Acetone Precipitation of Proteins and the Modification of Peptides

2010 ◽  
Vol 9 (1) ◽  
pp. 444-450 ◽  
Author(s):  
Deborah M. Simpson ◽  
Robert J. Beynon
Keyword(s):  
Acetone Precipitation

Preparation of a highly active form of nisin from Streptococcus lactis

1971 ◽  
Vol 17 (1) ◽  
pp. 61-67 ◽  
Author(s):  
F. J. Bailey ◽  
A. Hurst
Keyword(s):  
Active Form ◽  
Gradient Elution ◽  
Antibiotic Activity ◽  
Complex Medium ◽  
Main Peak ◽  
Acetone Precipitation ◽  
Highly Active ◽  
Streptococcus Lactis ◽  
Constant Ph ◽  
Ph Gradient Elution

Cells of Streptococcus lactis (354/07) synthesized and retained nisin when grown in a complex medium with 2.5% glucose at a constant pH of 6.7. Nisin was extracted from cells by a previously used method with hot 0.05 N HCl but milder methods of extraction from whole and broken cells using a variety of solvents were also tested. In the preferred method broken cells were extracted with 0.05 N HCl at 2 °C. The Cl− ions of the extract were exchanged for acetate on columns of the resin Amberlite CG 4B and the eluate was concentrated by acetone precipitation at −19 °C. The nisin was finally purified by pH gradient elution from CM cellulose columns. Three peaks with antibiotic activity were found, two of the peaks were minor and represented less than 5% of the nisin. The main peak gave a single band on electrophoresis. Electrophoresis of the material from the CM cellulose peaks revealed about 44 bands of basic proteins. Nisin made by the hot or cold HCl extraction behaved similarly in electrophoresis and CM cellulose chromatography but the antibiotic activity of the material isolated from the cold extract was nine times greater than that of the material isolated from the hot extract.

scholarly journals A method for the selective extraction of histone fractions f2(a)1 and f2(a)2 from calf thymus deoxyribonucleoprotein at pH7

1967 ◽  
Vol 105 (2) ◽  
pp. 611-614 ◽  
Author(s):  
E W Johns
Keyword(s):  
Acid Solution ◽  
Selective Extraction ◽  
New Method ◽  
Guanidinium Chloride ◽  
Histone Fraction ◽  
Acetone Precipitation ◽  
Calf Thymus

1. A new method has been developed for the specific extraction of histone fraction f2(a) from calf thymus deoxyribonucleoprotein at pH7 by using a mixture of ethanol and guanidinium chloride. 2. Fraction f2(a) has been separated into the subfractions f2(a)1 and f2(a)2 by acetone precipitation from acid solution, and at pH7. 3. Modifications of existing electrophoretic methods are described that enable these fractions to be more easily characterized.

Turbidimetric measurement of haze in canola oil by acetone precipitation

1996 ◽  
Vol 73 (11) ◽  
pp. 1557-1560 ◽  
Author(s):  
Hua Liu ◽  
Roman Przybylski ◽  
N. A. Michael Eskin
Keyword(s):  
Canola Oil ◽  
Acetone Precipitation ◽  
Turbidimetric Measurement

scholarly journals Identification of Phytochemicals from the Water Extract of Eurycoma longifolia Roots using Solid-Liquid and Liquid-Liquid Extraction Based Fractionation Techniques

2021 ◽  
Vol 68 (4) ◽  
pp. 765-772
Author(s):  
Lee Suan Chua ◽  
Abirame Segaran ◽  
Hoi Jin Wong
Keyword(s):  
Ethyl Acetate ◽  
Solid Phase ◽  
Water Extract ◽  
Reversed Phase ◽  
Liquid Extraction ◽  
Acetone Precipitation ◽  
Eurycoma Longifolia ◽  
Liquid Liquid Extraction ◽  
Saponin Content ◽  
Solid Liquid

Phytochemicals in the water extract of Eurycoma longofolia roots were identified using both solid-liquid and liquid-liquid extraction based fractionation techniques. A reversed phase C18 solid phase extraction (SPE) was used as solid-liquid extraction, whereas solvent partition was applied as liquid-liquid extraction. Total saponin was increased after fractionation. A few known quassinoids; eurycomanone, 13α(21)-epoxyeurycomanone, pasakbumin D, 13β,18-dihydroeurycomanol and 13β,21-dihydroxyeurycomanol were identified from the 40% and 60% methanol fractions of SPE. Solvent partition extract using ethyl acetate was found to have the highest saponin content compared to butanol and chloroform fractions. Subsequent acetone precipitation of the organic fractions recovered a formylated hexose trimer and other saccharide-containing compounds. Ethyl acetate effectively recovered saponins from E. longofolia water extract using liquid-liquid extraction followed by acetone precipitation.

scholarly journals Modified TCA/acetone precipitation of plant proteins for proteomic analysis

2018 ◽  
Author(s):  
Liangjie Niu ◽  
Hang Zhang ◽  
Zhaokun Wu ◽  
Yibo Wang ◽  
Hui Liu ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Protein Modification ◽  
Protein Extraction ◽  
Extended Period ◽  
Extraction Methods ◽  
Precipitation Method ◽  
Plant Proteins ◽  
Prolonged Incubation ◽  
Acetone Precipitation ◽  
Modified Method

AbstractProtein extracts obtained from cells or tissues often require removal of interfering substances for the preparation of high-quality protein samples in proteomic analysis. A number of protein extraction methods have been applied to various biological samples. TCA/acetone precipitation and phenol extraction, a common method of protein extraction, is thought to minimize protein degradation and activity of proteases as well as reduce contaminants like salts and polyphenols. However, the TCA/acetone precipitation method relies on the complete pulverization and repeated rinsing of tissue powder to remove the interfering substances, which is laborious and time-consuming. In addition, by prolonged incubation in TCA/acetone, the precipitated proteins are more difficult to re-dissolve. We have described a modified method of TCA/acetone precipitation of plant proteins for proteomic analysis. Proteins of cells or tissues were extracted using SDS-containing buffer, precipitated with equal volume of 20% TCA/acetone, and washed with acetone. Compared to classical TCA/acetone precipitation and simple acetone precipitation, this protocol generates comparable yields, spot numbers, and proteome profiling, but takes less time (ca. 45 min), thus avoiding excess protein modification and degradation after extended-period incubation in TCA/acetone or acetone. The modified TCA/acetone precipitation method is simple, fast, and suitable for proteomic analysis of various plant tissues in proteomic analysis.

scholarly journals Evaluation of female Aedes aegypti proteome via LC-ESI-MS/MS using two protein extraction methods

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e10863
Author(s):  
Abubakar Shettima ◽  
Intan H. Ishak ◽  
Syahirah Hanisah Abdul Rais ◽  
Hadura Abu Hasan ◽  
Nurulhasanah Othman
Keyword(s):  
Extraction Method ◽  
Protein Identification ◽  
Protein Extraction ◽  
Adaptor Protein ◽  
Control Measures ◽  
Protein Yield ◽  
Mass Spectrometry Analysis ◽  
Functional Protein ◽  
Acetone Precipitation ◽  
Esi Ms

Background Proteomic analyses have broadened the horizons of vector control measures by identifying proteins associated with different biological and physiological processes and give further insight into the mosquitoes’ biology, mechanism of insecticide resistance and pathogens-mosquitoes interaction. Female Ae. aegypti ingests human blood to acquire the requisite nutrients to make eggs. During blood ingestion, female mosquitoes transmit different pathogens. Therefore, this study aimed to determine the best protein extraction method for mass spectrometry analysis which will allow a better proteome profiling for female mosquitoes. Methods In this present study, two protein extractions methods were performed to analyze female Ae. aegyti proteome, via TCA acetone precipitation extraction method and a commercial protein extraction reagent CytoBusterTM. Then, protein identification was performed by LC-ESI-MS/MS and followed by functional protein annotation analysis. Results The CytoBusterTM reagent gave the highest protein yield with a mean of 475.90 µg compared to TCA acetone precipitation extraction showed 283.15 µg mean of protein. LC-ESI-MS/MS identified 1,290 and 890 proteins from the CytoBusterTM reagent and TCA acetone precipitation, respectively. When comparing the protein class categories in both methods, there were three additional categories for proteins identified using CytoBusterTM reagent. The proteins were related to scaffold/adaptor protein (PC00226), protein binding activity modulator (PC00095) and intercellular signal molecule (PC00207). In conclusion, the CytoBusterTM protein extraction reagent showed a better performance for the extraction of proteins in term of the protein yield, proteome coverage and extraction speed.

scholarly journals CHARACTERIZATION OF CELLULASE PREPARATION OF BACILLUS SP.G4 ISOLATED FROM THE TERMITES GUT

2018 ◽  
Vol 54 (4A) ◽  
pp. 115
Author(s):  
Đào Thị Thanh Xuân
Keyword(s):  
High Activity ◽  
Ammonium Sulfate ◽  
Rice Bran ◽  
Tween 80 ◽  
Bacillus Sp ◽  
High Concentration ◽  
Acetone Precipitation ◽  
Cmcase Activity ◽  
Purification Factor

The Bacillus sp. strain G4 isolated from termites gut can produce cellulase with high activity in rice bran medium after 72 h of incubation at 37 °C. The crude enzymewas collected by centrifugation at 6,000 rpmfor 15 minutes.  The precipitation of cellulasewith ammonium sulfate 60 – 90 % saturation and acetone at the concentration 60 % – 90 % was studied. Results showed that ammonium sulfate 90 % saturation gave cellulase with highest purification factor 8.87 but the yield was only 59.9 %, whereas acetone at 90 % concentration gave highest yield 83.57 % with purification factor 4.38. CMCase activity of cellulasepreparation obtained by acetone precipitation at 90 % was optimum at pH 7 and 60 °C. Furthermore, CMCase was stable at pH 6.0 - 7.0 and at temperature lower than 50 °C. The CMCase was activated by ion Ca2+ but inhibited by Co2+, Zn2+, Cu2+, Fe2+, and was not affected by Mg2+ and Ba2+. The CMCase was inhibited by high concentration of SDS while EDTA and Tween 80 played activated role.

scholarly journals Modified TCA/acetone precipitation of plant proteins for proteomic analysis

PLoS ONE ◽  
2018 ◽  
Vol 13 (12) ◽  
pp. e0202238 ◽  
Author(s):  
Liangjie Niu ◽  
Hang Zhang ◽  
Zhaokun Wu ◽  
Yibo Wang ◽  
Hui Liu ◽  
...  
Keyword(s):  
Proteomic Analysis ◽  
Plant Proteins ◽  
Acetone Precipitation
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