scholarly journals Optimization of Protein Isolation and Label-Free Quantitative Proteomic Analysis in Four Different Tissues of Korean Ginseng

Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1409
Author(s):  
Truong Van Nguyen ◽  
So-Wun Kim ◽  
Cheol-Woo Min ◽  
Ravi Gupta ◽  
Gi-Hyun Lee ◽  
...  
Keyword(s):  
Protein Extraction ◽  
Extraction Methods ◽  
Label Free ◽  
Protein Isolation ◽  
Mva Pathway ◽  
Ginsenoside Biosynthesis ◽  
Wide Range ◽  
Proteomics Approach ◽  
Reproducible Method ◽  
Korean Ginseng

Korean ginseng is one of the most valuable medicinal plants worldwide. However, our understanding of ginseng proteomics is largely limited due to difficulties in the extraction and resolution of ginseng proteins because of the presence of natural contaminants such as polysaccharides, phenols, and glycosides. Here, we compared four different protein extraction methods, namely, TCA/acetone, TCA/acetone–MeOH/chloroform, phenol–TCA/acetone, and phenol–MeOH/chloroform methods. The TCA/acetone–MeOH/chloroform method displayed the highest extraction efficiency, and thus it was used for the comparative proteome profiling of leaf, root, shoot, and fruit by a label-free quantitative proteomics approach. This approach led to the identification of 2604 significantly modulated proteins among four tissues. We could pinpoint differential pathways and proteins associated with ginsenoside biosynthesis, including the methylerythritol 4–phosphate (MEP) pathway, the mevalonate (MVA) pathway, UDP-glycosyltransferases (UGTs), and oxidoreductases (CYP450s). The current study reports an efficient and reproducible method for the isolation of proteins from a wide range of ginseng tissues and provides a detailed organ-based proteome map and a more comprehensive view of enzymatic alterations in ginsenoside biosynthesis.

scholarly journals Review: current trends in oat protein recovery and utilization in aqueous food systems

2021 ◽  
Author(s):  
Darius Sargautis ◽  
◽  
Tatjana Kince ◽  
Vanda Sargautiene ◽  
Keyword(s):  
Functional Properties ◽  
Food Systems ◽  
Protein Extraction ◽  
Protein Solubility ◽  
Extraction Methods ◽  
Scientific Data ◽  
Protein Recovery ◽  
Protein Isolation ◽  
Dry Fractionation ◽  
Isolation Methods

Oat protein itself, as a substance, has extensively been studied providing information on its nutritional value, some functional properties and possible applicability in food systems. Chosen protein isolation methods and technological aspects define final composition of obtained oat protein product, its concentration, nutrition value and its functionality in food industry. Scientific data on oat protein recovery methods, typically relying on protein solubility or dry fractionation, provides an insufficient knowledge about the success in commercialization of oat protein recovery technologies and their derivatives in form of oat protein. The aim of the study was to analyse and summarize the research findings on oat protein extraction methods and functional properties of oat protein. Semi-systematic, monographic methods were used to analyse the oat protein isolation techniques, functional properties of oat protein in aqueous food systems, covering the latest information on oat protein extraction methods. Wet and dry isolation methods were demonstrated as main methods in oat protein extraction. Functional properties of oat protein, such as thermal stability, solubility, emulsification, water hydration capacity and foaming were reviewed and evaluated, identifying limitations and protein alterations which occur through the oat protein extraction process. The study provides recent trends in oat protein recovery technologies, along with an overview of current and potential oat protein utilization in food systems.

scholarly journals Evaluation of protein extraction methods for enhanced proteomic analysis of tomato leaves and roots

2015 ◽  
Vol 87 (3) ◽  
pp. 1853-1863 ◽  
Author(s):  
MILCA B. VILHENA ◽  
MÔNICA R. FRANCO ◽  
DAIANA SCHMIDT ◽  
GISELLE CARVALHO ◽  
RICARDO A. AZEVEDO
Keyword(s):  
Gel Electrophoresis ◽  
Protein Extraction ◽  
Extraction Methods ◽  
Protein Yield ◽  
Crucial Aspect ◽  
Large Protein ◽  
Total Proteins ◽  
Solanum Lycopersicum L ◽  
Wide Range ◽  
Leaves And Roots

Proteomics is an outstanding area in science whose increasing application has advanced to distinct purposes. A crucial aspect to achieve a good proteome resolution is the establishment of a methodology that results in the best quality and wide range representation of total proteins. Another important aspect is that in many studies, limited amounts of tissue and total protein in the tissue to be studied are found, making difficult the analysis. In order to test different parameters, combinations using minimum amount of tissue with 4 protocols for protein extraction from tomato (Solanum lycopersicum L.) leaves and roots were evaluated with special attention to their capacity for removing interferents and achieving suitable resolution in bidimensional gel electrophoresis, as well as satisfactory protein yield. Evaluation of the extraction protocols revealed large protein yield differences obtained for each one. TCA/acetone was shown to be the most efficient protocol, which allowed detection of 211 spots for leaves and 336 for roots using 500 µg of leaf protein and 800 µg of root protein per gel.

scholarly journals Assessment and refinement of sample preparation methods for deep and quantitative plant proteome profiling

2018 ◽  
Author(s):  
Gaoyuan Song ◽  
Polly Y Yingshan Hsu ◽  
Justin W. Walley
Keyword(s):  
Mass Spectrometry ◽  
Sample Preparation ◽  
Protein Extraction ◽  
Leaf Tissue ◽  
Extraction Methods ◽  
Label Free ◽  
Preparation Methods ◽  
Proteome Profiling ◽  
Leaf Tissues ◽  
Sample Preparation Methods

SummaryA major challenge in the field of proteomics is obtaining high quality peptides for comprehensive proteome profiling by liquid chromatography mass spectrometry for many organisms. Here we evaluate and modify a range of sample preparation methods using photosynthetically active Arabidopsis leaf tissues from several developmental timepoints. We find that inclusion of FASP-based on filter digestion improves all protein extraction methods tested. Ultimately, we show that a detergent-free urea-FASP approach enables deep and robust quantification of leaf proteomes. For example, from 4-day-old leaf tissue we profiled up to 11,690 proteins from a single sample replicate. This method should be broadly applicable to researchers working on difficult to process samples from a range of plant and non-plant organisms.AbbreviationsChloroMethanol/Chloroform ExtractionFASPFilter Aided Sample PrepGOGene OntologyIAAIodoacetamideLFQLabel Free QuantificationMS/MSTandem mass spectrometryTFTranscription FactorUAUrea Extraction1D1 Dimensional2D2 Dimensional

Versatile Health Benefits of Catechin from Green Tea (Camellia sinensis)

2019 ◽  
Vol 15 (1) ◽  
pp. 3-10 ◽  
Author(s):  
Satheesh Babu Natarajan ◽  
Suriyakala Perumal Chandran ◽  
Sahar Husain Khan ◽  
Packiyaraj Natarajan ◽  
Karthiyaraj Rengarajan
Keyword(s):  
Green Tea ◽  
Camellia Sinensis ◽  
Health Benefits ◽  
Epigallocatechin Gallate ◽  
Extraction Methods ◽  
Major Constituent ◽  
Epicatechin Gallate ◽  
Wide Range ◽  
Green Tea Catechin ◽  
Anti Inflammatory

Background: Tea (Camellia sinensis, Theaceae) is the second most consumed beverage in the world. Green tea is the least processed and thus contain rich antioxidant level, and believed to have most of the health benefits. </p><p> Methods: We commenced to search bibliographic collection of peer reviewed research articles and review articles to meet the objective of this study. </p><p> Results: From this study, we found that the tea beverage contains catechins are believed to have a wide range of health benefits which includes neuroprotective, anti-inflammatory, antiulcer, antiviral, antibacterial, and anti-parasitic effects. The four major catechin compounds of green tea are epigallocatechin (EGC), epicatechin (EC), epigallocatechin gallate (EGCG), and epicatechin gallate (ECG), of which EGCG is the major constituent and representing 50-80% of the total catechin content. And also contain xanthine derivatives such as caffeine, theophylline, and theobromine, and the glutamide derivative theanine. It also contains many nutritional components, such as vitamin E, vitamin C, fluoride, and potassium. We sum up the various green tea phytoconstituents, extraction methods, and its medicinal applications. </p><p> Conclusion: In this review article, we have summarized the pharmacological importance of green tea catechin which includes antioxidant potential, anti-inflammatory, antimicrobial, anticancer, antidiabetic and cosmetic application.

scholarly journals Photonic Crystal Nanobeam Cavities for Nanoscale Optical Sensing: A Review

Micromachines ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 72 ◽  
Author(s):  
Da-Quan Yang ◽  
Bing Duan ◽  
Xiao Liu ◽  
Ai-Qiang Wang ◽  
Xiao-Gang Li ◽  
...  
Keyword(s):  
Photonic Crystal ◽  
Optical Waveguides ◽  
Optical Sensing ◽  
Early Stage ◽  
Disease Diagnosis ◽  
Label Free ◽  
Quality Factors ◽  
Integrated Sensor ◽  
Early Stage Disease ◽  
Wide Range

The ability to detect nanoscale objects is particular crucial for a wide range of applications, such as environmental protection, early-stage disease diagnosis and drug discovery. Photonic crystal nanobeam cavity (PCNC) sensors have attracted great attention due to high-quality factors and small-mode volumes (Q/V) and good on-chip integrability with optical waveguides/circuits. In this review, we focus on nanoscale optical sensing based on PCNC sensors, including ultrahigh figure of merit (FOM) sensing, single nanoparticle trapping, label-free molecule detection and an integrated sensor array for multiplexed sensing. We believe that the PCNC sensors featuring ultracompact footprint, high monolithic integration capability, fast response and ultrahigh sensitivity sensing ability, etc., will provide a promising platform for further developing lab-on-a-chip devices for biosensing and other functionalities.

scholarly journals Optimization of Protein Extraction Method for 2DE Proteomics of Goat’s Milk

Molecules ◽  
2020 ◽  
Vol 25 (11) ◽  
pp. 2625
Author(s):  
Muzammeer Mansor ◽  
Jameel R. Al-Obaidi ◽  
Nurain Nadiah Jaafar ◽  
Intan Hakimah Ismail ◽  
Atiqah Farah Zakaria ◽  
...  
Keyword(s):  
Extraction Method ◽  
Protein Extraction ◽  
Chloroform Solution ◽  
Milk Proteins ◽  
Peptide Mass Fingerprinting ◽  
Extraction Methods ◽  
Dairy Goats ◽  
Goat’S Milk ◽  
Peptide Mass ◽  
Goat's Milk

Two-dimensional electrophoretic (2DE)-based proteomics remains a powerful tool for allergenomic analysis of goat’s milk but requires effective extraction of proteins to accurately profile the overall causative allergens. However, there are several current issues with goat’s milk allergenomic analysis, and among these are the absence of established standardized extraction method for goat’s milk proteomes and the complexity of goat’s milk matrix that may hamper the efficacy of protein extraction. This study aimed to evaluate the efficacies of three different protein extraction methods, qualitatively and quantitatively, for the 2DE-proteomics, using milk from two commercial dairy goats in Malaysia, Saanen, and Jamnapari. Goat’s milk samples from both breeds were extracted by using three different methods: a milk dilution in urea/thiourea based buffer (Method A), a triphasic separation protocol in methanol/chloroform solution (Method B), and a dilution in sulfite-based buffer (Method C). The efficacies of the extraction methods were assessed further by performing the protein concentration assay and 1D and 2D SDS-PAGE profiling, as well as identifying proteins by MALDI-TOF/TOF MS/MS. The results showed that method A recovered the highest amount of proteins (72.68% for Saanen and 71.25% for Jamnapari) and produced the highest number of protein spots (199 ± 16.1 and 267 ± 10.6 total spots for Saanen and Jamnapari, respectively) with superior gel resolution and minimal streaking. Six milk protein spots from both breeds were identified based on the positive peptide mass fingerprinting matches with ruminant milk proteins from public databases, using the Mascot software. These results attest to the fitness of the optimized protein extraction protocol, method A, for 2DE proteomic and future allergenomic analysis of the goat’s milk.

scholarly journals Echocardiographic and invasive measurements of pulmonary artery pressure correlate closely at high altitude

2000 ◽  
Vol 279 (4) ◽  
pp. H2013-H2016 ◽  
Author(s):  
Yves Allemann ◽  
Claudio Sartori ◽  
Mattia Lepori ◽  
Sébastien Pierre ◽  
Christian Mélot ◽  
...  
Keyword(s):  
Pulmonary Hypertension ◽  
High Altitude ◽  
Doppler Echocardiography ◽  
Systolic Pulmonary Arterial Pressure ◽  
Wide Range ◽  
High Altitude Pulmonary Edema ◽  
Reproducible Method ◽  
Pulmonary Arterial ◽  
Spearman Test ◽  
Invasive Measurement

Exaggerated hypoxia-induced pulmonary hypertension is a hallmark of high-altitude pulmonary edema (HAPE) and plays a major role in its pathogenesis. Many studies of HAPE have estimated systolic pulmonary arterial pressure (SPAP) with Doppler echocardiography. Whereas at low altitude, Doppler echocardiographic estimation of SPAP correlates closely with its invasive measurement, no such evidence exists for estimations obtained at high altitude, where alterations of blood viscosity may invalidate the simplified Bernoulli equation. We measured SPAP by Doppler echocardiography and invasively in 14 mountaineers prone to HAPE and in 14 mountaineers resistant to this condition at 4,559 m. Mountaineers prone to HAPE had more pronounced pulmonary hypertension (57 ± 12 and 58 ± 10 mmHg for noninvasive and invasive determination, respectively; means ± SD) than subjects resistant to HAPE (37 ± 8 and 37 ± 6 mmHg, respectively), and the values measured in the two groups as a whole covered a wide range of pulmonary arterial pressures (30–83 mmHg). Spearman test showed a highly significant correlation ( r = 0.89, P < 0.0001) between estimated and invasively measured SPAP values. The mean difference between invasively measured and Doppler-estimated SPAP was 0.5 ± 8 mmHg. At high altitude, estimation of SPAP by Doppler echocardiography is an accurate and reproducible method that correlates closely with its invasive measurement.

Comparison of Two Protein Extraction Methods for Tea Residues

2011 ◽  
Vol 138-139 ◽  
pp. 933-936 ◽  
Author(s):  
Xuan Chen ◽  
Hong Yu Luo ◽  
Jun Yu ◽  
Peng Xiang Yue ◽  
Lin Zhou ◽  
...  
Keyword(s):  
High Temperature ◽  
Protein Extraction ◽  
Ethanol Concentration ◽  
Extraction Methods ◽  
Extraction Yield ◽  
Factorial Experiments ◽  
Extraction Technology ◽  
Digestion Method ◽  
Solid Ratio ◽  
Lower Temperature

Alcohol-alkali method and base digestion method were investigated to extract proteins from tea residues, respectively. According to single factorial experiments, results showed that the optimal extraction technology of alcohol-alkali method were pH 12, temperature of 80 °C, ethanol concentration of 60%, liquid-solid ratio of 40, 60 min, and the protein extraction rate reached 15.0%. And the optimal extract conditions of base digestion were pH 12, temperature of 80 °C, liquid-solid ratio of 50, 80 min, which made the protein yield reached 31.5%. Furthermore, alcohol-alkali method was more beneficial to protein extraction from tea residues under lower temperature and weak alkali condition (40-60 °C, pH 8-10). While base digestion had higher extraction yield under high temperature and strong alkali condition (60-80 °C, pH 11-12).

scholarly journals Evaluation of effective protein extraction procedure to profile petiole of Moringa oleifera

2020 ◽  
Vol 7 (2) ◽  
pp. 214
Author(s):  
Zetty Amirah Zulkifli ◽  
Zaidah Rahmat
Keyword(s):  
Moringa Oleifera ◽  
Protein Extraction ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Extraction Procedure ◽  
Electrophoretic Pattern ◽  
Antioxidant Property ◽  
Extraction Methods ◽  
Protein Profiling ◽  
Sds Page

Moringa oleifera is widely known as multipurpose tree since all of its parts confer multiple functions. The leaf is highly favourable among consumers while the petiole is mostly wasted. There are numerous studies on the flavonoid and antioxidant property of the stem and twig. However, study on the petiole has never been done. There-upon, this study was conducted to develop protein profiling of the petiole. In this study, 6 different protein extraction methods were tested on the fresh petiole before its protein quantity and quality were checked via Bradford assay and Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) respectively. The in-solution digestion was then done prior to LC-MS/MS analysis. The protein electrophoretic pattern from the SDS-PAGE proves that method 6 using Tris HCl buffer with incorporation of dithiothreitol (DTT) and phenylmethylsulfonyl fluoride (PMSF) confers the best quality of protein. It produced the highest number of visible individual bands compared to other methods. Meanwhile, 93 proteins were successfully identified via LCMS analysis where the protein, signal response and carbohydrate metabolism categories confer the highest percentage. High quality and content of the protein extracted from the petiole including the antioxidant, anticancer and antidiabetic protein identified suggested that consuming this part of the plant could enhance nutrients of human body.

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