Protein Extraction Efficiency and Selectivity of Esterified Styrene–Maleic Acid Copolymers in Thylakoid Membranes

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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Synthesis and Evaluation of a Library of Alternating Amphipathic Copolymers to Solubilize and Study Membrane Proteins

10.33774/chemrxiv-2021-c0clr-v2 ◽

2021 ◽

Author(s):

Adrian H. Kopf ◽

Odette Lijding ◽

Barend O. W. Elenbaas ◽

Martijn C. Koorengevel ◽

Justyna M. Dobruchowska ◽

...

Keyword(s):

Membrane Proteins ◽

Maleic Acid ◽

Rational Design ◽

Extraction Efficiency ◽

Yeast Mitochondria ◽

Electronic Effects ◽

Alternating Copolymers ◽

Polymer Properties

Amphipathic copolymers such as poly(styrene-maleic acid) (SMA) are promising tools for the facile extraction of membrane proteins (MPs) into native nanodiscs. Here, we designed and synthesized a library of well-defined alternating copolymers of SMA analogues in order to elucidate polymer properties that are important for MP solubilization and stability. MP extraction efficiency was determined using KcsA from E.coli membranes and general solubilization efficiency was investigated via turbidimetry experiments on membranes of E.coli, yeast mitochondria and synthetic lipids. Remarkably, halogenation of SMA copolymers dramatically improved solubilization efficiency in all systems, while substituents on the copolymer backbone improved resistance to Ca2+. Relevant polymer properties were found to include hydrophobic balance, size and positioning of substituents, rigidity and electronic effects. The library thus contributes to the rational design of copolymers for the study of MPs.

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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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Membrane protein extraction and purification using styrene–maleic acid (SMA) copolymer: effect of variations in polymer structure

Biochemical Journal ◽

10.1042/bcj20160723 ◽

2016 ◽

Vol 473 (23) ◽

pp. 4349-4360 ◽

Cited By ~ 55

Author(s):

Kerrie A. Morrison ◽

Aneel Akram ◽

Ashlyn Mathews ◽

Zoeya A. Khan ◽

Jaimin H. Patel ◽

...

Keyword(s):

Membrane Proteins ◽

Membrane Protein ◽

Maleic Acid ◽

Protein Extraction ◽

Affinity Purification ◽

Expression System ◽

Transmembrane Proteins ◽

Membrane Extraction ◽

Total Size ◽

Average Molecular Mass

The use of styrene–maleic acid (SMA) copolymers to extract and purify transmembrane proteins, while retaining their native bilayer environment, overcomes many of the disadvantages associated with conventional detergent-based procedures. This approach has huge potential for the future of membrane protein structural and functional studies. In this investigation, we have systematically tested a range of commercially available SMA polymers, varying in both the ratio of styrene and maleic acid and in total size, for the ability to extract, purify and stabilise transmembrane proteins. Three different membrane proteins (BmrA, LeuT and ZipA), which vary in size and shape, were used. Our results show that several polymers, can be used to extract membrane proteins, comparably to conventional detergents. A styrene:maleic acid ratio of either 2:1 or 3:1, combined with a relatively small average molecular mass (7.5–10 kDa), is optimal for membrane extraction, and this appears to be independent of the protein size, shape or expression system. A subset of polymers were taken forward for purification, functional and stability tests. Following a one-step affinity purification, SMA 2000 was found to be the best choice for yield, purity and function. However, the other polymers offer subtle differences in size and sensitivity to divalent cations that may be useful for a variety of downstream applications.

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Membrane Protein Extraction by Styrene-Maleic Acid Copolymers is Independent of Subcellular Localization but is Influenced by Membrane Organization

Biophysical Journal ◽

10.1016/j.bpj.2016.11.2094 ◽

2017 ◽

Vol 112 (3) ◽

pp. 385a

Author(s):

Jonas M. Doerr ◽

Juan J. Dominguez Pardo ◽

Marleen H. van Coevoorden-Hameete ◽

Casper C. Hoogenraad ◽

J. Antoinette Killian

Keyword(s):

Membrane Protein ◽

Subcellular Localization ◽

Maleic Acid ◽

Protein Extraction ◽

Membrane Organization

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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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