Improvement of protein extraction efficiency from defatted sesame meal with thermal and enzymatic treatments

Enzymatic hydrolysis has been widely applied for production of protein hydrolysate from shrimp waste and for purification of chitin. In the present study, shrimp (P. semisulcatus) head waste was hydrolysed, using a commercial proteolytic enzyme, Alcalase. In order to improve protein extraction efficiency, certain chemicals such as sodium sulphite and Triton x-100 were used along with the enzyme. When Alcalase (12 AU/kg) used alone, the yield of protein extraction was 45.1% and by using Triton x-100 (0.01 g/kg) together with Alcalase, the yield was decreased to 39%, whereas the presence of sodium sulphite (200 mmol/L) with the enzyme or with the enzyme and Triton x-100 increased the level of protein extraction to 62% and 65.1%, respectively. The resulting protein powder contained sufficient amounts of essential amino acids to be used in feed formulations. By precipitating proteins from the resulting protein extract at pH 3.1, the residual sulphite in protein powder was decreased by 97% and thus the powder can be considered suitable for animal and/or aquaculture feed formulations.

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EXPLORATION OF THE FACTORS AFFECTING THE SOLUBLE PROTEIN EXTRACTION FROM CULTURED SNAKEHEAD FISH (Channa striata) MUSCLE

Vietnam Journal of Science and Technology ◽

10.15625/2525-2518/55/5a/12181 ◽

2018 ◽

Vol 55 (5A) ◽

pp. 74

Author(s):

Vo Hoang Ngan

Keyword(s):

Soluble Protein ◽

Extraction Efficiency ◽

Protein Extraction ◽

Ph Value ◽

Fish Muscle ◽

Effect Of Temperature ◽

Extraction Temperature ◽

Solution Ph ◽

Factors Affecting ◽

Optimum Extraction

This study was conducted to identify the factors affecting the soluble protein extraction from cultured snakehead fish muscle, a protein having bioavailability and nutritional value. The study investigated the ratio of fish muscle to water (from 1:1 to 1:5 w/v) and salt (NaCl) concentration (from 0.05 M to 0.25 M) that are suitable for extracting soluble protein effectively. Research has applied a response surface methodology using a central composite design to optimize the effect of temperature and time extraction on soluble protein extraction efficiency. In addition, the study investigated the effect of pH of salt solution (pH 2 to pH 12) on the efficiency of soluble protein extraction from cultured snakehead fish muscle. As a result, the use of 0.15 M NaCl solution with 1:2 ratio of fish muscle to water gave high soluble protein extraction efficiency. The optimum extraction temperature and time were determined as 34.8 °C and 15.2 min respectively, and the appropriate pH value was also determined as pH 10. At the optimum extraction conditions, soluble protein recovery calculated by the amount of soluble protein extraction dividing total protein of fish muscle was 14.39 ± 0.18 %.

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Use of carbohydrase to enhance protein extraction efficiency and antioxidative properties of oat bran protein hydrolysates

Food Research International ◽

10.1016/j.foodres.2011.12.004 ◽

2012 ◽

Vol 46 (1) ◽

pp. 69-75 ◽

Cited By ~ 36

Author(s):

Sara Jodayree ◽

Jeffrey C. Smith ◽

Apollinaire Tsopmo

Keyword(s):

Extraction Efficiency ◽

Protein Extraction ◽

Protein Hydrolysates ◽

Antioxidative Properties ◽

Oat Bran

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Leaf protein analyses in order to utilise the leaf shoot of artichoke

Acta Agraria Debreceniensis ◽

10.34101/actaagrar/67/1751 ◽

2016 ◽

pp. 43-47

Author(s):

László Kaszás ◽

Miklós Fári ◽

Sándor Hodossi ◽

Éva Domokos-Szabolcsy

Keyword(s):

Protein Concentration ◽

Extraction Efficiency ◽

Protein Extraction ◽

Jerusalem Artichoke ◽

The Other ◽

Leaf Protein ◽

Alternative Source ◽

Feed Industry ◽

Alternative Protein

The constat growth of the Earth’s population brings with itself a higher demand for food and protein not only in human nutrition but also for the feeding of livestock. Currently, the feed industry is mainly built onseed-based protein, wherethebaseplant is soybean, which is large lycovered by imports in Hungary, similar toother European countries. However, the long-term economically sustainable lifestock breeding demandschanges which has also worked out strategies. An alternative protein sources could be green leafy plants. In current work the Jerusalem artichokes as an alternative source of protein was studied, compared to alfalfa as a valuable protein plant. Our results indicate that fiber fraction ofJerusalem artichoke shootswas 34 to 37% after pressing in the autumn period while alfalfa slightly lower values were obtained (30%). On the other hand extracted green leaf protein concentration was higher in alfalfa than in Jerusalem artichoke. Along with this higher protein content could be measured from the leaf protein concentration of alfalfa and almost each amino acids were more, as well comparing to Jerusalem artichoke. Overall, the alfalfa proved to be advantageous as expected both in leaf protein extraction efficiency, both regarding the content of the protein in the Jerusalem artichoke. However, considering aminoacid composition and green biomass production, Jerusalem artichokecould be a promisingplant species asplant protein sourceinthefuture.

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Hydrothermal pre-treatment coupled with urea solubilisation enables efficient protein extraction from microalgae

Green Chemistry ◽

10.1039/c9gc03120k ◽

2019 ◽

Vol 21 (23) ◽

pp. 6361-6371 ◽

Cited By ~ 4

Author(s):

Hakan Karan ◽

Renée de Boeck ◽

John Roles ◽

Ben Hankamer ◽

Ian L. Ross

Keyword(s):

Extraction Efficiency ◽

Protein Extraction ◽

Pre Treatment

Hydrothermal pre-treatment (HTP) allows microalgal protein extraction. Subsequent acetone and urea treatments improve the extraction efficiency.

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Effect of polyethylene glycol 20 000 on protein extraction efficiency of formalin-fixed paraffin-embedded tissues in South Africa

African Journal of Laboratory Medicine ◽

10.4102/ajlm.v10i1.1122 ◽

2021 ◽

Vol 10 (1) ◽

Author(s):

Sophia Rossouw ◽

Hocine Bendou ◽

Liam Bell ◽

Jonathan Rigby ◽

Alan Christoffels

Keyword(s):

Polyethylene Glycol ◽

Extraction Efficiency ◽

Protein Extraction ◽

Formalin Fixed Paraffin ◽

Liquid Chromatography Tandem Mass ◽

Formalin Fixed Paraffin Embedded ◽

Ffpe Tissues ◽

Protein Selection ◽

The Impact ◽

Formalin Fixed

Background: Optimal protocols for efficient and reproducible protein extraction from formalin-fixed paraffin-embedded (FFPE) tissues are not yet standardised and new techniques are continually developed and improved. The effect of polyethylene glycol (PEG) 20 000 on protein extraction efficiency has not been evaluated using human FFPE colorectal cancer tissues and there is no consensus on the protein extraction solution required for efficient, reproducible extraction.Objective: The impact of PEG 20 000 on protein extraction efficiency, reproducibility and protein selection bias was evaluated using FFPE colonic tissue via liquid chromatography tandem mass spectrometry analysis.Methods: This study was conducted from August 2017 to July 2019 using human FFPE colorectal carcinoma tissues from the Anatomical Pathology department at Tygerberg Hospital in South Africa. Samples were analysed via label-free liquid chromatography tandem mass spectrometry to determine the impact of using PEG 20 000 in the protein extraction solution. Data were assessed regarding peptide and protein identifications, method efficiency, reproducibility, protein characteristics and organisation relating to gene ontology categories.Results: Polyethylene glycol 20 000 exclusion increased peptides and proteins identifications and the method was more reproducible compared to the samples processed with PEG 20 000. However, no differences were observed with regard to protein selection bias. We found that higher protein concentrations ( 10 µg) compromised the function of PEG.Conclusion: This study indicates that protocols generating high protein yields from human FFPE tissues would benefit from the exclusion of PEG 20 000 in the protein extraction solution.

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Novel perspective on a conventional technique: Impact of ultra-low temperature on bacterial viability and protein extraction

PLoS ONE ◽

10.1371/journal.pone.0251640 ◽

2021 ◽

Vol 16 (5) ◽

pp. e0251640

Author(s):

Aditya Sarnaik ◽

Apurv Mhatre ◽

Muhammad Faisal ◽

Dylan Smith ◽

Ryan Davis ◽

...

Keyword(s):

Low Temperature ◽

Cell Viability ◽

Bacterial Cell ◽

Storage Time ◽

Extraction Efficiency ◽

Protein Extraction ◽

Fold Increase ◽

Cell Lysis ◽

Bacterial Cells ◽

The Impact

Ultra-low temperature (ULT) storage of microbial biomass is routinely practiced in biological laboratories. However, there is very little insight regarding the effects of biomass storage at ULT and the structure of the cell envelope, on cell viability. Eventually, these aspects influence bacterial cell lysis which is one of the critical steps for biomolecular extraction, especially protein extraction. Therefore, we studied the effects of ULT-storage (-80°C) on three different bacterial platforms: Escherichia coli, Bacillus subtilis and the cyanobacterium Synechocystis sp. PCC 6803. By using a propidium iodide assay and a modified MTT assay we determined the impact of ULT storage on cellular viability. Subsequently, the protein extraction efficiency was determined by analyzing the amount of protein released following the storage. The results successfully established that longer the ULT-storage time lower is the cell viability and larger is the protein extraction efficiency. Interestingly, E. coli and B. subtilis exhibited significant reduction in cell viability over Synechocystis 6803. This indicates that the cell membrane structure and composition may play a major role on cell viability in ULT storage. Interestingly, E. coli exhibited concomitant increase in cell lysis efficiency resulting in a 4.5-fold increase (from 109 μg/ml of protein on day 0 to 464 μg/ml of protein on day 2) in the extracted protein titer following ULT storage. Furthermore, our investigations confirmed that the protein function, tested through the extraction of fluorescent proteins from cells stored at ULT, remained unaltered. These results established the plausibility of using ULT storage to improve protein extraction efficiency. Towards this, the impact of shorter ULT storage time was investigated to make the strategy more time efficient to be adopted into protocols. Interestingly, E. coli transformants expressing mCherry yielded 2.7-fold increase (93 μg/mL to 254 μg/mL) after 10 mins, while 4-fold increase (380 μg/mL) after 120 mins of ULT storage in the extracted soluble protein. We thereby substantiate that: (1) the storage time of bacterial cells in -80°C affect cell viability and can alter protein extraction efficiency; and (2) exercising a simple ULT-storage prior to bacterial cell lysis can improve the desired protein yield without impacting its function.

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Development of Aqueous Two-Phase Systems Based on Deep Eutectic Solvents for Continuous Protein Extraction in a Microextractor

Engineering Proceedings ◽

10.3390/micromachines2021-09546 ◽

2021 ◽

Vol 4 (1) ◽

pp. 6

Author(s):

Anabela Ljubić ◽

Anita Šalić ◽

Bruno Zelić

Keyword(s):

Extraction Efficiency ◽

Protein Extraction ◽

Batch Reactor ◽

High Surface ◽

Deep Eutectic Solvents ◽

Thermomyces Lanuginosus ◽

Phase System ◽

Two Phase ◽

Promising Alternative ◽

The Impact

Currently, lipases are one of the most widely used enzymes, especially in catalysis, mostly due to their high activity in mild conditions and wide specificity. Therefore, obtaining the highest possible catalytic activity, which can be achieved through purification, is becoming more and more important. Since most of the purification techniques are time consuming, aqueous two-phase protein extraction is often investigated as a promising alternative. Additionally, this kind of extraction can be carried out in microextractors, which provides not only a continuous processing of raw materials, but also significantly higher efficiencies due to a high surface-to-volume ratio of microchannels. Extraction with deep eutectic solvents (DESs) fulfills all green chemistry principles, because DESs are biodegradable, non-toxic, and recyclable. In this research, the aqueous two-phase system based on natural DES for continuous protein extraction in a microextractor was investigated. The impact of salt concentration on extraction efficiency was investigated in batch experiments with six different previously characterized DESs. After determination of the optimal two-phase system features, the process was transferred to a microextractor. In addition, the selected DES was tested for recyclability while the developed extraction method was verified using raw lipase produced by Thermomyces lanuginosus solid-state cultivation on hull-less pumpkin oil pomace. The highest protein extraction efficiency achieved in a batch reactor was 94.70% for 30 min, while in a microextractor, the highest extraction efficiency obtained was 98.50% for 30 s. Obviously, the extraction process was significantly intensified by continuous microextraction. Additionally, the DES used in the microextraction experiments was efficiently reused in several extraction cycles.

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