysis showed that the methanol extract doesn't contain phenolic and flavonoid compounds or under detected limit. After silylation of methanol extract, three compounds namely 2-furancarboxaldehyde-5-(hydroxymethyl), furan-2-carboxylic acid-3-methyl- trimethylsilyl ester and D-erythro-pentofuranose-2-deoxy-1,3,5-tris-O-(trimethylsilyl) were identified by GC-MS analysis. These furan derivatives as they contain hydroxyl groups could be possessed antioxidant activities. The antioxidant enzymes were also detected in the miswak extract with high level of peroxidase and low level of catalase and polyphenoloxidase. CONCLUSIONS: The synergistic actions of antioxidant compounds and antioxidant enzymes make miswak is a good chewing stick for oral hygiene and food purposes.

Garden cress gum and maltodextrin as microencapsulation coats for entrapment of garden cress phenolic-rich extract: improved thermal stability, storage stability, antioxidant and antibacterial activities.

The obtained garden cress 6-day sprouts phenolic-rich extract (GCSP) contained efficient health-promoting antioxidant-phenolic compounds. To improve the stability, bioavailability, and functional properties of these valuable phenolic compounds, GCSP was encapsulated by freeze-drying technique using different ratios of garden cress gum (GG) and maltodextrin (M) in the absence and presence of sonication (S). The prepared S/GG-microcapsule retained the highest phenolic content (95%), antioxidant activity (141.6%), and encapsulation efficiency (98.2%). It displayed the highest bio-accessibility of GCSP-phenolic compounds in simulated intestine fluid (87%) and demonstrated the greatest storage-stability at 40 °C for 60 days. S/GG-microcapsule possessed better physical properties including moisture, solubility, swelling, and morphological structures using SEM. The main spectral features, crosslinking, and improved thermal stability were demonstrated for S/GG-microcapsule using FTIR and thermogravimetric analyses. S/GG-microcapsule demonstrated much greater antibacterial activity than GCSP against pathogenic bacteria. S/GG-microcapsule can be added to different food products to improve their antioxidant and antibacterial properties.

Statistical optimization, characterization, antioxidant and antibacterial properties of silver nanoparticle biosynthesized by saw palmetto seed phenolic extract.

On the global market, silver nanoparticles (Ag-NPs) are in high demand for their various applications in biomedicine, material engineering, and consumer products. This study highlighted the biosynthesis of the Ag-NPs using saw palmetto seed phenolic extract (SPS-phenolic extract), which contained vital antioxidant-phenolic compounds. Herein, central composite statistical design, response surface methodology, and sixteen runs were conducted to optimize Ag-NPs biosynthesis conditions for maximizing the production of Ag-NPs and their phenolic content. The best-produced SPS-Ag-NPs showed a surface plasmon resonance peak at 460 nm and nano-spherical sizes ranging from 11.17 to 38.32 nm using the UV spectrum analysis and TEM images, respectively. The produced SPS-Ag-NPs displayed a high negative zeta-potential value (- 32.8 mV) demonstrating their high stability. The FTIR analysis demonstrated that SPS-phenolic compounds were involved in sliver bio-reduction and in stabilizing, capping, and preventing Ag-NP aggregation. The thermogravimetric investigation revealed that the produced SPS-Ag-NPs have remarkable thermal stability. The produced SPS-Ag-NP exceeded total antioxidant activity (13.8 µmol Trolox equivalent) more than the SPS-phenolic extract (12.0 µmol Trolox equivalent). The biosynthesized SPS-Ag-NPs exhibited noticeably better antibacterial activity against multidrug-resistant Gram-negative E. coli and Gram-positive S. aureus compared to SPS-phenolic extract. Hence, the bio-synthesized SPS-Ag-NPs demonstrated great potential for use in biomedical and antimicrobial applications.

Antioxidant system of garden cress sprouts for using in bio-monitor of cadmium and lead contamination.

Based on garden cress significantly used for phytoremediation, the antioxidant system included antioxidant-phenolic compounds and antioxidant-enzymes of 6-day-garden cress sprouts (GCS) were assessed as potential bio-indicators for cadmium (Cd) and lead (Pb) contamination. Total phenolic and flavonoid contents of GCS germinated under Cd and Pb treatments (25-150 mg kg-1) gradually increased with increasing concentration of metals and peaked by 2.0, 2.6, and 2.5, 2.3 folds at 150 mg kg-1, respectively. By using DPPH, ABTS, and PMC antioxidant assays, the total antioxidant activity of phenolic compounds of GCS increased 6.1, 13.0, and 5.8-fold for Cd and 5.9, 14.6, and 8.2-fold for Pb at 150 mg kg-1, respectively. The antioxidant enzymes of GCS (POD, CAT, GR, and GST) were significantly activated in response to Cd and Pb stress, and two new electrophoretic POD bands were detected. GCS was absorbed 19.0% and 21.3% of Cd and Pb at 150 mg metal kg-1, respectively. In conclusion, the approaches of the antioxidant defense system of GSC could potentially be used as bio-indicator for monitoring Cd and Pb contamination in a short time of germination process.

Purification and characterization of cysteine protease of Sarcocystis fusiformis from infected Egyptian water buffaloes.

Sarcocystis spp. infects water buffaloes (Bubalus bubalis) causing sarcocystosis. In the present study, Sarcocystis fusiformis was recognized in Egyptian water buffaloes based on histological observation and molecular analysis of internal transcribed spacer 1 (ITS1), 18S ribosomal RNA (18S rRNA) and cytochrome c oxidase subunit I (COX-1) gene fragments. Chemotherapy and vaccines against Sarcocystis spp. could potentially target proteases because they may play a crucial role in the infection. Cysteine proteases are multifunctional enzymes involved in vital metabolic processes. However, the involvement of proteases in S. fusiform infection has not yet been characterized. Here, the purification and study on some biochemical properties of protease isolated from cysts of S. fusiform were carried out. Protease with a molecular weight of 100 kDa was purified. LC-MS/MS analyzed the protein sequence of purified protease and the data suggested that the enzyme might be related to the cysteine protease. The purified protease exhibited maximum activity at pH 6 and a temperature of 50 °C. The Michaelis-Menten constant (Km), the maximum velocity (Vmax), and the turnover number (Kcat) were determined. The complete inhibition effect of cysteine inhibitors indicated that the purified enzyme is a cysteine protease. The results suggested that S. fusiform proteolytic enzyme may be necessary for parasite survival in water buffaloes by digesting host tissues. Therefore, cysteine protease could be a suitable target for vaccinations.

Biotechnology approach using watermelon rind for optimization of α-amylase enzyme production from Trichoderma virens using response surface methodology under solid-state fermentation.

Production of amylases by fungi under solid-state fermentation is considered the best methodology for commercial scaling that addresses the ever-escalating needs of the worldwide enzyme market. Here response surface methodology (RSM) was used for the optimization of process variables for α-amylase enzyme production from Trichoderma virens using watermelon rinds (WMR) under solid-state fermentation (SSF). The statistical model included four variables, each detected at two levels, followed by model development with partial purification and characterization of α-amylase. The partially purified α-amylase was characterized with regard to optimum pH, temperature, kinetic constant, and substrate specificity. The results indicated that both pH and moisture content had a significant effect (P < 0.05) on α-amylase production (880 U/g) under optimized process conditions at a 3-day incubation time, moisture content of 50%, 30 °C, and pH 6.98. Statistical optimization using RSM showed R2 values of 0.9934, demonstrating the validity of the model. Five α-amylases were separated by using DEAE-Sepharose and characterized with a wide range of optimized pH values (pH 4.5-9.0), temperature optima (40-60 °C), low Km values (2.27-3.3 mg/mL), and high substrate specificity toward large substrates. In conclusion, this study presents an efficient and green approach for utilization of agro-waste for production of the valuable α-amylase enzyme using RSM under SSF. RSM was particularly beneficial for the optimization and analysis of the effective process parameters.

Egyptian chia seeds (Salvia hispanica L.) during germination: Upgrading of phenolic profile, antioxidant, antibacterial properties and relevant enzymes activities.

Little studies on chia sprouts were not deeply address the polyphenols profiles and their functional properties during long period of germination. This study aims to evaluate the impact of germination process on the phenolic profile, antioxidant and antibacterial properties and relevant enzymes activities of Egyptian chia seeds. The total phenolic and flavonoid contents of chia sprouts increased several times during ten days of germination and maximized on 7-day sprouts (6.4 and 11.5 folds, respectively). In HPLC analysis, seventeen phenolic compounds were detected on 7-day sprouts compared to fifteen in dry seeds, where two new phenolic compounds (p-coumaric acid and kaempferol) were detected. The concentrations of all the identified phenolic compounds increased several folds (1.8-27) on 7-day sprouts. The total antioxidant activity increased 10, 17, and 29 folds on 7-day sprouts using DPPH, ABTS and PMC antioxidant methods, respectively compared to the dry seeds. Both antioxidant and carbohydrate-cleaving enzymes increased in chia sprouts and correlated with their phenolic content and antioxidant activity. The phenolic content of 7-day sprouts showed a potent antibacterial activity against some human enteric pathogenic bacteria including Escherichia coli O157-H7, Salmonella typhi, Pseudomonas aeruginosa and Staphylococcus aureus with lower MIC values compared to the raw seeds.

Immobilization of horseradish peroxidase on cationic microporous starch: Physico-bio-chemical characterization and removal of phenolic compounds.

In the present study, two different modified starches; microporous starch (MPS) and cationic microporous starch (CMPS) were synthesized. The granules of MPS that distributed regularly were destroyed after the etherification reaction. The data depicted that the immobilization of horseradish peroxidase (HRP) on CMPS revealed highest immobilization efficiency (86%) at 100 mg of CMPS at pH = 6.0 and 100 units of enzyme. After 10 reuses of the CMPS-HRP, it retained 66% of initial activity. The soluble HRP showed broad pH optimum of 6.0-7.0, which changed to sharp pH = 6.0 for CMPS-HRP. Soluble-HRP and CMPS-HRP showed temperature optima at 30 °C and 40 °C, respectively. The CMPS-HRP showed high thermal stability up to 50 °C compared to the soluble HRP (40 °C). The Km values of soluble HRP and CMPS-HRP were 6.6 and 10.8 mM for H2O2 and 34 and 41.6 mM for guaiacol, respectively. CMPS-HRP showed higher affinity toward various substrates than the soluble-HRP. CMPS-HRP showed more resistance against heavy metals, urea, isopropanol, Triton X-100 and trypsin than soluble enzyme. The CMPS-HRP showed higher ability to remove phenol and p-chlorophenol compared to soluble-HRP.