Date nawah powder as a promising waste for ß-mannanase production from a new isolate Aspergillus niger MSSFW, statistically improving production and enzymatic characterization.

ß-Mannanase producing fungus was isolated from coffee powder waste and identified as Aspergillus niger MSSFW (Gen Bank accession number OR668928). Dates nawah powder as industrial and agricultural waste was the most inducer of ß-mannanase production. The Plackett-Burman and Central Composite designs were used to improve ß-mannanase titer. Optimization studies enhanced the enzyme yield with approximate 13.50-times. ß-Mannanase was purified by Sephadex G-150 gel filtration column and the molecular weight was estimated to be 60 kDa by SDS-PAGE. Crude and purified ß-mannanase displayed maximum activity at temperature 60 °C and 50 °C, respectively. Crude ß-mannanase showed an activation energy value 2.35-times higher than the purified enzyme. Activation energy for thermal denaturation of the purified ß-mannanase was 1.08-times higher than that of the crude enzyme. Purified ß-mannanase exhibited higher deactivation rate constant (Kd) and lower half-life (t0.5) and decimal reduction time (D-value) compared with the crude enzyme. Thermodynamic parameters of enthalpy, entropy, and free energy values for crude and purified ß-mannanase were calculated. Substrate kinetic parameters suggested that the purified ß-mannanase had a strong affinity toward locust bean gum by showing 3.44-times lower Km and 1.99-times higher Vmax compared to the crude enzyme.

Portulaca oleracea L seed extracts counteract diabetic nephropathy through SDF-1/IL10/PPARγ-mediated tuning of keap1/Nrf2 and NF-κB transcription in Sprague Dawley rats.

BACKGROUND & OBJECTIVE: While oxidative stress is the key player driving diabetic nephropathy (DN), firm glycemic control remains the pillar prophylactic measure. Purslane was extensively described as a potent hypoglycemic and hypolipidemic agent owing to its rich content of antioxidants. Therefore, this report aimed to assess the renoprotective potentials of methanol (MO) and methylene chloride (MC) fixed oil extracts of purslane seeds in a diabetic nephropathy (DN) model. METHODS: Purslane seeds were extracted using absolute methanol and methylene chloride, and type-1 diabetes was induced with a single 55 mg/kg dose of Streptozotocin (STZ) dissolved in 100 mmol/L citrate buffer (pH 4.5), and then diabetic animals were received MO, MC, for 42 consecutive days to compare their antidiabetic effect relative to the reference drug "Losartan". Renal functions and DN biomarkers were weekly assessed, and the relative expression of different oxido-inflammatory mediators was quantified in diabetic kidneys by RT-PCR. Data were statistically analyzed using GraphPad Prism 9.0.2. RESULTS: The oral administration of MO and MC extracts (250 mg/kg/day) significantly ameliorated the body weight loss (P < 0.0001 / each), fasting blood glucose levels (FBG) (P < 0.0001 / each), urine volume (P < 0.0001 / each), as well as serum creatinine (P < 0.0001 / each), uric acid (P = 0.0022, 0.0052), and blood urea nitrogen (BUN) (P = 0.0265, 0.0338); respectively, compared with the untreated diabetic rats. In addition, both extracts restored the effectuality of antioxidative machinery in diabetic kidneys as indicated by a significant reduction of ROS accumulation and lipid peroxidation; higher GSH content, and promoted activity of glutathione reductase and superoxide dismutase antioxidant enzymes (P < 0.0001 / each). Histologically, both extracts alleviated the DN-structural alterations including the glomerular congestion and tubular degeneration, with MC-treated kidneys showing near to normal architecture. The transcription profiles of all treated kidneys revealed a significantly downregulated expression of TNF-α, IL-6, Keap1 and NF-κB genes, concomitant with a significant upregulation of SDF-1, IL-10, Nrf2, HO-1, and PPARγ gene expression (P < 0.0001 / all). CONCLUSION: These findings highlight the remarkable DN-prophylactic potentials of purslane extracts mediated by neutralizing the hyperglycemia-induced ROS accumulation, and circumventing the downstream inflammatory cascades, surpassing the reference angiotensin receptor blocker; i.e. Losartan.

Evaluation of lupine seeds (Lupinus albus L.) neutral extract as a texture improver in low-fat yogurt production.

Aqueous lupine seeds (Lupinus albus L.) extracts were evaluated as a natural fat substitute in low-fat yogurt production. Thus, the chemical composition, particle size, molecular weight, total phenolic (TPC), and total flavonoids (TFC) of the selected extract were estimated. Also, the antimicrobial activity and antioxidant capacity of selected extract were investigated. Yogurt with neutral lupine extract (NeLP) had the highest all sensorial attributes compared to other extracts. Also, the incorporation of NeLP during low-fat yogurt processing increased the solid content, and viscosity, as well as improved the textural profile and sensorial attributes without any negative effect on the yogurt's color. SEM micrographs of NeLP-yogurt microstructure showed a matrix characterized by large fused casein micelles clusters with comparatively lower porosity compared to control yogurt (without NeLP). The chemical composition of NeLP indicated that the major sugar constituents are glucose and galactose with different molar fractions. The molecular weight of NeLP is 460.5 kDa with a particle size of 1519.9 nm. Also, IC50 of NeLP is 0.589 mg/ml, while TPC and TFC are 7.17, and 0.0137 g/100 g sample, respectively. Hence, lupine neutral extract (0.25%) could be used as a fat replacer or texture improver ingredient in such low-fat yogurt which led to improved its characteristics without any negative defect during 7 days at 5 °C.

Chemical modification of Aspergillus niger β-glucosidase and its catalytic properties

Aspergillus niger β-glucosidase was modified by covalent coupling to periodate activated polysaccharides (glycosylation). The conjugated enzyme to activated starch showed the highest specific activity (128.5 U/mg protein). Compared to the native enzyme, the conjugated form exhibited: a higher optimal reaction temperature, a lower Ea (activation energy), a higher Km (Michaelis constant) and Vmax (maximal reaction rate), and improved thermal stability. The calculated t1/2 (half-life) values of heat in-activation at 60 °C and 70 °C were 245.7 and 54.5 min respectively, whereas at these temperatures the native enzyme was less stable (t1/2 of 200.0 and 49.5 min respectively). The conjugated enzyme retained 32.3 and 29.7%, respectively from its initial activity in presence of 5 mM Sodium Dodecyl Sulphate (SDS) and p-Chloro Mercuri Benzoate (p-CMB), while the native enzyme showed a remarkable loss of activity (retained activity 1.61 and 13.7%, respectively). The present work has established the potential of glycosylation to enhance the catalytic properties of β-glucosidase enzyme, making this enzyme potentially feasible for biotechnological applications.