The effect of water content and carbon source addition on lipase production from Aspergillus aculeatus Ms.11 using spent coffee ground

Lipase is one of the industrially important enzymes, however, the production needs costly substrate. To overcome the problem, we developed an effective medium formulation to produce lipase from indigenous lipolytic mould Aspergillus aculeatus Ms.11 using spent coffee ground. We observed the effect of additional glucose and olive oil, and the influence of water content on lipase production. The experiments were done using the Solid-State Fermentation (SSF) method for 7 days. The results show that optimum lipase production on substrates with additional glucose and olive oil observed on day 4 with lipase activity of 16.296 U/mL and lipase productivity of 150.32 U/g/day. The optimum water content from the results is 50%. The highest lipase activity obtained using the water content is 291.80 U/mL, while the highest lipase productivity is 106.32 U/g/day. The results showed that water content as well as the addition of glucose and olive oil, affects lipase productivity of Aspergillus aculeatus Ms.11 on spent coffee ground. Further studies to optimise the production condition are suggested.

Ephedra alata extracts exerts anti-obesity, anti-hyperglycemia, anti-antipyretic and analgesic effects

Purpose This study paper aims to evaluate the Phytochemical Composition, anti-obesity, anti-antipyretic and analgesic effect of Ephedra alata (Ea) extracts. Design/methodology/approach Obesity was induced in male Wistar rats through a high-fat/fructose diet (HF/FD). Control rats received a standard diet. Findings Results of this study showed that the Ea methanol extract (MEEa) exhibited a prominent selective inhibitory effect against lipase activity (IC50 = 1.29 mg/ml) as compared to water and ethyl acetate extracts (with IC50 = 1.63 and 1.89, respectively). Also, MEEa exert antipyretic and analgesic activities. In high-fat-high-fructose diet rats, the administration of MEEa inhibited lipase activity in the intestine, pancreas and serum by 53%, 40% and 53%, respectively. It was found to significantly decrease body weight by 20% (p = 0.09) and delay the absorption of triglycerides (TG), total cholesterol (TC), LDL-cholesterol (LDL-C) and increase HDL-cholesterol (HDL-C). In addition, MEEa efficiently decreased a-amylase activity in the intestine, pancreas and serum by 43%, 26% and 46%, respectively, and blood glucose level by 35% (p = 0.06). Originality/value To the best of the authors’ knowledge, this study demonstrates for the first time that MEEa are efficient in preventing obesity and hyperglycemia, pain and fever.

MICROBIOLOGICAL APPROACH TO THE RECONSTRUCTION OF INITIAL PRESENCE OF FAT IN VESSELS FROM BURIALS OF ALANIC CULTURE

В статье представлены результаты определения липазной активности в образцах грунта из заполнений керамических сосудов из погребений в целях выявления исходного присутствия жира в составе ритуальной пищи. Уровень липазной активности в почве прямо зависит от количества поступающего субстрата (жира). После разложения органических остатков в почве формируется ферментный пул, который может сохраняться на протяжении неопределенно длительного периода времени. Предлагаемый нами новый методический подход позволил достоверно выявить сосуды, содержащие жир растительного и/или животного происхождения, что в первую очередь было характерно для образцов грунта из кружек. The article presents the results of determining lipase activity in soil samples from the filling of ceramic vessels from burials in order to identify the initial presence of fat as component of ritual food. The level of lipase activity in the soil directly depends on the amount of incoming materials (fats). After decomposition of organic residues in the soil, an enzyme pool is formed, which can be preserved for an indefinitely long period of time. Our new methodological approach made it possible to reliably identify vessels containing fat of vegetable and/or animal origin, which was primarily characteristic of soil samples from small jugs.

Silicon-based induced resistance in maize against fall armyworm [Spodoptera frugiperda (Lepidoptera: Noctuidae)]

The fall armyworm (Spodoptera frugiperda) is a major economic pest in the United States and has recently become a significant concern in African and Asian countries. Due to its increased resistance to current management strategies, including pesticides and transgenic corn, alternative management techniques have become more necessary. Currently, silicon (Si) is being used in many pest control systems due to its ability to increase plant resistance to biotic and abiotic factors and promote plant growth. The current experiments were carried out at the College of Plant Protection, Gansu Agricultural University, Lanzhou, China, to test the effect of Si on lifetable parameters and lipase activity of fall armyworm and vegetative and physiological parameters of maize plants. Two sources of Si (silicon dioxide: SiO2 and potassium silicate: K2SiO3) were applied on maize plants with two application methods (foliar application and soil drenching). The experiment results revealed that foliar applications of SiO2 and K2SiO3 significantly (P≤0.05) increased mortality percentage and developmental period and decreased larval and pupal biomass of fall armyworm. Similarly, both Si sources significantly (P≤0.05) reduced lipase activity of larvae, and fecundity of adults, whereas prolonged longevity of adults. Among plant parameters, a significant increase in fresh and dry weight of shoot, stem length, chlorophyll content, and antioxidant activity was observed with foliar applications of Si. Root fresh and dry weight was significantly (P ≤ 0.05) higher in plants treated with soil drenching of SiO2 and K2SiO3. Moreover, SiO2 performed better for all parameters as compared to K2SiO3 and control treatment. The study conclusively demonstrated a significant negative effect on various biological parameters of fall armyworm when plants were treated with Si, so it can be a promising strategy to control this pest.

The Agreement between Feline Pancreatic Lipase Immunoreactivity and DGGR-Lipase Assay in Cats—Preliminary Results

The colorimetric catalytic assay based on the use of 1,2-o-dilauryl-rac-glycero-3-glutaric acid-(6′-methylresorufin) (DGGR) ester as a substrate for pancreatic lipase activity is commonly used for the diagnosis of pancreatitis in dogs and cats. Even though the assay has generally been shown to yield consistent results with feline pancreatic lipase immunoreactivity (fPLI) assay, the agreement may vary between assays of different manufacturers. In this study, the chance-corrected agreement between a DGGR-lipase assay offered by one of the biggest providers of diagnostic solutions in Poland and fPLI assay was investigated. The study was carried out on 50 cats in which DGGR-lipase activity and fPLI were tested in the same blood sample. The chance-corrected agreement was determined using Gwet’s AC1 coefficient separately for the fPLI assay’s cut-off values of >3.5 μg/L and >5.3 μg/L. The DGGR-lipase activity significantly positively correlated with fPLI (Rs = 0.665; CI 95%: 0.451, 0.807, p < 0.001). The chance-corrected agreement between the fPLI assay and DGGR-lipase assay differed considerably depending on the cut-off values of the DGGR-lipase assay. When the cut-off value reported in the literature (>26 U/L) was used, it was poor to fair. It was moderate at the cut-off value recommended by the laboratory (>45 U/L), and good at the cut-off value recommended by the assay’s manufacturer (>60 U/L). The highest agreement was obtained between the fPLI assay at the cut-off value of 3.5 μg/L and the DGGR-lipase assay at the cut-off value of 55 U/L (AC1 = 0.725; CI 95%: 0.537, 0.914) and between the fPLI assay at the cut-off value of 5.3 μg/L and the DGGR-lipase assay at the cut-off value of 70 U/L (AC1 = 0.749; CI 95%: 0.577, 0.921). The study confirms that the chance-corrected agreement between the two assays is good. Prospective studies comparing both assays to a diagnostic gold standard are needed to determine which of them is more accurate.

Severe acquired hypertriglyceridemia following COVID-19

Severe hypertriglyceridemia is a major risk factor for acute pancreatitis. In exceptional cases, it is caused by plasma components inhibiting lipoprotein lipase activity. This phenomenon is predominantly associated with autoimmune diseases. Here, we report a case of severe hypertriglyceridemia due to a transient reduction in lipoprotein lipase activity following an episode of COVID-19 in an otherwise healthy 45-year-old woman. The lipoprotein lipase activity of the patient was markedly reduced compared with a healthy control and did recover to 20% of the healthy control’s lipoprotein lipase activity 5 months after the COVID-19 episode. Mixing tests substantiated reduced lipolytic capacity in the presence of the patient’s plasma at presentation compared with a homozygous lipoprotein lipase-deficient control, which was no longer present at follow-up. Western blotting confirmed that the quantity of lipoprotein lipase was not aberrant. Fibrate treatment and a strict hypolipidemic diet improved the patient’s symptoms and triglyceride levels.

Purification and Characterization of an Extracellular Lipase Produced by Aspergillus oryzae ST11 as a Potential Catalyst for an Organic Synthesis

The lipase producing Aspergillus sp. ST11 was identified by molecular and morphological methods. The primers ITS1/ITS4 were used for amplifying the ITS region. It showed that the strain was grouped with Aspergillus oryzae and Aspergillus flavus (98 % bootstrap value). The colony morphology of Aspergillus sp. ST11 on malt extract agar and Czapek yeast agar showed a characteristic of A. oryzae. Therefore, it was identified as Aspergillus oryzae ST11. The lipase produced by the strain was purified and characterized. The purification steps involved precipitation with chilled acetone and separation by column chromatography, with HiTrap® Q HP and Toyopearl Butyl-650M, respectively. After purification, the lipase activity was increased 13 fold and with 7.9 % yield. Its molecular mass was 25 kDa. The purified lipase was stable at a pH between 5.0 - 8.0 and had optimum activity at pH 7.5. It was stable at 30 °C and had optimum activity at 37 °C. Its activity was promoted in the presence of Mg2+ but it was greatly decreased in the presence of Co2+, Cu2+, Hg2+ and Zn2+. Surfactants (Triton X-100, Tween-80, Tween-20, arabic gum, and sodium dodecyl sulfate) showed negative effects on lipase activity, while inhibitors (PMSF, EDTA, and β-mercaptoethanol) did not reduce the activity significantly. Polar solvents, such as methanol and ethanol, had much negative effect on lipase activity compared to non-polar solvents, such as hexane and isooctane. The concentrated lipase from A. oryzae ST11 was used to catalyze the transesterification and gave the highest bioconversion (90 %) after 24 h. HIGHLIGHTS Extracellular lipase produced by Aspergillus oryzae ST11 could be applied in many applications which is more flexible compared with the use of whole-cell biocatalysts High stability toward a wide range of pH and temperature obtained from this extracellular lipase The concentrated lipase shows the capability to produce the high biodiesel from natural oil GRAPHICAL ABSTRACT

Application and characterization of crude fungal lipases used to degrade fat and oil wastes

Aspergillus niger MH078571.1 and A. niger MH079049.1 were identified previously as the two highest Aspergillus niger strains producing lipase. Biochemical characterizations of lipase activity and stability for these two strains were examined and revealed that the optimal temperature is 45 °C at pH 8for A. niger MH078571.1 and 55 °C for MH079049.1. The lipase production of both strains was studied on medium contains waste oil, as a cheap source to reduce the industrial cost, showed that the optimal incubation period for the enzyme production is 3 days. Moreover, an experiment on lipase activates in organic solvents demonstrated that 50% of acetone is the best solvent for the two strains. In the presence of surfactants, 0.1% of tween 80 surfactant showed the best lipase activities. Furthermore, Mg2+ and Zn2+ ions enhanced the lipase activity of A. niger MH078571.1, while Na2+ and Cu2+ enhanced the enzyme activity of A. niger MH079049.1. Lipase activity was also tested for industrial applications such as integrating it with different detergents. Maximum lipase activity was obtained with 1% of Omo as a powder detergent for both strains. In liquid detergent, 0.1% of Fairy showed maximum lipase activity in A. niger MH078571.1, while the lipase in A. niger MH079049.1 was more effective in 1% of Lux. Moreover, the degradation of natural animal fat with crude enzyme was tested using chicken and sheep fats. The results showed that more than 90% of fats degraded after 5 days of the incubation period.

In Silico Study, Physicochemical, and In Vitro Lipase Inhibitory Activity of α,β-Amyrenone Inclusion Complexes with Cyclodextrins

α,β-amyrenone (ABAME) is a triterpene derivative with many biological activities; however, its potential pharmacological use is hindered by its low solubility in water. In this context, the present work aimed to develop inclusion complexes (ICs) of ABAME with γ- and β-cyclodextrins (CD), which were systematically characterized through molecular modeling studies as well as FTIR, XRD, DSC, TGA, and SEM analyses. In vitro analyses of lipase activity were performed to evaluate possible anti-obesity properties. Molecular modeling studies indicated that the CD:ABAME ICs prepared at a 2:1 molar ratio would be more stable to the complexation process than those prepared at a 1:1 molar ratio. The physicochemical characterization showed strong evidence that corroborates with the in silico results, and the formation of ICs with CD was capable of inducing changes in ABAME physicochemical properties. ICs was shown to be a stronger inhibitor of lipase activity than Orlistat and to potentiate the inhibitory effects of ABAME on porcine pancreatic enzymes. In conclusion, a new pharmaceutical preparation with potentially improved physicochemical characteristics and inhibitory activity toward lipases was developed in this study, which could prove to be a promising ingredient for future formulations.