Optimization of the extraction and precipitation process of a leaf protein concentrate from Moringa oleifera Lam.

This study aimed to determine the best extraction and precipitation conditions of Moringa oleifera Lam. leaf protein. The influence of pH (10, 11, 12) and the concentration of NaCl (0, 0.25, 0.5) for the protein extraction process were studied through a Completely Randomized Design (CRD) with factorial arrange 32. The combination of pH 11 and 12 with 0 M NaCl had the best yield (P<0.05). The treatment of pH 11 without NaCl followed a precipitation stage for its purification, and the effect of different levels of pH (4, 4.5, 5) and temperature (40, 60, 80 °C) were evaluated using a CRD with factorial arrange 22 and 6 central points. The temperature did not affect the yield of the process in a significant way and the amount of precipitate was maximized at pH 4 and 4.5. From 100 g of the dry leaf, 7.26±0.19 g of protein was isolated with a recovery of 26.93±0.22 g 100 g-1 from the total protein. Due to their astringency and bitterness, consuming large amounts of Moringa oleifera Lam leaves is not a solution; therefore, obtaining a leaf proteinconcentrate could be useful for diverse applications in nutritional supplements, and as raw material for functional products development.

High throughput method development and optimised production of leaf protein concentrates with potential to support the agri-industry

Invasive plants offer an interesting and unconventional source of protein and the considerable investment made towards their eradication can potentially be salvaged through their revalorisation. To identify viable sources, effective and high-throughput screening methods are required, as well as efficient procedures to isolate these components. Rigorous assessment of low-cost, high-throughput screening assays for total sugar, phenolics and protein was performed, and ninhydrin, Lever and Fast Blue assays were found to be most suitable owing to high reliability scores and false positive errors less than 1%. These assays were used to characterise invasive Scottish plants such as Gorse (Ulex europeans), Broom (Cystisus scoparius) and Fireweed (Chamaenerion angustifolium). Protein extraction (alkali-, heat- and enzyme assisted) were tested on these plants, and further purification (acid and ethanol precipitation, as well as ultrafiltration) procedures were tested on Gorse, based on protein recovery values. Cellulase treatment and ethanol precipitation gave the highest protein recovery (64.0 ± 0.5%) and purity (96.8 ± 0.1%) with Gorse. The amino acid profile of the purified protein revealed high levels of essential amino acids (34.8 ± 0.0%). Comparison of results with preceding literature revealed a strong association between amino acid profiles and overall protein recovery with the extraction method employed. The final purity of the protein concentrates was closely associated to the protein content of the initial plant mass. Leaf protein extraction technology can effectively raise crop harvest indices, revalorise underutilised plants and waste streams.

Potato Response to Drought Stress: Physiological and Growth Basis

Drought poses a major challenge to the production of potatoes worldwide. Climate change is predicted to further aggravate this challenge by intensifying potato crop exposure to increased drought severity and frequency. There is an ongoing effort to adapt our production systems of potatoes through the development of drought-tolerant cultivars that are appropriately engineered for the changing environment. The breeding of drought-tolerant cultivars can be approached through the identification of drought-related physiological and biochemical traits and their deployment in new potato cultivars. Thus, the main objective of this study was to develop a method to identify and characterize the drought-tolerant potato genotypes and the related key traits. To achieve this objective, first we studied 56 potato genotypes including 54 cultivars and 2 advanced breeding lines to assess drought tolerance in terms of tuber yield in the greenhouse experiment. Drought differentially reduced tuber yield in all genotypes. Based on their capacity to maintain percent tuber yield under drought relative to their well-watered controls, potato genotypes differed in their ability to tolerate drought. We then selected six genotypes, Bannock Russet, Nipigon, Onaway, Denali, Fundy, and Russet Norkotah, with distinct yield responses to drought to further examine the physiological and biochemical traits governing drought tolerance. The drought-induced reduction in tuber yield was only 15–20% for Bannock Russet and Nipigon, 44–47% for Onaway and Denali, and 83–91% for Fundy and Russet Norkotah. The tolerant genotypes, Bannock Russet and Nipigon, exhibited about a 2–3-fold increase in instantaneous water-use efficiency (WUE) under drought as compared with their well-watered controls. This stimulation was about 1.8–2-fold for moderately tolerant genotypes, Onaway and Denali, and only 1.5-fold for sensitive genotypes, Fundy, and Russet Norkotah. The differential stimulation of instantaneous WUE of tolerant and moderately tolerant genotypes vs. sensitive genotypes was accounted for by the differential suppression of the rates of photosynthesis, stomatal conductance, and transpiration rates across genotypes. Potato genotypes varied in their response to leaf protein content under drought. We suggest that the rates of photosynthesis, instantaneous WUE, and leaf protein content can be used as the selection criteria for the drought-tolerant potato genotypes.

Characterization of radish leaf protein concentrates for biochemical, functional properties, antioxidant activity, mineral content and microbial stability

Leaf protein concentrate was extracted from radish leaves using heat coagulation and analyzed for its antioxidant capacity, mineral content, functional properties and microbial stability. Radish leaf protein concentrates (RLPC) constituted 48.3 % protein content and a yield of 38.51% (DW). Glutelins (42.27 %), prolamins (29.07%) and albumins (19.32 %) were found to be three major fractions of protein concentrate, while globulins (9.38%) was a minor component and their apparent molecular weights ranged between 12-60 kDa. Antioxidant activities (FRAP, ABTS and DPPH ) were higher in RLPC as compared to the isolated fractions. Among fractions, globulins and prolamins exhibited highest DPPH and FRAP activity while highest ABTS activity was associated with glutelins, respectively. Functional properties viz., water holding capacity, oil holding capacity, emulsifying capacity and emulsion stability of the RLPC were 545, 347, 51.8 and 49.4%, respectively. The maximum solubility of RLPC was observed at pH 12 (44.64%) and the minimum solubility was observed at pH 4 (28.24%). A considerable amount of minerals were present in the RLPC, Ca and Fe being the most abundant. Microbial load of RLPC remained in acceptable limits up to 35 and 21 days of storage under refrigerated and ambient conditions, respectively. These results indicated that LPC have desirable functional properties, a considerable mineral content, high antioxidant activity and sufficient microbial stability. Thus they could be used as a functional ingredient to be incorporated in food products to supplement diet and combat protein deficiency.

Effect of Protease Type and Peptide Size on the In Vitro Antioxidant, Antihypertensive and Anti-Diabetic Activities of Eggplant Leaf Protein Hydrolysates

Solanum macrocarpon (eggplant) leaf protein isolate (ELI) was hydrolyzed using four different enzymes to produce hydrolysates from alcalase (AH), chymotrypsin (CH) pepsin (PH) and trypsin (TH). CH had an overall stronger antioxidant property and was separated using ultrafiltration membranes into <1, 1–3 and 3–5 kDa peptide fractions. Gel-permeation chromatography confirmed conversion of the ELI (average of 22 kDa) into protein hydrolysates that contained smaller peptides (<6 kDa). A total of 23 peptides consisting of tri and tetrapeptides were identified from the CH, which is a wider spectrum when compared to seven for AH and four each for TH and PH. CH exhibited stronger scavenging activities against DPPH and hydroxyl radicals. CH and TH exhibited the strongest inhibitions against angiotensin-converting enzyme. In contrast, AH was the strongest inhibitor of α-amylase while AH and PH had strong inhibitory activities against α-glucosidase when compared with other hydrolysates. Ultrafiltration fractionation produced peptides that were stronger (p < 0.05) scavengers of DPPH, and hydroxyl radicals, in addition to better metal-chelating and enzyme inhibition agents. The study concluded that the eggplant protein hydrolysates and the UF fractions may find applications in tackling oxidative stress-related diseases and conditions involving excessive activities of the metabolic enzymes.