Optimization Of Protein Extraction Research Articles

Pomegranate seed as a novel source of plant protein: Optimization of protein extraction and evaluation of invitro digestibility, functional, and thermal properties.

This research was carried out to optimize the extraction process of proteins from pomegranate seeds and characterize their invitro digestibility as well as their thermal and functional properties. For this purpose, the study screened five parameters (liquid/solid ratio, pH, temperature, NaCl concentration, and time) that could potentially influence the extraction process. This screening was conducted using a two-level Placket-Burman design (PBD). The significant parameters (pH and NaCl concentration) were subsequently optimized using a three-level face-centered central composite design (FCCD) to determine the optimum extraction conditions. A maximum protein recovery of 83.8% was obtained at pH 11.0 and NaCl concentration of 0.0 M. Pomegranate seed protein isolate (PSPI) with a protein content of 92.4% (w/w) was obtained through the isoelectric precipitation of pomegranate seed protein extracted under the optimized conditions. An emulsifying activity index of 14.1 m2 g-1 was observed at the isoelectric pH, where the emulsion stability index was at 8.2%. PSPI also showed high water- and oil-holding capacities (3.7 and 4.3 g g-1, respectively). The essential amino acid levels in PSPI (except for valine and isoleucine) exceeded the recommended amounts set by WHO/FAO/UNU for adults, highlighting its high nutritional value. Based on thermal analysis data, denaturation of PSPI could occur at 89.5°C. The invitro digestibility of PSPI was found to be 74.3%. PSPI shows a potential as a novel ingredient for substituting animal-based proteins in various food applications.

Optimization of Protein Extraction from Halopteris scoparia Macroalgae by Ultrasonic-Assisted Enzymatic Extraction (UAEE): Bioactive, Chemical, and Technological Properties

Optimization of Protein Extraction from <i>Halopteris scoparia</i> Macroalgae by Ultrasonic-Assisted Enzymatic Extraction (UAEE): Bioactive, Chemical, and Technological Properties

Optimization of protein extraction from sesame seeds by using response surface methodology RSM

Optimization of protein extraction from sesame seeds by using response surface methodology RSM

Pomegranate seed as a novel source of plant protein: optimization of protein extraction and evaluation of in vitro digestibility, functional, and thermal properties

Pomegranate seed as a novel source of plant protein: optimization of protein extraction and evaluation of in vitro digestibility, functional, and thermal properties

Optimisation of protein extraction from selenium-enriched Brassicaceae leaves

Vegetable side-streams have gained attention for their potential as sustainable and nutritious food sources for human consumption. This study focused on the optimisation of leaf protein extraction from selenium (Se)-enriched leaf meals (LMs) derived from the Brassicaceae family, including broccoli (Se-BrLM), cabbage (Se-CbLM), radish (Se-RaLM), and Cardamine violifolia (Se-CmLM) leaf meals. The primary focus of the extraction process revolved around two key protein fractions: albumin and glutelin, both highly abundant in the LMs. The results highlighted the significance of ionic strength, extractant pH, and solid-to-liquid (S/L) ratio in leaf protein extraction, with sonication time exerting a comparatively less pronounced influence. Within the Brassicaceae species investigated, the optimal conditions included an extractant pH of 9–10, sonication duration of 30–45 s, an S/L ratio of 1:40, and the inclusion of salt at concentrations of 60 and 300 mM. Under these conditions, Se-CmLM achieved a significantly higher mass yield of 4.80 % (dry basis, db) and protein recovery of 17.55 % (db), with RuBisCo as the major protein within the resulting protein concentrates. The protein–phenolic complex content was minimal, with in vitro protein digestibility of 76.71–80.36 %. This study emphasises the potential of Brassicaceae leaves as sustainable protein sources.

Optimization of protein extraction from brewer’s spent grain and production of bioactive peptides

Optimization of protein extraction from brewer’s spent grain and production of bioactive peptides

What You Extract Is What You Get: Different Methods of Protein Extraction from Hemp Seeds

Cannabis sativa L. seeds are rich in essential polyunsaturated fatty acids and highly digestible proteins, with a good nutritional value. Proteomics studies on hempseed reported so far have mainly been conducted on processed seeds and, to our knowledge, no optimization of protein extraction from hemp seeds has been performed. This study investigates the SDS-PAGE profile of hempseed proteins comparing different methods of extraction, (Osborne sequential extraction, TCA/acetone, MTBE/methanol, direct protein solubilization of defatted hempseed flour), two conditions to keep low temperature during seed grinding (liquid nitrogen or ice) and two solubilization buffers (urea-based or Laemmli buffer). Among the tested conditions, the combination of liquid nitrogen + TCA/acetone + Laemmli buffer was not compatible with SDS-PAGE of proteins. On the other hand, urea-based buffer achieved more reproducible results if combined with all the other conditions. TCA/acetone, MTBE/methanol, and direct protein solubilization of defatted hempseed flour demonstrated a good overview of protein content, but less abundant proteins were poorly represented. The Osborne sequential separation was helpful in diluting abundant proteins thus enhancing the method sensitivity.

Optimization of Protein Extraction from Tuna Cooking Liquid and Its Property Analysis

In order to provide scientific basis for the comprehensive utilization of high nitrogen waste water from tuna processing industry. Besides, it can provide theoretical guidance for pollution reduction in the industry. On the basis of single factor experiment, the effects of liquid-to-material ratio, temperature, time, pH, and solid content on the ethanol extraction of tuna cooking liquid protein were studied by response surface test. Further, the structure of the extracted protein was characterized by Ultraviolet Scanning, Infrared Spectroscopy, Aamino Acid Analyzer and Circular Dichroism. The results showed that the optimal process conditions were temperature 36 ℃, time 2 h, liquid-to-material ratio 2.4, and the maximum protein extraction rate was 76.83%. The protein extracted by this method had the maximum absorption peak at about 218 nm, but no obvious absorption peak at 280 nm. The amino acid composition showed that the mass fraction of amino acids was 18.79%, and the content of hydroxyproline was 10.21%. The infrared spectrum had the characteristic absorption peak of gelatin. The above results indicate that the protein extracted from the cooking liquid by this process is gelatin.

Optimization of protein extraction from watermelon seeds by liquid-phase pulsed discharge based on energy input for scale-up application

The study is about the watermelon seeds protein extraction using the specific energy per pulse (EPP) (W/g) as the operation parameter for optimization by liquid-phase pulsed discharge (LPD) at various scales, in comparison with the extraction using discharge voltage (kV) as the operation parameter for optimization by LPD. The optimization based on the intrinsic parameters of EPP (W/g), liquid to solid (L/S) ratio (mL/g) and extraction time (min) were firstly implemented. The protein yield of 462.26±1.214 mg/g was obtained under optimal conditions (0.042 W/g EPP, 3.8 min extraction time and 40 mL/g L/S ratio), which was in accordance with the yield of 462.03 ± 1.061 mg/g obtained by traditional optimization based on discharge voltage, extraction time and L/S ratio. Then, the two optimization methods were compared at larger scale of 100–280 mL solvent loading. The protein yields of the EPP optimization were 461.313±0.990, 462.102±1.034, 462.009±0.872 and 460.319±0.981, respectively, which had no significant difference. However, the protein yields of traditional optimization method decreased rapidly. Thus, the results showed that the EPP optimization method had the favorable effect on the reliability and feasibility of the scale-up LPD extraction. These indicated that the EPP accurately reflected the LPD extraction mechanism, and could replace the discharge voltage.

Optimization of protein extraction and ELISA immunodetection from protein-based paint models with mesoporous silica nanoparticles and MCM41

Protein-based biological materials such as albumin, casein and collagen are found in various cultural heritage (CH) artefacts. This study focuses on the study of protein binders from easel paintings media. Proteins have complex structures which are difficult to identify with non-invasive spectroscopic methods (FT-IR, Raman, UV). Immunoassays such as ELISA determine the protein’s source of origin which is necessary for art objects. To increase the detection and identification of proteins by immunoassays, the efficiency of micro-extraction of proteins from heritage materials is a crucial step. Extractions mediated by cycles of orbital agitation and ultrasonic radiation give the possibility to extract proteins from easel painting sample. In this work, protein-based paint models coupled with silica nanoparticles were used for micro-extraction. Nanoparticles possess high surface-to-volume ratios that can attach bioactive molecules such as proteins and increase the total protein recovered from microsamples. Protein extracts were quantified with Bradford Assay in the presence of Coomassie blue. The protein recovery results were statistically computed, and the SPSS analysis shows significant (p < 0.05) increase in protein recovery, above 1.3 times for NPSiO2 and above 1.6 times for MCM-41. The statistical data shows evidence that silica nanoparticles intensify the total protein recovered from paint microsamples. Finally, ELISA was realized on the protein extracts to verify and compare the immunodetection of protein from the paint models with and without the use of silica nanoparticles.

Evaluation and Optimization of Protein Extraction From E. coli by Electroporation.

Growing diversity of protein-based technologies dictates further development of bio manufacturing to lower the cost of production and maximize yields. Intracellularly expressed recombinant proteins must be extracted from production host prior to purification. Use of electroporation to obtain proteins from bacteria and yeasts has been demonstrated in several studies for different modes of operation and formats. Here we tested various protocols for protein extraction from Escherichia coli by means of electroporation. The tested protocols were compared to established extraction methods of ultrasonication and glass-bead milling in terms of protein yields and content of impurities such as host cell DNA and endotoxins in the lysate. Protein extraction yield was maximal when exponentially growing bacteria were treated at 37°C, regardless of the electroporation mode of operation (batch or flow). We were unable to eliminate co-extraction of host DNA and endotoxins, but with 8 × 1 ms, 5 kV/cm, 1 Hz pulses they were minimized. Yields with optimized electroporation (up to 86 g protein/kg dry weight) were inferior to those in ultrasonication (up to 144 g protein/kg dry weight) and glass-bead milling (up to 280 g protein/kg dry weight). Nevertheless, electroporation largely avoids cell lysis and disintegration with which the extract is a mix of extracted proteins with debris of the bacterial envelope and bacterial DNA, which necessitates further purification.

Optimization of protein extraction from bamboo shoots and processing wastes using deep eutectic solvents in a biorefinery approach

Bamboo shoots are a popular food in Asia and throughout the world. As only the tip of the bamboo shoot is edible, significant quantities of processing byproducts are generated, with potential for valorization of these wastes. Deep eutectic solvents (DESs) are emerging as eco-friendly solvents for the extraction of bioactive compounds from natural products. This work investigated the extraction of protein from the tip of the bamboo shoot (TBS), as well as its processing byproducts (basal bamboo shoot (BBS) and sheath) using a DES comprised of compounds accepted as food additives: choline chloride (ChCl) as hydrogen bond acceptor (HBA) and levulinic acid as hydrogen bond donor (HBD). Central composite design (CCD) and Response Surface Methodology (RSM) were used to predict optimum extraction parameters for all bamboo shoot parts with HBA-to-HBD molar ratio of 6, temperature of 80 °C, time of 50 min, water content of 40% (v/v), and solid-to-liquid ratio of 30 mg/mL for TBS, 70 mg/mL for BBS and sheath. Under the optimal conditions, the protein extraction yields were 39.16 ± 1.22 mg/g dry weight (DW) for TBS, 15.46 ± 0.30 mg/g DW for BBS, and 9.54 ± 0.17 mg/g DW for sheath. Compared with conventional extraction using sodium hydroxide (NaOH) solution, significantly higher protein yield was recovered with DES for TBS, and comparable yields obtained for BBS and sheath. Results show the potential of this DES as a carrier for extracted bamboo proteins in functional food applications, as a biorefinery approach for bamboo shoot processing.

Optimization of Protein Extraction from the Low Organic Residual Municipal Sludge Waste

In this study, municipal sludge waste was used for protein extraction. Two pretreatment ways such as acid and alkaline hydrolysis were compared and the alkaline hydrolysis method was selected as the better way for protein extraction from the residual sludge. In order to obtain the optimal condition for protein extraction, the factors such as NaOH concentration, NaOH solution volume and pretreatment period were selected and optimized by response surface methodology, and the optimal condition for protein extraction was obtained: NaOH concentration was 3.85 mol/l , 24.0 g wet municipal sludge waste was added in 1 L of water, and the pretreatment period was of 12.16 h at 50 °C. Under the optimal extraction condition, the protein extraction yield could reach 5923±107 μg/g sludge, which were 2.07 folds than the initial production pretreated by NaOH.

Optimization of protein extraction from green algae Chaetomorpha sp. by response surface methodology

Green brackish algae Chaetomorpha sp. are easily found in shrimp ponds in Mekong Delta, Vietnam. They can also be co-cultured with shrimps in brackish water shrimp ponds to increase shrimp health and yield. Chaetomorpha sp. algae contain high amount of protein from 10 to 20% w/w db, including water soluble protein and alkaline-soluble protein with over 88% total protein. Dried material were used for protein extraction by using cellulase enzyme (Crestone Conc., Genecor) and NaOH solution. In this research, we optimize the extraction condition of protein from green algae Chaetomorpha sp. by using response surface methodology (RSM). At optimal extraction conditions, dried material was used for protein extraction by using cellulase enzyme (Crestone Conc., Genecor) with the enzyme dosage of 121 UI/g db at 400C during 90 mins. After extraction, the slurry was centrifuged to separate the algae biomass residue to extract the alkaline-soluble protein. The protein extraction yield by using cellulase enzyme was 38.921 mg/g db. After that the, algae biomass residue was extracted by a 1.2% NaOH solution for 78 mins at 500C. The protein extraction yield was 68.651 mg/g db. The total protein extraction yield was 105.755 mg/g db. The extraction yield was increased 10.33% when using the response surface methodology. Concentrated algae protein can be used as a good protein source for food and feed products.&#x0D;

Optimization of protein extraction and proteomic studies in Cenchrus polystachion (L.) Schult

Apomicts have been studied at their genetic levels, but there are no any direct evidence of its mechanism. In order to understand the mechanism involved, a close relative of Pennisetum, Cenchrus polystachion, an apomictic species was explored for more insights into protein expression in reproductive structures. Optimization of protein extraction was studied with the leaf tissue and optimized protocol was extrapolated to other five tissues. The phenol-based protein extraction emerged as the best method for plant leaf tissue providing a better protein yield, separation of bands, removal of non-protein components like polyphenolic compounds and nucleic acids. The proteome analysis of leaf, stigma, immature ovary, seed, anther sac and pollen tissues of Cenchrus polystachion were carried out identifying a total of 135407 proteins against the Poaceae database from UNIPROT/TrEMBL. The target candidate proteins found in all the tissues were identified and mainly comprised of Actin Protein, PIP, Starch Synthase, ATP Synthase, Glutathione S Transferase, Dehydroascorbate reductase, Ascorbate peroxidase and heat shock proteins. Visualization and descriptive statistics conveyed all the necessary information to understand the differential expression of proteins in Cenchrus polystachion. This study forms a base to understand the role of tissue specific expressed proteins in an apomictic plant.

Optimization of protein extraction from Chlorella Vulgaris via novel sugaring-out assisted liquid biphasic electric flotation system.

Microalgae biomass has been consumed as animal feed, fish feed and in human diet due to its high nutritional value. In this experiment, microalgae specie of Chlorella Vulgaris FSP-E was utilized for protein extraction via simple sugaring-out assisted liquid biphasic electric flotation system. The external electric force provided to the two-phase system assists in disruption of rigid microalgae cell wall and releases the contents of microalgae cell. This experiment manipulates various parameters to optimize the set-up. The liquid biphasic electric flotation set-up is compared with a control liquid biphasic flotation experiment without the electric field supply. The optimized separation efficiency of the liquid biphasic electric flotation system was 73.999±0.739% and protein recovery of 69.665±0.862% compared with liquid biphasic flotation, the separation efficiency was 61.584±0.360% and protein recovery was 48.779±0.480%. The separation efficiency and protein recovery for 5 × time scaled-up system was observed at 52.871±1.236% and 73.294±0.701%. The integration of simultaneous cell-disruption and protein extraction ensures high yield of protein from microalgae. This integrated method for protein extraction from microalgae demonstrated its potential and further research can lead this technology to commercialization.

Optimization of protein extraction from detoxified Jatropha seed cake using response surface methodology and amino acid analysis

Jatropha seed cake is a by-product of biodiesel industry which is rich in proteins and carbohydrates and contains many bioactive compounds. Besides having a good protein quality, the essential amino acid content (except lysine) is also high in Jatropha seed cake, and therefore, Jatropha proteins can be exploited for many technical applications. The major hindrance for its use as a protein supplement is the presence of anti-nutritional factors and toxins, especially the phorbol esters. The detoxification of Jatropha seed cake for the removal of phorbol esters has been shown in our earlier studies. Proteins were extracted from the detoxified Jatropha seed cake and analyzed for amino acid content. Various parameters such as temperature, solubilization pH, precipitation pH and extraction time were studied for their effect on protein content, and the process was optimized using response surface methodology among 5 best samples selected based on protein content and protein yield from dry concentrates. The amount of dry matter, protein content and protein yield of 17.1%, 41.98% and 38.66%, respectively, was obtained under the optimized conditions of 60 °C temperature, solubilization pH 11.0, precipitation pH 4.41 and extraction time of 0.78 h. Amino acid contents revealed that the extracted samples contained a higher concentration of essential amino acids, especially leucine (4.40 ± 0.98 g/100 g). The results indicated that the protein extraction procedure from Jatropha seed cake affected the amino acid content, yet the values were close to FAO/WHO reference protein, and thus, the Jatropha proteins can be explored further for their use as feed supplement.

Optimisation of Protein Extraction from Medicinal Cannabis Mature Buds for Bottom-Up Proteomics.

Medicinal cannabis is used to relieve the symptoms of certain medical conditions, such as epilepsy. Cannabis is a controlled substance and until recently was illegal in many jurisdictions. Consequently, the study of this plant has been restricted. Proteomics studies on Cannabis sativa reported so far have been primarily based on plant organs and tissues other than buds, such as roots, hypocotyl, leaves, hempseeds and flour. As far as we know, no optimisation of protein extraction from cannabis reproductive tissues has been attempted. Therefore, we set out to assess different protein extraction methods followed by mass spectrometry-based proteomics to recover, separate and identify the proteins of the reproductive organs of medicinal cannabis, apical buds and isolated trichomes. Database search following shotgun proteomics was limited to protein sequences from C. sativa and closely related species available from UniprotKB. Our results demonstrate that a buffer containing the chaotrope reagent guanidine hydrochloride recovers many more proteins than a urea-based buffer. In combination with a precipitation with trichloroacetic acid, such buffer proved optimum to identify proteins using a trypsin digestion followed by nano-liquid chromatography tandem mass spectrometry (nLC-MS/MS) analyses. This is validated by focusing on enzymes involved in the phytocannabinoid pathway.

Optimizing and predicting degree of hydrolysis of ultrasound assisted sodium hydroxide extraction of protein from tea (Camellia sinensis L.) residue using response surface methodology.

Response surface methodology was employed to investigate the effect of sodium hydroxide concentration (X 1: 0.05-0.15M), sonication time (X 2: 5-15min), ultrasonic power (X 3: 150-450W/L), and solid-liquid ratio (X 4: 20-60g/L) on the optimization of protein extraction from tea residue. Single frequency countercurrent ultrasound (SFCU) was employed to assist the extraction and subsequent hydrolysis of the protein. Optimal extraction conditions were established and response surfaces were generated using mathematical models. There were positive linear and negative quadratic effects of extraction variables on protein yield. The optimal predicted protein yield of 138.9mg/g was obtained under the optimum conditions of concentration of 0.13M, extraction time of 13min, ultrasonic power of 377W/L and solid-liquid ratio of 51.5g/L. A model for the degree of hydrolysis of the extraction process was also obtained which gave a predicted and experimental value of 8.4% and 7.5% respectively. Essential amino acid content of 36.7% was obtained under optimal conditions.

Optimization of protein extraction and two-dimensional gel electrophoresis profiles for the identification of Cordyceps sinensis and other similar species.

Given that Chinese materia medica (CMM) is expensive and rare, people take tremendous risk to adulterate and falsify Cordyceps sinensis with counterfeit species with similar morphological features. It is thus essential to develop new methods to identify the authenticity of Cordyceps sinensis. It is hypothesized in this study that Cordyceps sinensis possesses certain protein biomarkers distinct from its counterfeits, which can be identified by proteomic technologies for authentication purposes. This is the first study that aims to optimize the conditions for extracting proteins from Cordyceps sinensis, a hybrid of fungal-animal CMM, and to compare the two-dimensional gel electrophoresis (2-DE) profiles between different Cordyceps species. Two different protein extraction buffer systems, namely, phenol/sodium dodecyl sulfate (SDS) buffer or lysis buffer, were evaluated, where the preparation using lysis buffer yielded better protein content. The results also showed that extraction with lysis buffer without pre- or post-washing of samples was the most effective protocol, with over 220% of protein yield and 819 protein spots detected on a 2-DE gel. Moreover, the results demonstrated that Cordyceps sinensis possesses protein biomarkers distinct from its counterfeits, and these biomarkers are not source- or origin-dependent, strongly supporting the feasibility of using identified biomarkers as indicators for authentication of Cordyceps species. The findings of this study warrant further investigations on the structural identification of protein biomarkers of Cordyceps species.