Protein Extracts Research Articles

Snakehead Fish, Channa striata (Bloch, 1973), Protein Concentrate: Excellent Recovery of Fish-Based Albumin Source and Its Possible Application for Sperm Capacitation

Snakehead fish protein concentrate (SFC) shows promising recovery of excellent protein content that has broad potential applications, including the future use of its albumin content as a sperm capacitation promoter. However, the quantity of snakehead fish albumin recovered depends on several variables, including the habitat in which the fish thrives, extraction procedures, and the assay employed. To optimise the albumin content retrieved from snakehead fish flesh with more reliable results, multiple protein extraction methods were undertaken through the protein concentrate preparation and analysed using the Bromocresol purple (BCP) albumin assay. Total soluble protein (TSP), albumin level, and albumin proportion were recovered at a significant reduction in an acid solvent when exposed to up to 50 °C heating compared to extractions undertaken without heating. Moreover, the small number of protein bands identified indicates that the combination of acid solvent and heating treatment was similarly damaging. In this study, we detected the presence of albumin in SFC at 32 %, the highest percentage ever recorded employing the BCP assay, following extraction with water solvent without thermal treatment. Using SDS-PAGE and densitometric analysis, the identified putative albumin-related bands from the two major bands at 40 kDa and 47 kDa, containing respective proportions of 57 % and 16 % of the total protein loaded. Although this exploratory work requires additional analysis and purification steps, further research employing the SFC is worth identifying its ability and how this alternative albumin source can replace serum albumin to modulate the capacitation process in mammalian sperm.

곤충 단백질 추출물을 적용한 소재의 가공 특성 평가

곤충 단백질 추출물을 적용한 소재의 가공 특성 평가

Direct hydrolysis of einkorn whole grain flour proteins for the generation of bioactive peptides using various proteases

In this study, it was aimed to investigate the direct release of BAPs from einkorn flour in one-step process. Thus, the protein extraction step was eliminated, thereby reducing processing cost. Commercial proteases (Alcalase, Flavourzyme, Neutrase, and Trypsin), and crude enzyme from Bacillus mojavensis sp. EBTA7 were used for hydrolyzing einkorn flour (30 %, w/v) solutions at 50–60 °C. The supernatants after centrifugation were used for bioactivity and techno-functionality tests. All hydrolysates demonstrated significant antioxidant capacities, with values ranging from 17.7 to 33.0 μmol TE/g for DPPH, 107 to 190 μmol TE/g for ABTS, and 0.09 to 3.08 mg EDTA/g for ion-chelating activities. Alcalase and Flavourzyme hydrolysis had the highest DPPH activities, while Bacillus mojavensis sp. EBTA7 enzyme yielded relatively high ABTS and ion-chelating activities. Notably, Bacillus mojavensis sp. EBTA7 crude enzyme hydrolysates demonstrated higher oil absorption capacity (2.94 g oil/g hydrolysate), robust emulsion (227 min), and foam stability (94 %) compared to commercial enzymes. FTIR spectroscopy confirmed variations in the secondary structure of peptides. All hydrolysates exhibited negative zeta potentials. The SDS-PAGE showcased MW ranged from 14 to 70 kDa, which was influenced by both the enzyme type and the degree of hydrolysis. Overall, Bacillus mojavensis sp. EBTA7 hydrolysates revealed considerable bio and techno-functional characteristics.

Experimental evaluation of the process of protein removal from waste shrimp shells using papaya and pineapple juice solution

This research aims to improve the quality of chitin obtained from process protein extraction for shrimp shells. Previous studies showed that for the synthesis of chitosan, deproteinization, and demineralization is carried out by alkali and acid treatment, combined with heating, which affects the biosecurity of the material. Therefore, this study aims to remove protein from waste shrimp shells to purify chitin using a simple and safe green chemical approach (using pineapple and papaya juice solutions) and safe method. The raw shrimp shells were treated in steps: deproteinization with pineapple and papaya solutions for 12, 24, 36, and 48 h, varying immersion volumes from 10 mL to 60 mL. The experimental results showed that the protein in the shrimp shell was removed at 7.37% for freshwater shrimp (7.14%), higher than that of saltwater shrimp. The pineapple juice solution removed the protein better than the papaya juice.

Techno-functional properties of Camelina sativa cake proteins treated with the instant controlled pressure drop (DIC) technology

The instant controlled pressure drop (DIC) is one of the emerging technologies in food that can affect the extraction and techno-functional properties of proteins. Camelina sativa cake was subjected to various DIC treatments, with pressure (0.2–0.7 MPa) and exposure time (20–60 s) variables. Afterwards, protein extraction was conducted, and the influence of individual or combined variables on the techno-functional properties of the protein concentrate was analyzed, and each treatment was later analyzed against the control. Furthermore, DIC increased the protein extraction yield. The pressure, time, and their interaction variables influenced the oil absorption capacity (OAC), foaming capacity (FC), and stability (FS), while water-holding capacity (WHC) was solely affected by pressure. The results of comparing DIC against control showed significant differences in OAC, emulsifying activity index (EAI), FS, and FC. These findings strongly indicate that DIC enhances protein extraction yield and modifies the techno-functional properties of camelina proteins.

Propolis ethanol extract functionalized chitosan/Tenebrio molitor larvae protein film for sustainable active food packaging

The application of novel insect proteins as future food resources in the food field has attracted more and more attention. In this study, a biodegradable antibacterial food packaging material with beneficial mechanical properties was developed using Tenebrio molitor larvae protein (TMP), chitosan (CS) and propolis ethanol extract (PEE) as raw materials. PEE was uniformly dispersed in the film matrix and the composite films showed excellent homogeneity and compatibility. There are strong intermolecular hydrogen bond interactions between CS, TMP, and PEE in the films, which exhibit the structure characteristics of amorphous materials. Compared with CS/TMP film, the addition of 3 % PEE significantly enhanced the elongation at break (34.23 %), water vapor barrier property (22.94 %), thermal stability (45.84 %), surface hydrophobicity (20.25 %), and biodegradability of the composite film. The composite film has strong antioxidant and antimicrobial properties, which were enhanced with the increase of PEE content. These biodegradable films offer an eco-friendly end-of-life option when buried in soil. Composite films can effectively delay the spoilage of strawberries and extend the shelf life of strawberries. Biodegradable active packaging film developed with insect protein and chitosan can be used as a substitute for petroleum-based packaging materials, and has broad application prospects in the field of fruits preservation.

Application of Turbiscan Lab® to study the mechanism of fining in red wine using a specific yeast protein extract: A comparison with gelatine

Fining of wines using proteins is a standard practice in the winemaking process. Gelatine is a common fining agent used for red wines. However, the consumer demand for wines that are free of animal-derived products has reinforced the search for alternative protein sources such as yeast origin as the latter is originally present in wine. In 2012, the International Organization of Vine and Wine (OIV) authorized yeast protein extract (YPE) as a processing aid for the fining of musts and wines (OIV-Eno 452–2012). The present work aimed at better understanding the fining mechanisms of a specific YPE, to evaluate the part played by interactions in solution with polyphenols, but also at the interfaces with suspended particles. Polyphenols from the soluble phase of red wine were separated from the suspended particles by centrifugation, and these particles were resuspended in a wine model solution. Interactions with YPE were studied with each fraction separately through static multiple light scattering (SMLS) using the Turbiscan Lab® apparatus and optical microscopy. The impact on wine color and astringency was assessed by UV–visible spectrophotometry and BSA assay test. Faster aggregation and sedimentation kinetics were observed with gelatine than with YPE, but the levels of clarification and lees were similar for both fining agents after a couple of hours (4 h maximum). Gelatine resulted in higher tannin and pigment removal, likely to decrease astringency but also derived pigments involved in red wine color stability. Meanwhile, the impact of the YPE was moderate, likely to modulate astringency while preserving wine color and structure. In addition to interacting directly with polyphenols in solutions such as gelatine, YPE exhibited the additional ability to trap particles and promote their sedimentation in the absence of tannins. This double performance of the YPE used in this work, is likely to guarantee an effective fining of red wines as well as for low tannin content matrices such as rosé and white wines.

Contribution of sequential pretreatments and enzyme-assisted extraction to rice bran protein yield improvement

Alcalase is commonly used in enzyme-assisted extraction of rice bran protein. However, broad handling of rice bran can affect proteins before proteolysis. Various pretreatments, including thermal stabilization, washing, defatting, alkaline extraction, and enzymatic cellulosic degradation, and combinations were identified regarding the present and possible handling processes of rice bran protein extraction to investigate their effects on the proteolysis of the protein afterward. Then, the protein extraction yield and proteolytic abilities were compared. The thermal stabilization of the rice bran strongly affects enzymatic protein extraction, which more elaborate extraction protocols can mitigate. Rice bran oil can be extracted earlier without seriously affecting the protein extraction. Washing removes native protease inhibitors and hence speeds up the protein extraction. Alkaline extraction is highly efficient but can cause protein denaturation, generating a challenging protein separation from the gel-like structure. The protein extraction with enzymatic (Ultimase) pretreatment is an efficient method that allows high hydrolysis of intact rice bran protein during the protein extraction step.

Robust assessment of sample preparation protocols for proteomics of cells and tissues

In proteomic studies, the reliability and reproducibility of results hinge on well-executed protein extraction and digestion protocols. Here, we systematically compared three established digestion methods for macrophages, namely filter-assisted sample preparation (FASP), in-solution, and in-gel digestion protocols. We also compared lyophilization and manual lysis for liver tissue protein extraction, each of them tested using either sodium deoxycholate (SDC)- or RIPA-based lysis buffer. For the macrophage cell line, FASP using passivated filter units outperformed the other tested methods regarding the number of identified peptides and proteins. However, a careful standardization has shown that all three methods can yield robust results across a wide range of starting material (even starting with 1 μg of proteins). Importantly, inter and intra-day coefficients of variance (CVs) were determined for all sample preparation protocols. Thus, the median inter-day CVs for in-solution, in-gel and FASP protocols were respectively 10, 8 and 9%, very similar to the median CVs obtained for the intra-day analysis (9, 8 and 8%, respectively). Moreover, FASP digestion presented 80% of proteins with a CV lower than 25%, followed closely by in-gel digestion (78%) and in-solution sample preparation (72%) protocols. For tissue proteomics, both manual lysis and lyophilization presented similar proteome coverage and reproducibility, but the efficiency of protein extraction depended on the lysis buffer used, with RIPA buffer showing better results. In conclusion, although each sample preparation method has its own particularity, they are all suited for successful proteomic experiments if a careful standardization of the sample preparation workflow is carried out.

Molecular dynamics simulations of betaine-based deep eutectic solvents with varying iso-alcohols chain lengths

Betaine-based (BET) deep eutectic solvents (DESs) made with isoalcohols of varying hydrocarbon chain lengths, namely triethylene glycol (TEG), 1, 2-ethanediol (ETD), 1, 3-propanediol (PPD) and 1, 4-butanediol (BTD) were modelled. Their effects on the formation of aqueous biphasic system (ABS) with 2 M dipotassium phosphate salt solution (K2HPO4) as well as the extraction of common protein molecules, bovine serum albumin (BSA), were studied using molecular dynamics simulation with Gromacs. Qualitative and quantitative data were used to analyse the simulation results, which includes visualised systems in equilibrium, root mean square deviation (RMSD), root mean square fluctuation (RMSF), radius of gyration, radial distribution function (RDF) and hydrogen bond analysis. The results and literature research validated the role of inorganic salt solute concentration on the phase forming ability as well as the possibility of protein denaturation. More investigations and experimental works are to be carried out regarding the type of inorganic salt and its concentration for aqueous biphasic systems (ABSs) with alcohol based DESs and salt solution for protein extraction purposes. This is essential to understand the salting-out effect and to isolate the effects of hydrocarbon chain length from other influencing factors. Additionally, inorganic salt concentrations, presence of ether linkage as well as hydrocarbon chain length is shown to play a role in the dynamics between molecules in the system as well as protein conformational changes. Overall, ABS incorporating PPD and BTD with hydrocarbon chain lengths not exceeding C4 demonstrated superior performance in terms of protein conformational stability and interaction, showcasing their efficacy in protein extraction process.

Proteomic and metabolomic profiling of methicillin resistant versus methicillin sensitive Staphylococcus aureus using a simultaneous extraction protocol.

Understanding the biology of methicillin resistant Staphylococcus aureus (MRSA) is crucial to unlocking insights for new targets in our fight against this antimicrobial resistant priority pathogen. Although proteomics and metabolomic profiling offer the potential to elucidating such biological markers, reports of methodological approaches for carrying this out in S. aureus isolates remain limited. We describe the use of a dual-functionality methanol extraction method for the concurrent extraction of protein and metabolites from S. aureus and report on the comparative analysis of the proteomic and metabolomic profiles of MRSA versus methicillin sensitive S. aureus (MSSA). Bacterial reference strains MRSA ATCC43300 and MSSA ATCC25923 were used. The conventional urea methodology was used for protein extraction and a methanol based method was used for concurrent proteins and metabolites extraction. Proteomic and metabolomic profiling was carried out using TimsTOF mass spectrometry. Data processing was carried out using the MaxQuant version 2.1.4.0. This study represents the first report on the utilization of the methanol extraction method for concurrent protein and metabolite extraction in Gram positive bacteria. Our findings demonstrate good performance of the method for the dual extraction of proteins and metabolites from S. aureus with demonstration of reproducibility. Comparison of MRSA and MSSA strains revealed 407 proteins with significantly different expression levels. Enrichment analysis of those proteins revealed distinct pathways involved in fatty acid degradation, metabolism and beta-lactam resistance. Penicillin-binding protein PBP2a, the key determinant of MRSA resistance, exhibited distinct expression patterns in MRSA isolates. Metabolomic analysis identified 146 metabolites with only one exclusive to the MRSA. The enriched pathways identified were related to arginine metabolism and biosynthesis. Our findings demonstrate the effectiveness of the methanol-based dual-extraction method, providing simultaneous insights into the proteomic and metabolomic landscapes of S. aureus strains. These findings demonstrate the utility of proteomic and metabolomic profiling for elucidating the biological basis of antimicrobial resistance.

The Gliadin Analysis by Reversed-Phase High-Performance Liquid Chromatography of Traditional Fermented “Hamoum” Wheat in West Algeria

Our ancestors had always used food fermentation without having a scientific explanation. Their natural phenomena with therapeutic potential became interesting only in the early twentieth century. A comparative study between the electrophoretic profiles of wheat proteins of fermented wheat type Hamoum (FWH) and regular unfermented wheat (NUW) was performed to identify the effect of fermentation realized by the traditional technique using underground storage (locally named Matmora) for 12 months. SDS-PAGE electrophoresis was performed after the sequential extraction of different proteins (albumins/globulins, gliadins and glutenins). The results showed a total degradation of the regular wheat proteins after fermentation. Fermented wheat type Hamoum (FWH) could be a good alternative for people with disorders related to gluten ingestion as compared to regular unfermented wheat (NUW).

Sustainable extraction of proteins from lime peels using ultrasound, deep eutectic solvents, and pressurized liquids, as a source of bioactive peptides

The aim of this work was to develop, for the first time, sustainable strategies, based on the use of Ultrasound-Assisted Extraction, Natural Deep Eutectic Solvents, and Pressurized Liquid Extraction, to extract proteins from lime (Citrus x latifolia) peels and to evaluate their potential to release bioactive peptides. PLE showed the largest extraction of proteins (66–69%), which were hydrolysed using three different enzymes (Alcalase 2.4 L FG, Alcalase®PURE 2.4 L, and Thermolysin). The in vitro antioxidant and antihypertensive activities of released peptides were evaluated. Although all hydrolysates showed antioxidant and antihypertensive activity, the hydrolysate obtained with Thermolysin showed the most significant values. Since the Total Phenolic Content in all hydrolysates was low, peptides were likely the main contributors to these bioactivities. Hydrolysates were analyzed by UHPLC-QTOF-MS and a total of 98 different peptides were identified. Most of these peptides were rich in amino acids associated with antioxidant activity.

Extraction and characterization of mung bean proteins using different alkaline solutions

Extraction and characterization of mung bean proteins using different alkaline solutions

Molecular and functional characterization of ILYS-5, a major invertebrate lysozyme of Caenorhabditis elegans

To overcome bacterial invasion and infection, animals have evolved various antimicrobial effectors such as antimicrobial peptides and lysozymes. Although C. elegans is exposed to a variety of microbes due to its bacterivorous lifestyle, previous work on the components of its immune system mainly based on the description of transcriptional changes during bacterial challenges. Very few effector components of its immune system have been characterized so far. To investigate the role of lysozymes in terms of antibacterial defense and digestion, we studied a member of the widely neglected family of C. elegans invertebrate lysozymes (ILYS). We focused on the so far virtually undescribed ILYS-5, which we purified from protein extracts of C. elegans tracing its peptidoglycan-degrading activity and localized the tissue expression of the gene in vivo using a translational reporter construct. We recombinantly synthesized ILYS-5 and determined the physicochemical activity optimum and the antibacterial spectrum of a lysozyme from C. elegans for the first time. With an activity optimum at low ionic strength (≤100 mM) and at acidic pH (≤ pH 4.0), ILYS-5 is likely to be involved in killing and digestion of bacteria within acidified phagolysosomes and acidic regions of the gut, presumably secreted by lysosome-like vesicles. This notion is supported by potent activity against various live Gram-positive and Gram-negative bacteria. Notably, members of the natural associated microbiome of C. elegans are substantially less susceptible to ILYS-5. Ablation of the ilys-5 gene resulted in reduction of lifespan and fertility when cultured on the standard food bacterium Escherichia coli OP50, whereas exposure of the ilys-5 knock-out mutant to the host-associated bacterium Pseudomonas lurida MYb11 did not have a clear effect. These findings indicate a role of ILYS-5 in immunity and nutrition and a co-evolved adaptation of host and bacteria to the mutualistic nature of their interaction.

Recovery of Proteases and Protease Inhibitors from Ganoderma spp. Cultivated in Amazonian Lignocellulose Wastes.

Ganoderma spp. are a great source of bioactive molecules. The production and recovery of bioactive molecules vary according to strain, growth substrate, and extraction solution. Variations in protease and their inhibitors in basidiomata from a commercial strain (G. lingzhi) and an Amazonian isolate (Ganoderma sp.) cultivated in Amazonian lignocellulosic wastes and extracted with different solutions are plausible and were investigated in our study. Basidiomata from cultivation in substrates based on açaí seed, guaruba-cedro sawdust and three lots of marupá sawdust were submitted to extraction in water, Tris-HCl, and sodium phosphate. Protein content, proteases, and protease inhibitors were estimated through different assays. The samples were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR). Tris-HCl provided higher protein extraction from Ganoderma sp. and higher caseinolytic, gelatinolytic, and fibrinolytic activity for G. lingzhi cultivated in açaí. Water extracts of Ganoderma sp., in general, exhibited higher trypsin and papain inhibitor activities compared to G. lingzhi. Extracts in Tris-HCl and sodium phosphate showed more intense protein bands in SDS-- PAGE, highlighting bands of molecular weights around 100, 50, and 30 kDa. FTIR spectra showed patterns for proteins in all extracts, with variation in transmittance according to substrate and extractor. Water extract from Amazonian Ganoderma sp. cultivated in marupá wastes are promising as a source of protease inhibitors, while the Tris-HCL extract of G. lingzhi from açaí cultivation stands out as a source of proteases with fibrinolytic, caseinolytic, and gelatinolytic activities.

A Comparison of Phenolic, Flavonoid, and Amino Acid Compositions and In Vitro Antioxidant and Neuroprotective Activities in Thai Plant Protein Extracts.

The leaves of mulberry, Azolla spp., sunflower sprouts, cashew nut, and mung bean are considered rich sources of plant protein with high levels of branched-chain amino acids. Furthermore, they contain beneficial phytochemicals such as antioxidants and anti-inflammatory agents. Additionally, there are reports suggesting that an adequate consumption of amino acids can reduce nerve cell damage, delay the onset of memory impairment, and improve sleep quality. In this study, protein isolates were prepared from the leaves of mulberry, Azolla spp., sunflower sprouts, cashew nut, and mung bean. The amino acid profile, dietary fiber content, phenolic content, and flavonoid content were evaluated. Pharmacological properties, such as antioxidant, anticholinesterase, monoamine oxidase, and γ-aminobutyric acid transaminase (GABA-T) activities, were also assessed. This study found that concentrated protein from mung beans has a higher quantity of essential amino acids (52,161 mg/100 g protein) compared to concentrated protein from sunflower sprouts (47,386 mg/100 g protein), Azolla spp. (42,097 mg/100 g protein), cashew nut (26,710 mg/100 g protein), and mulberry leaves (8931 mg/100 g protein). The dietary fiber content ranged from 0.90% to 3.24%, while the phenolic content and flavonoid content ranged from 0.25 to 2.29 mg/g and 0.01 to 2.01 mg/g of sample, respectively. Sunflower sprout protein isolates exhibited the highest levels of dietary fiber (3.24%), phenolic content (2.292 ± 0.082 mg of GAE/g), and flavonoids (2.014 mg quercetin/g of sample). The biological efficacy evaluation found that concentrated protein extract from sunflower sprouts has the highest antioxidant activity; the percentages of inhibition of 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical were 20.503 ± 0.288% and 18.496 ± 0.105%, respectively. Five plant-based proteins exhibited a potent inhibition of acetylcholinesterase (AChE) enzyme activity, monoamine oxidase (MAO) inhibition, and GABA-T ranging from 3.42% to 24.62%, 6.14% to 20.16%, and 2.03% to 21.99%, respectively. These findings suggest that these plant protein extracts can be used as natural resources for developing food supplements with neuroprotective activity.

Evaluation of antioxidant, anti-inflammatory and antibacterial properties of protein extract isolated from Vigna trilobata

Evaluation of antioxidant, anti-inflammatory and antibacterial properties of protein extract isolated from Vigna trilobata

Evaluation of the functional properties of a protein isolate from Arthrospira maxima and its application in a meat sausage

Arthrospira maxima is a microalga that has been collected in Lake Texcoco in the Valley of Mexico since pre-Hispanic times and has been a traditional food source due to its high biomass production and protein content (50–60 %), making it promising for protein extraction. In this context, a protein isolate was obtained from powdered biomass of Arthrospira maxima (PbAm) by alkaline solubilization (pH 11) and isoelectric precipitation (pH 4.2). Arthrospira maxima protein isolate (AmPI) presented higher protein content (82.58 %) and total amino acids compared to PbAm. Functional properties of AmPI were evaluated in comparison with PbAm and soy protein isolate (SPI). Protein extraction resulted in a significant increase in protein solubility (PS) and foaming capacity (FC) of up to 87.78 % and 238.10 %, respectively. Emulsifying capacity (EC) of AmPI was superior to that of PbAm and SPI in pH range 5–7. Inclusion of AmPI as a partial substitute for SPI in the formulation of meat sausages was evaluated by implementing four treatments: T1 (15 % AmPI, 85 % SPI), T2 (10 % AmPI, 90 % SPI), T3 (5 % AmPI, 95 % SPI) and T4 (0 % AmPI, 100 % SPI). Although the texture attributes remained unchanged, a significant reduction in color parameters was observed as the concentration of AmPI increased. An inclusion of 15 % AmPI significantly enhanced the nutritional quality of meat sausages. Results highlight the excellent properties of AmPI, confirming Arthrospira maxima as a promising protein source in the food industry.

Valorization of Bombyx mori pupae and Moringa oleifera leaf as sources of alternative protein: Structural, functional and antioxidant properties

This study compared protein concentrates extracted from silkworm pupae (SPP) and moringa leaf (MLP) using alkaline extraction followed by isoelectric precipitation with commercial whey protein concentrate (WPC). SPP demonstrated higher protein extraction yield of 5.99 % and protein recovery of 66.02 % compared to MLP (4.39% and 55.27% of extraction yield and protein recovery, respectively). Proximate composition analysis revealed that SPP contained high protein of 65.40 % with ∼13 % fat content, while MLP had lower protein (48.70%) and fat content (0.33%). Particle size analysis showed small mean diameters of ∼123 nm for WPC, ∼192 nm for SPP, and ∼1485 nm for MLP with low polydispersity index (0.35 to 0.51). Secondary structure analysis highlighted β-sheet as predominant in WPC and MLP, while SPP exhibited a major β-turns structure. Solubility studies indicated that SPP had lowest solubility compared to others. In terms of functional properties, MLP exhibited the highest water and oil holding capacities, and foaming capacity, while SPP showed the greatest emulsifying capacity and stability. After passing simulated intestinal digestion, SPP exhibited the highest recovery of TPC, ABTS, DPPH, and FRAP. This study emphasizes the feasibility and advantages of utilizing SPP and MLP as food ingredients for various food applications.