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Process optimisation of defatted wheat germ protein extraction in a novel alkaline-based deep eutectic solvent (DES) via Box-Behnken experimental design (BBD)

Olalere OA, Gan CY

Food Chem, 2023 May 30;409:135224.
PMID: 36577323 DOI: 10.1016/j.foodchem.2022.135224

This is the first study to provide a preliminary investigation into the recovery of protein from wheat germ and the prediction of their extraction conditions in microwave cavity using a novel DES solvent. The response surface methodology was used to predict the microwaved protein extraction conditions of the DWG. The effects of DES buffer-concentration (X1: 0.01-1.00 g/mL), microwave power (X2: 50-250 W), irradiation time (X3: 1-5 min) and sample-to-buffer ratio (X4: 1:10-1:50) were tested using a single factor and Box-Behnken experimental design. Under the optimized conditions (X1 = 0.52 g/mL, X2 = 186 W, X3 = 3.28 min, and X4 = 1:39) protein yield and absorbed microwave were obtained at the optimal value of 33.00 % and 677 J/min, respectively with no denaturation of the protein as validated from the SDS-PAGE gel electrophoresis profile. Consequently, this investigation provides a practical approach for the extraction of bioactive protein from DWG using a novel deep eutectic solvent.
Fulltext Statistical Optimization of Flavonoid and Antioxidant Recovery from Macerated Chinese and Malaysian Lotus Root (Nelumbo nucifera) Using Response Surface Methodology

Tan SJ, Lee CK, Gan CY, Olalere OA

Molecules, 2021 Apr 01;26(7).
PMID: 33916148 DOI: 10.3390/molecules26072014

In this study, the combination of parameters required for optimal extraction of anti-oxidative components from the Chinese lotus (CLR) and Malaysian lotus (MLR) roots were carefully investigated. Box-Behnken design was employed to optimize the pH (X1: 2-3), extraction time (X2: 0.5-1.5 h) and solvent-to-sample ratio (X3: 20-40 mL/g) to obtain a high flavonoid yield with high % DPPHsc free radical scavenging and Ferric-reducing power assay (FRAP). The analysis of variance clearly showed the significant contribution of quadratic model for all responses. The optimal conditions for both Chinese lotus (CLR) and Malaysian lotus (MLR) roots were obtained as: CLR: X1 = 2.5; X2 = 0.5 h; X3 = 40 mL/g; MLR: X1 = 2.4; X2 = 0.5 h; X3 = 40 mL/g. These optimum conditions gave (a) Total flavonoid content (TFC) of 0.599 mg PCE/g sample and 0.549 mg PCE/g sample, respectively; (b) % DPPHsc of 48.36% and 29.11%, respectively; (c) FRAP value of 2.07 mM FeSO4 and 1.89 mM FeSO4, respectively. A close agreement between predicted and experimental values was found. The result obtained succinctly revealed that the Chinese lotus exhibited higher antioxidant and total flavonoid content when compared with the Malaysia lotus root at optimum extraction condition.
Optimisation of microwave-assisted extraction and functional elucidation of bioactive compounds from Cola nitida pod

Olalere OA, Gan CY, Akintomiwa OE, Adeyi O, Adeyi A

Phytochem Anal, 2021 Sep;32(5):850-858.
PMID: 33583076 DOI: 10.1002/pca.3030

INTRODUCTION: The quality characteristics and stability of phenolic by-products from Cola nitida wastes are critical factors for drug formulation and food nutraceutical applications.
OBJECTIVES: In this study, the effect of electromagnetic-based microwave-reflux extraction on the total phenolic content, antioxidant capacity, morphological characteristics, physisorption and chromatographic phenolic profiles were successfully investigated. These physicochemical analyses are often employed in the standardisation of dried herbal and food nutraceutical products.
MATERIAL AND METHODS: In this study, the electromagnetic-based extraction process was optimised using the Box-Behnken design. The oleoresin bio-products were subsequently characterised to determine the total phenolic content, morphological and microstructural degradation. These analyses were conducted to elucidate the effect of the microwave heating on the C. nitida pod powder.
RESULTS: From the predicted response, the optimal percentage yield was achieved at 26.20% under 5.39 min of irradiation time, 440 W microwave power and oven temperature of 55°C. Moreover, the rapid estimation of the phenolic content and antioxidant capacity were recorded at 124.84 ± 0.064 mg gallic acid equivalent (GAE)/g dry weight (d.w.) and 6.93 ± 0.34 μg/mL, respectively. The physicochemical characterisation results from the Fourier-transform infrared spectroscopy, field emission scanning electron microscopy and physisorption analyses showed remarkable changes in the micro-surface area (13.66%) characteristics.
CONCLUSION: The recorded optimal conditions established a basis for future scale-up of microwave extraction parameters with a potential for maximum yield. The physiochemical characterisation revealed the functional characteristics of C. nitida and their tolerance to microwave heating.
Fulltext Data article on elemental and metabolomic-based alkaloidal composition in black pepper oleoresin using a positive ESI-mode LC-QToF and ICP-mass spectroscopy

Olalere OA, Abdurahman NH, Yunus RBM, Alara OR

Data Brief, 2018 Aug;19:1627-1630.
PMID: 30229034 DOI: 10.1016/j.dib.2018.06.034

This paper contains data from the elemental and phytochemical profiling of black pepper oleoresin extracts using the LC-MS QToF and ICP-MS analysis. In recent years studies have shown the medicinal properties of extracts from these two cultivars of Piper nigrum. The medicinal properties are attributed to the presence of many secondary metabolites and mineral element in them. The phytochemical profiling was conducted using a Liquid Chromatography equipped with an electrospray time-of-flight mass spectrometer detectors. The mass spectrometer was equipped with an electrospray ionization sources operated in positive ion mode. The alkaloid compounds in the optimized black pepper extract were tentatively characterized in accordance with their ions׳ mass fragmentation.
Fulltext Holistic approach to microwave-reflux extraction and thermo-analytical fingerprints of under-utilized Artocarpus heterophyllus seed wastes

Olalere OA, Gan CY, Abdurahman HN, Adeyi O, Ahmad MM

Heliyon, 2020 Aug;6(8):e04770.
PMID: 32923719 DOI: 10.1016/j.heliyon.2020.e04770

The increase in wastes generated from jackfruit seeds has been largely under-utilized in Malaysia. Due to the high nutritional and medicinal content embedded in the cellulosic structure of jackfruit wastes, a need then arises for their physicochemical elucidations. In this study, the extraction of Artocarpus heterophyllus seed was carefully investigated using Taguchi orthogonal optimization design. Complete functional group characteristics and chemical profile of the A. heterophyllus seed extracts were obtained using different physicochemical characterization. The optimal conditions of the microwave extraction parameters were determined at 5 min of irradiation time, 450 W of power and 50 °C of temperature. Under this condition, the optimal yield of 17.34 (mg/g) % was achieved at an SNR ratio of 24.78. The mass spectrometry analysis tentatively identified a total of 90 and 148 secondary metabolites at positive and negative ESI modes, respectively. The chemical profile obtained provided a baseline reference for further investigation on the food and medicinal bioactive from Artocarpus heterophyllus seed oleoresins. The FT-infrared emission spectrum shows the presence of some specific carbohydrates and amide protein functional groups directly linked to C-O (1008 cm-1) the carbonyl (C=O) groups, respectively. Moreover, the morphological characteristics of the jackfruit raw and crude extracts conspicuously revealed large-sized globules which suggest the carbohydrates and protein contents. The result of this study indicates that the use of microwave extraction technology produced high-quality extracts with lower degradation of the thermal labile constituents. This will assist in determining the suitable conditions necessary for the total recovery of medicinal and nutritional constituents and conversion of agricultural waste products into useful products.
Novel antihypertensive peptides from lupin protein hydrolysate: An in-silico identification and molecular docking studies

Fadimu GJ, Gan CY, Olalere OA, Farahnaky A, Gill H, Truong T

Food Chem, 2023 May 01;407:135082.
PMID: 36493485 DOI: 10.1016/j.foodchem.2022.135082

Application of non-thermal treatment to proteins prior to enzymatic hydrolysis can facilitate the release of novel bioactive peptides (BPs) with unique biological activities. In this study, lupin protein isolate was pre-treated with ultrasound and hydrolysed using alcalase and flavourzyme to produce alcalase hydrolysate (ACT) and flavourzyme hydrolysate(FCT). These hydrolysates were fractionated into 1, 5, and 10 kDa molecular weight fractions using a membrane ultrafiltration technique. The in vitro angiotensin-converting enzyme (ACE) studies revealed that unfractionated ACT (IC50 = 3.21 mg mL-1) and FCT (IC50 = 3.32 mg mL-1) were more active inhibitors of ACE in comparison to their ultrafiltrated fractions with IC50 values ranging from 6.09 to 7.45 mg mL-1. Molecular docking analysis predicted three unique peptides from ACT (AIPPGIPY, SVPGCT, and QGAGG) and FCT (AIPINNPGKL, SGNQGP, and PPGIP) as potential ACE inhibitors. Thus, unique BPs with ACE inhibitory effects might be generated from ultrasonicated lupin protein.
Fulltext In-Silico Analysis and Antidiabetic Effect of α-Amylase and α-Glucosidase Inhibitory Peptides from Lupin Protein Hydrolysate: Enzyme-Peptide Interaction Study Using Molecular Docking Approach

Fadimu GJ, Farahnaky A, Gill H, Olalere OA, Gan CY, Truong T

Foods, 2022 Oct 26;11(21).
PMID: 36359988 DOI: 10.3390/foods11213375

The use of natural ingredients for managing diabetes is becoming more popular in recent times due to the several adverse effects associated with synthetic antidiabetic medications. In this study, we investigated the in vitro antidiabetic potential (through inhibition of α-glucosidase (AG) and α-amylase (AA)) of hydrolysates from lupin proteins pretreated with ultrasound and hydrolyzed using alcalase (ACT) and flavourzyme (FCT). We further fractionated ACT and FCT into three molecular weight fractions. Unfractionated ACT and FCT showed significantly (p < 0.05) higher AG (IC50 value = 1.65 mg/mL and 1.91 mg/mL) and AA (IC50 value = 1.66 mg/mL and 1.98 mg/mL) inhibitory activities than their ultrafiltrated fractions, where lower IC50 values indicate higher inhibitory activities. Then, ACT and FCT were subjected to peptide sequencing using LC-MS-QTOF to identify the potential AG and AA inhibitors. Molecular docking was performed on peptides with the highest number of hotspots and PeptideRanker score to study their interactions with AG and AA enzymes. Among the peptides identified, SPRRF, FE, and RR were predicted to be the most active peptides against AG, while AA inhibitors were predicted to be RPR, PPGIP, and LRP. Overall, hydrolysates prepared from lupin proteins using alcalase and flavourzyme may be useful in formulating functional food for managing diabetics.