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Sample records for optimized protein extraction

  1. Optimization of Protein Extraction for Lichen Thalli.

    Science.gov (United States)

    Kondratiuk, Anna S; Savchuk, Oleksiy M; Hur, Jae-Seoun

    2015-06-01

    Lichen-forming fungal proteins have been seldom searched due to many difficulties in their extraction. Phenols, quinones, proteases, and other components released during cell disruption have been known to be the greatest challenges related to protein extraction from lichens. To overcome these problems and maintain good electrophoretic resolution and high protein concentration, an extraction buffer containing polyvinylpolypyrrolidone, ascorbic acid, Triton X-100, polyethylene glycol, proteinase, and oxidase inhibitors in sodium phosphate buffer was developed. This extraction buffer showed high efficiency for all lichen species tested in the study.

  2. Investigation and optimization of the factors influencing sorghum protein extraction.

    Science.gov (United States)

    Park, Seok-Ho; Bean, Scott Roger

    2003-11-19

    To optimize the extraction of sorghum proteins, several variables were examined: sample-to-solvent ratio, detergent type and concentration, reducing agent type and concentration, extraction time, and buffer pH and concentration. Samples were quantified and characterized by RP-HPLC, FZCE, and nitrogen analysis. These studies revealed that pH, detergent type, reducing agent type, and sample-to-solvent ratio all had significant effects on the levels of protein extracted. Increasing SDS concentration (2%) and solvent-to-flour ratio (20:1) with multiple 5 min extracts reduced extraction time by 35-80% while still extracting the same levels of total protein relative to the control methodology. Reproducibility using the multiple extractions was found to be excellent with relative standard deviations of extractions.

  3. Optimal extraction and hydrolysis of Chlorella pyrenoidosa proteins.

    Science.gov (United States)

    Wang, Xiaoqin; Zhang, Xuewu

    2012-12-01

    In this study, for the first time, the applications of two new methods, ionic liquid and low-temperature high-pressure cell breakage methods, to the extraction of whole proteins in Chlorella pyrenoidosa cells were explored. Meanwhile, the comparison with three traditional methods was also made. The results indicated that the extraction rate for ionic liquid is only at moderate level, but the new low-temperature high-pressure cell breakage method can obviously increase the protein extraction rate up to 2- to 15-fold. Subsequently, the hydrolysis of the extracted proteins was conducted with three enzymes (papain, trypsin and alcalase). The data presented that the degree of hydrolysis for each enzyme under the optimal conditions is in the order of: alcalase (18.31%)>papain (14.33%)>trypsin (8.47%), demonstrating the potential of C. pyrenoidosa protein hydrolysates obtained here in nutritional supplement and medical foods.

  4. Optimized protein extraction methods for proteomic analysis of Rhizoctonia solani.

    Science.gov (United States)

    Lakshman, Dilip K; Natarajan, Savithiry S; Lakshman, Sukla; Garrett, Wesley M; Dhar, Arun K

    2008-01-01

    Rhizoctonia solani (Teleomorph: Thanatephorus cucumeris, T. praticola) is a basidiomycetous fungus and a major cause of root diseases of economically important plants. Various isolates of this fungus are also beneficially associated with orchids, may serve as biocontrol agents or remain as saprophytes with roles in decaying and recycling of soil organic matter. R. solani displays several hyphal anastomosis groups (AG) with distinct host and pathogenic specializations. Even though there are reports on the physiological and histological basis of Rhizoctonia-host interactions, very little is known about the molecular biology and control of gene expression early during infection by this pathogen. Proteamic technologies are powerful tools for examining alterations in protein profiles. To aid studies on its biology and host pathogen interactions, a two-dimensional (2-D) gel-based global proteomic study has been initiated. To develop an optimized protein extraction protocol for R. solani, we compared two previously reported protein extraction protocols for 2-D gel analysis of R. solani (AG-4) isolate Rs23. Both TCA-acetone precipitation and phosphate solubilization before TCA-acetone precipitation worked well for R. solani protein extraction, although selective enrichment of some proteins was noted with either method. About 450 spots could be detected with the densitiometric tracing of Coomassie blue-stained 2-D PAGE gels covering pH 4-7 and 6.5-205 kDa. Selected protein spots were subjected to mass spectrometric analysis with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Eleven protein spots were positively identified based on peptide mass fingerprinting match with fungal proteins in public databases with the Mascot search engine. These results testify to the suitability of the two optimized protein extraction protocols for 2-D proteomic studies of R. solani.

  5. Chemical optimization of protein extraction from sweet potato (Ipomoea batatas) peel

    Science.gov (United States)

    Proteins isolated from sweet potatoes (Ipomoea batatas) have been shown to possess antidiabetic, antioxidant, and antiproliferative properties. The objective of this study was to chemically optimize a process for extracting proteins from sweet potato peel. The extraction procedure involved mixing pe...

  6. Comparison and optimization of methods for the simultaneous extraction of DNA, RNA, proteins, and metabolites.

    Science.gov (United States)

    Vorreiter, Fränze; Richter, Silke; Peter, Michel; Baumann, Sven; von Bergen, Martin; Tomm, Janina M

    2016-09-01

    The challenge of performing a time-resolved comprehensive analysis of molecular systems has led to the quest to optimize extraction methods. When the size of a biological sample is limited, there is demand for the simultaneous extraction of molecules representing the four areas of "omics": genomics, transcriptomics, proteomics, and metabolomics. Here we optimized a protocol for the simultaneous extraction of DNA, RNA, proteins, and metabolites and compared it with two existing protocols. Our optimization comprised the addition of a methanol/chloroform metabolite purification before the separation of DNA/RNA and proteins. Extracted DNA, RNA, proteins, and metabolites were quantitatively and/or qualitatively analyzed. Of the three methods, only the newly developed protocol yielded all biomolecule classes of adequate quantity and quality. Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.

  7. Optimal protein extraction methods from diverse sample types for protein profiling by using Two-Dimensional Electrophoresis (2DE).

    Science.gov (United States)

    Tan, A A; Azman, S N; Abdul Rani, N R; Kua, B C; Sasidharan, S; Kiew, L V; Othman, N; Noordin, R; Chen, Y

    2011-12-01

    There is a great diversity of protein samples types and origins, therefore the optimal procedure for each sample type must be determined empirically. In order to obtain a reproducible and complete sample presentation which view as many proteins as possible on the desired 2DE gel, it is critical to perform additional sample preparation steps to improve the quality of the final results, yet without selectively losing the proteins. To address this, we developed a general method that is suitable for diverse sample types based on phenolchloroform extraction method (represented by TRI reagent). This method was found to yield good results when used to analyze human breast cancer cell line (MCF-7), Vibrio cholerae, Cryptocaryon irritans cyst and liver abscess fat tissue. These types represent cell line, bacteria, parasite cyst and pus respectively. For each type of samples, several attempts were made to methodically compare protein isolation methods using TRI-reagent Kit, EasyBlue Kit, PRO-PREP™ Protein Extraction Solution and lysis buffer. The most useful protocol allows the extraction and separation of a wide diversity of protein samples that is reproducible among repeated experiments. Our results demonstrated that the modified TRI-reagent Kit had the highest protein yield as well as the greatest number of total proteins spots count for all type of samples. Distinctive differences in spot patterns were also observed in the 2DE gel of different extraction methods used for each type of sample.

  8. Optimization of Protein Extraction and Two-Dimensional Electrophoresis Protocols for Oil Palm Leaf.

    Science.gov (United States)

    Daim, Leona Daniela Jeffery; Ooi, Tony Eng Keong; Yusof, Hirzun Mohd; Majid, Nazia Abdul; Karsani, Saiful Anuar Bin

    2015-08-01

    Oil palm (Elaeis guineensis) is an important economic crop cultivated for its nutritional palm oil. A significant amount of effort has been undertaken to understand oil palm growth and physiology at the molecular level, particularly in genomics and transcriptomics. Recently, proteomics studies have begun to garner interest. However, this effort is impeded by technical challenges. Plant sample preparation for proteomics analysis is plagued with technical challenges due to the presence of polysaccharides, secondary metabolites and other interfering compounds. Although protein extraction methods for plant tissues exist, none work universally on all sample types. Therefore, this study aims to compare and optimize different protein extraction protocols for use with two-dimensional gel electrophoresis of young and mature leaves from the oil palm. Four protein extraction methods were evaluated: phenol-guanidine isothiocyanate, trichloroacetic acid-acetone precipitation, sucrose and trichloroacetic acid-acetone-phenol. Of these four protocols, the trichloroacetic acid-acetone-phenol method was found to give the highest resolution and most reproducible gel. The results from this study can be used in sample preparations of oil palm tissue for proteomics work.

  9. Optimized extract preparation methods and reaction conditions for improved yeast cell-free protein synthesis.

    Science.gov (United States)

    Hodgman, C Eric; Jewett, Michael C

    2013-10-01

    Cell-free protein synthesis (CFPS) has emerged as a powerful platform technology to help satisfy the growing demand for simple, affordable, and efficient protein production. In this article, we describe a novel CFPS platform derived from the popular bio-manufacturing organism Saccharomyces cerevisiae. By developing a streamlined crude extract preparation protocol and optimizing the CFPS reaction conditions we were able to achieve active firefly luciferase synthesis yields of 7.7 ± 0.5 µg mL(-1) with batch reactions lasting up to 2 h. This duration of synthesis is the longest ever reported for a yeast CFPS batch reaction. Furthermore, by removing extraneous processing steps and eliminating expensive reagents from the cell-free reaction, we have increased relative product yield (µg protein synthesized per $ reagent cost) over an alternative commonly used method up to 2000-fold from ∼2 × 10(-4) to ∼4 × 10(-1)  µg $(-1) , which now puts the yeast CPFS platform on par with other eukaryotic CFPS platforms commercially available. Our results set the stage for developing a yeast CFPS platform that provides for high-yielding and cost-effective expression of a variety of protein therapeutics and protein libraries.

  10. Optimization of Urea Based Protein Extraction from Formalin-Fixed Paraffin-Embedded Tissue for Shotgun Proteomics

    OpenAIRE

    Luebker, Stephen A.; Koepsell, Scott A.

    2016-01-01

    Urea based protein extraction of formalin-fixed paraffin-embedded (FFPE) tissue provides the most efficient workflow for proteomics due to its compatibility with liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). This study optimizes the use of urea for proteomic analysis of clinical FFPE tissue. A series of protein extraction conditions manipulating temperature and buffer composition were compared to reduce carbamylation introduced by urea and increase pro...

  11. Optimization of Urea Based Protein Extraction from Formalin-Fixed Paraffin-Embedded Tissue for Shotgun Proteomics

    Science.gov (United States)

    Koepsell, Scott A.

    2016-01-01

    Urea based protein extraction of formalin-fixed paraffin-embedded (FFPE) tissue provides the most efficient workflow for proteomics due to its compatibility with liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). This study optimizes the use of urea for proteomic analysis of clinical FFPE tissue. A series of protein extraction conditions manipulating temperature and buffer composition were compared to reduce carbamylation introduced by urea and increase protein detection. Each extraction was performed on a randomized pair of serial sections of homogenous FFPE tissue and analyzed with LC-ESI-MS/MS. Results were compared in terms of yield, missed cleavages, and peptide carbamylation. Lowering extraction temperature to 60°C decreased carbamylation at the cost of decreased protein detection and yield. Protein extraction for at least 20 minutes at 95°C followed by 60°C for 2 hours maximized total protein yield while maintaining protein detection and reducing carbamylation by 7.9%. When accounting for carbamylation during analysis, this modified extraction temperature provides equivalent peptide and protein detection relative to the commercially available Qproteome® FFPE Tissue Kit. No changes to buffer composition containing 7 M urea, 2 M thiourea, and 1 M ammonium bicarbonate resulted in improvements to control conditions. Optimized urea in-solution digestion provides an efficient workflow with maximized yields for proteomic analysis of clinically relevant FFPE tissue. PMID:27660725

  12. Protein Extractability

    African Journals Online (AJOL)

    human nutrition, particularly in developing ... studied the effect of NaCl on the extractability of ... significant influence on the functional properties ..... Malaka, S. L. O. and Amund, O. O. Studies on the Life Cycle and Morphometrics of Honeybees,.

  13. Optimization of protein extraction process from jackfruit seed flour by reverse micelle system

    Directory of Open Access Journals (Sweden)

    Maycon Fagundes Teixeira Reis

    2016-06-01

    Full Text Available The extraction of protein from flour of jackfruit seeds by reverse micelles was evaluated. Reverse micelle system was composed of sodium dodecyl sulfate (SDS as surfactant, butanol as solvent, and water. The effects of stirring time, temperature, molar ratio H2O SDS-1, concentration of butanol (mass percentage and flour mass were tested in batch systems. Based on the adjusted linear regression model, only butanol concentration provided optimum extraction conditions (41.16%. Based on the analysis of surface response, the best extraction yield could be obtained at 25°C, stirring time of 120 min, mass of flour of 100 mg, and a ratio H2O SDS-1 of 50. Experimental results showed that a 79.00% extraction yield could be obtained.

  14. Optimization of Single Cell Protein Production by Candida utilis Using Juice Extracted from Pineapple Waste through Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Rosma, A.

    2005-01-01

    Full Text Available Response surface methodology was applied to optimize protein content in Candida utilis grown in pineapple waste medium. A three-level full factorial design was used to develop a quantitative interpretation of mathematical models between the two variables studied, inoculum size 2.0-10.0% (v/v and total soluble solids in medium (1-5 Brix at 30 h fermentation time. Yeast cells were harvested, ruptured mechanically and the soluble extract was freeze-dried for determination of protein, vitamin-B, 5'-ribonucleotide and total sugar content. Maximum protein content in the yeast 66.61% (w/w was obtained from the predicted optimum inoculum size of 7.83% (v/v and Brix level of 3.02. Highest level of biomass, vitamin-B, 5'-ribonucleotide and total sugar content within the experimental region increased 216.8%, 17.5%, 38.0% and 60.8% respectively after optimization. A verification experiment, conducted at optimized protein content conditions produced values that were close to the predicted values, indicating the reliability of the model used.

  15. Differential Proteome Analysis of the Preeclamptic Placenta Using Optimized Protein Extraction

    Directory of Open Access Journals (Sweden)

    Magnus Centlow

    2010-01-01

    Full Text Available The human placenta is a difficult tissue to work with using proteomic technology since it contains large amounts of lipids and glycogen. Both lipids and glycogen are known to interfere with the first step in the two-dimensional polyacrylamide gel electrophoresis (2D-PAGE, the isoelectric focusing. In order to gain the best possible protein separation on 2D-PAGE, an optimized sample preparation protocol for placental proteins was developed. Two different buffers, urea/CHAPS and Hepes, were used for solubilization in combination with six different precipitation methods. The removal of glycogen from the samples by centrifugation was crucial for the final proteome maps. Solubilization with urea/CHAPS in combination with dichloromethane/methanol or acidified acetone proved to be the best precipitation procedures. When applied to clinical placenta samples apolipoprotein A1 was found to be accumulated in the preeclamptic placenta, where it may either have a nutritional effect or act as a modifier of signal transduction.

  16. An Optimized Method for Protein Extraction from OCT-Embedded Human Kidney Tissue for Protein Quantification by LC-MS/MS Proteomics.

    Science.gov (United States)

    Vrana, Marc; Goodling, Anne; Afkarian, Maryam; Prasad, Bhagwat

    2016-10-01

    The existing biobanks of remnant tissue from clinically indicated kidney biopsies are attractive potential reservoirs for quantification of clinically relevant human tissue proteins by quantitative proteomics. However, a significant caveat of this strategy is that the tissues are often preserved in optimal cutting temperature (OCT) medium. Although OCT is an effective method of preserving the morphologic and immunohistological characteristics of tissues for later study, it significantly impacts efforts to quantify protein expression by liquid chromatography-tandem mass spectrometry methods. We report here a simple, reproducible, and cost-effective procedure to extract proteins from OCT-embedded tissue samples. Briefly, the excess frozen OCT medium was scraped before thawing from the tissue specimens stored at -80°C for ∼3 months. The tissue samples were homogenized and diethyl ether/methanol extraction was performed to remove the remaining OCT medium. The recovered protein was denatured, reduced, and alkylated. The second step of protein extraction and desalting was performed by chloroform/methanol/water extraction of denatured proteins. The resultant protein pellet was trypsin-digested and the marker proteins of various kidney cellular compartments were quantified by targeted selective reaction monitoring proteomics. Upon comparison of peptide signals from OCT-embedded tissue and flash-frozen tissue from the same donors, both individual protein quantities, and their interindividual variabilities, were similar. Therefore, the approach reported here can be applied to clinical reservoirs of OCT-preserved kidney tissue to be used for quantitative proteomics studies of clinically relevant proteins expressed in different parts of the kidney (including drug transporters and metabolizing enzymes).

  17. Optimal concentrations of N-decanoyl-N-methylglucamine and sodium dodecyl sulfate allow the extraction and analysis of membrane proteins.

    Science.gov (United States)

    Chuang, Jen-Hua; Kao, Yu-Jing; Ruderman, Neil B; Tung, Li-Chu; Lin, Yenshou

    2011-11-15

    We studied the extraction and analysis of integral membrane proteins possessing hydrophobic and hydrophilic domains and found that a nonionic detergent called MEGA-10, used in lysis buffers, had a superior extraction effect compared to most conventional detergents. A sodium dodecyl sulfate (SDS) concentration of >0.4% (w/v) in the sample buffer was crucial for those proteins to be clearly analyzed by electrophoresis and Western blotting. Furthermore, MEGA-10 had the tendency to maximally extract proteins around its critical micelle concentration (CMC) of 0.24% (w/v). These solutions can greatly assist functional investigations of membrane proteins in the proteomics era.

  18. Optimization of extraction of hemp protein by response surface methodology%响应面法优化火麻蛋白提取工艺研究

    Institute of Scientific and Technical Information of China (English)

    2016-01-01

    以火麻籽为原料,对碱提酸沉法提取火麻蛋白工艺参数进行研究,在单因素试验的基础上,采用响应面分析法对提取温度、提取时间、提取pH和料液比进行优化并得到回归模型.确定的碱提酸沉法提取火麻蛋白最佳工艺参数为:提取温度60℃,提取时间1 h,提取pH 10,料液比1:9.在最佳工艺条件下,火麻蛋白的提取率为63.5%.回归模型的预测值与实测值的相对误差为2.3%,该回归方程与实际情况拟合较好.DSC分析得出火麻蛋白的变性温度为83.0℃,纯化后的火麻蛋白相对分子质量分布均小于40 kDa.%With hemp seed as raw material,the process parameters of alkali extraction and acid precipita-tion of hemp protein were studied. On the basis of single factor experiment,the extraction temperature,ex-traction time,extraction pH and ratio of material to liquid were optimized by response surface methodolo-gy, and the regression model was finally established. The results showed that the optimal parameters of al-kali extraction and acid precipitation of hemp protein were obtained as follows:extraction temperature 60℃,extraction time 1 h,extraction pH 10 and ratio of material to liquid 1:9. Under the optimal condi-tions,the extraction rate of hemp protein was 63. 5%. The relative error between predictive value and ex-perimental value of the regression model was 2. 3%,indicating a good fit between regression equation and actual situation. The DSC analysis results showed that the denaturation temperature of hemp protein was 83. 0℃, and the relative molecular weight distributions of purified hemp protein were less than 40 kDa.

  19. Optimizing Sustainable Geothermal Heat Extraction

    Science.gov (United States)

    Patel, Iti; Bielicki, Jeffrey; Buscheck, Thomas

    2016-04-01

    Geothermal heat, though renewable, can be depleted over time if the rate of heat extraction exceeds the natural rate of renewal. As such, the sustainability of a geothermal resource is typically viewed as preserving the energy of the reservoir by weighing heat extraction against renewability. But heat that is extracted from a geothermal reservoir is used to provide a service to society and an economic gain to the provider of that service. For heat extraction used for market commodities, sustainability entails balancing the rate at which the reservoir temperature renews with the rate at which heat is extracted and converted into economic profit. We present a model for managing geothermal resources that combines simulations of geothermal reservoir performance with natural resource economics in order to develop optimal heat mining strategies. Similar optimal control approaches have been developed for managing other renewable resources, like fisheries and forests. We used the Non-isothermal Unsaturated-saturated Flow and Transport (NUFT) model to simulate the performance of a sedimentary geothermal reservoir under a variety of geologic and operational situations. The results of NUFT are integrated into the optimization model to determine the extraction path over time that maximizes the net present profit given the performance of the geothermal resource. Results suggest that the discount rate that is used to calculate the net present value of economic gain is a major determinant of the optimal extraction path, particularly for shallower and cooler reservoirs, where the regeneration of energy due to the natural geothermal heat flux is a smaller percentage of the amount of energy that is extracted from the reservoir.

  20. 葵花籽粕蛋白质提取工艺参数优化%Parameters Optimization of the Extraction Technology of Protein from Sunflower Meal

    Institute of Scientific and Technical Information of China (English)

    加列西·马那甫; 景伟文; 吐力吾汗·阿米汗

    2014-01-01

    To study the extraction technology of protein from sunflower meal using salt extraction. Based on the single-factor test,the extraction factors of NaCl concentration,extraction temperature and water/dry ratio were optimized using the response surface methodology with the extraction rate of protein as the response value. As a result ,the optimum extraction conditions of protein from sunflower meal were 1.51 mol/L of NaCl concentration , 1∶12.88 (g/mL) of dry/water ratio,60.27℃of extraction temperature. Under these conditions,the extraction rate of chlorogenic acid was 36.64%.%研究葵花籽粕蛋白质盐提法提取最佳工艺。在单因素试验的基础上,选取NaCl浓度、提取温度和料液比为自变量,以蛋白质提取率为响应值,利用响应面法对葵花籽粕蛋白质的提取工艺进行优化,得到回归方程的预测模型,该模型能较好的反映各因素与响应值之间的关系。最佳提取工艺条件为,NaCl浓度1.51 mol/L、料液比1∶12.88(g/mL)、提取温度60.27℃。在此条件下,蛋白质的提取率为36.64%。

  1. Optimal Extraction of Geothermal Energy

    Energy Technology Data Exchange (ETDEWEB)

    Golabi, Kamal; Scherer, Charles, R.

    1977-06-01

    This study is concerned with the optimal extraction of energy from a hot water geothermal field. In view of the relative "commercial" availability of the many energy sources alternative to geothermal, it is possible that a socially "best" extraction policy may not include producing geothermal energy as fast as the current technology will permit. Rather, a truly "optimal" policy will depend on, among other things, the costs and value of geothermal energy in the future and the analogous values of other energy sources. Hence, a general approach to this problem would make the policy contingent on pertinent information on alternative sources. A good example of this approach is given in Manne's (1976) Energy Technology Assessment Model, where he points out that "Each energy source has its own cost parameters and introduction date, but is interdependent with other components of the energy sector." (Manne (1976), p. 379). But by their large dimensions, such relativity macro-analyses tend to preclude a close look at the specific technology of a process is important in developing meaningful resource management models, we substitute for a macro model the increasing value over time of the energy extracted. In this contact we seek an extraction rate (and an economic life) that maximizes the net discounted value of the energy extracted. [DJE-2005

  2. Optimization of an Efficient Protein Extraction Protocol Compatible with Two-Dimensional Electrophoresis and Mass Spectrometry from Recalcitrant Phenolic Rich Roots of Chickpea (Cicer arietinum L.

    Directory of Open Access Journals (Sweden)

    Moniya Chatterjee

    2012-01-01

    Full Text Available Two-dimensional electrophoresis and mass spectrometry are undoubtedly two essential tools popularly used in proteomic analyses. Utilization of these techniques however largely depends on efficient and optimized sample preparation, regarded as one of the most crucial steps for recovering maximum amount of reliable information. The present study highlights the optimization of an effective and efficient protocol, capable of extraction of root proteins from recalcitrant phenolic rich tissues of chickpea. The widely applicable TCA-acetone and phenol-based methods have been comparatively evaluated, amongst which the latter appeared to be better suited for the sample. The phenol extraction-based method further complemented with sodium dodecyl sulphate (SDS and pulsatory treatments proved to be the most suitable method represented by greatest spot number, good resolution, and spot intensities. All the randomly selected spots showed successful identification when subjected to further downstream MALDI-TOF and MS/MS analyses. Hence, the information obtained collectively proposes the present protein extraction protocol to be an effective one that could be applicable for recalcitrant leguminous root samples.

  3. Optimization of an Efficient Protein Extraction Protocol Compatible with Two-Dimensional Electrophoresis and Mass Spectrometry from Recalcitrant Phenolic Rich Roots of Chickpea (Cicer arietinum L.).

    Science.gov (United States)

    Chatterjee, Moniya; Gupta, Sumanti; Bhar, Anirban; Das, Sampa

    2012-01-01

    Two-dimensional electrophoresis and mass spectrometry are undoubtedly two essential tools popularly used in proteomic analyses. Utilization of these techniques however largely depends on efficient and optimized sample preparation, regarded as one of the most crucial steps for recovering maximum amount of reliable information. The present study highlights the optimization of an effective and efficient protocol, capable of extraction of root proteins from recalcitrant phenolic rich tissues of chickpea. The widely applicable TCA-acetone and phenol-based methods have been comparatively evaluated, amongst which the latter appeared to be better suited for the sample. The phenol extraction-based method further complemented with sodium dodecyl sulphate (SDS) and pulsatory treatments proved to be the most suitable method represented by greatest spot number, good resolution, and spot intensities. All the randomly selected spots showed successful identification when subjected to further downstream MALDI-TOF and MS/MS analyses. Hence, the information obtained collectively proposes the present protein extraction protocol to be an effective one that could be applicable for recalcitrant leguminous root samples.

  4. Optimal and Sustainable Groundwater Extraction

    Directory of Open Access Journals (Sweden)

    Christopher A. Wada

    2010-08-01

    Full Text Available With climate change exacerbating over-exploitation, groundwater scarcity looms as an increasingly critical issue worldwide. Minimizing the adverse effects of scarcity requires optimal as well as sustainable patterns of groundwater management. We review the many sustainable paths for groundwater extraction from a coastal aquifer and show how to find the particular sustainable path that is welfare maximizing. In some cases the optimal path converges to the maximum sustainable yield. For sufficiently convex extraction costs, the extraction path converges to an internal steady state above the level of maximum sustainable yield. We describe the challenges facing groundwater managers faced with multiple aquifers, the prospect of using recycled water, and the interdependence with watershed management. The integrated water management thus described results in less water scarcity and higher total welfare gains from groundwater use. The framework also can be applied to climate-change specifications about the frequency, duration, and intensity of precipitation by comparing before and after optimal management. For the case of South Oahu in Hawaii, the prospect of climate change increases the gains of integrated groundwater management.

  5. 马鹿茸水溶性总蛋白提取工艺研究%Optimization of Extraction Technology of Soluble Proteins from Velvet Antler of Cervus elaphus Linnaeus

    Institute of Scientific and Technical Information of China (English)

    周杰

    2016-01-01

    目的:优选鹿茸水溶性蛋白的最佳提取工艺.方法:以马鹿茸为原料,采用组织匀浆结合浸提的方法提取可溶性蛋白,以鹿茸可溶性蛋白得率和含量为评价指标,通过单因素实验研究匀浆方法、匀浆次数、提取溶液浓度和pH对鹿茸可溶性蛋白得率的影响,优化提取工艺.结果:鹿茸可溶性蛋白的最佳提取工艺为,以pH为7,50mmol?L-1的Tris-HCl缓冲溶液为提取溶剂,采用电动匀浆机,匀浆次数为3次.结论:优化得到的提取工艺稳定、可行,可为进一步研究鹿茸蛋白的药理药效提供参考.%Objective To optimize the appropriate extracting technology for soluble proteins from velvet antler.Methods Extracted the soluble proteins with homogenate method, studied the effect of homogenate method, homogenate times, concentration and pH of extraction solvent, using soluble proteins extraction rates and content as the indices. Results: The optimal extraction technology for the soluble proteins was as follows: electric homogenate, the extraction solvent of 50 mmol·L-1, pH of 7.The extraction rate of he soluble proteins from velvet antler was and the content of the soluble proteins was .Conclusion The optimal extraction technology is stable and feasible, and can provide reference for further study on velvet antler.

  6. Vessel tree extraction using locally optimal paths

    DEFF Research Database (Denmark)

    Lo, Pechin Chien Pau; van Ginneken, Bram; de Bruijne, Marleen

    2010-01-01

    This paper proposes a method to extract vessel trees by continually extending detected branches with locally optimal paths. Our approach uses a cost function from a multi scale vessel enhancement filter. Optimal paths are selected based on rules that take into account the geometric characteristics...... of the vessel tree. Experiments were performed on 10 low dose chest CT scans for which the pulmonary vessel trees were extracted. The proposed method is shown to extract a better connected vessel tree and extract more of the small peripheral vessels in comparison to applying a threshold on the output...

  7. Study of total dry matter and protein extraction from canola meal as affected by the pH, salt addition and use of zeta-potential/turbidimetry analysis to optimize the extraction conditions.

    Science.gov (United States)

    Gerzhova, Alina; Mondor, Martin; Benali, Marzouk; Aider, Mohammed

    2016-06-15

    Total dry matter and proteins were differentially and preferentially extracted from canola meal (CM) under different conditions. The effect of the extraction medium pH, CM concentration and salt concentrations were found to have different influences on the extractability of total dry matter and proteins from CM. The pH of the extracting medium had the most significant effect. The maximal total dry matter (42.8±1.18%) extractability was obtained with 5% CM at pH 12 without salt addition, whereas the maximal for total protein (58.12±1.47%) was obtained with 15% CM under the same conditions. The minimal extractability for the dry matter (26.63±0.67%) was obtained with 5% CM at pH 10 without salt added and the minimal protein extractability was observed in a 10% CM at pH 10, in 0.01 NaCl. Turbidity and ζ-potential measurements indicated that pH 5 was the optimum condition for the highest protein extraction yield. SDS-PAGE analysis showed that salt addition contributes to higher solubility of canola proteins specifically cruciferin fraction, although it reduces napin extraction.

  8. Extraction Optimization of Polysaccharides from Pitaya Stems

    Institute of Scientific and Technical Information of China (English)

    HE Cong-fen; LI Peng; ZHAO Hua; SONG Li-ya; ZHU Jun; DONG Yin-mao

    2011-01-01

    [Objective] The aim was to describe the extraction of polysaccharides from pitaya stems.[Method] The hot water,enzyme-assisted and microwave-assisted methods were used,with the microwave-assisted extraction being deemed optimal by general evaluation.[Result] The main factors affecting the yield of polysaccharides in the microwave-assisted extraction,by order of magnitude,were as follows:timemicrowave powertemperature;additionally,optimal conditions included a 10 min extraction time,an 80 ℃ extraction temperature and a microwave setting of 200 W.Using these optimal conditions,the yield of PSPS(Polysaccharides from Pitaya Stems) was 1.42%.After purification,the yield of PSPS was 0.74%.[Conclusion] The PSPS was analyzed by IR,MALDI-TOF-MS and an element analysis technique.It was shown to be a polysaccharide mixture,and the molecular weight was between 3 900 and 4 300 Da.

  9. Quantitative and qualitative optimization of allergen extraction from peanut and selected tree nuts. Part 1. Screening of optimal extraction conditions using a D-optimal experimental design.

    Science.gov (United States)

    L'Hocine, Lamia; Pitre, Mélanie

    2016-03-01

    A D-optimal design was constructed to optimize allergen extraction efficiency simultaneously from roasted, non-roasted, defatted, and non-defatted almond, hazelnut, peanut, and pistachio flours using three non-denaturing aqueous (phosphate, borate, and carbonate) buffers at various conditions of ionic strength, buffer-to-protein ratio, extraction temperature, and extraction duration. Statistical analysis showed that roasting and non-defatting significantly lowered protein recovery for all nuts. Increasing the temperature and the buffer-to-protein ratio during extraction significantly increased protein recovery, whereas increasing the extraction time had no significant impact. The impact of the three buffers on protein recovery varied significantly among the nuts. Depending on the extraction conditions, protein recovery varied from 19% to 95% for peanut, 31% to 73% for almond, 17% to 64% for pistachio, and 27% to 88% for hazelnut. A modulation by the buffer type and ionic strength of protein and immunoglobuline E binding profiles of extracts was evidenced, where high protein recovery levels did not always correlate with high immunoreactivity.

  10. Automatic Extraction of Protein Interaction in Literature

    OpenAIRE

    Liu, Peilei; Wang, Ting

    2014-01-01

    Protein-protein interaction extraction is the key precondition of the construction of protein knowledge network, and it is very important for the research in the biomedicine. This paper extracted directional protein-protein interaction from the biological text, using the SVM-based method. Experiments were evaluated on the LLL05 corpus with good results. The results show that dependency features are import for the protein-protein interaction extraction and features related to the interaction w...

  11. Implantable defibrillator lead extraction with optimized standard extraction techniques

    Institute of Scientific and Technical Information of China (English)

    Xian-Ming Chu; Xue-Bin Li; Ping Zhang; Yi An; Jiang-Bo Duan; Long Wang; Ding Li; Bing Li; Ji-Hong Guo

    2013-01-01

    Background Implantable cardioverter-defibrillator (ICD) leads might not be extracted especially in developing countries because of the high cost and lack of specialized tools. We aimed to evaluate transvenous extraction of ICD leads using optimized standard techniques. Methods We prospectively analyzed clinical characteristics, optimized extraction techniques and the feasibility of extraction for 40 patients (33 males; mean age 47.9 ± 16.1 years) with 42 ICD leads. Results Complete procedural success rate was 95.2% (40/42), and the clinical success rate was 97.6% (41/42). One ICD lead required cardiothoracic surgery. Minor complications occurred in three cases (7.5%), and no major complications or death occurred. Locking stylets were used to extract most leads (34, 81.0%) and almost half of the leads (20, 47.6%) required mechanical dilatation to free fibrotic adhesions; these leads had been implanted for a longer period of time than the others (43.7 ± 18.2 vs. 18.4 ± 13.4 months, P < 0.05). Three-quarters of the leads (30, 71.4%) were extracted with locking stylets plus manual traction (12, 28.6%), or mechanical dilatation with counter-traction (18, 42.8%) by the superior vena cava approach and one-quarter of the leads (11, per lead. Linear regression analysis showed that the extraction time was significantly correlated with implant duration (r = 0.70, P < 0.001). optimized procedure for transvenous extraction of ICD leads provides a practical and low-cost method for standard procedures.

  12. Optimal Extraction of Fibre Optic Spectroscopy

    CERN Document Server

    Sharp, R

    2009-01-01

    We report an optimal extraction methodology, for the reduction of multi-object fibre spectroscopy data, operating in the regime of tightly packed (and hence significantly overlapping) fibre profiles. The routine minimises crosstalk between adjacent fibres and statistically weights the extraction to reduce noise. As an example of the process we use simulations of the numerous modes of operation of the AAOmega fibre spectrograph and observational data from the SPIRAL Integral Field Unit at the Anglo-Australian Telescope.

  13. Airway Tree Extraction with Locally Optimal Paths

    DEFF Research Database (Denmark)

    Lo, Pechin Chien Pau; Sporring, Jon; Pedersen, Jesper Johannes Holst

    2009-01-01

    This paper proposes a method to extract the airway tree from CT images by continually extending the tree with locally optimal paths. This is in contrast to commonly used region growing based approaches that only search the space of the immediate neighbors. The result is a much more robust method...... for tree extraction that can overcome local occlusions. The cost function for obtaining the optimal paths takes into account of an airway probability map as well as measures of airway shape and orientation derived from multi-scale Hessian eigen analysis on the airway probability. Significant improvements...

  14. Response Surface Method Optimizing Ultrasonic to Assist the Extraction of Tea Seed Protein Technology%响应面法优化超声波辅助提取茶叶籽蛋白工艺

    Institute of Scientific and Technical Information of China (English)

    苏超; 冯磊; 黄伟; 向小乐; 麻成金; 向勇平

    2012-01-01

    Response surface method (RSM) was applied to optimize ultrasonic so as to assist the extraction of tea seed protein technology. On the basis of a single factor experiment, choosing the solid-liquid ratio, extraction pH, ultrasound temperature, ultrasound time as influencing factors, the tea seed protein extraction yield as response value and applying Box-behnken central combination experimental design to set a mathematical model to analysis the response surface and ridge analysis. The result showed that, the conditions of optimizing the extraction of tea seed protein liquid were: feed liquid ratio 1:21, extraction pH 9.6, ultrasonic temperature 45℃, ultrasonic time 40min. In this optimum condition, the extraction yield of tea seed protein could reach to 80.1%.%利用响应面法(RSM)优化超声波辅助提取茶叶籽蛋白工艺条件,在单因素试验基础上,选取料液比、浸提液pH、超声温度、超声时间为影响因子,以茶叶籽蛋白提取率为响应值,应用Box-behnken中心组合试验设计建立数学模型,进行响应面分析以及岭嵴分析.结果表明,超声波辅助提取茶叶籽蛋白优化工艺条件为:料液比1∶21、浸提液pH9.6、超声温度45℃、超声时间40min,在此最佳条件下,茶叶籽蛋白提取率可达80.1%.

  15. An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis.

    Science.gov (United States)

    Mansour, Anthony G; Khalil, Pamela Abou; Bejjani, Noha; Chatila, Rajaa; Dagher-Hamalian, Carole; Faour, Wissam H

    2017-03-01

    Deparaffinization of formalin-fixed paraffin embedded (FFPE) tissues with xylene currently remains a major challenge to the biomedical community. We developed an efficient xylene-free protocol to isolate proteins from archived FFPE human tissue sections. A total of 79 different types of FFPE tissue sections of 8 µm thickness were obtained from various archived FFPE specimens. Deparaffinization was conducted by gently washing each section with around 1 ml of hot distilled water (≈80°C). The deparaffinized tissues were homogenized in lysis buffer, and the isolated proteins were quantified and efficiently resolved using western blot analysis for the presence of Protein kinase B (PKB/AKT) and β-actin. Moreover, a significant amount of proteins was successfully isolated with an average of 2.31 µg/µl. The migration pattern of AKT and β-actin obtained from the specimens was similar to the positive control obtained from protein lysates prepared from in vitro cultured MDA231 cancer cell lines. AKT was successfully identified in all specimens, and β-actin protein was resolved with an efficiency higher than 80%. The entire extraction procedure requires only 20 minutes. This newly developed technique is an efficient, safe, cost-effective, and rapid method to isolate proteins from FFPE tissue sections adequate for molecular analysis.

  16. Study on Optimization of Extraction of Yellow Pigment from Corn Protein Powder%玉米黄色素萃取工艺优化研究

    Institute of Scientific and Technical Information of China (English)

    孙强; 黄纪念; 宋国辉; 芦鑫; 张丽霞

    2011-01-01

    以玉米淀粉副产物玉米黄浆粉为原料,采用不同的有机溶剂从玉米黄浆粉中提取玉米黄色素,并利用单因素和正交试验来确定最佳因素和工艺条件。结果表明,温度对玉米黄色素提取率的影响最大,其次是提取时间,而料液比的影响较小。其最佳工艺条件为:以无水乙醇为提取剂,料液比为1 15,提取温度为70℃,提取时间为1.5h,玉米黄色素的提取率为56.84%。%The corn yellow pigment was extracted from corn protein powder,which was the by-product in starch plant.This paper studied a technological route of extracting corn pigment from corn protein powder with different organic solvent,and the optimum factor and the technological conditions were determined by the mono-factor and the orthogonal experiment.The results showed that the temperature could have a greater impact on extraction of the corn pigment than time and ratio of material to liquid.The optimum technological condition by solvent was that alcohol was used as the extraction reagent,the extraction temperature was 70℃,the time was 1.5h,the ratio of material to liquid is 1:15,and the yield of yellow pigment was 56.84%.

  17. 丰年虫蛋白质的分类提取工艺优化及抗氧化活性%Extraction Optimization and Antioxidant Activity of Water-Soluble Protein from Chirocephalus diaphanous

    Institute of Scientific and Technical Information of China (English)

    聂路; 张建新; 彭鑫华; 王欣; 黄晓燕

    2011-01-01

    目的:对丰年虫蛋白质进行分类提取和抗氧化活性比较,并对抗氧化活性最强的蛋白质进行提取工艺优化。方法:从丰年虫脱脂粉中分离出水溶性、碱溶性、酸溶性和醇溶性蛋白,并对其进行体外抗氧化活性研究,选择一种抗氧化活性最强的蛋白质,利用中心组合试验设计对其提取工艺进行优化。结果:丰年虫脱脂粉中最主要的蛋白是水溶性蛋白和碱溶性蛋白,其次是醇溶性蛋白,最少的是酸溶性蛋白,该4类分别占脱脂粉的25.03%、26.98%、1.83%、0.53%;通过体外抗氧化研究发现水溶性蛋白抗氧化活性最强;得到提取水溶性蛋白的最优工艺条件是料液比1:7.5、提取温度49.8℃、提取时间2.8h,预测值和实测值的偏差为0.88%。结论:丰年虫水溶性蛋白的抗氧化活性最强,优化的提取工艺条件具有可行性。%Objective: To compare the antioxidant activity of different protein fractions from Chirocephalus diaphanous and optimize the extraction of the fraction with the strongest antioxidant activity.Methods: The residue left after water-soluble protein extraction of Chirocephalus diaphanous was extracted with 0.1 mol/L aqueous NaOH solution,0.1 mol/L aqueous acetic acid solution and 95% ethanol,respectively.The antioxidant activity of the water-soluble,acid-soluble protein,alkaline-soluble protein and alcohol-soluble proteins obtained was comparatively assessed based on their DPPH,hydroxyl and superoxide anion radical activities and total reducing power to identify the protein fraction with the most powerful antioxidant activity among them.Moreover,response surface methodology based on central composite design was used to optimize the extraction of the most antioxidant protein fraction from Chirocephalus diaphanous.Results: The contents of water-soluble,alkaline-soluble,alcohol-soluble and acid-soluble proteins were 25.03%,26.98%,1.83% and 0.53%,respectively

  18. OPTIMAL MAINTENANCE AND REPLACEMENT OF EXTRACTION MACHINERY

    Institute of Scientific and Technical Information of China (English)

    Suresh P.SETHI; Hong-Mo YEH; Rong ZHANG; Andrew K.S.JARDINE

    2008-01-01

    This paper considers a problem of optimal preventive maintenance and replacement schedule ofequipment devoted to extracting resources from known deposits. Typical examples are oil drills, mine shovels, etc. At most one replacement of the existing machinery by a new one is allowed. The problem is formulated as an optimal control problem subject to the state constraint that the remaining deposit at any given time is nonnegative. We show that the optimal preventive maintenance, production rates, and the replacement and salvage times of the existing machinery and the new one, if required, can be obtained by solving sequentially a series of free-end-point optimal control problems. Moreover, an algorithm based on this result is developed and used to solve two illustrative examples.

  19. Optimal Management of Geothermal Heat Extraction

    Science.gov (United States)

    Patel, I. H.; Bielicki, J. M.; Buscheck, T. A.

    2015-12-01

    Geothermal energy technologies use the constant heat flux from the subsurface in order to produce heat or electricity for societal use. As such, a geothermal energy system is not inherently variable, like systems based on wind and solar resources, and an operator can conceivably control the rate at which heat is extracted and used directly, or converted into a commodity that is used. Although geothermal heat is a renewable resource, this heat can be depleted over time if the rate of heat extraction exceeds the natural rate of renewal (Rybach, 2003). For heat extraction used for commodities that are sold on the market, sustainability entails balancing the rate at which the reservoir renews with the rate at which heat is extracted and converted into profit, on a net present value basis. We present a model that couples natural resource economic approaches for managing renewable resources with simulations of geothermal reservoir performance in order to develop an optimal heat mining strategy that balances economic gain with the performance and renewability of the reservoir. Similar optimal control approaches have been extensively studied for renewable natural resource management of fisheries and forests (Bonfil, 2005; Gordon, 1954; Weitzman, 2003). Those models determine an optimal path of extraction of fish or timber, by balancing the regeneration of stocks of fish or timber that are not harvested with the profit from the sale of the fish or timber that is harvested. Our model balances the regeneration of reservoir temperature with the net proceeds from extracting heat and converting it to electricity that is sold to consumers. We used the Non-isothermal Unconfined-confined Flow and Transport (NUFT) model (Hao, Sun, & Nitao, 2011) to simulate the performance of a sedimentary geothermal reservoir under a variety of geologic and operational situations. The results of NUFT are incorporated into the natural resource economics model to determine production strategies that

  20. Watermark Extraction Optimization Using PSO Algorithm

    Directory of Open Access Journals (Sweden)

    Mohammad Dehghani Soltani

    2013-04-01

    Full Text Available In this study we propose an improved method for watermarking based on ML detector that in comparison with similar methods this scheme has more robustness against attacks, with the same embedded length of logo. Embedding the watermark will perform in the low frequency coefficients of wavelet transform of high entropy blocks (blocks which have more information. Then in the watermark extraction step by using PSO algorithm in a way that maximum quality in comparison with previous methods obtain, by optimizing the Lagrange factor in the Neyman-Peyrson test, we extract the watermark. Finally, performance of proposed scheme has been investigated and accuracy of results are shown by simulation.

  1. Detergent selection for enhanced extraction of membrane proteins.

    Science.gov (United States)

    Arachea, Buenafe T; Sun, Zhen; Potente, Nina; Malik, Radhika; Isailovic, Dragan; Viola, Ronald E

    2012-11-01

    Generating stable conditions for membrane proteins after extraction from their lipid bilayer environment is essential for subsequent characterization. Detergents are the most widely used means to obtain this stable environment; however, different types of membrane proteins have been found to require detergents with varying properties for optimal extraction efficiency and stability after extraction. The extraction profiles of several detergent types have been examined for membranes isolated from bacteria and yeast, and for a set of recombinant target proteins. The extraction efficiencies of these detergents increase at higher concentrations, and were shown to correlate with their respective CMC values. Two alkyl sugar detergents, octyl-β-d-glucoside (OG) and 5-cyclohexyl-1-pentyl-β-d-maltoside (Cymal-5), and a zwitterionic surfactant, N-decylphosphocholine (Fos-choline-10), were generally effective in the extraction of a broad range of membrane proteins. However, certain detergents were more effective than others in the extraction of specific classes of integral membrane proteins, offering guidelines for initial detergent selection. The differences in extraction efficiencies among this small set of detergents supports the value of detergent screening and optimization to increase the yields of targeted membrane proteins.

  2. Optimization of the enzymatic extraction process of protein in tea residue by response surface methodology%响应面优化酶法提取茶渣蛋白的工艺研究

    Institute of Scientific and Technical Information of China (English)

    李圆圆; 王洪新; 殷飞; 兰海崇; 佘世凤

    2013-01-01

    To optimize the extraction parameters of protein in tea residue,the Central Composite Design was used to analyze the factors that affect the protein extraction rate.Alcalase was selected to extract the protein in tea residue. Based on single factor experiment results,enzyme hydrolysis temperature,reaction time,the ratio of liquid to solid, enzyme amount and enzyme hydrolysis pH were chosen as influencing factors and the protein yield was selected as the response value. The results showed that the optimal extraction conditions were: enzyme hydrolysis temperature 60℃,reaction time 1.5h,the ratio of liquid to solid 20:1,enzyme amount 2.5% and enzyme hydrolysis pH9.5.ln this optimal process condition,the yield of protein in tea residue was 56.95%.Compared with the previous report,extraction rate of protein in tea residue increased significantly with the application of response surface analysis methodology.%以茶渣为原料,选用碱性蛋白酶,利用中心组合实验设计对影响茶渣蛋白提取率的因素水平进行综合研究,优化茶渣蛋白的提取工艺参数根据单因素实验结果,以提取温度、提取时间、液固比、加酶量和pH为影响因子,茶渣蛋白提取率为响应值,确定最优提取条件为:提取温度60℃,提取时间1.5h,液固比20∶1,加酶量2.5%,pH9.5,此时茶渣蛋白提取率为56.95% 与之前报道相比,应用响应面分析法研究酶法提取茶渣蛋白,提取率有显著提高.

  3. 响应面优化花生分离蛋白提取工艺的研究%Optimization of processing parameters for extraction of peanut protein isolate by response surface methodology

    Institute of Scientific and Technical Information of China (English)

    章宝; 单杨; 李高阳

    2012-01-01

    Peanut protein isolate was prepared through alkaline extraction and acid precipitation from low variability peanut meal. The optimal values of theoretical parameters including extraction temperature, ratio of liquid and material,extraction time and extraction pH were investigated by Box-Behnken surface response design method. The optimum conditions of alkaline extraction were as followed =extraction temperature 57.5℃, ratio of liquid to material 12:1,extraction time 100min and pH 9.2. And based on the single-factor test,the optimum pH4.5 of acid precipitation was determined. Under these conditions,the maximum protein extraction rate of 87.57% was obtained. The relative error with predicted value was 0.4%.%采用碱提酸沉法从低变性花生粕中提取花生分离蛋白,以花生分离蛋白提取率为响应值,应用Box-BehnkenDesign(BBD)对碱提温度、时间、pH和液料比4因素进行响应面分析,确定碱提的最佳工艺条件为:碱提温度57.5℃、时间100min、pH9.2和液料比12∶1;通过单因素实验确定最佳酸沉pH为4.5。在此条件下,花生分离蛋白最高提取率为87.57%,与预测值87.88%相对误差约为0.4%。

  4. Nucleic acid protocols: Extraction and optimization

    Directory of Open Access Journals (Sweden)

    Saeed El-Ashram

    2016-12-01

    Full Text Available Yield and quality are fundamental features for any researchers during nucleic acid extraction. Here, we describe a simplified, semi-unified, effective, and toxic material free protocol for extracting DNA and RNA from different prokaryotic and eukaryotic sources exploiting the physical and chemical properties of nucleic acids. Furthermore, this protocol showed that DNA and RNA are under triple protection (i.e. EDTA, SDS and NaCl during lysis step, and this environment is improper for RNase to have DNA liberated of RNA and even for DNase to degrade the DNA. Therefore, the complete removal of RNA under RNase influence is achieved when RNase is added after DNA extraction, which gives optimal quality with any protocols. Similarly, DNA contamination in an isolated RNA is degraded by DNase to obtain high-quality RNA. Our protocol is the protocol of choice in terms of simplicity, recovery time, environmental safety, amount, purity, PCR and RT-PCR applicability.

  5. [Efficient extraction of transmembrane proteins using ProteoExtract Transmembrane Protein Extraction Kit].

    Science.gov (United States)

    Błachnio, Karina

    2010-01-01

    Detergents commonly used for solubilization of membrane proteins may be ionic or non-ionic. Exposing membrane proteins to detergents, however, can adversely affect their native structure, which can be a major hindrance for functional studies. This is especially true for proteins with multiple transmembrane domains. The ProteoExtract Transmembrane Protein Extraction Kit (TM-PEK), offered by Merck, provides a detergent-free novel reagents to enable the mild and efficient extraction of proteins containing seven transmembrane domains, such as GPCRs (G-Protein Coupled Receptors) e.g.: Frizzled-4 and CELSR-3, from mammalian cells. The fraction enriched in transmembrane proteins using TM-PEK is directly compatible with enzyme assays, non-denaturing gel electrophoresis, 1- and 2-D SDS-PAGE, MS analysis, Western blotting, immunoprecipitation and ELISA. Unlike many alternatives, TM-PEK extraction procedure does not require sonication, extended rigorous vortexing, ultracentrifugation, or incubation of samples at elevated temperatures--thus minimizing the risk of post-extraction degradation or modifications.

  6. Protein structure refinement by optimization.

    Science.gov (United States)

    Carlsen, Martin; Røgen, Peter

    2015-09-01

    Knowledge-based protein potentials are simplified potentials designed to improve the quality of protein models, which is important as more accurate models are more useful for biological and pharmaceutical studies. Consequently, knowledge-based potentials often are designed to be efficient in ordering a given set of deformed structures denoted decoys according to how close they are to the relevant native protein structure. This, however, does not necessarily imply that energy minimization of this potential will bring the decoys closer to the native structure. In this study, we introduce an iterative strategy to improve the convergence of decoy structures. It works by adding energy optimized decoys to the pool of decoys used to construct the next and improved knowledge-based potential. We demonstrate that this strategy results in significantly improved decoy convergence on Titan high resolution decoys and refinement targets from Critical Assessment of protein Structure Prediction competitions. Our potential is formulated in Cartesian coordinates and has a fixed backbone potential to restricts motions to be close to those of a dihedral model, a fixed hydrogen-bonding potential and a variable coarse grained carbon alpha potential consisting of a pair potential and a novel solvent potential that are b-spline based as we use explicit gradient and Hessian for efficient energy optimization.

  7. How biomass composition determines protein extractability

    NARCIS (Netherlands)

    Sari, Y.W.; Syafitri, U.; Sanders, J.P.M.; Bruins, M.E.

    2015-01-01

    Biomass consists of a complex mixture of different components, of which protein potentially has a high added value for biorefinery. In this study, protein extractability of different types of biomass, mostly by-products, was analyzed. Protein yield obtained from a three step extraction using alkali

  8. How biomass composition determines protein extractability

    NARCIS (Netherlands)

    Sari, Y.W.; Syafitri, U.; Sanders, J.P.M.; Bruins, M.E.

    2015-01-01

    Biomass consists of a complex mixture of different components, of which protein potentially has a high added value for biorefinery. In this study, protein extractability of different types of biomass, mostly by-products, was analyzed. Protein yield obtained from a three step extraction using alkali

  9. 响应面法优化苦瓜籽蛋白质提取工艺%Optimization of protein extraction from defatted bitter melon seed using response surface methodology

    Institute of Scientific and Technical Information of China (English)

    张治强; 成兰英

    2013-01-01

    The optimum extracting conditions of protein from bitter melon seeds by the salt solvent method were studied .The effects of concentration of NaCI,pH and extraction time on the yield of protein were studied based on the extraction rate.According to the single factor experiments, the optimum extract process for bitter melon seed protein was determined through Box-Benhnken central composite design and response surface methodology.The results showed that the optimized extract process for bitter gourd seed protein was that the material/liquid ratio was 1:6 and the temperature was at 4℃,the concentration of NaCI was 1.3mol/L and pH8.8,at a extract time of 3. 6h,the extraction rate of bitter gourd seed protein was 23.80% ,3,24 times higher than that before the optimization. In addition, a quadratic equation model for protein extraction was set up and it played an important role in predicting the extraction of protein.Furthermore,the protein content of bitter melon seeds was 32.13% determined by Kjeldahl method.%以苦瓜籽为原料,采用紫外光谱和响应面法研究盐溶法提取苦瓜籽蛋白质的最佳工艺.以苦瓜籽蛋白质的提取率为指标,考察NaCl浓度、pH、提取时间对苦瓜籽蛋白质提取率的影响.在单因素实验的基础上,通过Box-Benhnken中心组合设计和响应面分析法,确定苦瓜籽蛋白质的最佳提取工艺.结果表明:在料液比为1:6,提取温度为4℃下,NaCl浓度1.3mol/L,pH8.8,提取时间3.6h,苦瓜籽蛋白质的提取率为23.80%,较优化前提高了3.24倍,同时建立了盐提蛋白质的二次多项数学模型,对苦瓜籽蛋白质的提取具有很好的预测作用.此外,通过凯氏定氮法确定苦瓜籽中蛋白质含量为32.13%.

  10. Optimization of RNA Extraction from Rat Pancreatic Tissue

    Directory of Open Access Journals (Sweden)

    Sanaz Dastgheib

    2014-05-01

    Full Text Available Background: Optimized RNA extraction from tissues and cell lines consists of four main stages regardless of the method of extraction: 1 homogenizing, 2 effective denaturation of proteins from RNA, 3 inactivation of ribonuclease, and 4 removal of any DNA, protein, and carbohydrate contamination. Isolation of undamaged intact RNA is challenging when the related tissue contains high levels of RNase. Various technical difficulties occur during extraction of RNA from pancreatic tissue due to spontaneous autolysis. Since standard routine protocols yield unacceptable results in pancrease, we have designed a simple method for RNA extraction by comparing different protocols. Methods: We obtained 20-30 mg pancreatic tissues in less than 2 min from 30 rats. Several methods were performed to extract RNA from pancreatic tissue and evaluate its integrity. All methods were performed three times to obtain reproducible results. Results: Immersing pancreatic tissue in RNA-later for 24 h at -80ºC yielded high quality RNA by using the TriPure reagent which was comparable to the commercial RNeasy Micro Kit. The quality of RNA was evaluated by spectrophotometer, electrophoresis and RT-PCR. We separated intact 28S and 18S ribosomal RNA (rRNA when our procedure was compared with the RNeasy Micro Kit. Finally, full length of the actin gene was amplified by RT-PCR. Conclusion: We designed a simple, fast, cost-effective method for complete RNA extraction from the least amount of quantitatively intact pancreatic tissue

  11. Techno-economical evaluation of protein extraction for microalgae biorefinery

    Science.gov (United States)

    Sari, Y. W.; Sanders, J. P. M.; Bruins, M. E.

    2016-01-01

    Due to scarcity of fossil feedstocks, there is an increasing demand for biobased fuels. Microalgae are considered as promising biobased feedstocks. However, microalgae based fuels are not yet produced at large scale at present. Applying biorefinery, not only for oil, but also for other components, such as carbohydrates and protein, may lead to the sustainable and economical microalgae-based fuels. This paper discusses two relatively mild conditions for microalgal protein extraction, based on alkali and enzymes. Green microalgae (Chlorella fusca) with and without prior lipid removal were used as feedstocks. Under mild conditions, more protein could be extracted using proteases, with the highest yields for microalgae meal (without lipids). The data on protein extraction yields were used to calculate the costs for producing 1 ton of microalgal protein. The processing cost for the alkaline method was € 2448 /ton protein. Enzymatic method performed better from an economic point of view with € 1367 /ton protein on processing costs. However, this is still far from industrially feasible. For both extraction methods, biomass cost per ton of produced product were high. A higher protein extraction yield can partially solve this problem, lowering processing cost to €620 and 1180 /ton protein product, using alkali and enzyme, respectively. Although alkaline method has lower processing cost, optimization appears to be better achievable using enzymes. If the enzymatic method can be optimized by lowering the amount of alkali added, leading to processing cost of € 633/ton protein product. Higher revenue can be generated when the residue after protein extraction can be sold as fuel, or better as a highly digestible feed for cattle.

  12. A Cooperative Approach for the Extraction of Protein Motifs

    Institute of Scientific and Technical Information of China (English)

    Chao CHEN; Yuan Xin TIAN; Xiao Yong ZOU; Pei Xiang CAI; Jin Yuan MO

    2006-01-01

    By integrating the concept of cooperative approach, an extension of the fast annealing coevolutionary algorithm is presented in this paper. It outperformed the original algorithm in the domain of function optimization, especially in terms of convergence rate. It was also applied to a real optimization problem, protein motif extraction. And a satisfactory result has been obtained with the accuracy of prediction achieving 67.0%, which is in agreement with the result in the PROSITE database.

  13. Towards plant protein refinery: Review on protein extraction using alkali and potential enzymatic assistance.

    Science.gov (United States)

    Sari, Yessie W; Mulder, Wilhelmus J; Sanders, Johan P M; Bruins, Marieke E

    2015-08-01

    The globally increasing protein demands require additional resources to those currently available. Furthermore, the optimal usage of protein fractions from both traditional and new protein resources, such as algae and leaves, is essential. Here, we present an overview on alkaline plant protein extraction including the potentials of enzyme addition in the form of proteases and/or carbohydrolases. Strategic biomass selection, combined with the appropriate process conditions can increase protein yields after extraction. Enzyme addition, especially of proteases, can be useful when alkaline protein extraction yields are low. These additions can also be used to enable processing at a pH closer to 7 to avoid the otherwise severe conditions that denature proteins. Finally, a protein biorefinery concept is presented that aims to upcycle residual biomass by separating essential amino acids to be used for food and feed, and non-essential amino acids for production of bulk chemicals.

  14. Effect of Ultrasound in Soybean Protein Extraction

    Science.gov (United States)

    Fukase, Hirokazu; Ohdaira, Etsuzo; Masuzawa, Nobuyoshi; Ide, Masao

    1994-05-01

    Application of ultrasound for accelerating the extraction of nutriments in food processing has been attempted. However, conditions of exposure to ultrasound were not clear in previous studies. This paper reports on the relationship between the ultrasonic pressure and the amount of extracted protein from soybeans. Experiments were conducted using a beaker, in which the ultrasonic fields were precisely measured. Soybean flakes suspended in water were put in the beaker and placed in a water tank. The amount of extracted protein in water upon ultrasonic exposure was calculated by the Kjeldahl method. It was found that the amount of extracted protein increased in proportion to ultrasonic pressure up to the total amount of soybean protein soluble in water. Furthermore, this paper describes the denaturation of the protein produced by the ultrasonic cavitation.

  15. Direct Cellular Lysis/Protein Extraction Protocol for Soil Metaproteomics

    Energy Technology Data Exchange (ETDEWEB)

    Chourey, Karuna [ORNL; Jansson, Janet [Lawrence Berkeley National Laboratory (LBNL); Verberkmoes, Nathan C [ORNL; Shah, Manesh B [ORNL; Chavarria, Krystle L. [Lawrence Berkeley National Laboratory (LBNL); Tom, Lauren M [Lawrence Berkeley National Laboratory (LBNL); Brodie, Eoin L. [Lawrence Berkeley National Laboratory (LBNL); Hettich, Robert {Bob} L [ORNL

    2010-01-01

    We present a novel direct protocol for deep proteome characterization of microorganisms in soil. The method employs thermally assisted detergent-based cellular lysis (SDS) of soil samples, followed by TCA precipitation for proteome extraction/cleanup prior to liquid chromatography-mass spectrometric characterization. This approach was developed and optimized using different soils inoculated with genome-sequenced bacteria (Gram-negative Pseudomonas putida or Gram-positive Arthrobacter chlorophenolicus). Direct soil protein extraction was compared to protein extraction from cells isolated from the soil matrix prior to lysis (indirect method). Each approach resulted in identification of greater than 500 unique proteins, with a wide range in molecular mass and functional categories. To our knowledge, this SDS-TCA approach enables the deepest proteome characterizations of microbes in soil to date, without significant biases in protein size, localization, or functional category compared to pure cultures. This protocol should provide a powerful tool for ecological studies of soil microbial communities.

  16. Direct cellular lysis/protein extraction protocol for soil metaproteomics.

    Science.gov (United States)

    Chourey, Karuna; Jansson, Janet; VerBerkmoes, Nathan; Shah, Manesh; Chavarria, Krystle L; Tom, Lauren M; Brodie, Eoin L; Hettich, Robert L

    2010-12-03

    We present a novel direct protocol for deep proteome characterization of microorganisms in soil. The method employs thermally assisted detergent-based cellular lysis (SDS) of soil samples, followed by TCA precipitation for proteome extraction/cleanup prior to liquid chromatography-mass spectrometric characterization. This approach was developed and optimized using different soils inoculated with genome-sequenced bacteria (Gram-negative Pseudomonas putida or Gram-positive Arthrobacter chlorophenolicus). Direct soil protein extraction was compared to protein extraction from cells isolated from the soil matrix prior to lysis (indirect method). Each approach resulted in identification of greater than 500 unique proteins, with a wide range in molecular mass and functional categories. To our knowledge, this SDS-TCA approach enables the deepest proteome characterizations of microbes in soil to date, without significant biases in protein size, localization, or functional category compared to pure cultures. This protocol should provide a powerful tool for ecological studies of soil microbial communities.

  17. 超声破碎变异链球菌后蛋白质提取条件的优化研究%Optimization study of streptococcus mutans proteins extraction condition

    Institute of Scientific and Technical Information of China (English)

    刘果; 何永红; 万呼春; 钟晓波; 齐进

    2012-01-01

    Objective To investigate the optimal standing time during Streptococcus mutans (one of caries-related bacterium) proteins extraction after the cells broken by ultrasonic. Methods S. mutans splitted under ultrasonic after suspending in cell lysis buffer. Contrast the forms of S. mutans before and after ultrasonic splitting. And the proteins were collected after bacterio-suspen-sion were standing for 0,10,30,50,70 min. Then the influence of standing time on extraction efficiency was studied by using SDS-PAGE and measuring the proteins concentration. Results Few cells broke incompletely after splitting. The protein concentration after standing for 30 min was higher than that of the others(P<0. 05). While the result of SDS-PAGE showed the abundance and distribution of protein bands were basically the same. It s supposed that the release and solubility of protein need necessary time to complete. Conclusion Standing for 30 min after S. mutans cells splitting could raise the proteins extraction efficiency.%目的 寻找超声破碎后高效率提取致龋菌变异链球菌蛋白质的最佳放置时间.方法 以裂解液为提取介质,采用超声法破碎细菌.对比破碎前、后菌形态并将菌提取液分别放置0、10、30、50、70 min收集蛋白上清液,通过RC DC法测定蛋白质浓度及蛋白质SDS-PAGE电泳,比较放置不同时间后的蛋白质提取效果.结果 超声破碎后,少部分细菌破碎不完全.RC DC法测定结果显示,超声破碎后放置30 min时提取的蛋白质浓度高于其他各组(P<0.05),但SDS-PAGE电泳显示蛋白质的条带分布和丰度无明显差别,推测蛋白质的释放和溶解需要一定时间完成.结论 超声法提取变异链球菌蛋白质后放置30 min可显著提高蛋白质提取效率.

  18. Evaluation of the arsenic binding capacity of plant proteins under conditions of protein extraction for gel electrophoretic analysis.

    Science.gov (United States)

    Schmidt, Anne-Christine; Steier, Sandra; Otto, Matthias

    2009-03-15

    As prerequisite for the investigation of arsenic-binding proteins in plants, the general influence of different extraction parameters on the binding behaviour of arsenic to the plant protein pool was investigated. The concentration of the extraction buffer affected the extraction yield both for proteins and for arsenic revealing an optimal buffer concentration of 5mM Tris/HCl, pH 8. The addition of 1 or 2% (w/v) SDS to the extraction buffer produced a two- to threefold enhancement of the total protein extraction yield but strongly suppressed the simultaneous extraction of arsenic from 80+/-8% extraction yield obtained without SDS to 48+/-2% in presence of 2% (w/v) SDS. The arsenic binding capacity of the protein fraction obtained after extraction with Tris buffer and protein precipitation by trichloroacetic acid in acetone was estimated to be 1.4+/-0.6% independently on the original spiking concentration of arsenic provided in the form of monomethylarsonate to the extracts. Due to the low total protein concentrations of the plant extracts that varied in the range from 75 to 412 microgmL(-1) depending on the extraction parameters, high arsenic concentrations of 263-1001 mg (kgproteinmass)(-1) resulted for spiking concentrations of 10 mgAsL(-1). The optimized protein isolation procedure was applied to plants grown under arsenic exposure and revealed a similar arsenic binding capacity as for the spiked protein extracts.

  19. 响应面法超声波辅助提取核桃蛋白工艺优化%Optimization of the preparation of walnut protein with ultrasonic-assisted extraction method

    Institute of Scientific and Technical Information of China (English)

    敬思群

    2012-01-01

    Walnut protein was obstained by ultrasonic-assisted extraction method with the defatted walnut meal as raw material. The preparation process of walnut protein was optimized according to Box- Behnken central composition design principle on foundation of single factor experiment, the method of response surface analysis with 3 factors and 3 levels combined with Minitab15.0-statistical data analysis software was adopted with the leaching rate of walnut protein as the response value. The effect of extraction time, temperature, liquid to solid ratio and pH on the leaching rate of walnut protein were studied in this paper. According to the above results, the optimum preparation conditions were obtained as follows: extraction time 19 min, reaction time 90 min, temperature 46 %, ratio of solvent: meal at 20:1 and pHS.6. Under these conditions, the leaching rate of walnut protein reached 18.66%.%以脱脂核桃粕为原料,利用超声波辅助提取制备核桃蛋白。在单因素实验的基础上,根据Box-Behnken的中心组合实验设计原理,运用Minitab15.0数据统计分析软件,采用3因素3水平的响应面分析法,以核桃蛋白浸出率为响应值,研究了超声时间、超声温度、液料比和pH对核桃蛋白浸出率的影响,并优化了提取工艺。确定了超声辅助提取核桃蛋白的最佳工艺条件为:超声时间为19 min,超声温度为46℃,液料比为20:1,pH为8.6,在此条件下核桃蛋白的浸出率达到68.98%。

  20. 复合反胶束萃取花生蛋白的工艺优化%Optimization of composite reverse micelles extraction of protein from peanut by orthogonal design

    Institute of Scientific and Technical Information of China (English)

    郭珍; 陈复生; 方志锋

    2013-01-01

    Peanut protein was extracted using bis (2-ethylhexyl) sodium sulfos-succinate(AOT)、sodium dodecyl sulfonate (SDS)/isooctane -heptylane capryl alcohol with sonication. The effects of ultrasound time, peanut concentration, ultrasonic power, pH,KCI concentration, temperature, Wo, AOT : SDS, the concentration of surfactant on the forward extraction rates were investigated. Orthogonal experiment was designed to optimize the main seven factors,the results showed that the optimum conditions of forward extraction were: temperature 35℃ , KCI concentration 0 mol/L, pH 8, the concentration of surfactant 0.08 g/mL, AOT : SDS 4 : 3,ultrasonic power 180 W, Wo 15,the extraction rate was 93.33%.%采用AOT(丁二酸二异辛酯磺酸钠)、SDS(十二烷基硫酸钠)/异辛烷—正辛醇复合反胶束体系,超声辅助萃取花生蛋白,研究超声时间、花生浓度、超声功率、pH、离子浓度、温度、W0(反胶束溶液增溶水与表面活性剂摩尔比)、AOT(g)∶SDS(g)、表面活性剂浓度对蛋白萃取率的影响.结果表明,最佳提取工艺为温度35℃、KCl浓度0 mol/L,pH值8,表面活性剂浓度0.08 g/mL,AOT(g)∶SDS(g)为4∶3,超声功率180 W,W0值15,该工艺条件下,花生蛋白的萃取率为93.33%.

  1. Sentence Simplification Aids Protein-Protein Interaction Extraction

    CERN Document Server

    Jonnalagadda, Siddhartha

    2010-01-01

    Accurate systems for extracting Protein-Protein Interactions (PPIs) automatically from biomedical articles can help accelerate biomedical research. Biomedical Informatics researchers are collaborating to provide metaservices and advance the state-of-art in PPI extraction. One problem often neglected by current Natural Language Processing systems is the characteristic complexity of the sentences in biomedical literature. In this paper, we report on the impact that automatic simplification of sentences has on the performance of a state-of-art PPI extraction system, showing a substantial improvement in recall (8%) when the sentence simplification method is applied, without significant impact to precision.

  2. 白鲢中盐溶蛋白提取工艺优化及其加工特性研究%Optimization of extraction conditions of silver carp salt soluble proteins and their processing

    Institute of Scientific and Technical Information of China (English)

    雷颂; 窦建洲; 梅志方; 杨品红; 王伯华

    2015-01-01

    Silver carp,one of the four famous freshwater fish,has limited commercial value due to its strong earthy taste and lots of fish bones.However,its salt soluble proteins can be extracted and has higher economic value.Therefore,optimization of extraction conditions of silver carp salt soluble protein and their processing properties are studied in this paper,The best extraction conditions are 0.8 mol/L NaCl concentration,pH 6.0,extraction time 30 h and solid-liquid ratio 1∶5.Under the above conditions,salt soluble protein concentration is 2.602 4 g/100 g and extraction yield is about 80%.Silver carp thermal induced salt soluble protein gel has good water retention,water holding capacity can be up to 98.24%,the best inducing temperature is 80 ℃.Phosphate can help water retention of the gel.Silver carp salt soluble protein has good emulsification ability and its emulsifying activity (EAI) is 19.47 m2/g,emulsion stability (ESI) is 89.45%.Therefore,the gel can effectively improve the quality of meat products.The study provides a new application of silver carp.%对白鲢盐溶蛋白的提取工艺优化及其加工特性的研究结果显示:NaCl浓度为0.8 mol/L、pH值为6.0、浸提时间为30 h、固液比为1∶5(g∶ mL),在此条件下鲢鱼鱼背肉盐溶蛋白提取浓度为2.6024 g/100 g,盐溶蛋白的提取率最高,为80%左右.白鲢盐溶蛋白热诱导凝胶具有较好保水性,可达98.24%左右,其最佳热诱导凝胶温度为80℃,添加磷酸盐对白鲢盐溶蛋白热诱导凝胶保水性具有一定提升作用.白鲢盐溶蛋白具有较好乳化能力,其乳化活性(EAI)为19.47 m2/g,乳化稳定性(ESI)为89.45%,可有效改善肉糜制品加工品质,为白鲢精深加工提供了新思路.

  3. A Novel Approach for Protein-Named Entity Recognition and Protein-Protein Interaction Extraction

    OpenAIRE

    Meijing Li; Tsendsuren Munkhdalai; Xiuming Yu; Keun Ho Ryu

    2015-01-01

    Many researchers focus on developing protein-named entity recognition (Protein-NER) or PPI extraction systems. However, the studies about these two topics cannot be merged well; then existing PPI extraction systems’ Protein-NER still needs to improve. In this paper, we developed the protein-protein interaction extraction system named PPIMiner based on Support Vector Machine (SVM) and parsing tree. PPIMiner consists of three main models: natural language processing (NLP) model, Protein-NER mod...

  4. 酶法提取玉米胚芽蛋白的工艺优化%Corn germ protein enzymatic extraction process optimization

    Institute of Scientific and Technical Information of China (English)

    皮钰珍; 刘彦妮; 岳喜庆; 尹伊

    2011-01-01

    玉米胚芽蛋白是一种优质植物蛋白,其必需氨基酸种类齐全,营养价值高,同时具有多种生理功能,在食品加工中可作为一种理想的植物蛋白质添加物,研究了使用酶法提取玉米胚芽蛋白的工艺参数,通过单因素实验和响应曲面法确定了提取玉米胚芽蛋白的最佳工艺条件:料液比1:6.8g/mL,加酶量3000u/g料,酶解温度60℃,酶解时间180min,pH7.0,此条件玉米胚芽蛋白的提取率可达到31.06%。%Corn germ protein is a high quality vegetable protein,essential amino acid full range of its high nutritional value,but also has a variety of physiological functions;in food processing plants can be used as an ideal protein supplement.In this paper,the us

  5. Polysaccharides in Sipunculus nudus: Extraction condition optimization and antioxidant activities

    Science.gov (United States)

    Zhang, Qin; Dong, Lanfang; Tong, Tong; Wang, Qingchao; Xu, Mingzhu

    2017-02-01

    Marine organisms constitute unlimited resource of bioactive substances due to their high biodiversity and represent a valuable source of new compounds. This study optimized the alkali-extraction conditions and antioxidant activities of soluble polysaccharides from the body wall of Sipunculus nudus. The effects of solid-liquid ratio, extraction duration, extraction temperature, and alkali concentration on the yield of S. nudus polysaccharides (SNP) were examined, according to which the optimal combination of extraction parameters was obtained by an orthogonal test. The relative influencing importance of different extraction parameters on the yield of SNP followed the order as solid-liquid ratio > extraction temperature > alkali concentration > extraction duration. The highest extraction yield, 1.98%, was achieved under an optimal extraction condition: temperature 60°C, solid-liquid ratio 1:6 g mL-1, duration 5 h, and alkali (NaOH) mass fraction 6%. The in vitro antioxidant activities examination showed that extracted SNP under this optimized condition had strong power in reducing certain hydroxyl and superoxide radical scavenging abilities. The promising results showed that extracted SNP could be a potent natural antioxidant.

  6. Optimization of extraction of soybean protein by mixed reverse micelles composing of SDS-tween 60/isooctane-octanol%SDS-Tween60混合反胶束体系萃取大豆蛋白工艺优化

    Institute of Scientific and Technical Information of China (English)

    李润洁; 布冠好; 陈复生; 徐卫河

    2013-01-01

    soybean protein was extracted by SDS(sodium dodecyl sulfonate)-tween 60 (polyoxyethylene(60)sorbitan monostearate)/isooctane-octanol mixed reverse micelles system with ultrasonic assisted. The effects of the concentration of SDS and tween 60,the ratio of SDS to tween 60,KCl concentration and pH of buffer,water content(W0),extraction temperature and time, adding amount of soybean powder on the forward extraction rates were investigated. Orthogonal experiment was designed to optimize the extraction process,the results showed that the optimum conditions of forward extraction were:the concentration of SDS and tween 60 was 0.10 g/mL,the ratio of SDS to tween 60 was 7∶3,KCl concentration 0.10 mol/L,pH 7,water content(W0)20, extraction temperature 35℃,the forward extraction rate was 89.46%±1.21%. Then,the effect of KCl concentration,pH,ethanol concentration on backward also be studied and the suitable condition is KCl concentration 1.4 mol/L,pH 8.5,the concentration of ethanol 15%,the backward extraction rate was 87.37%±1.64%,so,the whole extraction rate was 78.16%±1.98%.%以全脂大豆粉为原料,采用SDS(十二烷基硫酸钠)-Tween 60(聚氧乙烯失水山梨醇硬脂酸酯)/异辛烷-正辛醇形成混合反胶束体系,辅以超声波萃取大豆蛋白;研究两种表面活性剂总浓度和质量比、缓冲溶液KCl浓度和pH、W0、萃取温度和时间、大豆粉加入量等因素对大豆蛋白前萃率影响,及缓冲溶液KCl浓度、pH和乙醇加入量对大豆蛋白后萃率影响。通过正交试验优化萃取工艺,得到最优前萃取条件为:表面活性剂总浓度0.10 g/mL,质量比为7∶3、KCl浓度0.10 mol/L,pH 7、W020、萃取温度35℃;在此条件下,大豆蛋白前萃率为89.46%±1.21%。最优后萃取条件为:缓冲溶液KCl浓度1.4 mol/L、pH 8.5、乙醇加入量15%,大豆蛋白后萃率为87.37%±1.64%;总萃取率为78.16%±1.98%。

  7. Simulating Protein Conformations through Global Optimization

    CERN Document Server

    Mucherino, A; Pardalos, P M

    2008-01-01

    Many researches have been working on the protein folding problem from more than half century. Protein folding is indeed one of the major unsolved problems in science. In this work, we discuss a model for the simulation of protein conformations. This simple model is based on the idea of imposing few geometric requirements on chains of atoms representing the backbone of a protein conformation. The model leads to the formulation of a global optimization problem, whose solutions correspond to conformations satisfying the desired requirements. The global optimization problem is solved by the recently proposed Monkey Search algorithm. The simplicity of the optimization problem and the effectiveness of the used meta-heuristic search allowed the simulation of a large set of high-quality conformations. We show that, even though only few geometric requirements are imposed, some of the simulated conformation results to be similar (in terms of RMSD) to conformations real proteins actually have in nature.

  8. Extracting knowledge from protein structure geometry

    DEFF Research Database (Denmark)

    Røgen, Peter; Koehl, Patrice

    2013-01-01

    to minima of their free energy surfaces. It is well known however that the situation is more complicated as the current force fields used for molecular simulations fail to recognize native states from misfolded structures. In an attempt to solve this problem, we follow an alternate approach and derive a new...... potential from geometric knowledge extracted from native and misfolded conformers of protein structures. This new potential, Metric Protein Potential (MPP), has two main features that are key to its success. Firstly, it is composite in that it includes local and nonlocal geometric information on proteins...

  9. Optimization of enzyme assisted extraction of polysaccharides from Ganoderma lucidum

    Institute of Scientific and Technical Information of China (English)

    Xu Chunhua; Tian Zhenle; Zhang Chenju; Yu Xiaobing; Zheng Huihua

    2014-01-01

    In the present work,an enzyme assisted extraction method is used to isolate crude polysaccharides from Ganoderma lucidum. The isolating effect was optimized with orthographic graph statistic method with three levels and four independent variables. Complex enzyme,extraction temperature,extraction time and ex-traction pH were combined to obtain the best possible combination to get maximum amount of extract and crude polysaccharides yield. The optimum extraction conditions were:complex enzyme amount of 3%(w/v),extrac-tion temperature at 45℃,extraction time of 3 h and extraction pH at 7. Under these conditions,the experimen-tal amount of extract is 8.9%and the yield of crude polysaccharides is 1.1%,which are in close agreement with the value predicted by the model.

  10. Optimization of buffer solutions for protein crystallization.

    Science.gov (United States)

    Gosavi, Rajendrakumar A; Mueser, Timothy C; Schall, Constance A

    2008-05-01

    Increasing the solubility of protein stock solutions to above that in a standard chromatography buffer (50 mM Tris-HCl pH 7.5, 100 mM NaCl) led to an increase in the number of crystallization conditions for ten globular proteins subjected to two crystal screens: the Index and Precipitant/Precipitant-Additive (P/PA) Screens. Solubility enhancement of protein stock solutions was achieved through screening and selection of buffer components to formulate an optimal buffer. Relative improvements in solubility were estimated through protection against the precipitation of protein by polyethylene glycol 8000. Proteins with limited solubility improvement in optimal buffer showed an enhancement in solubility on addition of glycerol. Maximum solubility was then determined by the concentration of optimized solutions until precipitate formed. The supernatant concentration then provided an estimate of the upper limit of protein solubility. This 'solubility' estimate is used to specify the initial concentration of the protein used in the screening experiments and is an important step in successful crystallization. Buffer optimization and establishment of initial protein concentration for crystal screening based on solubility estimates provides a methodology for improved crystal screening results.

  11. Optimization of aqueous extraction process to enhance the ...

    African Journals Online (AJOL)

    Kumar Sudhir

    2014-02-12

    Feb 12, 2014 ... Aqueous extraction process was optimized to reduce endotoxins from mixed substrate (1:1) for further phytase .... Microorganism and chemicals ..... The experimental design output (Table 2) was analyzed .... synthesis.

  12. Optimization of acidic extraction of astaxanthin from Phaffia rhodozyma

    Institute of Scientific and Technical Information of China (English)

    Hui NI; Qi-he CHEN; Guo-qing HE; Guang-bin WU; Yuan-fan YANG

    2008-01-01

    Optimization of a process for extracting astaxanthin from Phaffia rhodozyma by acidic method was investigated,regarding several extraction factors such as acids, organic solvents, temperature and time. Fractional factorial design, central composite design and response surface methodology were used to derive a statistically optimal model, which corresponded to the following optimal condition: concentration of lactic acid at 5.55 mol/L, ratio of ethanol to yeast dry weight at 20.25 ml/g, temperature for cell-disruption at 30 ℃, and extraction time for 3 min. Under this condition, astaxanthin and the total carotenoids could be extracted in amounts of 1294.7 μg/g and 1516.0 μg/g, respectively. This acidic method has advantages such as high extraction efficiency, low chemical toxicity and no special requirement of instruments. Therefore, it might be a more feasible and practical method for industrial practice.

  13. Optimizing the extraction of antibacterial compounds from pineapple leaf fiber

    Directory of Open Access Journals (Sweden)

    Zhuang Zhikai

    2016-01-01

    Full Text Available Five different solvents (petroleum ether, chloroform, ethyl acetate, acetone, and distilled water were used to extract antibacterial compounds from pineapple leaf fiber. Compounds extracted using acetone showed the greatest antibacterial effect against Escherichia coli, measured by inhibition zone diameter. Three extraction parameters including temperature, time and solid-liquid ratio were optimized through orthogonal experiment based on single factor investigations for achieving maximum active substance extraction rate and bacteriostatic effect. Results showed that using acetone, the optimum extraction conditions for temperature, time and solid-liquid ratio were 45°C, 8 h, and 1:40 (g/ml, respectively.

  14. Optimization of the extraction of curcumin from Curcuma longa rhizomes

    Directory of Open Access Journals (Sweden)

    Viviane P. Paulucci

    2012-01-01

    Full Text Available The aim of this work was to study the effect of dynamic maceration factors upon the curcumin content of Curcuma longa L., Zingiberaceae, extracts and to determine the optimum set of parameters for the extraction of curcumin using a 2(5 full factorial design and the response surface methodology. Under the established conditions, the content of soluble solids and curcumin in the extracts ranged from 0.8 to 3.4%, and from 0.1 to 1.8%, respectively. The most influential variable observed for the extraction was the ethanolic strength of the solvent. The optimized condition involves an extraction time of 12 h, agitation speed of 30 rpm, drug to solvent ratio of 1/6, extraction temperature of 80 ºC and the solvent with ethanolic strength of 70%. The data reported herein are useful for further developments of curcuma phytopharmaceutical intermediate products with optimized characteristics.

  15. Protein Structure Refinement by Optimization

    DEFF Research Database (Denmark)

    Carlsen, Martin

    on whether the three-dimensional structure of a homologous sequence is known. Whether or not a protein model can be used for industrial purposes depends on the quality of the predicted structure. A model can be used to design a drug when the quality is high. The overall goal of this project is to assess...... of the performance of the resulting potential and find a significant improvement by using a metric based on intrinsic geometry. It is well-known that energy minimization of a potential that is efficient in ordering a fixed set of decoys need not bring the decoys closer to the native state. The next part of the work...

  16. Influence of extraction method on protein profile of soybeans

    OpenAIRE

    Pavlićević Milica Ž.; Stanojević Slađana P.; Vucelić-Radović Biljana V.

    2013-01-01

    Comparison between protein profiles of soybean obtained by commonly used methods of extraction (Tris buffer and Tris-urea buffer) with methods used for extraction of plant proteins for 2D PAGE analysis (direct solubilization in IEF buffer, acetone extraction, phenol extraction, extraction with urea solubilization buffer and thiourea-urea extraction) was investigated. 2D profiles of samples extracted directly in IEF buffer, in urea solubilization buffer and in acetone were characterized ...

  17. Optimal Extraction Conditions of Anti-obesity Lipase Inhibitor from Phellinus linteus and Nutritional Characteristics of the Extracts.

    Science.gov (United States)

    Lee, Jong-Kug; Song, Jung-Hwa; Lee, Jong-Soo

    2010-03-01

    In an effort to develop novel mushroom-derived anti-obesity nutraceuticals, water and ethanol extracts containing the lipaseinhibitory compound from Phellinus linteus were prepared, and their nutritional components were determined. The optimal conditions for the extraction of P. linteus lipase inhibitor involved the treatment of the fruiting bodies with distilled water at 80℃ for 72 hr and 80% ethanol at 100℃ for 60 hr, respectively. The distilled water extract and ethanol extract contained 10.9% and 6.11% of crude protein, and 0.96% and 15.86% of crude fat, respectively. Additionally, the distilled water extract contained a large quantity of minerals, including 239.5 mg of K, 39.3 mg of Mg, and 39.3 mg of Na. The free amino acid content of the distilled water extracts was also higher than that of the ethanol extracts, and in particular, the distilled water extracts contained 5,139 mg of asparagine, 3,891 mg of tryptophan, 2,598 mg of alanine, and 2,066 mg of serine in 100 g of the distilled water extracts. 100 g of the distilled water and ethanol extracts were found to contain 12.31 g and 8.16 g of malic acid, respectively.

  18. Otimização da extração enzimática da proteína do feijão = Optimization of enzymatic protein extraction from beans

    Directory of Open Access Journals (Sweden)

    Carlos de Oliveira Lopes Junior

    2010-07-01

    Full Text Available Considerando-se a importância do feijão na dieta do brasileiro e, ainda, que este alimento não é permitido na alimentação de fenilcetonúricos, é fundamental promover a extração de suas proteínas, visando à posterior remoção de fenilalanina. No intuito de se otimizar a extração proteica do feijão, dois métodos enzimáticos foram, inicialmente, empregados. Para aquele que produziu o maior rendimento de extração proteica (REP, foi realizado um estudo do efeito de diversos parâmetros, tais como velocidade de centrifugação, tempo de reação, pH inicial e tipo de enzima (uma de Bacillus licheniformis, duas de Bacillus subtilis, uma de Aspergillus sojae e uma de Papaya carica. Os resultados indicaram que o melhor método foi o que empregou uma concentração de matéria-prima de 1:10 (p v-1, pH inicial de 10,5, temperatura de 50ºC, e ausência de agitação emultraturrax. Com relação aos parâmetros testados, o maior REP (93,1% foi obtido ao se utilizar a enzima de Bacillus licheniformis, na velocidade de centrifugação de 10.640 x g, tempo de reação de 3h e pH inicial de 10,5. Considering the importance of beans in the Brazilian diet and the fact that it is forbidden in phenylketonuric feeding, it is relevant to promote the extraction of its proteins, for phenylalanine removal. Aiming for the optimization of protein extraction from beans, twoenzymatic methods were initially employed. For the one that produced the highest protein extract yield (PEY, a study was conducted to assess de effect of several parameters, such as centrifugation velocity, reaction time, initial pH and type of enzyme (one of Bacillus licheniformis, two of Bacillus subtilis, one of Aspergillus sojae and one of Papaya carica. The results showed that the best method was the one that employed a concentration of raw material of 1:10 (w v-1, initial pH of 10.5, temperature of 50°C, in the absence of ultraturrax treatment. With regard to the parameters

  19. Optimal separation of jojoba protein using membrane processes

    Energy Technology Data Exchange (ETDEWEB)

    Nabetani, Hiroshi; Abbott, T.P.; Kleiman, R. [National Center for Agricultural Utilization Research, Peoria, IL (United States)

    1995-05-01

    The efficiency of a pilot-scale membrane system for purifying and concentrating jojoba protein was estimated. In this system, a jojoba extract was first clarified with a microfiltration membrane. The clarified extract was diafiltrated and the protein was purified with an ultrafiltration membrane. Then the protein solution was concentrated with the ultrafiltration membrane. Permeate flux during microfiltration was essentially independent of solids concentration in the feed, in contrast with the permeate flux during ultrafiltration which was a function of protein concentration. Based on these results, a mathematical model which describes the batchwise concentration process with ultrafiltration membranes was developed. Using this model, the combination of batchwise concentration with diafiltration was optimized, and an industrial-scale process was designed. The effect of ethylenediaminetetraacetic acid (EDTA) on the performance of the membrane system was also investigated. The addition of EDTA increased the concentration of protein in the extract and improved the recovery of protein in the final products. The quality of the final product (color and solubility) was also improved. However, EDTA decreased permeate flux during ultrafiltration.

  20. Evaluation of Protein Extraction Methods for Proteomic Analysis of Non-Model Recalcitrant Plant Tissues

    OpenAIRE

    2012-01-01

    Plant tissues contain relatively low amounts of proteins whose extraction is often difficult due to the presence of interfering compounds such as rigid cellulosic cell wall, storage polysaccharides, lipids and other contaminants that can cause protein degradation or modification. Therefore it is important to optimize protein extraction and to establish a robust protocol for two-dimensional gel electrophoresis (2-DE) and downstream processing. In this study, acetone, trichloroacetic acid/aceto...

  1. 糯米蛋白提取工艺优化及其水解物性质的测定%Optimization of alkaline protease extraction of sticky rice protein from sticky rice and properties mensuration of hydrolysates of sticky rice protein

    Institute of Scientific and Technical Information of China (English)

    王国泽; 袁怀波; 徐建文

    2012-01-01

    以糯米为原料,采用碱性蛋白酶水解法提取了糯米蛋白。就影响糯米蛋白提取率的5个因素:料液比(g/mL)、加酶量(E/S)、pH、温度和提取时间进行单因素实验和正交实验。研究确立了提取糯米蛋白的最佳工艺条件为料液比1:12,加酶量2%,pH9,温度45℃,提取时间3h。在此条件下,糯米蛋白的提取率可达8.529%。实验表明,糯米蛋白水解物是一类天然的抗氧化氨基酸及其聚合物。%In order to obtain the extraction of sticky rice protein, sticky rice was used as raw materials by alkaline protease .The influence factors, amount of solid to liquid ratio, enzyme dosage, pH, temperature and time of the alkaline protease,which could affect the efficiency of extracting sticky rice protein,were investigated by singlefactor experiment and orthogonal test.The research results showed that the optimal technical parameters could be obtained as following : solid to liquid ratio 1: ]2, enzyme dosage 2%, pH = 9, temperature 45 ℃ and time 3 h.The max extraction rate of sticky rice protein reached to 8.529%.Antioxidant tests of protein hydrolysates by papain showed that hydrolyzate from sticky rice protein serves as amino acids or their polymer.

  2. comparison of protein extraction methods for the leaves of ficus ...

    African Journals Online (AJOL)

    F. I. Abdullah

    2017-05-01

    May 1, 2017 ... degradation. Based on the results of ... Eze and Dumboff [30] reported that the addition of chlorophyll extract to the standard protein .... protein extraction from recalcitrant plants such as banana (Musa spp.), apple (Malus ...

  3. Protein extraction from formalin-fixed, paraffin-embedded tissue sections: quality evaluation by mass spectrometry.

    Science.gov (United States)

    Shi, Shan-Rong; Liu, Cheng; Balgley, Brian M; Lee, Cheng; Taylor, Clive R

    2006-06-01

    A satisfactory protocol of protein extraction has been established based on the heat-induced antigen retrieval (AR) technique widely applied in immunohistochemistry for archival formalin-fixed, paraffin-embedded (FFPE) tissue sections. Based on AR, an initial serial experiment to identify an optimal protocol of heat-induced protein extraction was carried out using FFPE mouse tissues. The optimal protocol for extraction of proteins was then performed on an archival FFPE tissue of human renal carcinoma. FFPE sections were boiled in a retrieval solution of Tris-HCl containing 2% SDS, followed by incubation. Fresh tissue taken from the same case of renal carcinoma was processed for extraction of proteins by a conventional method using radioimmunoprecipitation assay solution, to compare the efficiency of protein extraction from FFPE tissue sections with extraction from fresh tissue. As a control, further sections of the same FFPE sample were processed by the same procedure without heating treatment. Evaluation of the quality of protein extracted from FFPE tissue was done using gel electrophoresis and mass spectrometry, showing most identified proteins extracted from FFPE tissue sections were overlapped with those extracted from fresh tissue.

  4. Enzymatic Pre-Treatment Increases the Protein Bioaccessibility and Extractability in Dulse (Palmaria palmata

    Directory of Open Access Journals (Sweden)

    Hanne K. Mæhre

    2016-10-01

    Full Text Available Several common protein extraction protocols have been applied on seaweeds, but extraction yields have been limited. The aims of this study were to further develop and optimize existing extraction protocols and to examine the effect of enzymatic pre-treatment on bioaccessibility and extractability of seaweed proteins. Enzymatic pre-treatment of seaweed samples resulted in a three-fold increase in amino acids available for extraction. Combining enzymatic pre-treatment with alkaline extraction resulted in a 1.6-fold increase in the protein extraction yield compared to a standard alkaline extraction protocol. A simulated in vitro gastrointestinal digestion model showed that enzymatic pre-treatment of seaweed increased the amount of amino acids available for intestinal absorption 3.2-fold. In conclusion, enzymatic pre-treatment of seaweeds is effective for increasing the amount of amino acids available for utilization and may thus be an effective means for increasing the utilization potential of seaweed proteins. However, both the enzymatic pre-treatment protocol and the protein extraction protocol need further optimization in order to obtain optimal cost-benefit and results from the in vitro gastrointestinal digestion model need to be confirmed in clinical models.

  5. Soil metaproteomics – Comparative evaluation of protein extraction protocols

    OpenAIRE

    Keiblinger, Katharina M.; Wilhartitz, Inés C.; Schneider, Thomas; Roschitzki, Bernd; Schmid, Emanuel; Eberl, Leo; Riedel, Kathrin; Zechmeister-Boltenstern, Sophie

    2012-01-01

    Metaproteomics and its potential applications are very promising to study microbial activity in environmental samples and to obtain a deeper understanding of microbial interactions. However, due to the complexity of soil samples the exhaustive extraction of proteins is a major challenge. We compared soil protein extraction protocols in terms of their protein extraction efficiency for two different soil types. Four different protein extraction procedures were applied based on (a) SDS extractio...

  6. Optimized microbial DNA extraction from diarrheic stools

    Directory of Open Access Journals (Sweden)

    Donatin Emilie

    2012-12-01

    Full Text Available Abstract Background The detection of enteropathogens in stool specimens increasingly relies on the detection of specific nucleic acid sequences. We observed that such detection was hampered in diarrheic stool specimens and we set-up an improved protocol combining lyophilization of stools prior to a semi-automated DNA extraction. Findings A total of 41 human diarrheic stool specimens comprising of 35 specimens negative for enteropathogens and six specimens positive for Salmonella enterica in culture, were prospectively studied. One 1-mL aliquot of each specimen was lyophilised and total DNA was extracted from lyophilised and non-lyophilised aliquots by combining automatic and phenol-chloroform DNA extraction. DNA was incorporated into real-time PCRs targeting the 16S rRNA gene of Bacteria and the archaea Methanobrevibacter smithii and the chorismate synthase gene of S. enterica. Whereas negative controls consisting in DNA-free water remained negative, M. smithii was detected in 26/41 (63.4% non-lyophilised (Ct value 28.78 ± 9.1 versus 39/41 (95.1% lyophilised aliquots (Ct value 22.04 ± 5.5; bacterial 16S rRNA was detected in 33/41 (80.5% non-lyophilised (Ct value 28.11 ± 5.9 versus 40/41 (97.6% lyophilised aliquots (Ct value 24.94 ± 6.6; and S. enterica was detected in 6/6 (100% non-lyophilized and lyophilized aliquots (Ct value 26.98 ± 4.55 and 26.16 ± 4.97, respectively. S. enterica was not detected in the 35 remaining diarrheal-stool specimens. The proportion of positive specimens was significantly higher after lyophilization for the detection of M. smithii (p = 0.00043 and Bacteria (p = 0.015. Conclusion Lyophilization of diarrheic stool specimens significantly increases the PCR-based detection of microorganisms. The semi-automated protocol described here could be routinely used for the molecular diagnosis of infectious diarrhea.

  7. Optimization of energy extraction in transverse galloping

    Science.gov (United States)

    Sorribes-Palmer, F.; Sanz-Andres, A.

    2013-11-01

    A numerical method to analyse the stability of transverse galloping based on experimental measurements, as an alternative method to polynomial fitting of the transverse force coefficient Cz, is proposed in this paper. The Glauert-Den Hartog criterion is used to determine the region of angles of attack (pitch angles) prone to present galloping. An analytic solution (based on a polynomial curve of Cz) is used to validate the method and to evaluate the discretization errors. Several bodies (of biconvex, D-shape and rhomboidal cross sections) have been tested in a wind tunnel and the stability of the galloping region has been analysed with the new method. An algorithm to determine the pitch angle of the body that allows the maximum value of the kinetic energy of the flow to be extracted is presented.

  8. Microwave optimization of mucilage extraction from Opuntia ficus indica Cladodes.

    Science.gov (United States)

    Felkai-Haddache, Lamia; Dahmoune, Farid; Remini, Hocine; Lefsih, Khalef; Mouni, Lotfi; Madani, Khodir

    2016-03-01

    In this study, microwave-assisted extraction (MAE) of polysaccharides from Opuntia ficus indica Cladodes were investigated using response surface methodology (RSM). The effects of three extraction factors on the yield of mucilage were examined. The results indicated that the optimum extraction conditions were determined as follows: microwave power X1, 700 W; extraction time X2, 5.15 minand ratio water/raw material X3, 4.83 mL/g at fixed pH 11. Under these optimal extraction conditions, mucilage yield was found to be Y, 25.6%. A comparison between the model results and experimental data gave a high correlation coefficient (R(2)=0.88), adjusted coefficient (Radj=0.83) and low root mean square error (RMSE=2.45) and showed that the two models were able to predict a mucilage yield by green extraction microwave process.

  9. Optimal protein intake in the elderly.

    Science.gov (United States)

    Wolfe, Robert R; Miller, Sharon L; Miller, Kevin B

    2008-10-01

    The recommended dietary allowance (RDA) for protein, as promulgated by the Food and Nutrition Board of the United States National Academy of Science, is 0.8 g protein/kg body weight/day for adults, regardless of age. This value represents the minimum amount of protein required to avoid progressive loss of lean body mass in most individuals. There is an evidence that the RDA for elderly may be greater than 0.8 g/kg/day. Evidence indicates that protein intake greater than the RDA can improve muscle mass, strength and function in elderly. In addition, other factors, including immune status, wound healing, blood pressure and bone health may be improved by increasing protein intake above the RDA. Furthermore, the RDA does not address the recommended intake of protein in the context of a balanced diet. Concerns about potential detrimental effects of increased protein intake on bone health, renal function, neurological function and cardiovascular function are generally unfounded. In fact, many of these factors are improved in elderly ingesting elevated quantities of protein. It appears that an intake of 1.5 g protein/kg/day, or about 15-20% of total caloric intake, is a reasonable target for elderly individuals wishing to optimize protein intake in terms of health and function.

  10. Optimization study of Chromalaena odorata essential oil extracted using solventless extraction technique

    Science.gov (United States)

    Nasshorudin, Dalila; Ahmad, Muhammad Syarhabil; Mamat, Awang Soh; Rosli, Suraya

    2015-05-01

    Solventless extraction process of Chromalaena odorata using reduced pressure and temperature has been investigated. The percentage yield of essential oil produce was calculated for every experiment with different experimental condition. The effect of different parameters, such as temperature and extraction time on the yield was investigated using the Response Surface Methodology (RSM) through Central Composite Design (CCD). The temperature and extraction time were found to have significant effect on the yield of extract. A final essential oil yield was 0.095% could be extracted under the following optimized conditions; a temperature of 80 °C and a time of 8 hours.

  11. Optimization of Extraction Technology of Lotus Seed Total Protein by Response Surface Methodology%响应面法优化莲子总蛋白提取工艺的研究

    Institute of Scientific and Technical Information of China (English)

    夏琨; 郑铁松

    2012-01-01

    The extraction of total protein from lotus seed was studied. Tris-HC1 buffer was selected as the best extract. According to the single-factor experiments, three levels of the factors (the ratio of buffer to solid, extraction time and temperature) were selected for the Box-Behnken factorial design to establish a quadric regression equation for describing the yield of total protein. By response surface analysis and the actual operation, the optimum extraction conditions were obtained as follows: the ratio of buffer to solid of 7.2:1, extraction time of 2.4 h and extraction temperature of 11.1℃, the extraction yield of total protein was 81.35%, resenting a relative error of less than 0.5 %.%为优化莲子总蛋白的提取工艺,首先确定了最佳提取液为Tris-Hcl缓冲液;在单因素试验的基础上,采用Box-Behnken试验设计方法,研究液料比、提取时间和提取温度对莲子总蛋白提取率的影响,建立总蛋白提取率的二次回归方程,并对其工艺参数进行优化。结合实际操作的研究结果表明:提取莲子总蛋白的最佳工艺参数为液料比7.2:1,提取时间2.4h,提取温度11.1℃,总蛋白的提取率达到81.35%,与回归模型的相对误差小于0.5%。

  12. Optimization of ultrasonic extraction of 23 elements from cotton.

    Science.gov (United States)

    Rezić, I

    2009-01-01

    Optimization of ultrasonic extraction of 23 elements from cotton was performed with different solvent volume ratios. For this purpose nitric acid, hydrochloric acid and water were mixed and applied in a mixture for the extraction of elements adsorbed on cotton material. The elements chosen for the extraction procedure (Al, As, Be, Bi, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Mg, Mn, Mo, Na, Ni, Pb, Sb, Si, Sn, Tl and Zn) were those that are important in textile processing. Some of them cause problems during fiber processing, dyeing or bleaching. The removal of elements from the processed fabric can be successfully done with ultrasonic extraction in the ultrasonic bath. Extraction procedure was optimized by software package Design Expert 6 (DX6) and the optimum of ultrasonic extraction was achieved with the mixture of 1M HCl-1M HNO(3)-H(2)O=3.32/2.83/93.85 (v/v). Ultrasonic extraction was a fast and efficient extraction procedure easily applied on cotton textile material.

  13. Extraction of plant proteins for two-dimensional electrophoresis

    OpenAIRE

    Granier, Fabienne

    1988-01-01

    Three different extraction procedures for two-dimensional electrophoresis of plant proteins are compared: (i) extraction of soluble proteins with a nondenaturing Tris-buffer, (ii) denaturing extraction in presence of sodium dodecyl sulfate at elevated temperature allowing the solubilization of membrane proteins in addition to a recovery of soluble proteins, and (iii) a trichloroacetic acid-acetone procedure allowing the direct precipitation of total proteins.

  14. Optimal geothermal resource extraction for electric power applications

    Energy Technology Data Exchange (ETDEWEB)

    Blair, P.D.; Cassel, T.A.V.

    1979-06-27

    A phase of ongoing work at the University or Pennsylvania concerning the analysis of capital investments in the development of geothermal electric power facilities is reported. The phase being addressed deals with determining the optimal rate of extraction of geothermal resources for producing electric power. The mathematical approach for estimating an optimal time-path of production to maximize the net present value of a rate sensitive reservoir is formulated.

  15. Pectin from Abelmoschus esculentus: optimization of extraction and rheological properties.

    Science.gov (United States)

    Chen, Yi; Zhang, Jian-Guo; Sun, Han-Ju; Wei, Zhao-Jun

    2014-09-01

    Response surface methodology (RSM) was applied to optimize the parameters of pectin extraction from okra pods. The extracted okra pectin was then investigated by steady-shear and oscillatory rheological measurements. Statistical analysis showed that the linear term of the liquid-solid ratio, the quadratic term of the pH, and the linear term of the extraction time showed highly significant effects on pectin yield. The optimal extraction conditions that maximized the pectin yield within the experimental range of the variables researched were a pH of 3.9, an extraction time of 64 min, an extraction temperature of 60°C, and a liquid-solid ratio of 42:1. Under these conditions, the pectin yield was predicted to be 2.71%. At a liquid-solid ratio less than 2.5% w/w in aqueous solution, the pectin extracted from okra presented non-Newtonian shear-thinning behavior and could be well described by the Cross model. The okra pectin showed predominantly viscous responses (G'

  16. Linguistic feature analysis for protein interaction extraction

    Directory of Open Access Journals (Sweden)

    Cornelis Chris

    2009-11-01

    Full Text Available Abstract Background The rapid growth of the amount of publicly available reports on biomedical experimental results has recently caused a boost of text mining approaches for protein interaction extraction. Most approaches rely implicitly or explicitly on linguistic, i.e., lexical and syntactic, data extracted from text. However, only few attempts have been made to evaluate the contribution of the different feature types. In this work, we contribute to this evaluation by studying the relative importance of deep syntactic features, i.e., grammatical relations, shallow syntactic features (part-of-speech information and lexical features. For this purpose, we use a recently proposed approach that uses support vector machines with structured kernels. Results Our results reveal that the contribution of the different feature types varies for the different data sets on which the experiments were conducted. The smaller the training corpus compared to the test data, the more important the role of grammatical relations becomes. Moreover, deep syntactic information based classifiers prove to be more robust on heterogeneous texts where no or only limited common vocabulary is shared. Conclusion Our findings suggest that grammatical relations play an important role in the interaction extraction task. Moreover, the net advantage of adding lexical and shallow syntactic features is small related to the number of added features. This implies that efficient classifiers can be built by using only a small fraction of the features that are typically being used in recent approaches.

  17. A cooperative fast annealing coevolutionary algorithm for protein motif extraction

    Institute of Scientific and Technical Information of China (English)

    CHEN Chao; TIAN YuanXin; ZOU XiaoYong; CAI PeiXiang; MO JinYuan

    2007-01-01

    By integrating the cooperative approach with the fast annealing coevolutionary algorithm (FAEA), a so-called cooperative fast annealing coevolutionary algorithm (CFACA) is presented in this paper for the purpose of solving high-dimensional problems. After the partition of the search space in CFACA, each smaller one is then searched by a separate FAEA. The fitness function is evaluated by combining sub-solutions found by each of the FAEAs. It demonstrates that the CFACA outperforms the FAEA in the domain of function optimization, especially in terms of convergence rate. The current algorithm is also applied to a real optimization problem of protein motif extraction. And a satisfactory result has been obtained with the accuracy of prediction achieving 67.0%, which is in agreement with the result in the PROSITE database.

  18. An efficient procedure for protein extraction from formalin-fixed, paraffin-embedded tissues for reverse phase protein arrays

    Directory of Open Access Journals (Sweden)

    Guo Huifang

    2012-09-01

    Full Text Available Abstract Introduction Protein extraction from formalin-fixed paraffin-embedded (FFPE tissues is challenging due to extensive molecular crosslinking that occurs upon formalin fixation. Reverse-phase protein array (RPPA is a high-throughput technology, which can detect changes in protein levels and protein functionality in numerous tissue and cell sources. It has been used to evaluate protein expression mainly in frozen preparations or FFPE-based studies of limited scope. Reproducibility and reliability of the technique in FFPE samples has not yet been demonstrated extensively. We developed and optimized an efficient and reproducible procedure for extraction of proteins from FFPE cells and xenografts, and then applied the method to FFPE patient tissues and evaluated its performance on RPPA. Results Fresh frozen and FFPE preparations from cell lines, xenografts and breast cancer and renal tissues were included in the study. Serial FFPE cell or xenograft sections were deparaffinized and extracted by six different protein extraction protocols. The yield and level of protein degradation were evaluated by SDS-PAGE and Western Blots. The most efficient protocol was used to prepare protein lysates from breast cancer and renal tissues, which were subsequently subjected to RPPA. Reproducibility was evaluated and Spearman correlation was calculated between matching fresh frozen and FFPE samples. The most effective approach from six protein extraction protocols tested enabled efficient extraction of immunoreactive protein from cell line, breast cancer and renal tissue sample sets. 85% of the total of 169 markers tested on RPPA demonstrated significant correlation between FFPE and frozen preparations (p Conclusions With optimized protein extraction methods, FFPE tissues can be a valuable source in generating reproducible and biologically relevant proteomic profiles using RPPA, with specific marker performance varying according to tissue type.

  19. Efficient extraction of olive pulp and stone proteins by using an enzyme-assisted method.

    Science.gov (United States)

    Vergara-Barberán, María; Lerma-García, María Jesús; Herrero-Martínez, José Manuel; Simó-Alfonso, Ernesto Francisco

    2014-07-01

    An efficient protein extraction protocol for proteins from olive pulp and stone by using enzymes was developed. For this purpose, different parameters that affect the extraction process, such as enzyme type and content, pH, and extraction temperature and time, were tested. The influence of these factors on protein recovery was examined using the standard Bradford assay, while the extracted proteins were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The best extraction conditions were achieved at pH 7.0 and 5% (v/v) Palatase® 20000 L (lipase) for pulp and Lecitase® Ultra (phospholipase) for stone proteins. The optimal extraction temperature and time were 30 and 40 °C for 15 min for pulp and stone tissues, respectively. Under these conditions, several protein extracts coming from olive fruits of different genetic variety were analyzed, their profiles being compared by SDS-PAGE. The developed enzyme-assisted extraction method showed faster extraction, higher recovery, and reduced solvent usage than the nonenzymatic methods previously described in the literature. In the case of stone proteins, different electrophoretic profiles and band intensities were obtained that could be helpful to distinguish samples according to their genetic variety.

  20. Soil metaproteomics - Comparative evaluation of protein extraction protocols.

    Science.gov (United States)

    Keiblinger, Katharina M; Wilhartitz, Inés C; Schneider, Thomas; Roschitzki, Bernd; Schmid, Emanuel; Eberl, Leo; Riedel, Kathrin; Zechmeister-Boltenstern, Sophie

    2012-11-01

    Metaproteomics and its potential applications are very promising to study microbial activity in environmental samples and to obtain a deeper understanding of microbial interactions. However, due to the complexity of soil samples the exhaustive extraction of proteins is a major challenge. We compared soil protein extraction protocols in terms of their protein extraction efficiency for two different soil types. Four different protein extraction procedures were applied based on (a) SDS extraction without phenol, (b) NaOH and subsequent phenol extraction, (c) SDS-phenol extraction and (d) SDS-phenol extraction with prior washing steps. To assess the suitability of these methods for the functional analysis of the soil metaproteome, they were applied to a potting soil high in organic matter and a forest soil. Proteins were analyzed by two-dimensional liquid chromatography/tandem mass spectrometry (2D-LC-MS/MS) and the number of unique spectra as well as the number of assigned proteins for each of the respective protocols was compared. In both soil types, extraction with SDS-phenol (c) resulted in "high" numbers of proteins. Moreover, a spiking experiment was conducted to evaluate protein recovery. To this end sterilized forest soil was amended with proteins from pure cultures of Pectobacterium carotovorum and Aspergillus nidulans. The protein recovery in the spiking experiment was almost 50%. Our study demonstrates that a critical evaluation of the extraction protocol is crucial for the quality of the metaproteomics data, especially in highly complex samples like natural soils.

  1. Pressurized water extraction of isoflavones by experimental design from soybean flour and Soybean Protein Isolate.

    Science.gov (United States)

    Moras, Benjamin; Rey, Stéphane; Vilarem, Gérard; Pontalier, Pierre-Yves

    2017-01-01

    A Doehlert experimental design was conducted and surface response methodology was used to determine the effect of temperature, contact time and solid liquid ratio on isoflavone extraction from soybean flour or Soybean Protein Isolate in pressurized water system. The optimal conditions conducted gave an extraction yield of 85% from soybean flour. For Soybean Protein Isolate compared to soybean flour, the isoflavone extraction yield is 61%. This difference could be explained by higher aglycon content, while aglycon appears to be the least extracted isoflavone by pressurized water. The solid liquid ratio in the ASE cell was the overriding factor in obtaining high yields with both soybean products, while temperature has less influence. A high temperature causes conversion of the malonyls-glucosides and glucosides isoflavone derivatives into glucosides or aglycons forms. pressurized water extraction showed a high solubilization of protein material up to 95% of inserted Soybean Protein Isolate.

  2. 响应面法优化酶法提取小麦麸皮蛋白%Optimization of extraction for wheat bran protein by protease based on response surface methology

    Institute of Scientific and Technical Information of China (English)

    王步江; 袁玮; 刘金福

    2011-01-01

    On the basis of single factor tests and the Box-Benhnken center united experimental design principle,the method of response surface analysis with 3-factors and 3-levels was adopted.A second order quadratic equation for extraction of the protein was built.Response surface and contour were graphed with the extraction rate of the protein,the extraction rate as the response value.Based on the analysis of the response surface plots and their corresponding contour plots,effects of the temperature,enzyme concentration,and time were explored.By using this new method,the optimum extraction condition was obtained as follows:the temperature was 54.3 ℃,enzyme concentration was 0.86%,soild to liquid ratio was 1:10,time was 2.56 h,the extraction rate was 30.45%.%根据Box-Benhnken的中心组合试验设计原理,在单因素试验的基础上,采用3因素3水平的响曲面分析法,建立小麦麸皮蛋白提取的二次多项式模型,以蛋白提取率为响应值作响应面和等高线,考察酶解时间、温度和加酶量对蛋白提取率的影响。结果表明,提取小麦麸皮蛋白的工艺为:酶解时间2.56h,酶解温度为54.3℃,加酶量为0.86%,此时麸皮蛋白的最佳提取率为30.45%。

  3. Selective extraction of isolated mitotic apparatus. Evidence that typical microtubule protein is extracted by organic mercurial.

    Science.gov (United States)

    Bibring, T; Baxandall, J

    1971-02-01

    Mitotic apparatus isolated from sea urchin eggs has been treated with meralluride sodium under conditions otherwise resembling those of its isolation. The treatment causes a selective morphological disappearance of microtubules while extracting a major protein fraction, probably consisting of two closely related proteins, which constitutes about 10% of mitotic apparatus protein. Extraction of other cell particulates under similar conditions yields much less of this protein. The extracted protein closely resembles outer doublet microtubule protein from sea urchin sperm tail in properties considered typical of microtubule proteins: precipitation by calcium ion and vinblastine, electrophoretic mobility in both acid and basic polyacrylamide gels, sedimentation coefficient, molecular weight, and, according to a preliminary determination, amino acid composition. An antiserum against a preparation of sperm tail outer doublet microtubules cross-reacts with the extract from mitotic apparatus. On the basis of these findings it appears that microtubule protein is selectively extracted from isolated mitotic apparatus by treatment with meralluride, and is a typical microtubule protein.

  4. Optimization of enzyme assisted extraction of polysaccharides from Astragalus membranaceus.

    Science.gov (United States)

    Chen, Huaguo; Zhou, Xin; Zhang, Junzeng

    2014-10-13

    Astragalus polysaccharide (APS) is known to have a variety of pharmacological activities. In the present study, enzyme assisted extraction of APS from Astragalus mongholicus using various enzymes were examined. Research found that glucose oxidase offered a better performance in enhancement of extraction yields of APS than other ones. Glucose oxidase assisted extraction process was further optimized by using response surface method (RSM) to obtain maximum yield of crude APS. The optimized extraction conditions were as follows: enzyme amount of 3.0%, enzyme treated time of 3.44 d, enzyme treated temperature of 56.9 °C and extraction solvent pH of 7.8. Under these conditions, the experimental yield was 29.96 ± 0.14%, which was well in close agreement with the value (30.19%) predicted by RSM model and increased more than 250% compared with none enzyme treated ones. Pharmacological test showed that enzyme assisted APS had a better antioxidant activity (about 2 times higher) than none enzyme treated ones.

  5. Production of surgical gloves from low extractable protein RVNRL

    Energy Technology Data Exchange (ETDEWEB)

    Marga, Utama; Yanti, S.; Made, Sumarti; Marsongko; Tita, Puspitasari; Dian, Iramani [Center for Research and Development of Isotopes and Radiation Technology, National Nuclear Energy Agency, Jakarta (Indonesia); Makuuchi, K. [EB System Cooperation, Takasaki, Gunma (Japan); Yoshii, F. [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment; Siswanto [Research Unit for Biotechnology of Estate Crop (Indonesia)

    2001-03-01

    Study on the production of surgical gloves from low extractable protein PVNRL (Radiation Vulcanization of Natural Rubber Latex) in home industry scale with normal butyl acrylate as sensitizer has been carried out. The variation of dipping speed, concentration of coagulant agent and selection of antioxidant for producing good quality of surgical gloves were evaluated. The water-extractable protein and PBS (Phosphate Buffer Saline) - extractable protein content, the physical and mechanical properties of gloves were measured. The results show that for producing a good quality of surgical gloves from low extractable protein RVNRL, the concentration of latex is 50% with calcium nitrate as coagulant agent between 15-20%. By using this condition the physical and mechanical properties of surgical gloves is required to ASTM standard such as tensile strength more than 24 MPa, PBS-extractable protein is around 41-68 ug/g and water-extractable protein contents is around 23-35 ug/g. (author)

  6. PROTEIN EXTRACTION OF MAIZE (ZEA MAYS FOR PROTEOMIC 2 – DE ANALYSIS

    Directory of Open Access Journals (Sweden)

    Edita Gregová

    2014-12-01

    Full Text Available A strength of two-dimensional polyacrylamide gel electrophoresis (2D PAGE is its ability to resolve and investigate the abundance of several thousand proteins in a single sample. Two different extraction procedures for two-dimensional electrophoresis of plant proteins are compared in this work. Phenol-based extraction methods have been mainly used to extract proteins from different organs or tissues on many species. We wanted to determine which of these protocols was optimal for starch plants in order to achieve both efficient protein extraction and high spot resolution on 2-D gels. The phenol-based protocol was superior to the sodium phosphatase methods, showing larger protein yields and greater spot resolution on 2-D gels.

  7. Integration of galacturonic acid extraction with alkaline protein extraction from green tea leaf residue

    NARCIS (Netherlands)

    Zhang, Chen; Bozileva, Elvira; Klis, van der Frits; Dong, Yiyuan; Sanders, Johan P.M.; Bruins, Marieke E.

    2016-01-01

    Leaf pectin can be used as a feedstock for galacturonic acid (GA) production, but high extraction costs limit economic feasibility. To improve the extraction efficiency, leaf pectin extraction was integrated with an already cost-effective alkaline protein extraction, focusing on high yield of GA

  8. Mitotic apparatus: the selective extraction of protein with mild acid.

    Science.gov (United States)

    Bibring, T; Baxandall, J

    1968-07-26

    The treatment of isolated mitotic apparatus with mild (pH 3) hydrochloric acid results in the extraction of less than 10 percent of its protein, accompanied by the selective morphological disappearance of the microtubules. The same extraction can be shown to dissolve outer doublet microtubules from sperm flagella. A protein with points of similarity to the flagellar microtubule protein is the major component of the extract from mitotic apparatus.

  9. Anthelmintic activity of Leucaena leucocephala protein extracts on Haemonchus contortus

    Directory of Open Access Journals (Sweden)

    Alexandra Martins dos Santos Soares

    Full Text Available Abstract The objective of this study was to evaluate the effects of protein extracts obtained from the plant Leucaena leucocephala on the nematode parasite Haemonchus contortus. The seeds, shell and cotyledon of L. leucocephala were separated and their proteins extracted using a sodium phosphate buffer, and named as TE (total seed extract, SE (shell extract and CE (cotyledon extract. Soluble protein content, protease, protease inhibitory and chitinase activity assays were performed. Exsheathment inhibition of H. contortus larvae were performed at concentrations of 0.6 mg mL–1, and egg hatch assays were conducted at protein concentrations of 0.8, 0.4, 0.2, 0.1 and 0.05 mg mL–1. The effective concentration for 50% hatching inhibition (EC50 was estimated by probit. Different proportions of soluble proteins, protease and chitinase were found in TE and CE. Protease inhibitory activity was detected in all extracts. The EC50 of the CE and TE extracts were 0.48 and 0.33 mg mL–1, respectively. No ovicidal effects on H. contortus were detected in SE extracts, and none of the protein extracts demonstrated larvicidal effects on H. contortus. We therefore conclude that protein extracts of L. leucocephala had a detrimental effect on nematode eggs, which can be correlated with the high protease and chitinase activity of these extracts.

  10. Anthelmintic activity of Leucaena leucocephala protein extracts on Haemonchus contortus.

    Science.gov (United States)

    Soares, Alexandra Martins dos Santos; de Araújo, Sandra Alves; Lopes, Suzana Gomes; Costa Junior, Livio Martins

    2015-01-01

    The objective of this study was to evaluate the effects of protein extracts obtained from the plant Leucaena leucocephala on the nematode parasite Haemonchus contortus. The seeds, shell and cotyledon of L. leucocephala were separated and their proteins extracted using a sodium phosphate buffer, and named as TE (total seed extract), SE (shell extract) and CE (cotyledon extract). Soluble protein content, protease, protease inhibitory and chitinase activity assays were performed. Exsheathment inhibition of H. contortus larvae were performed at concentrations of 0.6 mg mL-1, and egg hatch assays were conducted at protein concentrations of 0.8, 0.4, 0.2, 0.1 and 0.05 mg mL-1. The effective concentration for 50% hatching inhibition (EC50) was estimated by probit. Different proportions of soluble proteins, protease and chitinase were found in TE and CE. Protease inhibitory activity was detected in all extracts. The EC50 of the CE and TE extracts were 0.48 and 0.33 mg mL-1, respectively. No ovicidal effects on H. contortus were detected in SE extracts, and none of the protein extracts demonstrated larvicidal effects on H. contortus. We therefore conclude that protein extracts of L. leucocephala had a detrimental effect on nematode eggs, which can be correlated with the high protease and chitinase activity of these extracts.

  11. Effective protein extraction protocol for proteomics studies of Jerusalem artichoke leaves.

    Science.gov (United States)

    Zhang, Meide; Shen, Shihua

    2013-07-01

    Protein extraction is a crucial step for proteomics studies. To establish an effective protein extraction protocol suitable for two-dimensional electrophoresis (2DE) analysis in Jerusalem artichoke (Helianthus tuberosus L.), three different protein extraction methods-trichloroacetic acid/acetone, Mg/NP-40, and phenol/ammonium acetate-were evaluated using Jerusalem artichoke leaves as source materials. Of the three methods, trichloroacetic acid/acetone yielded the best protein separation pattern and highest number of protein spots in 2DE analysis. Proteins highly abundant in leaves, such as Rubisco, are typically problematic during leaf 2DE analysis, however, and this disadvantage was evident using trichloroacetic acid/acetone. To reduce the influence of abundant proteins on the detection of low-abundance proteins, we optimized the trichloroacetic acid/acetone method by incorporating a PEG fractionation approach. After optimization, 363 additional (36.2%) protein spots were detected on the 2DE gel. Our results suggest that trichloroacetic acid/acetone method is a better protein extraction technique than Mg/NP-40 and phenol/ammonium acetate in Jerusalem artichoke leaf 2DE analysis, and that trichloroacetic acid/acetone method combined with PEG fractionation procedure is the most effective approach for leaf 2DE analysis of Jerusalem artichoke.

  12. Optimal processes for probabilistic work extraction beyond the second law.

    Science.gov (United States)

    Cavina, Vasco; Mari, Andrea; Giovannetti, Vittorio

    2016-07-05

    According to the second law of thermodynamics, for every transformation performed on a system which is in contact with an environment of fixed temperature, the average extracted work is bounded by the decrease of the free energy of the system. However, in a single realization of a generic process, the extracted work is subject to statistical fluctuations which may allow for probabilistic violations of the previous bound. We are interested in enhancing this effect, i.e. we look for thermodynamic processes that maximize the probability of extracting work above a given arbitrary threshold. For any process obeying the Jarzynski identity, we determine an upper bound for the work extraction probability that depends also on the minimum amount of work that we are willing to extract in case of failure, or on the average work we wish to extract from the system. Then we show that this bound can be saturated within the thermodynamic formalism of quantum discrete processes composed by sequences of unitary quenches and complete thermalizations. We explicitly determine the optimal protocol which is given by two quasi-static isothermal transformations separated by a finite unitary quench.

  13. Optimal processes for probabilistic work extraction beyond the second law

    Science.gov (United States)

    Cavina, Vasco; Mari, Andrea; Giovannetti, Vittorio

    2016-07-01

    According to the second law of thermodynamics, for every transformation performed on a system which is in contact with an environment of fixed temperature, the average extracted work is bounded by the decrease of the free energy of the system. However, in a single realization of a generic process, the extracted work is subject to statistical fluctuations which may allow for probabilistic violations of the previous bound. We are interested in enhancing this effect, i.e. we look for thermodynamic processes that maximize the probability of extracting work above a given arbitrary threshold. For any process obeying the Jarzynski identity, we determine an upper bound for the work extraction probability that depends also on the minimum amount of work that we are willing to extract in case of failure, or on the average work we wish to extract from the system. Then we show that this bound can be saturated within the thermodynamic formalism of quantum discrete processes composed by sequences of unitary quenches and complete thermalizations. We explicitly determine the optimal protocol which is given by two quasi-static isothermal transformations separated by a finite unitary quench.

  14. Optimization of microwave assisted extraction of pectin from orange peel.

    Science.gov (United States)

    Prakash Maran, J; Sivakumar, V; Thirugnanasambandham, K; Sridhar, R

    2013-09-12

    In this study, microwave-assisted extraction was applied for pectin extraction from the dried orange peel and Box-Behnken response surface design was used to study and optimize the effects of processing variables (microwave power, irradiation time, pH and solid-liquid ratio) on the yield of pectin. The amount of pectin extracted increased with increasing microwave power, while it reduces as the time, pH and solid-liquid ratio increased. From the results, second order polynomial model was developed and it adequately explained the data variation and significantly represented the actual relationship between independent variables and the response. An optimization study using Derringer's desired function methodology was performed and optimal conditions based on both individual and combinations of all independent variables (microwave power of 422W, irradiation time of 169 s, pH of 1.4 and solid-liquid ratio of 1:16.9 g/ml) were determined with maximum pectin yield of 19.24%, which was confirmed through validation experiments.

  15. Optimization of pectin extraction and antioxidant activities from Jerusalem artichoke

    Science.gov (United States)

    Liu, Shengyi; Shi, Xuejie; Xu, Lanlan; Yi, Yuetao

    2016-03-01

    Jerusalem artichoke is an economic crop widely planted in saline-alkaline soil. The use of Jerusalem artichoke is of great significance. In this study, the response surface method was employed to optimize the effects of processing variables (extraction temperature, pH, extraction time, and liquid-to-solid ratio) on the yield of Jerusalem artichoke pectin. Under the optimal extraction conditions: pH 1.52, 63.62 min, 100°C and a liquid-to-solid ratio of 44.4 mL/g, the maximum pectin yield was predicted to be 18.76%. Experiments were conducted under these optimal conditions and a pectin yield of 18.52±0.90% was obtained, which validated the model prediction. The effects of diff erent drying methods (freeze drying, spray drying and vacuum drying) on the properties of Jerusalem artichoke pectin were evaluated and they were compared with apple pectin. FTIR spectral analysis showed no major structural diff erences in Jerusalem artichoke pectin samples produced by various drying treatments. The antioxidant activities of pectin dried by diff erent methods were investigated using in vitro hydroxyl and DPPH radical scavenging systems. The results revealed that the activities of spray dried pectin (SDP) and apple pectin (AP) were stronger than those of vacuum oven dried pectin (ODP) and vacuum freeze dried pectin (FDP). Therefore compared with the other two drying methods, the spray drying method was the best.

  16. Yellow Mealworm Protein for Food Purposes - Extraction and Functional Properties.

    Science.gov (United States)

    Zhao, Xue; Vázquez-Gutiérrez, José Luis; Johansson, Daniel P; Landberg, Rikard; Langton, Maud

    2016-01-01

    A protocol for extraction of yellow mealworm larvae proteins was established, conditions were evaluated and the resulting protein extract was characterised. The freeze-dried yellow mealworm larvae contained around 33% fat, 51% crude protein and 43% true protein on a dry matter basis. The true protein content of the protein extract was about 75%, with an extraction rate of 70% under optimised extraction conditions using 0.25 M NaOH, a NaOH solution:ethanol defatted worm ratio of 15:1 mL/g, 40°C for 1 h and extraction twice. The protein extract was a good source of essential amino acids. The lowest protein solubility in distilled water solution was found between pH 4 and 5, and increased with either increasing or decreasing pH. Lower solubility was observed in 0.5 M NaCl solution compared with distilled water. The rheological tests indicated that temperature, sample concentration, addition of salt and enzyme, incubation time and pH alterations influenced the elastic modulus of yellow mealworm protein extract (YMPE). These results demonstrate that the functional properties of YMPE can be modified for different food applications.

  17. Yellow Mealworm Protein for Food Purposes - Extraction and Functional Properties.

    Directory of Open Access Journals (Sweden)

    Xue Zhao

    Full Text Available A protocol for extraction of yellow mealworm larvae proteins was established, conditions were evaluated and the resulting protein extract was characterised. The freeze-dried yellow mealworm larvae contained around 33% fat, 51% crude protein and 43% true protein on a dry matter basis. The true protein content of the protein extract was about 75%, with an extraction rate of 70% under optimised extraction conditions using 0.25 M NaOH, a NaOH solution:ethanol defatted worm ratio of 15:1 mL/g, 40°C for 1 h and extraction twice. The protein extract was a good source of essential amino acids. The lowest protein solubility in distilled water solution was found between pH 4 and 5, and increased with either increasing or decreasing pH. Lower solubility was observed in 0.5 M NaCl solution compared with distilled water. The rheological tests indicated that temperature, sample concentration, addition of salt and enzyme, incubation time and pH alterations influenced the elastic modulus of yellow mealworm protein extract (YMPE. These results demonstrate that the functional properties of YMPE can be modified for different food applications.

  18. Optimization and orthogonal design of an ultrasonic-assisted aqueous extraction process for extracting chlorogenic acid from dry tobacco leaves

    Institute of Scientific and Technical Information of China (English)

    Martin Tongai Mazvimba; YU Ying; CUI Zhi-Qin; ZHANG Ying

    2012-01-01

    Processing parameters for heat reflux and ultrasonic-assisted extraction techniques were optimized.Optimal operating conditions,extraction solvents and extraction yields for both methods were established.Although methanol showed high extraction efficiency in heat reflux extraction,residual amounts of methanol caused adulteration of extracts.To circumvent this drawback,a novel ultrasonic-assisted aqueous extraction process was optimized and orthogonally designed to pave the way for replacing the toxic organic solvent,methanol with water.A new approach which utilizes fractional volumes of an extraction solvent was developed to minimize solvent consumption,improve chlorogenic acid solubility in water and enhance its aqueous extraction from dried tobacco leaves.Desired trajectories for the new ultrasonic assisted aqueous extraction process were found.

  19. Optimized Reputable Sensing Participants Extraction for Participatory Sensor Networks

    Directory of Open Access Journals (Sweden)

    Weiwei Yuan

    2014-01-01

    Full Text Available By collecting data via sensors embedded personal smart devices, sensing participants play a key role in participatory sensor networks. Using information provided by reputable sensing participants ensures the reliability of participatory sensing data. Setting a threshold for the reputation, and those whose reputations are bigger than this value are regarded as reputable. The bigger the threshold value is, the more reliable the extracted reputable sensing participant is. However, if the threshold value is too big, only very limited participatory sensing data can be involved. This may cause unexpected bias in information collection. Existing works did not consider the relationship between the reliability of extracted reputable sensing participants and the ratio of usable participatory sensing data. In this work, we propose a criterion for optimized reputable sensing participant extraction in participatory sensor networks. This is achieved based on the mathematical analysis on the ratio of available participatory sensing data and the reliability of extracted reputable sensing participants. Our suggested threshold value for reputable sensing participant extraction is only related to the power of sensing participant’s reputation distribution. It is easy to be applied in real applications. Simulation results tested on real application data further verified the effectiveness of our proposed method.

  20. Comparison of Methods for Protein Extraction from Pine Needles

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    Extraction of proteins from pine needles for proteomic analysis has long been a challenge for scientists. We compared three different protein extraction methods including sucrose, Tris-HCl and trichloroacetic acid (TCA)/acetone (TCA method) to determine their efficiency in separating pine needle proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional PAGE (2D-PAGE). Proteins were then separated by SDS-PAGE. Among three methods the method using sucrose extraction buffer showed the highest efficiency and highest quality in separating proteins. In addition, clearer and more stable strips were detected by SDS-PAGE using sucrose extraction buffer. When the proteins extracted using sucrose extraction buffer were separated by 2D-PAGE, more than 300 protein spots, with isoelectric points (PI) ranging from 4.0 to 7.0 and molecular weights (MW) from 6.5 to 97.4 kD, were observed. This confirmed that the method with sucrose extraction buffer was an efficient and reliable method for extracting proteins from pine needles.

  1. A comprehensive benchmark of kernel methods to extract protein-protein interactions from literature.

    Science.gov (United States)

    Tikk, Domonkos; Thomas, Philippe; Palaga, Peter; Hakenberg, Jörg; Leser, Ulf

    2010-07-01

    The most important way of conveying new findings in biomedical research is scientific publication. Extraction of protein-protein interactions (PPIs) reported in scientific publications is one of the core topics of text mining in the life sciences. Recently, a new class of such methods has been proposed - convolution kernels that identify PPIs using deep parses of sentences. However, comparing published results of different PPI extraction methods is impossible due to the use of different evaluation corpora, different evaluation metrics, different tuning procedures, etc. In this paper, we study whether the reported performance metrics are robust across different corpora and learning settings and whether the use of deep parsing actually leads to an increase in extraction quality. Our ultimate goal is to identify the one method that performs best in real-life scenarios, where information extraction is performed on unseen text and not on specifically prepared evaluation data. We performed a comprehensive benchmarking of nine different methods for PPI extraction that use convolution kernels on rich linguistic information. Methods were evaluated on five different public corpora using cross-validation, cross-learning, and cross-corpus evaluation. Our study confirms that kernels using dependency trees generally outperform kernels based on syntax trees. However, our study also shows that only the best kernel methods can compete with a simple rule-based approach when the evaluation prevents information leakage between training and test corpora. Our results further reveal that the F-score of many approaches drops significantly if no corpus-specific parameter optimization is applied and that methods reaching a good AUC score often perform much worse in terms of F-score. We conclude that for most kernels no sensible estimation of PPI extraction performance on new text is possible, given the current heterogeneity in evaluation data. Nevertheless, our study shows that three

  2. A comprehensive benchmark of kernel methods to extract protein-protein interactions from literature.

    Directory of Open Access Journals (Sweden)

    Domonkos Tikk

    Full Text Available The most important way of conveying new findings in biomedical research is scientific publication. Extraction of protein-protein interactions (PPIs reported in scientific publications is one of the core topics of text mining in the life sciences. Recently, a new class of such methods has been proposed - convolution kernels that identify PPIs using deep parses of sentences. However, comparing published results of different PPI extraction methods is impossible due to the use of different evaluation corpora, different evaluation metrics, different tuning procedures, etc. In this paper, we study whether the reported performance metrics are robust across different corpora and learning settings and whether the use of deep parsing actually leads to an increase in extraction quality. Our ultimate goal is to identify the one method that performs best in real-life scenarios, where information extraction is performed on unseen text and not on specifically prepared evaluation data. We performed a comprehensive benchmarking of nine different methods for PPI extraction that use convolution kernels on rich linguistic information. Methods were evaluated on five different public corpora using cross-validation, cross-learning, and cross-corpus evaluation. Our study confirms that kernels using dependency trees generally outperform kernels based on syntax trees. However, our study also shows that only the best kernel methods can compete with a simple rule-based approach when the evaluation prevents information leakage between training and test corpora. Our results further reveal that the F-score of many approaches drops significantly if no corpus-specific parameter optimization is applied and that methods reaching a good AUC score often perform much worse in terms of F-score. We conclude that for most kernels no sensible estimation of PPI extraction performance on new text is possible, given the current heterogeneity in evaluation data. Nevertheless, our study

  3. Metaproteomics: Evaluation of protein extraction from activated sludge.

    Science.gov (United States)

    Hansen, Susan Hove; Stensballe, Allan; Nielsen, Per Halkjaer; Herbst, Florian-Alexander

    2014-11-01

    Metaproteomic studies of full-scale activated sludge systems require reproducible protein extraction methods. A systematic evaluation of three different extractions protocols, each in combination with three different methods of cell lysis, and a commercial kit were evaluated. Criteria used for comparison of each method included the extracted protein concentration and the number of identified proteins and peptides as well as their phylogenetic, cell localization and functional distribution and quantitative reproducibility. Furthermore, the advantage of using specific metagenomes and a 2-step database approach was illustrated. The results recommend a protocol for protein extraction from activated sludge based on the protein extraction reagent B-Per and bead beating. The data have been deposited to the ProteomeXchange with identifier PXD000862 (http://proteomecentral.proteomexchange.org/dataset/PXD000862).

  4. Preparation of protein samples for gel electrophoresis by sequential extraction

    Institute of Scientific and Technical Information of China (English)

    钟伯雄; 翁宏飚; 等

    2002-01-01

    Since preparation and solubilization of protein samples are crucial factors in proteome research,the authors established a sequential extraction technique to prepare protein samples from the body wall of the 5th instar larvae of silkworm.Bombyx mori.Two kinds of protein samples were obtained from the body wall using the method.Between the two types of samples only about 15% proteins were identical;the majority were different,indicating that more species of proteins could be obtained with the sequential extraction method;which will be useful for preparation of protein samples for proteome study.

  5. Preparation of protein samples for gel electrophoresis by sequential extraction

    Institute of Scientific and Technical Information of China (English)

    钟伯雄; 翁宏飚; 方维焕

    2002-01-01

    Since preparation and solubilization of protein samples are crucial factors in proteome research, the authors established a sequential extraction technique to prepare protein samples from the body wall of the 5th instar larvae of silkworm, Bombyx mori. Two kinds of protein samples were obtained from the body wall using the method. Between the two types of samples only about 15% proteins were identical; the majority were different, indicating that more species of proteins could be obtained with the sequential extraction method; which will be useful for preparation of protein samples for proteome study.

  6. Perceptual Dominant Color Extraction by Multidimensional Particle Swarm Optimization

    Science.gov (United States)

    Kiranyaz, Serkan; Uhlmann (Eurasip Member), Stefan; Ince, Turker; Gabbouj, Moncef

    2010-12-01

    Color is the major source of information widely used in image analysis and content-based retrieval. Extracting dominant colors that are prominent in a visual scenery is of utmost importance since the human visual system primarily uses them for perception and similarity judgment. In this paper, we address dominant color extraction as a dynamic clustering problem and use techniques based on Particle Swarm Optimization (PSO) for finding optimal (number of) dominant colors in a given color space, distance metric and a proper validity index function. The first technique, so-called Multidimensional (MD) PSO can seek both positional and dimensional optima. Nevertheless, MD PSO is still susceptible to premature convergence due to lack of divergence. To address this problem we then apply Fractional Global Best Formation (FGBF) technique. In order to extract perceptually important colors and to further improve the discrimination factor for a better clustering performance, an efficient color distance metric, which uses a fuzzy model for computing color (dis-) similarities over HSV (or HSL) color space is proposed. The comparative evaluations against MPEG-7 dominant color descriptor show the superiority of the proposed technique.

  7. Perceptual Dominant Color Extraction by Multidimensional Particle Swarm Optimization

    Directory of Open Access Journals (Sweden)

    Moncef Gabbouj

    2009-01-01

    Full Text Available Color is the major source of information widely used in image analysis and content-based retrieval. Extracting dominant colors that are prominent in a visual scenery is of utmost importance since the human visual system primarily uses them for perception and similarity judgment. In this paper, we address dominant color extraction as a dynamic clustering problem and use techniques based on Particle Swarm Optimization (PSO for finding optimal (number of dominant colors in a given color space, distance metric and a proper validity index function. The first technique, so-called Multidimensional (MD PSO can seek both positional and dimensional optima. Nevertheless, MD PSO is still susceptible to premature convergence due to lack of divergence. To address this problem we then apply Fractional Global Best Formation (FGBF technique. In order to extract perceptually important colors and to further improve the discrimination factor for a better clustering performance, an efficient color distance metric, which uses a fuzzy model for computing color (dis- similarities over HSV (or HSL color space is proposed. The comparative evaluations against MPEG-7 dominant color descriptor show the superiority of the proposed technique.

  8. Soil Vapor Extraction System Optimization, Transition, and Closure Guidance

    Energy Technology Data Exchange (ETDEWEB)

    Truex, Michael J.; Becker, Dave; Simon, Michelle A.; Oostrom, Martinus; Rice, Amy K.; Johnson, Christian D.

    2013-02-08

    Soil vapor extraction (SVE) is a prevalent remediation approach for volatile contaminants in the vadose zone. A diminishing rate of contaminant extraction over time is typically observed due to 1) diminishing contaminant mass, and/or 2) slow rates of removal for contamination in low-permeability zones. After a SVE system begins to show indications of diminishing contaminant removal rate, SVE performance needs to be evaluated to determine whether the system should be optimized, terminated, or transitioned to another technology to replace or augment SVE. This guidance specifically addresses the elements of this type of performance assessment. While not specifically presented, the approach and analyses in this guidance could also be applied at the onset of remediation selection for a site as a way to evaluate current or future impacts to groundwater from vadose zone contamination. The guidance presented here builds from existing guidance for SVE design, operation, optimization, and closure from the U.S. Environmental Protection Agency, U.S. Army Corps of Engineers, and the Air Force Center for Engineering and the Environment. The purpose of the material herein is to clarify and focus on the specific actions and decisions related to SVE optimization, transition, and/or closure.

  9. Enzyme-Enhanced Extraction of Phenolic Compounds and Proteins from Flaxseed Meal

    OpenAIRE

    Bernardo Dias Ribeiro; Daniel Weingart Barreto; Maria Alice Zarur Coelho

    2013-01-01

    Flaxseed (Linum usitatissimum) meal, the main byproduct of the flaxseed oil extraction process, is composed mainly of proteins, mucilage, and phenolic compounds. The extraction methods of phenolics either commonly employed the use of mixed solvents (dioxane/ethanol, water/acetone, water/methanol, and water/ethanol) or are done with the aid of alkaline, acid, or enzymatic hydrolysis. This work aimed at the study of optimal conditions for a clean process, using renewable solvents and enzymes, f...

  10. Optimizing conditions for enzymatic extraction of sunflower oil

    Directory of Open Access Journals (Sweden)

    Badr, F. H.

    1992-10-01

    Full Text Available Sunflower seed oil was extracted with an enzymatic processes using different hydrolytic enzymes: cellulase, hemicellulase, animal proteinase, acid proteinase, pectinase and pectinex, as compared to enzyme - free aqueous extraction. All the hydrolytic enzymes enhanced oil extraction from sunflower seeds. The most optimal conditions for oil extraction from sunflower seeds were: 2% enzyme concentration, 30% substrate concentration and 3 hrs period. Using Boganov and Buchkov equation showed that time must be prolonged to get higher yields. The maximum yield during 3 hrs extraction with enzymatic process ranged between 44,5%-57,1% of the soxhlet extractable oil. The potency of the investigated enzymes in extracting oil was in the following order: acid proteinase > cellulase > hemicellulase > animal proteinase > pectinex > pectinase when compared at the previous optimal conditions.

    Aceite de semilla de girasol fue extraído mediante un proceso enzimático usando diferentes enzimas hidrolíticos: celulasa, hemicelulasa, proteinasa animal, proteinasa acida, pectinasa y pectinex, comparando con la extracción acuosa libre de enzima. Todos los enzimas hidrolíticos incrementan la extracción de aceites de semilla de girasol. Las condiciones óptimas para la extracción de aceite a partir de semillas de girasol fueron: 2% de concentración de enzima, 30% de concentración de sustrato y un período de 3 horas. La ecuación de Boganov y Buchkov mostró que el tiempo debe ser prolongado para alcanzar altos rendimientos. El máximo rendimiento durante tres horas de extracción con proceso enzimático osciló entre el 44,5%-57,1% del aceite extraído con soxhlet. La potencia de los enzimas investigados en la extracción de aceite siguió el orden: proteinasa acida > celulasa > hemicelulasa > proteinasa animal > pectinex > pectinasa cuando fue previamente comparado con las condiciones óptimas.

  11. Non-conventional approaches to food processing in CELSS, 1. Algal proteins: Characterization and process optimization

    Science.gov (United States)

    Nakhost, Z.; Karel, M.; Krukonis, V. J.

    1987-01-01

    Protein isolate obtained from green algae cultivated under controlled conditions was characterized. Molecular weight determination of fractionated algal proteins using SDS-polyacrylamide gel electrophoresis revealed a wide spectrum of molecular weights ranging from 15,000 to 220,000. Isoelectric points of dissociated proteins were in the range of 3.95 to 6.20. Amino acid composition of protein isolate compared favorably with FAO standards. High content of essential amino acids leucine, valine, phenylalanine and lysine make algal protein isolate a high quality component of closed ecological life support system diets. To optimize the removal of algal lipids and pigments supercritical carbon dioxide extraction (with and without ethanol as a co-solvent) was used. Addition of ethanol to supercritical carbon dioxide resulted in more efficient removal of algal lipids and produced protein isolate with a good yield and protein recovery. The protein isolate extracted by the above mixture had an improved water solubility.

  12. Non-conventional approaches to food processing in CELSS. I - Algal proteins: Characterization and process optimization

    Science.gov (United States)

    Nakhost, Z.; Karel, M.; Krukonis, V. J.

    1987-01-01

    Protein isolate obtained from green algae (Scenedesmus obliquus) cultivated under controlled conditions was characterized. Molecular weight determination of fractionated algal proteins using SDS-polyacrylamide gel electrophoresis revealed a wide spectrum of molecular weights ranging from 15,000 to 220,000. Isoelectric points of dissociated proteins were in the range of 3.95 to 6.20. Amino acid composition of protein isolate compared favorably with FAO standards. High content of essential amino acids leucine, valine, phenylalanine and lysine makes algal protein isolate a high quality component of CELSS diets. To optimize the removal of algal lipids and pigments supercritical carbon dioxide extraction (with and without ethanol as a co-solvent) was used. Addition of ethanol to supercritical CO2 resulted in more efficient removal of algal lipids and produced protein isolate with a good yield and protein recovery. The protein isolate extracted by the above mixture had an improved water solubility.

  13. Non-conventional approaches to food processing in CELSS. I - Algal proteins: Characterization and process optimization

    Science.gov (United States)

    Nakhost, Z.; Karel, M.; Krukonis, V. J.

    1987-01-01

    Protein isolate obtained from green algae (Scenedesmus obliquus) cultivated under controlled conditions was characterized. Molecular weight determination of fractionated algal proteins using SDS-polyacrylamide gel electrophoresis revealed a wide spectrum of molecular weights ranging from 15,000 to 220,000. Isoelectric points of dissociated proteins were in the range of 3.95 to 6.20. Amino acid composition of protein isolate compared favorably with FAO standards. High content of essential amino acids leucine, valine, phenylalanine and lysine makes algal protein isolate a high quality component of CELSS diets. To optimize the removal of algal lipids and pigments supercritical carbon dioxide extraction (with and without ethanol as a co-solvent) was used. Addition of ethanol to supercritical CO2 resulted in more efficient removal of algal lipids and produced protein isolate with a good yield and protein recovery. The protein isolate extracted by the above mixture had an improved water solubility.

  14. A green deep eutectic solvent-based aqueous two-phase system for protein extracting

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Kaijia; Wang, Yuzhi, E-mail: wyzss@hnu.edu.cn; Huang, Yanhua; Li, Na; Wen, Qian

    2015-03-15

    Highlights: • A strategy for the protein purification with a deep eutectic solvent(DES)-based aqueous two-phase system. • Choline chloride-glycerin DES was selected as the extraction solvent. • Bovine serum albumin and trypsin were used as the analytes. • Aggregation phenomenon was detected in the mechanism research. - Abstract: As a new type of green solvent, deep eutectic solvent (DES) has been applied for the extraction of proteins with an aqueous two-phase system (ATPS) in this work. Four kinds of choline chloride (ChCl)-based DESs were synthesized to extract bovine serum albumin (BSA), and ChCl-glycerol was selected as the suitable extraction solvent. Single factor experiments have been done to investigate the effects of the extraction process, including the amount of DES, the concentration of salt, the mass of protein, the shaking time, the temperature and PH value. Experimental results show 98.16% of the BSA could be extracted into the DES-rich phase in a single-step extraction under the optimized conditions. A high extraction efficiency of 94.36% was achieved, while the conditions were applied to the extraction of trypsin (Try). Precision, repeatability and stability experiments were studied and the relative standard deviations (RSD) of the extraction efficiency were 0.4246% (n = 3), 1.6057% (n = 3) and 1.6132% (n = 3), respectively. Conformation of BSA was not changed during the extraction process according to the investigation of UV–vis spectra, FT-IR spectra and CD spectra of BSA. The conductivity, dynamic light scattering (DLS) and transmission electron microscopy (TEM) were used to explore the mechanism of the extraction. It turned out that the formation of DES–protein aggregates play a significant role in the separation process. All the results suggest that ChCl-based DES-ATPS are supposed to have the potential to provide new possibilities in the separation of proteins.

  15. Evaluation of extraction procedures for 2-DE analysis of aphid proteins.

    Science.gov (United States)

    Yiou, Pan; Shaoli, An; Kebin, Li; Tao, Wang; Kui, Fang; Hua, Zhang; Yu, Sun; Xun, Yang; Jinghui, Xi

    2013-02-01

    Protein sample preparation is a crucial step in a 2-DE proteomics approach. In order to establish a routine protocol for the application of proteomics analysis to aphids, this study focuses on the specific protein extraction problems in insect tissues and evaluates four methods to bypass them. The approaches of phenol extraction methanol/ammonium acetate precipitation (PA), TCA/acetone precipitation, PEG precipitation, and no precipitation were evaluated for proteins isolation and purification from apterous adult aphids, Sitobion avenae. For 2-DE, the PA protocol was optimal, resulting in good IEF and clear spots. PA method yielded the greatest amount of protein and displayed most protein spots in 2-DE gels, as compared with the TCA/acetone precipitation, PEG precipitation and no precipitation protocols. Analysis of protein yield, image quality and spot numbers demonstrate that the TCA/acetone precipitation protocol is a reproducible and reliable method for extracting proteins from aphids. The PEG precipitation approach is a newly developed protein extraction protocol for aphids, from which more unique protein spots can be detected, especially for detection of acid proteins. These protocols are expected to be applicable to other insects or could be of interest to laboratories involved in insect proteomics, despite the amounts and types of interfering compounds vary considerably in different insects.

  16. Comparison of Different Protein Extraction Methods for Gel-Based Proteomic Analysis of Ganoderma spp.

    Science.gov (United States)

    Al-Obaidi, Jameel R; Saidi, Noor Baity; Usuldin, Siti Rokhiyah Ahmad; Hussin, Siti Nahdatul Isnaini Said; Yusoff, Noornabeela Md; Idris, Abu Seman

    2016-04-01

    Ganoderma species are a group of fungi that have the ability to degrade lignin polymers and cause severe diseases such as stem and root rot and can infect economically important plants and perennial crops such as oil palm, especially in tropical countries such as Malaysia. Unfortunately, very little is known about the complex interplay between oil palm and Ganoderma in the pathogenesis of the diseases. Proteomic technologies are simple yet powerful tools in comparing protein profile and have been widely used to study plant-fungus interaction. A critical step to perform a good proteome research is to establish a method that gives the best quality and a wide coverage of total proteins. Despite the availability of various protein extraction protocols from pathogenic fungi in the literature, no single extraction method was found suitable for all types of pathogenic fungi. To develop an optimized protein extraction protocol for 2-DE gel analysis of Ganoderma spp., three previously reported protein extraction protocols were compared: trichloroacetic acid, sucrose and phenol/ammonium acetate in methanol. The third method was found to give the most reproducible gels and highest protein concentration. Using the later method, a total of 10 protein spots (5 from each species) were successfully identified. Hence, the results from this study propose phenol/ammonium acetate in methanol as the most effective protein extraction method for 2-DE proteomic studies of Ganoderma spp.

  17. Algal Proteins: Extraction, Application, and Challenges Concerning Production

    Directory of Open Access Journals (Sweden)

    Stephen Bleakley

    2017-04-01

    Full Text Available Population growth combined with increasingly limited resources of arable land and fresh water has resulted in a need for alternative protein sources. Macroalgae (seaweed and microalgae are examples of under-exploited “crops”. Algae do not compete with traditional food crops for space and resources. This review details the characteristics of commonly consumed algae, as well as their potential for use as a protein source based on their protein quality, amino acid composition, and digestibility. Protein extraction methods applied to algae to date, including enzymatic hydrolysis, physical processes, and chemical extraction and novel methods such as ultrasound-assisted extraction, pulsed electric field, and microwave-assisted extraction are discussed. Moreover, existing protein enrichment methods used in the dairy industry and the potential of these methods to generate high value ingredients from algae, such as bioactive peptides and functional ingredients are discussed. Applications of algae in human nutrition, animal feed, and aquaculture are examined.

  18. Algal Proteins: Extraction, Application, and Challenges Concerning Production

    Science.gov (United States)

    Bleakley, Stephen; Hayes, Maria

    2017-01-01

    Population growth combined with increasingly limited resources of arable land and fresh water has resulted in a need for alternative protein sources. Macroalgae (seaweed) and microalgae are examples of under-exploited “crops”. Algae do not compete with traditional food crops for space and resources. This review details the characteristics of commonly consumed algae, as well as their potential for use as a protein source based on their protein quality, amino acid composition, and digestibility. Protein extraction methods applied to algae to date, including enzymatic hydrolysis, physical processes, and chemical extraction and novel methods such as ultrasound-assisted extraction, pulsed electric field, and microwave-assisted extraction are discussed. Moreover, existing protein enrichment methods used in the dairy industry and the potential of these methods to generate high value ingredients from algae, such as bioactive peptides and functional ingredients are discussed. Applications of algae in human nutrition, animal feed, and aquaculture are examined. PMID:28445408

  19. Small-scale extraction of recombinant proteins from bacteria.

    Science.gov (United States)

    Simpson, Richard J

    2010-09-01

    Bacteria are particularly convenient for producing recombinant proteins for purification purposes. To monitor induction as well as the levels of recombinant protein expression, it is important to have a rapid, simple method for estimating bacterial protein expression. This protocol describes the preparation of small-scale bacterial extracts using cell lysis with 0.5% Triton X-100.

  20. Protein extraction from xylem and phloem sap.

    Science.gov (United States)

    Kehr, Julia; Rep, Martijn

    2007-01-01

    It is well known that phloem and xylem vessels transport small nutrient molecules over long distances in higher plants. The finding that proteins also occur in both transport fluids was unexpected, and the function of most of these proteins is not yet well understood. This chapter outlines how proteins can be obtained and purified from xylem and phloem saps to perform subsequent proteomic analyses.

  1. Multiplex protein pattern unmixing using a non-linear variable-weighted support vector machine as optimized by a particle swarm optimization algorithm.

    Science.gov (United States)

    Yang, Qin; Zou, Hong-Yan; Zhang, Yan; Tang, Li-Juan; Shen, Guo-Li; Jiang, Jian-Hui; Yu, Ru-Qin

    2016-01-15

    Most of the proteins locate more than one organelle in a cell. Unmixing the localization patterns of proteins is critical for understanding the protein functions and other vital cellular processes. Herein, non-linear machine learning technique is proposed for the first time upon protein pattern unmixing. Variable-weighted support vector machine (VW-SVM) is a demonstrated robust modeling technique with flexible and rational variable selection. As optimized by a global stochastic optimization technique, particle swarm optimization (PSO) algorithm, it makes VW-SVM to be an adaptive parameter-free method for automated unmixing of protein subcellular patterns. Results obtained by pattern unmixing of a set of fluorescence microscope images of cells indicate VW-SVM as optimized by PSO is able to extract useful pattern features by optimally rescaling each variable for non-linear SVM modeling, consequently leading to improved performances in multiplex protein pattern unmixing compared with conventional SVM and other exiting pattern unmixing methods.

  2. Intracellular coagulation inhibits the extraction of proteins from Prochloron

    Science.gov (United States)

    Fall, R.; Lewin, R. A.; Fall, L. R.

    1983-01-01

    Protein extraction from the prokaryotic alga Prochloron LP (isolated from the ascidian host Lissoclinum patella) was complicated by an irreversible loss of cell fragility in the isolated algae. Accompanying this phenomenon, which is termed intracellular coagulation, was a redistribution of thylakoids around the cell periphery, a loss of photosynthetic O2 production, and a drastic decrease in the extractability of cell proteins. Procedures are described for the successful preparation and transport of cell extracts yielding the enzymes glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase as well as other soluble proteins.

  3. Intracellular coagulation inhibits the extraction of proteins from Prochloron

    Science.gov (United States)

    Fall, R.; Lewin, R. A.; Fall, L. R.

    1983-01-01

    Protein extraction from the prokaryotic alga Prochloron LP (isolated from the ascidian host Lissoclinum patella) was complicated by an irreversible loss of cell fragility in the isolated algae. Accompanying this phenomenon, which is termed intracellular coagulation, was a redistribution of thylakoids around the cell periphery, a loss of photosynthetic O2 production, and a drastic decrease in the extractability of cell proteins. Procedures are described for the successful preparation and transport of cell extracts yielding the enzymes glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase as well as other soluble proteins.

  4. Fish Muscle Proteins: Extraction, Quantitation, and Electrophoresis

    Science.gov (United States)

    Smith, Denise

    Electrophoresis can be used to separate and visualize proteins. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), proteins are separated based on size. When protein samples are applied to such gels, it is usually necessary to know the protein content of the sample. This makes it possible to apply a volume of sample to the gel such that samples have a comparable amount of total protein. While it is possible to use an official method of protein analysis (e.g., Kjeldahl, N combustion) for such an application, it often is convenient to use a rapid spectroscopic protein analysis that requires only a small amount of sample. The bicinchoninic acid (BCA) assay method will be used for this purpose.

  5. Microbial protein in soil: influence of extraction method and C amendment on extraction and recovery.

    Science.gov (United States)

    Taylor, Erin B; Williams, Mark A

    2010-02-01

    The capacity to study the content and resolve the dynamics of the proteome of diverse microbial communities would help to revolutionize the way microbiologists study the function and activity of microorganisms in soil. To better understand the limitations of a proteomic approach to studying soil microbial communities, we characterized extractable soil microbial proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Two methods were utilized to extract proteins from microorganisms residing in a Quitman and Benfield soil: (1) direct extraction of bulk protein from soil and (2) separation of the microorganisms from soil using density gradient centrifugation and subsequent extraction (DGC-EXT) of microbial protein. In addition, glucose and toluene amendments to soil were used to stimulate the growth of a subset of the microbial community. A bacterial culture and bovine serum albumin (BSA) were added to the soil to qualitatively assess their recovery following extraction. Direct extraction and resolution of microbial proteins using SDS-PAGE generally resulted in smeared and unresolved banding patterns on gels. DGC-EXT of microbial protein from soil followed by separation using SDS-PAGE, however, did resolve six to 10 bands in the Benfield but not the Quitman soil. DGC-EXT of microbial protein, but not direct extraction following the addition of glucose and toluene, markedly increased the number of bands (approximately 40) on the gels in both Benfield and Quitman soils. Low recoveries of added culture and BSA proteins using the direct extraction method suggest that proteins either bind to soil organic matter and mineral particles or that partial degradation takes place during extraction. Interestingly, DGC may have been preferentially selected for actively growing cells, as gauged by the 10-100x lower cy19:0/18:1omega7 ratio of the fatty acid methyl esters in the isolated community compared to that for the whole soil. DGC can be used to

  6. A comprehensive factorial design study of variables affecting protein extraction from formalin-fixed kidney tissue samples.

    Science.gov (United States)

    Araújo, J E; Oliveira, E; Otero-Glez, A; Santos Nores, J; Igrejas, G; Lodeiro, C; Capelo, J L; Santos, H M

    2014-02-01

    Formalin-fixed tissues are an important source of biological samples for biomedical research. However, proteomics analysis of formalin-fixed tissues has been set aside by formalin-induced protein modifications, which reduce protein extraction efficiency. In this study, a two level full factorial experimental design (2(4)) was used to determine the effects of the extracting conditions in the efficiency of protein recovery from formalin-fixed kidney samples. The following variables were assessed: temperature of extraction, pH of extraction, composition of the extracting buffer and the use ultrasonic energy applied with probe. It is clearly demonstrated that when hating and ultrasonic energy are used in conjunction, a 7-fold increase (p protein extraction is obtained if compared to extracting conditions for which neither heating nor ultrasonic energy are used. The optimization study was done following the amount of protein extracted by UV (Nanodrop(®) technology, protein ABS at 280 nm) and by 1D SDS-PAGE. Extracts obtained with the optimized conditions were subjected to LC-MALDI MS/MS. A total of 112 proteins were identified.

  7. [Optimizing the ultrasonic extraction of tannin in Elaeagnus angustifolia L. by uniform design].

    Science.gov (United States)

    Jiang, Fashou; Dan, Jianming; Wang, Hangyu; Wang, Jin

    2002-11-01

    The influences of ultrasonic frequency and ultrasonic time on the extraction rate of Tannin in Elaeagnus angustifolia L. were studied. The optimizing had been got. Results showed that the ultrasonic extraction method of Tannin in Elaeagnus angustifolia L. could save time, improve extraction rates, and need not be hot. Compared with traditional extraction, the ultrasonic extraction method is a good way.

  8. Extraction optimization of polyphenols, antioxidant and xanthine oxidase inhibitory activities from Prunus salicina Lindl.

    OpenAIRE

    LI,Yibin; LAI,Pufu; CHEN,Junchen; SHEN,Hengsheng; TANG,Baosha; Wu, Li; WENG,Minjie

    2016-01-01

    Abstract Optimization of polyphenols extraction from plum (Prunus salicina Lindl.) was evaluated using response surface methodology. The Box-Behnken experimental results showed the optimal conditions involved an extraction temperature of 59 °C, a sonication time of 47 min, and an ethanol concentration of 61% respectively. The maximum extraction yield of total polyphenols was 44.74 mg gallic acid equivalents per gram of dried plum at optimal conditions. Polyphenol extracts exhibited stronger a...

  9. Extractable proteins from field radiation vulcanized natural rubber latex

    Energy Technology Data Exchange (ETDEWEB)

    Parra, Duclerc F. [Chemical and Environmental Centre, Nuclear Energy Research Institute, Av. Lineu Prestes, 2242-CEP Sao Paulo (Brazil)]. E-mail: dfparra@ipen.br; Pinto Martins, Carlos Felipe [Chemical and Environmental Centre, Nuclear Energy Research Institute, Av. Lineu Prestes, 2242-CEP Sao Paulo (Brazil); Collantes, Hugo D.C. [Chemical and Environmental Centre, Nuclear Energy Research Institute, Av. Lineu Prestes, 2242-CEP Sao Paulo (Brazil); Lugao, Ademar B. [Chemical and Environmental Centre, Nuclear Energy Research Institute, Av. Lineu Prestes, 2242-CEP Sao Paulo (Brazil)

    2005-07-01

    The type I allergy associated with the use of natural rubber latex (NRL) products is caused by the NRL proteins leached by the sweat or other body fluids. Makuuchi's group proposed for the first time the proteins removal by the addition of water-soluble polymers (WSP) on radiation vulcanization of natural rubber latex (RVNRL) that is a promising process under development in many countries. In this study, Brazilian field natural rubber was irradiated with a {sup 60}Co gamma source to reduce the content of WSP in the final product. WSP was used as additive to improve the extraction of protein. After irradiation the RVNRL was centrifuged to extract the WSP and proteins. The analytical methodology for protein content was based on the modified Lowry method according to ASTM D5712. Protein determination was carried out in serum of latex and in the extracts of the gloves. The concentration of extractable water-soluble proteins in serum of irradiated field NRL (NRL1), not irradiated one (NRL2); of twice centrifuged sample with polymer additive NRL (NRL3) and of the glove manufactured (NRLG) are compared with commercial glove (CG). The irradiation process increases the extractable water-soluble proteins, EP, as reported in the literature. In this study the use of polymeric additive on the bi-centrifugation process to remove protein was successful and the EP of the glove obtained in NRL3 was at around 40% of the commercial glove.

  10. Extraction of intracellular protein from Glaciozyma antarctica for proteomics analysis

    Science.gov (United States)

    Faizura, S. Nor; Farahayu, K.; Faizal, A. B. Mohd; Asmahani, A. A. S.; Amir, R.; Nazalan, N.; Diba, A. B. Farah; Muhammad, M. Nor; Munir, A. M. Abdul

    2013-11-01

    Two preparation methods of crude extracts of psychrophilic yeast Glaciozyma antarctica were compared in order to obtain a good recovery of intracellular proteins. Extraction with mechanical procedures using sonication was found to be more effective for obtaining good yield compare to alkaline treatment method. The procedure is simple, rapid, and produce better yield. A total of 52 proteins were identified by combining both extraction methods. Most of the proteins identified in this study involves in the metabolic process including glycolysis pathway, pentose phosphate pathway, pyruyate decarboxylation and also urea cyle. Several chaperons were identified including probable cpr1-cyclophilin (peptidylprolyl isomerase), macrolide-binding protein fkbp12 and heat shock proteins which were postulate to accelerate proper protein folding. Characteristic of the fundamental cellular processes inferred from the expressed-proteome highlight the evolutionary and functional complexity existing in this domain of life.

  11. Optimization of Extraction Parameters for Enhanced Production of Ovotransferrin from Egg White for Antimicrobial Applications

    Directory of Open Access Journals (Sweden)

    Eyad M. A. Alshammari

    2015-01-01

    Full Text Available Ovotransferrin is the second most abundant protein (~12-13% of the total egg protein in egg white after ovalbumin. Ovotransferrin is a potent natural antimicrobial agent as it possesses antibacterial, antifungal, and antiviral properties and is also the major metal binding protein found in egg, which makes it an industrially important protein. Ovotransferrin was extracted from egg white using its metal (iron binding properties. In the present study, eggs from two different sources were used (fresh local eggs from domestic household source and poultry eggs from shops to compare the results and Response Surface Methodology was used for the experiment design and data analysis. The following extraction conditions were optimized so as to maximize the yield of ovotransferrin from egg white: ethanol % (v/v and pH and volume (mL of 25 mM FeCl3/50 mL of egg white. A maximum yield of ~85 ± 2.5% was obtained near the optimum extraction conditions. The yield was calculated based on the theoretical value (934 mg of ovotransferrin in 100 mL of 1.5x diluted egg white solution. Our results suggest that efficient downstream processing may reduce the cost of overall production process of this promising enzyme, making it a natural and cost-effective alternative to the existing chemically synthesized antimicrobial agents.

  12. DNA extraction from benthic Cyanobacteria: comparative assessment and optimization.

    Science.gov (United States)

    Gaget, V; Keulen, A; Lau, M; Monis, P; Brookes, J D

    2017-01-01

    Benthic Cyanobacteria produce toxic and odorous compounds similar to their planktonic counterparts, challenging the quality of drinking water supplies. The biofilm that benthic algae and other micro-organisms produce is a complex and protective matrix. Monitoring to determine the abundance and identification of Cyanobacteria, therefore, relies on molecular techniques, with the choice of DNA isolation technique critical. This study investigated which DNA extraction method is optimal for DNA recovery in order to guarantee the best DNA yield for PCR-based analysis of benthic Cyanobacteria. The conventional phenol-chloroform extraction method was compared with five commercial kits, with the addition of chemical and physical cell-lysis steps also trialled. The efficacy of the various methods was evaluated by measuring the quantity and quality of DNA by UV spectrophotometry and by quantitative PCR (qPCR) using Cyanobacteria-specific primers. The yield and quality of DNA retrieved with the commercial kits was significantly higher than that of DNA obtained with the phenol-chloroform protocol. Kits including a physical cell-lysis step, such as the MO BIO Power Soil and Biofilm kits, were the most efficient for DNA isolation from benthic Cyanobacteria. These commercial kits allow greater recovery and the elimination of dangerous chemicals for DNA extraction, making them the method of choice for the isolation of DNA from benthic mats. They also facilitate the extraction of DNA from benthic Cyanobacteria, which can help to improve the characterization of Cyanobacteria in environmental studies using qPCRs or population composition analysis using next-generation sequencing. © 2016 The Society for Applied Microbiology.

  13. Optimization of Phenolic Antioxidant Extraction from Wuweizi (Schisandra chinensis Pulp Using Random-Centroid Optimazation Methodology

    Directory of Open Access Journals (Sweden)

    Xiong Yu

    2011-09-01

    Full Text Available The extraction optimization and composition analysis of polyphenols in the fresh pulp of Wuweizi (Schisandra chinensis have been investigated in this study. The extraction process of polyphenols from Wuweizi pulp was optimized using Random-Centroid Optimization (RCO methodology. Six factors including liquid and solid ratio, ethanol concentration, pH, temperature, heating time and extraction times, and three extraction targets of polyphenol content, antioxidant activity and extract yield were considered in the RCO program. Three sets of optimum proposed factor values were obtained corresponding to three extraction targets respectively. The set of optimum proposed factor values for polyphenol extraction given was chosen in further experiments as following: liquid and solid ratio (v/w 8, ethanol 67.3% (v/v, initial pH 1.75, temperature 55 °C for 4 h and extraction repeated for 4 times. The Wuweizi polyphenol extract (WPE was obtained with a yield of 16.37 mg/g and composition of polyphenols 1.847 mg/g, anthocyanins 0.179 mg/g, sugar 9.573 mg/g and protein 0.327 mg/g. The WPE demonstrated high scavenging activities against DPPH radicals.

  14. comparison of protein extraction methods for the leaves of ficus ...

    African Journals Online (AJOL)

    F. I. Abdullah

    2017-05-01

    May 1, 2017 ... This study investigated several extraction methods for proteins from the leaves of Ficus .... found to be effective to remove plant lipids and pigments [22]. ... The addition of SDS as a solubilizing agent in the Tris buffered phenol.

  15. [The optimation of extraction technology of chuanxiong and xiangfu compatibility by SFE-CO2].

    Science.gov (United States)

    Lin, Qing; Li, Yuan; Li, Mei; Huang, Kai-Ming

    2013-02-01

    To optimize the extraction conditions of Chuanxiong and Xiangfu compatibility by supercritical CO2 and control the quality of the extract. The orthogonal design experiments were used to select the optimal conditions and the factors were extraction pressure, extraction temperature, extraction time and flow rate of CO2. The extraction rate was used as inspection index. The contents of alpha-cyperolone and butenyl were detected by HPLC. The optimal conditions were as follows: extraction pressure 25 Mpa, extraction temperature 45 degrees C, extraction time 2 h, flow rate of CO2 20 Kg/h. The average content of alpha-cyperolone, butenyl was 3.26%, 1.50% respectively. The method of supercritical CO2 extraction is reliable and feasible and can be used in industry. The quality of the extract is stable and controlled.

  16. [Optimization of extraction process for tannins from Geranium orientali-tibeticum by supercritical CO2 method].

    Science.gov (United States)

    Xie, Song; Tong, Zhi-Ping; Tan, Rui; Liu, Xiao-Zhen

    2014-08-01

    In order to optimize extraction process conditions of tannins from Geranium orientali-tibeticum by supercritical CO2, the content of tannins was determined by phosphomolybdium tungsten acid-casein reaction, with extraction pressure, extraction temper- ature and extraction time as factors, the content of tannins from extract of G. orientali-tibeticum as index, technology conditions were optimized by orthogonal test. Optimum technology conditions were as follows: extraction pressure was 25 MPa, extraction temperature was 50 °C, extracted 1.5 h. The content of tannins in extract was 12.91 mg x g(-1), extract rate was 3.67%. The method established could be used for assay the contents of tannin in G. orientali-tibeticum. The circulated extraction was an effective extraction process that was stable and feasible, and that provides a way of the extraction process conditions of tannin from G. orientali-tibeticum.

  17. [Optimization of extraction process for tannins from Geranium orientali-tibeticum by supercritical CO2 method].

    Science.gov (United States)

    Xie, Song; Tong, Zhi-Ping; Tan, Rui; Liu, Xiao-Zhen

    2014-08-01

    In order to optimize extraction process conditions of tannins from Geranium orientali-tibeticum by supercritical CO2, the content of tannins was determined by phosphomolybdium tungsten acid-casein reaction, with extraction pressure, extraction temper- ature and extraction time as factors, the content of tannins from extract of G. orientali-tibeticum as index, technology conditions were optimized by orthogonal test. Optimum technology conditions were as follows: extraction pressure was 25 MPa, extraction temperature was 50 °C, extracted 1.5 h. The content of tannins in extract was 12.91 mg x g(-1), extract rate was 3.67%. The method established could be used for assay the contents of tannin in G. orientali-tibeticum. The circulated extraction was an effective extraction process that was stable and feasible, and that provides a way of the extraction process conditions of tannin from G. orientali-tibeticum.

  18. Optimization of the Phenol -Chloroform Silica DNA Extraction Method in Ancient Bones DNA Extraction

    Directory of Open Access Journals (Sweden)

    Morteza Sadeghi

    2014-04-01

    Full Text Available Introduction: DNA extraction from the ancient bones tissues is currently very difficult. Phenol chloroform silica method is one of the methods currently used for this aim. The purpose of this study was to optimize the assessment method. Methods: DNA of 62 bone tissues (average 3-11 years was first extracted with phenol chloroform silica methods and then with changing of some parameters of the methods the extracted DNA was amplified in eight polymorphisms area including FES, F13, D13S317, D16, D5S818, vWA and CD4. Results from samples gained by two methods were compared in acrylamide gel. Results: The average of PCR yield for new method and common method in eight polymorphism regions was 75%, 78%, 81%, 76%, 85%, 71%, 89%, 86% and 64%, 39%, 70%, 49%, 68%, 76%, 71% and 28% respectively. The average of DNA in optimized (in 35l silica density and common method were 267.5 µg/ml with 1.12 purity and 192.76 g/ml with 0.84 purity respectively. Conclusions: According to the findings of this study, it is estimated that longer EDTA attendance is an efficient agent in removing calcium and also adequate density of silica particles can be efficient in removal of PCR inhibitors.

  19. Optimization of capillary electrophoresis-inductively coupled plasma mass spectrometry for species analysis of metallothionein-like proteins extracted from liver tissues of Elbe-bream and Roe deer

    Energy Technology Data Exchange (ETDEWEB)

    Proefrock, Daniel; Prange, Andreas E-mail: andreas.prange@gkss.de; Schaumloeffel, Dirk; Ruck, Wolfgang

    2003-08-15

    Species analysis of metallothionein-like proteins (MLP) in liver tissues from Elbe-Bream (Abramis brama L.) and Roe Deer (Capreolus capreolus L.) using capillary electrophoresis (CE) combined with inductively coupled plasma mass spectrometry detection is described. In order to allow systematic development of the method, commercially available metallothionein (MT) preparations of rabbit liver were used. Optimum separation efficiency was obtained by investigating the influence of parameters such as voltage, capillary temperature, buffer concentration, buffer pH and the use of different buffer systems. Instrumental parameters such as CE capillary position, interface adjustment and contamination problems are also discussed. Separation was performed using uncoated fused silica capillaries with 75 {mu}m i.d. and 70 cm length. The optimum conditions were found to be: Separation voltage 30 kV, positive polarity, capillary temperature 288.15 K and a buffer concentration of 100 mmol l{sup -1} Tricine-NH{sub 3} adjusted to pH 7.2. Sample preparation was performed so as to minimize oxidation and heavy metal contamination of the samples. The high molecular mass protein matrix was reduced by acetonitrile precipitation. For commercial MT preparations the relative standard deviations (R.S.D) in the retention times were 0.9% for MT-1 and 1.9% for MT-2; the R.S.D.'s in the peak areas were less than 6% for MT-1 and 16% for MT-2, respectively. Under optimized conditions the MLPs in the real samples could be separated efficiently in less than 10 min. By comparison with the migration times of commercially available MT preparations, two of the observed peaks could be assigned to MT-1 and MT-2.

  20. Extraction and fractionation of wheat flour proteins

    NARCIS (Netherlands)

    Graveland, A.; Bosveld, P.; Lichtendonk, W.J.; Moonen, H.H.E.; Scheepstra, A.

    1982-01-01

    Extraction of wheat flour with 1.5% sodium dodecyl sulphate (SDS) solution dissolved 65–67% of the total flour nitrogen. The SDS‐insoluble proteinaceous material was separated into glycoproteins‐I, II and III by ultracentrifugation. Part of the SDS‐soluble proteinaceous material was precipitated by

  1. Optimization of extraction and antioxidant activity of polysaccharides from Salvia miltiorrhiza Bunge residue.

    Science.gov (United States)

    Jiang, Yuanyuan; Wang, Long; Zhang, Li; Wang, Tao; Zhou, Yonghong; Ding, Chunbang; Yang, Ruiwu; Wang, Xiaoli; Yu, Lin

    2015-08-01

    In this study, the process of extracting polysaccharides from Salvia miltiorrhiza Bunge residue was optimized by using a Box-Behnken design. Statistical analysis of the results showed that the linear and quadratic terms of the three variables of the extraction process had significant effects. The optimal conditions are as follows: extracting time of 2.6 h, extraction temperature of 89 °C, and ratio of water to raw material of 32 mL/g. Moreover, a new polysaccharide with antioxidant activity [i.e., SMWP-1 (∼5.27×10(5) Da)] was isolated from S. miltiorrhiza residue. The carbohydrate, uronic acid, and protein contents of SMWP-1 were 90.11%, 0.13%, and 0.53%, respectively. The SMWP-1 is composed of glucose, xylose, mannose, and galactose. The preliminary structural characterization of SMWP-1 was determined via Fourier transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM) analyses. This polysaccharide exhibited strong reducing power and free-radical scavenging activities in vitro against 2,2-diphenyl-1-picrylhydrazyl, superoxide anion, and hydroxyl. Therefore, SMWP-1 can be investigated further as a novel natural antioxidant.

  2. Optimization of the trienzyme extraction for the microbiological assay of folate in vegetables.

    Science.gov (United States)

    Chen, Liwen; Eitenmiller, Ronald R

    2007-05-16

    Response surface methodology was applied to optimize the trienzyme digestion for the extraction of folate from vegetables. Trienzyme extraction is a combined enzymatic digestion by protease, alpha-amylase, and conjugase (gamma-glutamyl hydrolase) to liberate the carbohydrate and protein-bound folates from food matrices for total folate analysis. It is the extraction method used in AOAC Official Method 2004.05 for assay of total folate in cereal grain products. Certified reference material (CRM) 485 mixed vegetables was used to represent the matrix of vegetables. Regression and ridge analysis were performed by statistical analysis software. The predicted second-order polynomial model was adequate (R2 = 0.947), without significant lack of fit (p > 0.1). Both protease and alpha-amylase have significant effects on the extraction. Ridge analysis gave an optimum trienzyme digestion time: Pronase, 1.5 h; alpha-amylase, 1.5 h; and conjugase, 3 h. The experimental value for CRM 485 under this optimum digestion was close to the predicted value from the model, confirming the validity and adequacy of the model. The optimized trienzyme digestion condition was applied to five vegetables and yielded higher folate levels than the trienzyme digestion parameters employed in AOAC Official Method 2004.05.

  3. Optimization of extraction method of the water-soluble proteins of barley and malt and construction of two-dimensional electrophoretic map%大麦和麦芽水溶性蛋白提取方案的优化及双向电泳图谱的建立

    Institute of Scientific and Technical Information of China (English)

    夏先锋; 朴永哲; 沈海萍; 翟明昌; 成建国; 赵长新

    2012-01-01

    To develop an efficient and high resolution 2-DE procedure for barley and malt research, in this study, the method of extraction and sample preparation for two dimensional gel electrophoresis (2-DE) were optimized using high quality barley cultivars Gairdner and by changing the type and amount of extraction reagents as well as extraction time. By using the optimized method, protein extraction and sample preparation methods could be achieved. The protein sample of barley was obtained after grinding, buffer-phenol extraction, ethanol washing, and was freeze-dried. Protein spots were separated by 2-DE technique, stained with silver nitrate, recorded and detected by PDQuest software. The results showed that over 700 spots of barley and about 500 spots of malt were separated by 2-DE. The established of those methods may provide powerful tools for developmental proteomics and quality improvement of barley in the future.%为了建立一套适用于大麦和麦芽的双向电泳分析方法,得到更加清晰、全面的双向电泳图谱。以高产优质大麦品种Gairdner为材料,对提取方法以及提取过程中所用的试剂的种类、用量和处理时间等进行比较分析和优化,运用蛋白定量法测定提取样品的蛋白质含量,摸索出一套完整的用于双向电泳分析的大麦蛋白提取和样品制备方法。大麦或麦芽经研磨,缓冲液-苯酚提取,醇洗,冻干后得到的蛋白质样品,双向电泳分离,硝酸银染色,PDQuest分析,图谱结果显示大麦检测到700个蛋白质点,麦芽检测到500个蛋白质点。这一方法的构建可作为大麦蛋白质组研究的重要工具,为今后大麦蛋白质组学以及对大麦品质改良的研究奠定了一定的基础。

  4. Comparison of Protein Extracts from Various Unicellular Green Sources

    Science.gov (United States)

    2017-01-01

    Photosynthetic unicellular organisms are considered as promising alternative protein sources. The aim of this study is to understand the extent to which these green sources differ with respect to their gross composition and how these differences affect the final protein isolate. Using mild isolation techniques, proteins were extracted and isolated from four different unicellular sources (Arthrospira (spirulina) maxima, Nannochloropsis gaditana, Tetraselmis impellucida, and Scenedesmus dimorphus). Despite differences in protein contents of the sources (27–62% w/w) and in protein extractability (17–74% w/w), final protein isolates were obtained that had similar protein contents (62–77% w/w) and protein yields (3–9% w/w). Protein solubility as a function of pH was different between the sources and in ionic strength dependency, especially at pH < 4.0. Overall, the characterization and extraction protocol used allows a relatively fast and well-described isolation of purified proteins from novel protein sources. PMID:28701042

  5. Optimized Feature Extraction for Temperature-Modulated Gas Sensors

    Directory of Open Access Journals (Sweden)

    Alexander Vergara

    2009-01-01

    Full Text Available One of the most serious limitations to the practical utilization of solid-state gas sensors is the drift of their signal. Even if drift is rooted in the chemical and physical processes occurring in the sensor, improved signal processing is generally considered as a methodology to increase sensors stability. Several studies evidenced the augmented stability of time variable signals elicited by the modulation of either the gas concentration or the operating temperature. Furthermore, when time-variable signals are used, the extraction of features can be accomplished in shorter time with respect to the time necessary to calculate the usual features defined in steady-state conditions. In this paper, we discuss the stability properties of distinct dynamic features using an array of metal oxide semiconductors gas sensors whose working temperature is modulated with optimized multisinusoidal signals. Experiments were aimed at measuring the dispersion of sensors features in repeated sequences of a limited number of experimental conditions. Results evidenced that the features extracted during the temperature modulation reduce the multidimensional data dispersion among repeated measurements. In particular, the Energy Signal Vector provided an almost constant classification rate along the time with respect to the temperature modulation.

  6. [Optimization for supercritical CO2 extraction with response surface methodology of Prunus armeniaca oil].

    Science.gov (United States)

    Chen, Fei-Fei; Wu, Yan; Ge, Fa-Huan

    2012-03-01

    To optimize the extraction conditions of Prunus armeniaca oil by Supercritical CO2 extraction and identify its components by GC-MS. Optimized of SFE-CO extraction by response surface methodology and used GC-MS to analysis Prunus armeniaca oil compounds. Established the model of an equation for the extraction rate of Prunus armeniaca oil by supercritical CO2 extraction, and the optimal parameters for the supercritical CO2 extraction determined by the equation were: the extraction pressure was 27 MPa, temperature was 39 degrees C, the extraction rate of Prunus armeniaca oil was 44.5%. 16 main compounds of Prunus armeniaca oil extracted by supercritical CO2 were identified by GC-MS, unsaturated fatty acids were 92.6%. This process is simple, and can be used for the extraction of Prunus armeniaca oil.

  7. Extraction optimization and bioactivity of exopolysaccharides from Agaricus bisporus.

    Science.gov (United States)

    Mao, Yong; Mao, Jian; Meng, Xiangyong

    2013-02-15

    Response surface methodology was used to optimize the extraction parameters of EPS produced by Agaricus bisporus MJ-0811 in submerged culture. The optimal levels for ethanol concentration (85%, v/v), pH (8) and precipitation time (22 h) were determined, and EPS production was estimated at 2.71 g/L. The actual yield of EPS under these conditions was 2.69 g/L. In addition, the antioxidant activity of EPS was investigated by measuring its scavenging ability on superoxide radicals and hydroxyl radicals in vitro. Furthermore, the hypoglycemic activity of EPS was investigated by measuring its effects on body weights and blood glucose of diabetic mice. The study suggests that EPS has beneficial antioxidant activities (hydroxyl radical-scavenging activities, superoxide radical-scavenging activities) in vitro, anti-diabetic effect on alloxan induced diabetic mice. The EPS from A. bisporus may be a novel resource of natural antioxidants and anti-diabetic agents for use in the functional food or medicine.

  8. Enzyme assisted protein extraction from rapeseed, soybean, and microalgae meals

    NARCIS (Netherlands)

    Sari, Y.W.; Bruins, M.E.; Sanders, J.P.M.

    2013-01-01

    Oilseed meals that are by-products from oil production are potential resources for protein. The aim of this work is to investigate the use of enzymes in assisting in the extraction of protein from different oilseed meals, namely rapeseed, soybean, and microalgae meals. In addition, microalgae withou

  9. Enzyme assisted protein extraction from rapeseed, soybean, and microalgae meals

    NARCIS (Netherlands)

    Sari, Y.W.; Bruins, M.E.; Sanders, J.P.M.

    2013-01-01

    Oilseed meals that are by-products from oil production are potential resources for protein. The aim of this work is to investigate the use of enzymes in assisting in the extraction of protein from different oilseed meals, namely rapeseed, soybean, and microalgae meals. In addition, microalgae withou

  10. Enzyme assisted protein extraction from rapeseed, soybean, and microalgae meals

    NARCIS (Netherlands)

    Sari, Y.W.; Bruins, M.E.; Sanders, J.P.M.

    2013-01-01

    Oilseed meals that are by-products from oil production are potential resources for protein. The aim of this work is to investigate the use of enzymes in assisting in the extraction of protein from different oilseed meals, namely rapeseed, soybean, and microalgae meals. In addition, microalgae

  11. Optimization and comparison of ultrasound/microwave assisted extraction (UMAE) and ultrasonic assisted extraction (UAE) of lycopene from tomatoes.

    Science.gov (United States)

    Lianfu, Zhang; Zelong, Liu

    2008-07-01

    The extracting technology including ultrasonic and microwave assisted extraction (UMAE) and ultrasonic assisted extraction (UAE) of lycopene from tomato paste were optimized and compared. The results showed that the optimal conditions for UMAE were 98 W microwave power together with 40 KHz ultrasonic processing, the ratio of solvents to tomato paste was 10.6:1 (V/W) and the extracting time should be 367 s; as for UAE, the extracting temperature was 86.4 degrees C, the ratio of the solvents to tomato paste was 8.0:1 (V/W) and the extracting time should be 29.1 min, while the percentage of lycopene yield was 97.4% and 89.4% for UMAE and UAE, respectively. These results implied that UMAE was far more efficient extracting method than UAE.

  12. Nutritional quality of sunflower seed protein fraction extracted with isopropanol.

    Science.gov (United States)

    Sen, M; Bhattacharyya, D K

    2000-01-01

    This study investigated the nutritional effect of sunflower seed protein fraction (SSPF) extracted with isopropanol on growth, plasma and tissue lipid profile, protein content and erythrocyte membrane lipid profile of rats. Dehulled sunflower seeds were extracted with isopropanol at 50 +/- 1 degree C resulting in a protein fraction (71.5%) with low residual chlorogenic acid (0.07%) and fiber (3.3%) contents. Rats fed the sunflower seed protein fraction had a similar body weight gain and food efficiency ratios in comparison to those fed casein. Rats fed SSPF in contrast had a significantly higher growth and food efficiency ratio than the rats fed sunflower meal (SM), extracted with hexane. However, dietary proteins exerted a separate effect on plasma total cholesterol, low density lipoprotein (LDL)-cholesterol, low density lipoprotein to high density lipoprotein cholesterol (LDL-C/HDL-C) ratio and triglyceride content. Sunflower seed protein fraction resulted in a significant decrease in plasma cholesterol (p < 0.05) and LDL-cholesterol (p < 0.02) levels compared to the casein fed rats. Membrane phospholipid profile also showed a marked variation with the type of dietary protein. Rats fed SSPF and SM did not show much variation in plasma lipids, plasma proteins, liver and brain lipids and membrane phospholipid concentrations. Protein content, liver and brain lipid profile of the groups fed SSPF and casein were comparable, suggesting that the nutritional value of SSPF is better than SM and equivalent to that of casein.

  13. Optimization of extraction of apple aroma by dynamic headspace and influence of saliva on extraction of volatiles.

    Science.gov (United States)

    Mehinagic, Emira; Prost, Carole; Demaimay, Michel

    2004-08-11

    The dynamic headspace procedure of aroma extraction was optimized on Gala apples (Malus domestica). Two parameters affecting the extractability of compounds were studied: temperature and purge time. The influence of artificial saliva was also included. An increase in purge time and temperature caused an increase in the extraction of volatiles from the apple matrix. The optimum point of extraction was 40 degrees C and 70 min of purge. The study also showed that the addition of saliva influenced the extraction of volatile compounds, but this effect was different from one compound to another. To verify that the headspace extracts presented a global odor representativeness of fresh apple under these conditions of extraction, eight assessors compared the odor of extracts with fresh fruit odor for three different cultivars. With regard to the sensory profiles of extracts, the optimal conditions of extraction were suitable for extraction of volatile compounds, even if cooked apple odor appeared in some extracts. The similarity marks of extracts were low but acceptable.

  14. Comparison of five methods for direct extraction of surface proteins from Listeria monocytogenes for proteomic analysis by orbitrap mass spectrometry.

    Science.gov (United States)

    Tiong, Hung King; Hartson, Steven; Muriana, Peter M

    2015-03-01

    Extracts of surface proteins, with minimal artifacts from contaminating cytosolic components, are highly desirable for investigating surface factors involved in the attachment and formation of biofilms by bacteria that are problematic in commercial food processing facilities. In this study, we compared the protein profiles of the food pathogen, Listeria monocytogenes, recovered after applying different surface protein extraction methods compiled from the literature: trypsin-enzymatic shaving with BICAM/sucrose or Tris/sucrose buffers (Tryp B+S, Tryp T+S), Tris-buffered urea (UB), lithium chloride (LiCl) and Tris-buffered urea applied with hypotonic-stressed cells (UB-Ghost), and subjected them to liquid chromatography tandem mass spectrometry and protein identification. The data indicate that the UB-Ghost extraction method provides a cleaner extract of surface proteins including the predicted (this study and the literature) or validated members (literature) from L. monocytogenes. This was determined by an accumulative lower unique peptide number exhibited by mass spectrometry for total cytoplasmic proteins among different surface extracts, with a majority of proteins demonstrating hydrophilic properties. The extracted proteins were from different functional categories and have associations with the cell surface, intermediary metabolism, information pathways, or functionally unknown proteins as suggested by in silico analyses performed by other groups (Leger and ListiList). The utilization of an optimized method for surface protein extraction should greatly facilitate identification by LC-MS/MS that could be useful to anyone working on molecular proteomics of bacterial surfaces.

  15. Optimization on Extraction Engineering of the Anti - inflammatory Bioactive Materials from Ainsliaea Fragrans Champ

    Directory of Open Access Journals (Sweden)

    Yang Jie

    2016-01-01

    Full Text Available Ainsliaea fragrans Champ.(A.fragrans is a traditional Chinese herbal, phenolic compounds was the major anti - inflammatory bioactive constituents. To improve the bioavailability and enhanced the curative effect of A.fragrans, the anti - inflammatory effect of phenolic acids and the “non-active” group of control vectors constitute a new biomedical material, which is of great significance to the treatment of diseases inflammation. Hence, in this thesis, regarding the total phenolic acid transfer rate as the indicator, L9(34 orthogonal design was used to optimize the extraction process of total Phenolic acid from A.fragrans by reflux extraction method on solvent dosage, extraction times and extraction time.The optimal extraction technology was as follows: 15 times of water volume, reflux extraction 3 times, extraction time 60 min. The result of pharmacological activity indicated anti-inflammatory effect: 95% ethanol extraction > water extraction > 30% ethanol extraction > 60% ethanol extraction.

  16. Optimizing the extraction of antibacterial compounds from pineapple leaf fiber

    National Research Council Canada - National Science Library

    Zhikai Zhuang; Jin Zhang; Mingfu Li; Ganran Deng; Zhongqing Ou; Wenwei Lian; Junyan He; Tao Huang

    2016-01-01

    ...) were used to extract antibacterial compounds from pineapple leaf fiber. Compounds extracted using acetone showed the greatest antibacterial effect against , measured by inhibition zone diameter...

  17. Optimization of Ultrasonic Extraction of Phenolic Antioxidants from Green Tea Using Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Lan-Sook Lee

    2013-10-01

    Full Text Available Response surface methodology (RSM has been used to optimize the extraction conditions of antioxidants with relatively low caffeine content from green tea by using ultrasonic extraction. The predicted optimal conditions for the highest antioxidant activity and minimum caffeine level were found at 19.7% ethanol, 26.4 min extraction time, and 24.0 °C extraction temperature. In the predicted optimal conditions, the experimental values were very close to the predicted values. Moreover, the ratio of (EGCg + ECg/EGC was identified a major factor contributing to the antioxidant activity of green tea extracts. In this study, ultrasonic extraction showed that the ethanol concentration and extraction time used for antioxidant extraction could be remarkably reduced without a decrease in antioxidant activity compared to the conventional extraction conditions.

  18. Towards plant protein refinery: Review on protein extraction using alkalo and potential enzymatic assistance

    NARCIS (Netherlands)

    Sari, Y.W.; Mulder, W.J.; Sanders, J.P.M.; Bruins, M.E.

    2015-01-01

    The globally increasing protein demands require additional resources to those currently available. Furthermore, the optimal usage of protein fractions from both traditional and new protein resources, such as algae and leaves, is essential. Here, we present an overview on alkaline plant protein extra

  19. Towards plant protein refinery: Review on protein extraction using alkalo and potential enzymatic assistance

    NARCIS (Netherlands)

    Sari, Y.W.; Mulder, W.J.; Sanders, J.P.M.; Bruins, M.E.

    2015-01-01

    The globally increasing protein demands require additional resources to those currently available. Furthermore, the optimal usage of protein fractions from both traditional and new protein resources, such as algae and leaves, is essential. Here, we present an overview on alkaline plant protein

  20. Extractable protein of radiation vulcanized natural rubber latex

    Energy Technology Data Exchange (ETDEWEB)

    Soebianto, Y.S. [Center for Research and Development of Isotopes and Radiation Technology, BATAN, Jakarta (Indonesia); Upul, R.M. [Rubber Research Institute of Sri Lanka, Ratmalana (Sri Lanka); Makuuchi, K.; Yoshii, F.; Kume, T. [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-03-01

    A new method to reduce the protein level in the latex products by irradiation is reported. Water soluble protein (WSP) solution (10%) was added into radiation vulcanized NR latex (RVNRL) as much as 3 phr in three different processes: added to RVNRL, added to re-centrifuged RVNRL, and added to RVNRL followed by centrifugation. The protein content was determined by enhanced BCA method, and identified by SDS-PAGE analysis. Addition of WSP followed by centrifugation reduces EP up to the minimum protein detection, and shortens the leaching time to 20-30 min. SDS-PAGE analysis confirms the reduction of soluble protein in the serum phase, and disappearance of protein bands in the rubber extract. Protein-WSP interaction produces water soluble complex, and removed by centrifugation. The molecular weight of WSP dictates the efficiency of protein removal. (author)

  1. Optimization of protein buffer cocktails using Thermofluor.

    Science.gov (United States)

    Reinhard, Linda; Mayerhofer, Hubert; Geerlof, Arie; Mueller-Dieckmann, Jochen; Weiss, Manfred S

    2013-02-01

    The stability and homogeneity of a protein sample is strongly influenced by the composition of the buffer that the protein is in. A quick and easy approach to identify a buffer composition which increases the stability and possibly the conformational homogeneity of a protein sample is the fluorescence-based thermal-shift assay (Thermofluor). Here, a novel 96-condition screen for Thermofluor experiments is presented which consists of buffer and additive parts. The buffer screen comprises 23 different buffers and the additive screen includes small-molecule additives such as salts and nucleotide analogues. The utilization of small-molecule components which increase the thermal stability of a protein sample frequently results in a protein preparation of higher quality and quantity and ultimately also increases the chances of the protein crystallizing.

  2. Impurities Removal in Seawater to Optimize the Magnesium Extraction

    Science.gov (United States)

    Natasha, N. C.; Firdiyono, F.; Sulistiyono, E.

    2017-02-01

    Magnesium extraction from seawater is promising way because magnesium is the second abundant element in seawater and Indonesia has the second longest coastline in the world. To optimize the magnesium extraction, the impurities in seawater need to be eliminated. Evaporation and dissolving process were used in this research to remove the impurities especially calcium in seawater. Seawater which has been evaporated from 100 ml to 50 ml was dissolved with variations solution such as oxalic acid and ammonium bicarbonate. The solution concentration is 100 g/l and it variations are 2 ml, 4 ml, 6 ml, 8 ml, 10 ml, 20 ml, 30 ml, 40 ml and 50 ml. This step will produce precipitate and filtrate then it will be analysed to find out the result of this process. The precipitate was analysed by X-ray Diffraction (XRD) and Scanning Electron Microscope (SEM) but the filtrate was analysed by Inductively Coupled Plasma (ICP). XRD analysis shows that calcium oxalate and calcium carbonate were formed and ICP analysis shows that the remaining calcium in seawater using oxalic acid is about 0.01% and sodium 0.14% but when using ammonium bicarbonate the remaining calcium is 2.5% and sodium still more than 90%. The results show that both oxalic acid and ammonium bicarbonate can remove the impurities but when using oxalic acid, not only the impurities but also magnesium was precipitated. The conclusion of this research is the best solution to remove the impurities in seawater without precipitate the magnesium is using ammonium bicarbonate.

  3. Optimization and evaluation of wheat germ oil extracted by supercritical CO{sub 2}

    Energy Technology Data Exchange (ETDEWEB)

    Jiang, S. T.; Niu, L.

    2011-07-01

    A Box-Behnken design combined with response surface methodology (RSM) was used to optimize the parameters of supercritical CO{sub 2} extraction (SFE) of wheat germ oil. The quality of the oil and residual meal obtained by SFE and solvent extraction (SE) were evaluated from proximate analysis, fatty acid composition and antioxidant activity. A maximum oil yield of 10.46% was achieved under the optimal conditions of wheat germ particle size 60-80 mesh; water content 4.37%; pressure 30MPa; temperature 40 degree centigrade extraction time 1.7h. The oil obtained by SFE showed stronger DPPH radical scavenging ability than SE oil at the same concentration. The fatty acid composition of SFE oil was similar to SE oil. Higher contents of protein (34.3%) and lysine (2.47g/100g) were found in the residual meal obtained by SFE. The results show that oil and defatted meal obtained by SFE can be promising nutritional sources for food. (Author) 20 refs.

  4. Optimizing Fungal DNA Extraction Methods from Aerosol Filters

    Science.gov (United States)

    Jimenez, G.; Mescioglu, E.; Paytan, A.

    2016-12-01

    Fungi and fungal spores can be picked up from terrestrial ecosystems, transported long distances, and deposited into marine ecosystems. It is important to study dust-borne fungal communities, because they can stay viable and effect the ambient microbial populations, which are key players in biogeochemical cycles. One of the challenges of studying dust-borne fungal populations is that aerosol samples contain low biomass, making extracting good quality DNA very difficult. The aim of this project was to increase DNA yield by optimizing DNA extraction methods. We tested aerosol samples collected from Haifa, Israel (polycarbonate filter), Monterey Bay, CA (quartz filter) and Bermuda (quartz filter). Using the Qiagen DNeasy Plant Kit, we tested the effect of altering bead beating times and incubation times, adding three freeze and thaw steps, initially washing the filters with buffers for various lengths of time before using the kit, and adding a step with 30 minutes of sonication in 65C water. Adding three freeze/thaw steps, adding a sonication step, washing with a phosphate buffered saline overnight, and increasing incubation time to two hours, in that order, resulted in the highest increase in DNA for samples from Israel (polycarbonate). DNA yield of samples from Monterey (quart filter) increased about 5 times when washing with buffers overnight (phosphate buffered saline and potassium phophate buffer), adding a sonication step, and adding three freeze and thaw steps. Samples collected in Bermuda (quartz filter) had the highest increase in DNA yield from increasing incubation to 2 hours, increasing bead beating time to 6 minutes, and washing with buffers overnight (phosphate buffered saline and potassium phophate buffer). Our results show that DNA yield can be increased by altering various steps of the Qiagen DNeasy Plant Kit protocol, but different types of filters collected at different sites respond differently to alterations. These results can be used as

  5. Optimization of Enzymatic Process for Vanillin Extraction Using Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Qinghuang Wang

    2012-07-01

    Full Text Available Vanillin was extracted from vanilla beans using pretreatment with cellulase to produce enzymatic hydrolysis, and response surface methodology (RSM was applied to optimize the processing parameters of this extraction. The effects of heating time, enzyme quantity and temperature on enzymatic extraction of vanillin were evaluated. Extraction yield (mg/g was used as the response value. The results revealed that the increase in heating time and the increase in enzyme quantity (within certain ranges were associated with an enhancement of extraction yield, and that the optimal conditions for vanillin extraction were: Heating time 6 h, temperature 60 °C and enzyme quantity 33.5 mL. Calculated from the final polynomial functions, the optimal response of vanillin extraction yield was 7.62 mg/g. The predicted results for optimal reaction conditions were in good agreement with experimental values.

  6. Preliminary study on total protein extraction methods from Enterococcus faecalis biofilm.

    Science.gov (United States)

    Chen, W; Liang, J; He, Z; Jiang, W

    2016-08-30

    Enterococcus faecalis is the major pathogen of post-endodontic disease and refractory periapical periodontitis, and recent research on this species has focused on its pathogenicity. E. faecalis most often causes disease in the form of a biofilm, and total protein expression shows a strong association with its virulence. Therefore, the purpose of our study was to explore different methods of extracting the total proteins of the E. faecalis (ATCC 33186 standard strain) biofilm. The total proteins in the biofilm were extracted using an ultrasonication method with varied parameters, including duration, amplitude setting, period, and duty cycle. After the optimal conditions of ultrasonication were determined based on the protein profile from sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, the total protein content in the biofilm was detected using the bicinchoninic acid assay, Bradford Coomassie brilliant blue assay, and Lowry assay, and the results were compared and analyzed. The parameters for the optimal conditions of ultrasonication were as follows: a processing duration of 2 min, amplitude setting of 20%, and ultrasonication period of 4 s at a 50% duty cycle. The total protein content was 2299.1 mg/dish when measured by the bicinchoninic assay, 3793.8 mg/dish when measured by the Bradford Coomassie brilliant blue assay, and 1858.0 mg/dish when measured by the Lowry assay. These results demonstrate that the Bradford Coomassie brilliant blue assay is a simple and feasible method for use in detecting the total protein content in a bacterial biofilm.

  7. Optimization study of fructans extraction from Agave tequilana Weber azul variety

    Directory of Open Access Journals (Sweden)

    Jesús Aarón SALAZAR-LEYVA

    Full Text Available Abstract Agave tequilana Weber azul variety is a valuable source of fructans. In this study, a response surface methodology was employed to optimize the extraction yield of fructans from agave pines. A Box Behnken factorial design was applied to evaluate the effects of several conditions such as extraction temperature, water raw material ratio and extraction time on extraction yield. Under the optimum extraction conditions (extraction temperature of 79.1 °C, water raw material ratio of 5.13 mL/g, and extraction time of 1.48 h fructans extraction yield was 83.12%. The chromatographic profile of the agave crude extract presented four peaks, out of which fructans were the most preponderant (~87%. The obtained results suggest that the response surface methodology is adequate to optimize fructans extraction from Agave tequilana Weber azul variety.

  8. A comparison of protein extraction methods suitable for gel-based proteomic studies of aphid proteins.

    Science.gov (United States)

    Cilia, M; Fish, T; Yang, X; McLaughlin, M; Thannhauser, T W; Gray, S

    2009-09-01

    Protein extraction methods can vary widely in reproducibility and in representation of the total proteome, yet there are limited data comparing protein isolation methods. The methodical comparison of protein isolation methods is the first critical step for proteomic studies. To address this, we compared three methods for isolation, purification, and solubilization of insect proteins. The aphid Schizaphis graminum, an agricultural pest, was the source of insect tissue. Proteins were extracted using TCA in acetone (TCA-acetone), phenol, or multi-detergents in a chaotrope solution. Extracted proteins were solubilized in a multiple chaotrope solution and examined using 1-D and 2-D electrophoresis and compared directly using 2-D Difference Gel Electrophoresis (2-D DIGE). Mass spectrometry was used to identify proteins from each extraction type. We were unable to ascribe the differences in the proteins extracted to particular physical characteristics, cell location, or biological function. The TCA-acetone extraction yielded the greatest amount of protein from aphid tissues. Each extraction method isolated a unique subset of the aphid proteome. The TCA-acetone method was explored further for its quantitative reliability using 2-D DIGE. Principal component analysis showed that little of the variation in the data was a result of technical issues, thus demonstrating that the TCA-acetone extraction is a reliable method for preparing aphid proteins for a quantitative proteomics experiment. These data suggest that although the TCA-acetone method is a suitable method for quantitative aphid proteomics, a combination of extraction approaches is recommended for increasing proteome coverage when using gel-based separation techniques.

  9. Simulation and optimization of continuous extractive fermentation with recycle system

    Science.gov (United States)

    Widjaja, Tri; Altway, Ali; Rofiqah, Umi; Airlangga, Bramantyo

    2017-05-01

    Extractive fermentation is continuous fermentation method which is believed to be able to substitute conventional fermentation method (batch). The recovery system and ethanol refinery will be easier. Continuous process of fermentation will make the productivity increase although the unconverted sugar in continuous fermentation is still in high concentration. In order to make this process more efficient, the recycle process was used. Increasing recycle flow will enhance the probability of sugar to be re-fermented. However, this will make ethanol enter fermentation column. As a result, the accumulated ethanol will inhibit the growth of microorganism. This research aims to find optimum conditions of solvent to broth ratio (S:B) and recycle flow to fresh feed ratio in order to produce the best yield and productivity. This study employed optimization by Hooke Jeeves method using Matlab 7.8 software. The result indicated that optimum condition occured in S: B=2.615 and R: F=1.495 with yield = 50.2439 %.

  10. OPTIMIZATION OF DYEING PARAMETERS TO DYE COTTON WITH CARROT EXTRACTION

    Directory of Open Access Journals (Sweden)

    MIRALLES Verónica

    2017-05-01

    Full Text Available Natural dyes derived from flora and fauna are believed to be safe because of non-toxic, non-carcinogenic and biodegradable nature. Furthermore, natural dyes do not cause pollution and waste water problems. Natural dyes as well as synthetic dyes need the optimum parameters to get a good dyeing. On some occasions, It is necessary the use of mordants to increase the affinity between cellulose fiber and natural dye, but there are other conditions to optimize in the dyeing process, like time, temperature, auxiliary porducts, etc. In addition, the optimum conditions are different depends on the type of dye and the fiber nature. The aim of this work is the use of carrot extract to dye cotton fabric by exhaustion at diverse dyeing conditions. Diffferent dyeing processes were carried out to study the effect of pH condition and the temperature, using 7, 6 and 4 pH values and 95 ºC and 130ºC for an hour. As a result some images of dyed samples are shown. Moreover, to evaluate the colour of each sample CIELAB parameters are analysed obtained by reflexion spectrophotometre. The results showed that the temperature used has an important influence on the colour of the dyed sample.

  11. A computational framework for the design of optimal protein synthesis.

    Science.gov (United States)

    Racle, Julien; Overney, Jan; Hatzimanikatis, Vassily

    2012-08-01

    Despite the establishment of design principles to optimize codon choice for heterologous expression vector design, the relationship between codon sequence and final protein yield remains poorly understood. In this work, we present a computational framework for the identification of a set of mutant codon sequences for optimized heterologous protein production, which uses a codon-sequence mechanistic model of protein synthesis. Through a sensitivity analysis on the optimal steady state configuration of protein synthesis we are able to identify the set of codons, that are the most rate limiting with respect to steady state protein synthesis rate, and we replace them with synonymous codons recognized by charged tRNAs more efficient for translation, so that the resulting codon-elongation rate is higher. Repeating this procedure, we iteratively optimize the codon sequence for higher protein synthesis rate taking into account multiple constraints of various types. We determine a small set of optimized synonymous codon sequences that are very close to each other in sequence space, but they have an impact on properties such as ribosomal utilization or secondary structure. This limited number of sequences can then be offered for further experimental study. Overall, the proposed method is very valuable in understanding the effects of the different properties of mRNA sequences on the final protein yield in heterologous protein production and it can find applications in synthetic biology and biotechnology.

  12. [Extraction and application of Perna viridis foot protein as bioadhesive].

    Science.gov (United States)

    Jiang, Zhen; Liu, Jiapeng; Jin, Lihua; Zhang, Qiqing

    2010-12-01

    Mussel adhesive proteins have attracted increasing interests for their potential use as environmentally friendly bioadhesives in medicine and aqueous conditions. In this study, surface coating analysis, quartz crystal microbalance (QCM), cell and bone tissue adhesion and cytotoxicity assay were used to study the properties of the Perna viridis foot proteins (Pvfp) extract as bioadhesive. The results of coating ability on various materials and QCM analysis revealed that Pvfp extract has comparable or superior adsorbtion ability to that of Cell-Tak (the naturally extracted MAP mixture from Mytilus edulis, and has been commercialized), and also, the cell adhesion ability of Pvfp extract was stronger than that of Cell-Tak and poly-L-lysine. No cytotoxicity was detected using human HeLa and 293T cells. Furthermore, broken bones of mouse could be stuck together by use of Pvfp extract. In bulk-scale adhesion tests, Pvfp extract showed much greater tensile strength than did fibrin glue for conglutinating poly (vinl chloride) sticks and for binding together pig's femur segments. These results suggested that Pvfp extract be an efficient cell and tissue adhesive in biotechnological application and it might be a potential bioadhesive in medical practice.

  13. BIOMARKERS TO DEFINE OPTIMAL PROTEIN REQUIREMENT

    OpenAIRE

    Di Girolamo, Filippo Giorgio

    2015-01-01

    Dietary proteins are the source of the amino acids required by the body for tissue growth and maintenance. The Population Reference Intake (PRI) for proteins, as defined by the European Food Safety Authority (EFSA) for healthy adults, including the elderly, is 0.83 g/kg body weight/day. This amount is defined on the net balance of body protein (or “nitrogen balance”, given by the difference between dietary nitrogen intake and losses) equivalent to 0.66 g/kg/day plus a safety factor for interp...

  14. Structuring detergents for extracting and stabilizing functional membrane proteins.

    Directory of Open Access Journals (Sweden)

    Rima Matar-Merheb

    Full Text Available BACKGROUND: Membrane proteins are privileged pharmaceutical targets for which the development of structure-based drug design is challenging. One underlying reason is the fact that detergents do not stabilize membrane domains as efficiently as natural lipids in membranes, often leading to a partial to complete loss of activity/stability during protein extraction and purification and preventing crystallization in an active conformation. METHODOLOGY/PRINCIPAL FINDINGS: Anionic calix[4]arene based detergents (C4Cn, n=1-12 were designed to structure the membrane domains through hydrophobic interactions and a network of salt bridges with the basic residues found at the cytosol-membrane interface of membrane proteins. These compounds behave as surfactants, forming micelles of 5-24 nm, with the critical micellar concentration (CMC being as expected sensitive to pH ranging from 0.05 to 1.5 mM. Both by 1H NMR titration and Surface Tension titration experiments, the interaction of these molecules with the basic amino acids was confirmed. They extract membrane proteins from different origins behaving as mild detergents, leading to partial extraction in some cases. They also retain protein functionality, as shown for BmrA (Bacillus multidrug resistance ATP protein, a membrane multidrug-transporting ATPase, which is particularly sensitive to detergent extraction. These new detergents allow BmrA to bind daunorubicin with a Kd of 12 µM, a value similar to that observed after purification using dodecyl maltoside (DDM. They preserve the ATPase activity of BmrA (which resets the protein to its initial state after drug efflux much more efficiently than SDS (sodium dodecyl sulphate, FC12 (Foscholine 12 or DDM. They also maintain in a functional state the C4Cn-extracted protein upon detergent exchange with FC12. Finally, they promote 3D-crystallization of the membrane protein. CONCLUSION/SIGNIFICANCE: These compounds seem promising to extract in a functional state

  15. Designed Proteins as Optimized Oxygen Carriers for Artificial Blood

    Science.gov (United States)

    2014-02-01

    transport throughout the body. In year two, we developed a new model for oxyferrous state lifetimes, including an equation which predicts an O2...chain four helix bundle. Table 1 demonstrates that the addition of the optimized binding site to both ligating helices of the full chain more than... triples the lifetime. Table 1. Oxyferrous lifetime for single chain proteins with the optimal binding site Protein ligation Rair(s-1) Kd,O2 (mM) kox

  16. Pectin from Opuntia ficus indica: Optimization of microwave-assisted extraction and preliminary characterization.

    Science.gov (United States)

    Lefsih, Khalef; Giacomazza, Daniela; Dahmoune, Farid; Mangione, Maria Rosalia; Bulone, Donatella; San Biagio, Pier Luigi; Passantino, Rosa; Costa, Maria Assunta; Guarrasi, Valeria; Madani, Khodir

    2017-04-15

    Optimization of microwave-assisted extraction (MAE) of water-soluble pectin (WSP) from Opuntia ficus indica cladodes was performed using Response Surface Methodology. The effect of extraction time (X1), microwave power (X2), pH (X3) and solid-to-liquid ratio (X4) on the extraction yield was examined. The optimum conditions of MAE were as follows: X1=2.15min; X2=517W; X3=2.26 and X4=2g/30.6mL. The maximum obtained yield of pectin extraction was 12.57%. Total carbohydrate content of WSP is about 95.5% including 34.4% of Galacturonic acid. Pectin-related proteins represent only the 0.66% of WSP mass. HPSEC and light scattering analyses reveal that WSP is mostly constituted of high molecular pectin and FTIR measurements show that the microwave treatment does not alter the chemical structure of WSP, in which Galacturonic acid content and yield are 34.4% and 4.33%, respectively. Overall, application of MAE can give rise to high quality pectin.

  17. Codon optimizing for increased membrane protein production

    DEFF Research Database (Denmark)

    Mirzadeh, K.; Toddo, S.; Nørholm, Morten

    2016-01-01

    . As demonstrated with two membrane-embedded transporters in Escherichia coli, the method was more effective than optimizing the entire coding sequence. The method we present is PCR based and requires three simple steps: (1) the design of two PCR primers, one of which is degenerate; (2) the amplification...

  18. Extraction optimization of water-extracted mycelial polysaccharide from endophytic fungus Fusarium oxysporum Dzf17 by response surface methodology.

    Science.gov (United States)

    Li, Peiqin; Lu, Shiqiong; Shan, Tijiang; Mou, Yan; Li, Yan; Sun, Weibo; Zhou, Ligang

    2012-01-01

    Water-extracted mycelial polysaccharide (WPS) from the endophytic fungus Fusarium oxysporum Dzf17 isolated from Dioscorea zingiberensis was found to be an efficient elicitor to enhance diosgenin accumulation in D. zingigerensis cultures, and also demonstrated antioxidant activity. In this study, response surface methodology (RSM) was employed to optimize the extraction process of WPS from F. oxysporum Dzf17 using Box-Behnken design (BBD). The ranges of the factors investigated were 1-3 h for extraction time (X(1)), 80-100 °C for extraction temperature (X(2)), and 20-40 (v/w) for ratio of water volume (mL) to raw material weight (g) (X(3)). The experimental data obtained were fitted to a second-order polynomial equation using multiple regression analysis. Statistical analysis showed that the polynomial regression model was in good agreement with the experimental results with the determination coefficient (R(2)) of 0.9978. By solving the regression equation and analyzing the response surface contour plots, the extraction parameters were optimized as 1.7 h for extraction time, 95 °C for extraction temperature, 39 (v/w) for ratio of water volume (mL) to raw material weight (g), and with 2 extractions. The maximum value (10.862%) of WPS yield was obtained when the WPS extraction process was conducted under the optimal conditions.

  19. Dynamic optimization of the benzene extractive distillation unit

    National Research Council Canada - National Science Library

    Ghaee, A; Sotudeh-Gharebagh, R; Mostoufi, N

    2008-01-01

    A mathematical model has been developed for describing the dynamic operation of the N-formylmorpholine extractive distillation column and the corresponding solvent recovery column in the benzene extraction plant...

  20. Optimizing an emperical scoring function for transmembrane protein structure determination.

    Energy Technology Data Exchange (ETDEWEB)

    Young, Malin M.; Sale, Kenneth L.; Gray, Genetha Anne; Kolda, Tamara Gibson

    2003-10-01

    We examine the problem of transmembrane protein structure determination. Like many other questions that arise in biological research, this problem cannot be addressed by traditional laboratory experimentation alone. An approach that integrates experiment and computation is required. We investigate a procedure which states the transmembrane protein structure determination problem as a bound constrained optimization problem using a special empirical scoring function, called Bundler, as the objective function. In this paper, we describe the optimization problem and some of its mathematical properties. We compare and contrast results obtained using two different derivative free optimization algorithms.

  1. Optimizing an emperical scoring function for transmembrane protein structure determination.

    Energy Technology Data Exchange (ETDEWEB)

    Young, Malin M.; Sale, Kenneth L.; Gray, Genetha Anne; Kolda, Tamara Gibson

    2003-10-01

    We examine the problem of transmembrane protein structure determination. Like many other questions that arise in biological research, this problem cannot be addressed by traditional laboratory experimentation alone. An approach that integrates experiment and computation is required. We investigate a procedure which states the transmembrane protein structure determination problem as a bound constrained optimization problem using a special empirical scoring function, called Bundler, as the objective function. In this paper, we describe the optimization problem and some of its mathematical properties. We compare and contrast results obtained using two different derivative free optimization algorithms.

  2. Optimization of membrane protein overexpression and purification using GFP fusions

    NARCIS (Netherlands)

    Drew, David; Lerch, Mirjam; Kunji, Edmund; Slotboom, Dirk-Jan; de Gier, Jan-Willem

    2006-01-01

    Optimizing conditions for the overexpression and purification of membrane proteins for functional and structural studies is usually a Laborious and time-consuming process. This process can be accelerated using membrane protein-GFP fusions(1-3), which allows direct monitoring and visualization of mem

  3. Algorithm Engineering for Optimal Alignment of Protein Structure Distance Matrices

    NARCIS (Netherlands)

    Wohlers, I.; Andonov, R.; Klau, G.W.

    2011-01-01

    Protein structural alignment is an important problem in computational biology. In this paper, we present first successes on provably optimal pairwise alignment of protein inter-residue distance matrices, using the popular DALI scoring function. We introduce the structural alignment problem formally,

  4. Depletion mapping and constrained optimization to support managing groundwater extraction

    Science.gov (United States)

    Fienen, Michael N.; Bradbury, Kenneth R.; Kniffin, Maribeth; Barlow, Paul M.

    2017-01-01

    Groundwater models often serve as management tools to evaluate competing water uses including ecosystems, irrigated agriculture, industry, municipal supply, and others. Depletion potential mapping—showing the model-calculated potential impacts that wells have on stream baseflow - can form the basis for multiple potential management approaches in an oversubscribed basin. Specific management approaches can include scenarios proposed by stakeholders, systematic changes in well pumping based on depletion potential, and formal constrained optimization, which can be used to quantify the tradeoff between water use and stream baseflow. Variables such as the maximum amount of reduction allowed in each well and various groupings of wells using, for example, K-means clustering considering spatial proximity and depletion potential are considered. These approaches provide a potential starting point and guidance for resource managers and stakeholders to make decisions about groundwater management in a basin, spreading responsibility in different ways. We illustrate these approaches in the Little Plover River basin in central Wisconsin, United States—home to a rich agricultural tradition, with farmland and urban areas both in close proximity to a groundwater-dependent trout stream. Groundwater withdrawals have reduced baseflow supplying the Little Plover River below a legally established minimum. The techniques in this work were developed in response to engaged stakeholders with various interests and goals for the basin. They sought to develop a collaborative management plan at a watershed scale that restores the flow rate in the river in a manner that incorporates principles of shared governance and results in effective and minimally disruptive changes in groundwater extraction practices.

  5. Continuous protein recovery from whey using liquid-solid circulating fluidized bed ion-exchange extraction.

    Science.gov (United States)

    Lan, Qingdao; Bassi, Amarjeet; Zhu, Jing-Xu Jesse; Margaritis, Argyrios

    2002-04-20

    A liquid-solid circulating fluidized bed (LSCFB) continuous ion-exchange extraction system has been investigated for total protein recovery from whey solutions under various operating conditions. The effectiveness of a dynamic seal was evaluated between the riser and the downcomer, and the best conditions for the establishment of this seal were established. Start-up studies indicated that the system is robust and stable. Under optimal conditions, a productivity of 8.2 g of total protein removed per hour per kilogram of resin was achieved with a protein removal efficiency of 78.4%. However, higher overall protein recovery of up to 90% was also achieved under other conditions, with lower protein concentration in the effluent and a lower overall productivity.

  6. Sequential extractions: A new way for protein quantification-data from peanut allergens.

    Science.gov (United States)

    Zhou, Ningling; Li, Wenying; Wu, Zhihua; Li, Xin; Yang, Anshu; Tong, Ping; Chen, Hongbing

    2015-09-01

    Quantification of certain protein contents in the matrix is essential in protein analyses. The amount of total protein in the matrix can be determined by the Kjeldahl method. However, few methods can quantify certain protein contents in the matrix without extracting all of them in solution. Extracting all of the contents is difficult for proteins, especially relatively insoluble ones. A five-step sequential extraction method was developed for the quantification of certain proteins in defatted peanut flour based on the relationship between the extracted protein contents and the extraction times. The extracted proteins (i.e., total protein, Ara h 1, and Ara h 2) were quantitatively analyzed in each extraction of the same condition. An exponential equation was obtained between the extraction times and the respective amount of extracted protein as well as both the total protein and a particular protein. In particular, the amount of protein extracted each time can be a geometric sequence. If all proteins can be extracted with sufficient extraction times, the protein contents in the peanut matrix can be calculated using a mathematical summation formula. This sum should be all proteins in the matrix. The five-step sequential extraction method can provide a means to quantify certain proteins in the matrix.

  7. A Shortest Dependency Path Based Convolutional Neural Network for Protein-Protein Relation Extraction

    OpenAIRE

    2016-01-01

    The state-of-the-art methods for protein-protein interaction (PPI) extraction are primarily based on kernel methods, and their performances strongly depend on the handcraft features. In this paper, we tackle PPI extraction by using convolutional neural networks (CNN) and propose a shortest dependency path based CNN (sdpCNN) model. The proposed method (1) only takes the sdp and word embedding as input and (2) could avoid bias from feature selection by using CNN. We performed experiments on sta...

  8. Topology optimization of pulse shaping filters using the Hilbert transform envelope extraction

    DEFF Research Database (Denmark)

    Lazarov, Boyan Stefanov; Matzen, René; Elesin, Yuriy

    2011-01-01

    Time domain topology optimization is applied to design pulse shaping filters. The objective function depends on the pulse envelope, which is extracted by utilizing the Hilbert transform. The gradients with respect to the topology optimization variables are derived, and the optimization methodology...

  9. Recent Advances in Protein Extraction and Chiral Separation of Biomolecules

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Reverse micelles create unique environment in organic media. They are capable of solubilizing hydrophilic biomolecules (e.g., proteins, peptides, amino acids, and DNAs) in their aqueous interior. This feature brings about the practical use of biomaterials in organic media because reverse micelles solubilize them with the intrinsic activity. In this paper, we focus on recent two topics concerning protein extraction and chiral separation of biomolecules using liquid membranes. In the first topic, we present recent attempts to extract proteins from an aqueous solution into isooctane using reverse micelles, and some important operational parameters to achieve an efficient protein transfer are discussed. Furthermore, novel function of reverse micelles as a protein activation medium is introduced. In the reverse micellar phase, denatured proreins were completely reactivated in the reverse micellar solution. The reverse micellar technique is found to be a useful tool not only for protein separation but also for protein refolding. Furthermore, we found that a cyclic ligand carixarene has an extraction ability to set up optimum conditions for protein transfer. In the second topic, we have found that a supported liquid membrane (SLM) encapsulating enzymes shows high enantioselectivity (enantioselective excess value is over 96%) in the transport of racemic pharmaceutical compound ibuprofen. A different experiment also suggests that the α-chymotrypsin-catalyzed reactions droved the enantioselective transport of L-phenylalanine based on the enantioselectivity of the enzyme. The SLM encapsulating the surfactant-enzyme complex enabled the highly enantioselective separation of racemic mixtures. It can be envisioned that arrangement of appropriate enzymes in the SLM system will allow enantioselective separation of various useful organic compounds.

  10. Combining optimal control theory and molecular dynamics for protein folding.

    Science.gov (United States)

    Arkun, Yaman; Gur, Mert

    2012-01-01

    A new method to develop low-energy folding routes for proteins is presented. The novel aspect of the proposed approach is the synergistic use of optimal control theory with Molecular Dynamics (MD). In the first step of the method, optimal control theory is employed to compute the force field and the optimal folding trajectory for the Cα atoms of a Coarse-Grained (CG) protein model. The solution of this CG optimization provides an harmonic approximation of the true potential energy surface around the native state. In the next step CG optimization guides the MD simulation by specifying the optimal target positions for the Cα atoms. In turn, MD simulation provides an all-atom conformation whose Cα positions match closely the reference target positions determined by CG optimization. This is accomplished by Targeted Molecular Dynamics (TMD) which uses a bias potential or harmonic restraint in addition to the usual MD potential. Folding is a dynamical process and as such residues make different contacts during the course of folding. Therefore CG optimization has to be reinitialized and repeated over time to accomodate these important changes. At each sampled folding time, the active contacts among the residues are recalculated based on the all-atom conformation obtained from MD. Using the new set of contacts, the CG potential is updated and the CG optimal trajectory for the Cα atoms is recomputed. This is followed by MD. Implementation of this repetitive CG optimization-MD simulation cycle generates the folding trajectory. Simulations on a model protein Villin demonstrate the utility of the method. Since the method is founded on the general tools of optimal control theory and MD without any restrictions, it is widely applicable to other systems. It can be easily implemented with available MD software packages.

  11. Optimization of Extraction Parameters for Natural Dye from Pterocarpus santalinus by using Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Hemanthraj.K.P.M

    2014-09-01

    Full Text Available Pterocarpus species has been admired for centuries for its dye, beautiful color, hardness and durability. The present study deals with the extraction of natural dye from Pterocarpus wood materials. Response surface methodology was used to study the optimal conditions for the extraction of dye. Factors such as extraction temperature, extraction time, and solid to liquid ratio were identified to be significantly affecting natural dye extraction efficiency. By using three-level three-factor Box-Behnken design, the optimized conditions for dye extraction by different techniques such as Solvent, Ultrasonic and Microwave extraction method. Microwave assisted extraction method showed the highest natural dye yield percentage which is 50.0 for ethyl acetate solvent and 50.2 for methanol solvent.

  12. Response Surface Optimized Infrared-Assisted Extraction and UHPLC Determination of Flavonoid Types from Flos Sophorae.

    Science.gov (United States)

    Mou, Qianqian; He, Jingxia; Yin, Rongli; Yang, Bin; Fu, Meihong; Fang, Jing; Li, Hua

    2017-06-15

    Single-factor experiment and Box-Behnken design were applied to optimize the infrared-assisted extraction (IRAE) of rutin, quercetin, kaempferol, and isorhamnetin from Flos sophorae. Four factors (extract solvent, solid-liquid ration, extraction time, infrared power) affecting the extraction yield of flavonoids were studied. Under optimized conditions the extraction yield was 33.199 ± 0.24 mg/g, which substantially improved, compared with heating reflux extraction (HRE) and ultrasonic-assisted extraction (UAE), while extraction time was only 9 min. The eluents were rich in 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azobis (2-methyl-propionamidine) dihydrochloride radical scavenging potential (IC50 of DPPH: 53.44 ± 0.01 μg/mL, oxygen radical absorbance capacity (ORAC): 3785.83 ± 52 μmol/g) than the extracts obtained by HRE and UAE. In addition, an ultra-high performance liquid chromatography method was optimized for the identification and quantification of the tested flavonoids, and the method was validated based on its correlation coefficient (r), reproducibility (RSD, n = 5), and recovery values, which were 0.9994-0.9998, 0.74-1.83%, and 97.78-102.94%, respectively. These results confirmed that high extraction yield of flavonoids results in stronger antioxidant values and response surface methodology optimization of IRAE is a promising alternative to traditional extraction techniques for flavonoids from medicinal plants.

  13. PRELIMINARY STUDY OF EXTRACTABLE PROTEIN BINDING USING MALEIC ANHYDRIDE COPOLYMER

    Institute of Scientific and Technical Information of China (English)

    Thirawan Nipithakul; Ladawan Watthanachote; Nanticha Kalapat

    2012-01-01

    A preliminary study of using maleic anhydride copolymer for protein binding has been carried out.The polymeric films were prepared by compression of the purified resin and annealing the film to induce efficient back formation of the anhydride groups.The properties of the film surface were analyzed by attenuated total reflection Fourier transforms infrared spectroscopy and water contact angle measurements.The protein content was determined by Bradford assay.To obtain optimum conditions,immersion time for protein binding was examined.Results revealed that proteins can be successfully immobilized onto the film surface via covalent linkage.The efficiency of the covalent binding of the extractable protein to maleic anhydride-polyethylene film was estimated at 69.87 μtg/cm2,although the film had low anhydride content (3%) on the surface.

  14. Proteins identified from care solution extractions of silicone hydrogels.

    Science.gov (United States)

    Emch, Andrew J; Nichols, Jason J

    2009-02-01

    The purpose of this study was to investigate the quantity and identify the proteins extracted from two different types of silicone hydrogel contact lenses by several multipurpose care solutions after 1 day of wear. Ten subjects were recruited to wear galyfilcon A lenses (Acuvue Advance, Vistakon) followed by lotrafilcon B lenses (O2 Optix, CIBA Vision) each for four consecutive days. Each day, subjects inserted a new pair of lenses for 8 h of wear after which both lenses were removed using forceps (lenses were not rubbed or rinsed after removal). Lenses were pooled in one of four commercially available care solutions for a 24-h soak followed by precipitation, resuspension in water, and quantification by Bradford assay and identification by mass spectrometry. Protein recovery from care solutions was as follows (quantities are in microg/lens): AQuify (galyfilcon A: 0.56, lotrafilcon B: 1.24), Complete MoisturePlus (galyfilcon A: 1.44, lotrafilcon B: 1.47), Opti-Free Express (galyfilcon A: 2.31, lotrafilcon B: 5.67), and ReNu MoistureLoc (galyfilcon A: 1.17, lotrafilcon B: 4.38). For each care solution, greater quantities of protein were removed from lotrafilcon B (3.19 +/- 2.19 microg/lens) than from galyfilcon A (1.37 +/- 0.72 microg/lens). Lactoferrin, lysozyme, and lipocalin were the most commonly identified, whereas various keratin compounds and other unique proteins were also detected. Opti-Free Express was consistently associated with the more efficient removal of proteins from these silicone hydrogels. More total protein was removed from lotrafilcon B than from galyfilcon A (approximately 2 x more protein) for all four care solutions, and 12 total unique protein species were recovered from galyfilcon A, whereas only 10 were recovered from lotrafilcon B. The higher quantities of protein extracted from lotrafilcon B may be due to stronger protein binding with this material and/or to differences in solution efficacy.

  15. Optimization of Enzymatic Process for Vanillin Extraction Using Response Surface Methodology

    OpenAIRE

    Qinghuang Wang; Zhong Chu; Huasong Wu; Lehe Tan; Fei Xu; Fenglin Gu

    2012-01-01

    Vanillin was extracted from vanilla beans using pretreatment with cellulase to produce enzymatic hydrolysis, and response surface methodology (RSM) was applied to optimize the processing parameters of this extraction. The effects of heating time, enzyme quantity and temperature on enzymatic extraction of vanillin were evaluated. Extraction yield (mg/g) was used as the response value. The results revealed that the increase in heating time and the increase in enzyme quantity (within certain ran...

  16. Optimization of the Ocimum basilicum L. extraction process regarding the antioxidant activity

    OpenAIRE

    Vidović Senka S.; Zeković Zoran P.; Lepojević Žika D.; Radojković Marija M.; Jokić Stela D.; Anačkov Goran T.

    2012-01-01

    The levels of input variables (temperature and extraction solvent) that optimize a particular response (total phenols content, total flavonoids content and antioxidant activity) of the Ocimum basilicum L. extraction process were determined by the response surface methodology (RSM). The influence of theextraction temperature on extraction process was investigated in the range from 33.8ºC to 76.2ºC, as well as of extraction solvent ethanol, in the range of concentrations from 21.7% to 78....

  17. Optimization of Process Parameters and Kinetic Model of Enzymatic Extraction of Polyphenols from Lonicerae Flos

    OpenAIRE

    Kong,Fansheng; Yu, Shujuan; Bi, Yongguang; Huang, Xiaojun; Huang, Mengqian

    2016-01-01

    Objective: To optimize and verify the cellulase extraction of polyphenols from honeysuckle and provide a reference for enzymatic extracting polyphenols from honeysuckle. Materials and Methods: The uniform design was used According to Fick's first law and kinetic model, fitting analysis of the dynamic process of enzymatic extracting polyphenols was conducted. Results: The optimum enzymatic extraction parameters for polyphenols from honeysuckle are found to be 80% (v/v) of alcohol, 35:1 (mL/g) ...

  18. Deep eutectic solvents as novel extraction media for protein partitioning.

    Science.gov (United States)

    Zeng, Qun; Wang, Yuzhi; Huang, Yanhua; Ding, Xueqin; Chen, Jing; Xu, Kaijia

    2014-05-21

    Four kinds of green deep eutectic solvent (DES) were synthesized, including choline chloride (ChCl)-urea, tetramethylammonium chloride (TMACl)-urea, tetrapropylammonium bromide (TPMBr)-urea and ChCl-methylurea. An aqueous two-phase system (ATPS) based ChCl-urea DES was studied for the first time for the extraction of bovine serum albumin (BSA). Single factor experiments proved that the extraction efficiency of BSA was influenced by the mass of the DES, concentration of K2HPO4 solution, separation time and extraction temperature. The optimum conditions were determined through an orthogonal experiment with the four factors described above. The results showed that under the optimum conditions, the average extraction efficiency could reach up to 99.94%, 99.72%, 100.05% and 100.05% (each measured three times). The relative standard deviations (RSD) of extraction efficiencies in precision, repeatability and stability experiments were 0.5533% (n = 5), 0.8306% (n = 5) and 0.9829% (n = 5), respectively. UV-vis and FT-IR spectra confirmed that there were no chemical interactions between BSA and the DES in the extraction process, and the CD spectra proved that the conformation of BSA did not change after extraction. The conductivity, DLS and TEM were combined to investigate the microstructure of the top phase and the possible mechanism for the extraction. The results showed that hydrophobic interactions, hydrogen bonding interactions and the salting-out effect played important roles in the transfer process, and the aggregation and surrounding phenomenon were the main driving forces for the separation. All of these results proved that ionic liquid (IL)-based ATPSs could potentially be substituted with DES-based ATPSs to offer new possibilities in the extraction of proteins.

  19. Improved hybrid optimization algorithm for 3D protein structure prediction.

    Science.gov (United States)

    Zhou, Changjun; Hou, Caixia; Wei, Xiaopeng; Zhang, Qiang

    2014-07-01

    A new improved hybrid optimization algorithm - PGATS algorithm, which is based on toy off-lattice model, is presented for dealing with three-dimensional protein structure prediction problems. The algorithm combines the particle swarm optimization (PSO), genetic algorithm (GA), and tabu search (TS) algorithms. Otherwise, we also take some different improved strategies. The factor of stochastic disturbance is joined in the particle swarm optimization to improve the search ability; the operations of crossover and mutation that are in the genetic algorithm are changed to a kind of random liner method; at last tabu search algorithm is improved by appending a mutation operator. Through the combination of a variety of strategies and algorithms, the protein structure prediction (PSP) in a 3D off-lattice model is achieved. The PSP problem is an NP-hard problem, but the problem can be attributed to a global optimization problem of multi-extremum and multi-parameters. This is the theoretical principle of the hybrid optimization algorithm that is proposed in this paper. The algorithm combines local search and global search, which overcomes the shortcoming of a single algorithm, giving full play to the advantage of each algorithm. In the current universal standard sequences, Fibonacci sequences and real protein sequences are certified. Experiments show that the proposed new method outperforms single algorithms on the accuracy of calculating the protein sequence energy value, which is proved to be an effective way to predict the structure of proteins.

  20. Optimization of ultrasonic cell grinder extraction of anthocyanins from blueberry using response surface methodology.

    Science.gov (United States)

    Jiang, Hai-Long; Yang, Jun-Li; Shi, Yan-Ping

    2017-01-01

    Ultrasonic cell grinder extraction (UCGE), using water as the solvent, was firstly applied to extract anthocyanins from blueberry. Extraction yield was related with four variables, including ratio of solution to solid, extraction power, buffer time, and extraction time. On the basis of response surface methodology (RSM), the optimal conditions were determined to be the ratio of solution to solid as 25:1(mL/g), the extraction power as 1500W, the buffer time as 3.0s, and the extraction time as 40min. The experimental yield of anthocyanins using UCGE was 2.89mg/g higher than that of conventional ultrasound-assisted extraction (CUAE). This study indicated that UCGE was an innovative, efficient, and environment friendly method in ultrasonic extraction fields, and had a potential to effectively extract other bioactive constituents.

  1. Improved protein extraction and protein identification from archival formalin-fixed paraffin-embedded human aortas.

    Science.gov (United States)

    Fu, Zongming; Yan, Kun; Rosenberg, Avraham; Jin, Zhicheng; Crain, Barbara; Athas, Grace; Heide, Richard S Vander; Howard, Timothy; Everett, Allen D; Herrington, David; Van Eyk, Jennifer E

    2013-04-01

    Evaluate combination of heat and elevated pressure to enhance protein extraction and quality of formalin-fixed (FF), and FF paraffin-embedded (FFPE) aorta for proteomics. Proteins were extracted from fresh frozen aorta at room temperature (RT). FF and FFPE aortas (3 months and 15 years) were extracted at RT, heat alone, or a combination of heat and high pressure. Protein yields were compared, and digested peptides from the extracts were analyzed with MS. Combined heat and elevated pressure increased protein yield from human FF or FFPE aorta compared to matched tissues with heat alone (1.5-fold) or at RT (8.3-fold), resulting in more proteins identified and with more sequence coverage. The length of storage did adversely affect the quality of proteins from FF tissue. For long-term storage, aorta was preserved better with FFPE than FF alone. Periostin and MGF-E8 were demonstrated suitable for MRM assays from FFPE aorta. Combination of heat and high pressure is an effective method to extract proteins from FFPE aorta for downstream proteomics. This method opens the possibility for use of archival and often rare FFPE aortas and possibly other tissues available to proteomics for biomarker discovery and quantification. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. Large-scale extraction of recombinant proteins from bacteria.

    Science.gov (United States)

    Simpson, Richard J

    2010-09-01

    Bacteria are particularly convenient for producing recombinant proteins for purification purposes. Suitable extraction methods for bacterial cells include sonication, glass bead milling, grinding with alumina or sand, high-pressure shearing using the French pressure cell (French Press), and lysozyme treatment. These procedures are applicable for preparing extracts from a variety of gram-negative bacteria such as Escherichia coli and Klebsiella pneumoniae, and gram-positive bacteria such as Bacillus subtilis. Disruption of bacterial cells by enzymatic means is commonly used because a relatively uniform treatment is obtained when cells are in suspension. This protocol describes the enzymatic disruption of E. coli.

  3. Extraction optimization of polyphenols, antioxidant and xanthine oxidase inhibitory activities from Prunus salicina Lindl.

    Directory of Open Access Journals (Sweden)

    Yibin LI

    2016-01-01

    Full Text Available Abstract Optimization of polyphenols extraction from plum (Prunus salicina Lindl. was evaluated using response surface methodology. The Box-Behnken experimental results showed the optimal conditions involved an extraction temperature of 59 °C, a sonication time of 47 min, and an ethanol concentration of 61% respectively. The maximum extraction yield of total polyphenols was 44.74 mg gallic acid equivalents per gram of dried plum at optimal conditions. Polyphenol extracts exhibited stronger antioxidant activities than Vc by evaluating of 1,1-diphenyl-2-picrylhydrazyl (DPPH and hydroxyl radical scavenging activity. Furthermore, polyphenol extracts (IC50 = 179 g/mL showed obvious inhibitory effects on xanthine oxidase. These findings suggest that polyphenol extracts from P. salicina can be potentially used as natural antioxidant and xanthine oxidase inhibitory agents.

  4. Process Optimization of Tempeh Protein Isolate from Soybean (Glycine Max Merr and Cowpea (Vigna Unguiculata Mixture

    Directory of Open Access Journals (Sweden)

    Asrul Bahar

    2015-04-01

    Full Text Available Production of ‘tempeh’ (fermented soybean protein isolates in Indonesia is still very low. It is necessary to increase the production with the alternative materials which is make by soybean and cowpea mixture. The utilization of this soybean cake (tempeh as protein isolate raw material is expecting benefits both of soybean and cowpea component which can complement each other. Protein’s content of protein isolates should be a minimum of 90% (db. Therefore, it is necessary to find the precipitation pH and the optimum level of purification in order to get tempeh protein isolate with high protein content. The analytical method used descriptive analysis. The optimal process of tempeh protein isolates from soybean and cowpea mixture conducted with pH 5 and pH 4 precipitation, the oil extraction was carried out at the beginning of the process (before extraction of protein and before the drying stage. The result showed that the tempeh protein isolate of soybean and cowpea mixing have 75.12% (db protein content. It was increasing in 20.67% to 50.16% from previous research.

  5. [Technical condition of semi-bionic extraction in optimizing formula of Rhizoma Cyperi that had been extracted by SFE-CO2 through homogeneous design].

    Science.gov (United States)

    Sun, Xiumei; Shi, Haiyan; Zhang, Zhaowang; Wang, Yingzi

    2009-11-01

    To optimize the extraction condtions in the modification of Rhizoma Cyperi, which had been extracted by the technique of supercritical CO2 extraction. The semi-bionic extraction (SBE) conditions were optimized through homogeneous design while the extracts (extracts were adopted as markers. The optimized SBE extraction conditions are the following: pH in first extraction was adjusted to 2.005 3. And then pH in second and third extraction adjusted to 6.508 2 and 8.945 6, time of the whole extracted process was 3.912 7 h. Combine the faction of production,we make sure the conclusion is pH in first extraction was 2.00, pH in second and third extraction adjusted to 6.50 and 9.00, duration of run was 2.0 h, 1.0 h and 1.0 h, respectively.

  6. The Optimization of Milk Protein Using Different Breeding Systems

    Directory of Open Access Journals (Sweden)

    Ramona Iancu

    2011-10-01

    Full Text Available The purpose of this study is to increase the intensive production systems in dairy goat flocks necessity specifications on feeding strategies and housing structures to optimize milk production and animal welfare and to control the impact of animal husbandry on the environment. Milk serves as the most excellent sources of protein with rich quality protein content. It doesn’t contain a large quantity of proteins, but those are containing all the essential amino acids that the body requires. The nutritional and processing properties and there by the market value of milk, largely depends on its protein content.

  7. Protein Extraction Methods for Two-Dimensional Electrophoresis from Baphicacanthus cusia (Nees) Bremek Leaves-A Medicinal Plant with High Contents of Interfering Compounds

    Institute of Scientific and Technical Information of China (English)

    XIANG Xiao-liang; NING Shu-ju; JIANG Xia; GONG Xiao-gui; ZHU Ren-lei; WEI Dao-zhi

    2010-01-01

    Protein extraction is a critical step for two-dimensional electrophoresis (2-DE). Different plant samples require different and adaptive protein extraction protocols. The leaves of medicinal plant, Baphicacanthus cusia (Nees) Bremek are notoriously recalcitrant to common protein extraction methods due to high levels of interfering compounds (especially the secondary metabolites and pigments). This study was aimed to establish a routine procedure for the proteomic analysis of B. cusia leaves, and a new protocol for the protein extraction was developed by optimizing tfichloroacetic acid (TCA)/acetone extraction method. The efficiency of this protocol was demonstrated by comparison with 3 published protein extraction methods (chloroform/acetone, Mg/NP-40, Tris-base/acetone). The results showed that the optimized TCA/acetone precipitation extraction method gave a relatively high protein yield (9.263 mg g fresh weight), high-resolution separation, clear protein profiles, the highest proteins spots (1 311 protein spots), and displayed less contamination in 2DE gels. Therefore, the results suggested that the optimized TCA/acetone method was the most effective among the 4 methods for B. cusia leaves.

  8. Response surface optimization of microwave-assisted extraction for HPLC-fluorescence determination of puerarin and daidz

    Institute of Scientific and Technical Information of China (English)

    Ying-Kun Liu; E Yan; Han-Ying Zhan; Zhi-Qi Zhang

    2011-01-01

    Microwave-assisted extraction was optimized with response surface methodology for HPLC-fluorescence determination of puerarin and daidzein in Radix Puerariae thomsonii.The optimized extraction procedure was achieved by soaking the sample with 70% methanol

  9. Optimization of carotenoids extraction from Penaeus semisulcatus shrimp wastes

    Institute of Scientific and Technical Information of China (English)

    Gholamreza jahed Khaniki; Parisa Sadighara; Ramin Nabizadeh Nodehi; Mahmood Alimohammadi; Naiema Vakili Saatloo

    2013-01-01

    Objective: To find effective method for carotenoids extraction from shrimp waste which is one of the important sources of natural carotenoids and produced in large quantities in Iran. Methods: Two methods of carotenoids extraction, enzymatic and alkaline (NaOH 1 normal) treatment, were assayed. About 5 g of gritted shrimp wastes were used at each stage. For alkaline treatment, sodium hydroxide were added to shrimp waste. After 48 h, the mixture was filtered and centrifuged.Results:Alcalase extraction produced (234.00±2.00) mg/L carotenoid and NaOH extraction produced (170.00±1.53) mg/L carotenoid. Based on the samples analyzed, alcalase enzyme showed more efficiency than NaOH extraction to achieve carotenoids from shrimp waste.Conclusions:It can be concluded that using alcalase enzyme for carotenoids extraction can produce higher carotenoids concentration than NaOH extraction method. So alcalase enzyme method can be used for achieving this kind of antioxidant.

  10. Studies on Optimal Extraction Conditions of Flavonoid in Leaves of Black Currnt

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Employing the orthogonal design,the optimal extraction conditions for flavonoid in leaves of black currant were determined by colorimetric estimation,which were extraction agent of 75% EtOH-H2O,the material ratio (ratio of weight of fresh leaves to volume of the solvent)of 1:16, the temperature of 35 ℃, the time of 1 h and times of three. A total extraction rate of over 97% and the fiavonoid contents of 179. 33 mg/100 g were obtained under the optimal extraction conditions.

  11. Process Optimization of Ultrasonic Extraction of Puerarin Based on Support Vector Machine

    Institute of Scientific and Technical Information of China (English)

    Juan Chen; Xiaoyi Huang; Yanlei Qi; Xin Qi; Qing Guo

    2014-01-01

    In ultrasonic extraction technology, optimization of technical parameters often considers extraction medium only, without including ultrasonic parameters. This paper focuses on controlling the ultrasonic extraction process of puerarin, investigating the influence of ultrasonic parameters on extraction rate, and empirical y analyzing the main components of Pueraria, i.e., isoflavone compounds. A method is presented combining orthogonal experi-mental design with a support vector machine and a predictive model is established for optimization of technical parameters. From the analysis with the predictive model, appropriate process parameters are achieved for higher extraction rate. With these parameters in the ultrasonic extraction of puerarin, the experimental result is satisfactory. This method is of significance to the study of extracting root-stock plant medicines.

  12. Statistical optimization of aqueous extraction of pectin from waste durian rinds.

    Science.gov (United States)

    Maran, J Prakash

    2015-02-01

    The objectives of this present study was to investigate and optimize the aqueous extraction conditions such as solid-liquid (SL) ratio (1:5-1:15 g/ml), pH (2-3), extraction time (20-60 min) and extraction temperature (75-95 °C) on maximum extraction of pectin from durian rinds using four factors, three levels Box-Behnken response design. The experimental data obtained were fitted to a second-order polynomial equation using multiple regression analysis and analyzed by analysis of variance (ANOVA). The optimum extraction condition was found to be as follows: SL ratio of 1:10 g/ml, pH of 2.8, extraction time of 43 min and extraction temperature of 86 °C respectively. Under the optimal conditions, the experimental pectin yield (9.1%) was well correlated with predicted yield (9.3%).

  13. Optimization of extraction time and temperature on antioxidant activity of Schizophyllum commune aqueous extract using response surface methodology.

    Science.gov (United States)

    Yim, Hip Seng; Chye, Fook Yee; Rao, Vigneswara; Low, Jia Yin; Matanjun, Patricia; How, Siew Eng; Ho, Chun Wai

    2013-04-01

    Central composite design of response surface methodology (RSM) was employed to optimize the extraction time (X 1 : 99.5-290.5 min) and temperature (X 2 : 30.1-54.9 °C) of Schizophyllum commune aqueous extract with high antioxidant activities and total phenolic content (TPC). Results indicated that the data were adequately fitted into four second-order polynomial models. The extraction time and temperature were found to have significant linear, quadratic and interaction effects on antioxidant activities and TPC. The optimal extraction time and temperature were: 290.5 min and 35.7 °C (DPPH(•) scavenging ability); 180.7 min and 41.7 °C (ABTS(•+) inhibition ability); 185.2 min and 42.4 °C (ferric reducing antioxidant power, FRAP); 290.5 min and 40.3 °C (TPC). These optimum conditions yielded 85.10%; 94.31%; 0.74 mM Fe(2+) equivalent/100 g; 635.76 mg gallic acid equivalent/100 g, respectively. The yields of antioxidant activities and TPC obtained experimentally were close to its predicted values. The establishment of such model provides a good experimental basis employing RSM for optimizing the extraction time and temperature on antioxidants from S. commune aqueous extract.

  14. Experimental study on the optimization of extraction process of garlic oil and its antibacterial effects.

    Science.gov (United States)

    Guo, Yajie

    2014-01-01

    Garlic oil which is the main active constituent of garlic has a wide range of pharmacological activities, and a broad antibacterial spectrum. It also has a strong anti-cancer activity, and can significantly inhibit a variety of tumors such as liver cancer, gastric cancer and colon cancer. The objective is to study the extraction process of garlic oil and its antibacterial effects. CO2 Supercritical extraction was used to investigate the optimal processing conditions for garlic oil extraction; filter paper test and suspension dilution test were applied to determine the bacteriostatic action of garlic oil. In the CO2 supercritical extraction experiment, factors influencing the yield of garlic oil were: extraction pressure > extraction temperature > extraction time in descending order. Range analysis showed that the optimal experimental conditions for CO2 supercritical extraction of garlic oil were extraction pressure of 15 Mpa, temperature of 40 °C, and duration of 1 h. Different concentrations of garlic oil could all inhibit the growth of Staphylococcus aureus, Escherichia coli and Bacillus subtilis, suggesting that garlic oil has an antibacterial effect. The optimal experimental conditions for CO2 supercritical extraction of garlic oil were: extraction pressure of 15 Mpa, temperature of 40 °C, and duration of 1 h; garlic oil has an antibacterial effect.

  15. Optimization of Subcritical Water Extraction of Resveratrol from Grape Seeds by Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Yajie Tian

    2017-03-01

    Full Text Available The subcritical water extraction (SWE is a high-efficiency and environment-friendly extraction method. The extraction of resveratrol (RES of grape seeds obtained from the wine production process was proposed using subcritical water extraction (SWE. The effects of different extraction process parameters on RES yield were investigated by single factors. Extraction optimization was conducted using response surface methodology (RSM. Extraction temperature was proven to be the most significant factor influencing RES yield. The optimal conditions was as follows: extraction pressure of 1.02 MPa, temperature of 152.32 °C, time of 24.89 min, and a solid/solvent ratio of 1:15 g/mL. Under these optimal conditions, the predicted extraction RES yield was 6.90 μg/g and the recoveries was up to 91.98%. Compared to other previous studies, this method required less pollution and less treatment time to extract RES from grape seeds. From these results, added economic value to this agroindustrial residue is proposed using environmentally friendly extraction techniques.

  16. Extraction optimization of mucilage from Basil (Ocimum basilicum L.) seeds using response surface methodology.

    Science.gov (United States)

    Nazir, Sadaf; Wani, Idrees Ahmed; Masoodi, Farooq Ahmad

    2017-05-01

    Aqueous extraction of basil seed mucilage was optimized using response surface methodology. A Central Composite Rotatable Design (CCRD) for modeling of three independent variables: temperature (40-91 °C); extraction time (1.6-3.3 h) and water/seed ratio (18:1-77:1) was used to study the response for yield. Experimental values for extraction yield ranged from 7.86 to 20.5 g/100 g. Extraction yield was significantly (P Temperature and water/seed ratio were found to have pronounced effect while the extraction time was found to have minor possible effects. Graphical optimization determined the optimal conditions for the extraction of mucilage. The optimal condition predicted an extraction yield of 20.49 g/100 g at 56.7 °C, 1.6 h, and a water/seed ratio of 66.84:1. Optimal conditions were determined to obtain highest extraction yield. Results indicated that water/seed ratio was the most significant parameter, followed by temperature and time.

  17. Optimizing Standard Sequential Extraction Protocol With Lake And Ocean Sediments

    Science.gov (United States)

    The environmental mobility/availability behavior of radionuclides in soils and sediments depends on their speciation. Experiments have been carried out to develop a simple but robust radionuclide sequential extraction method for identification of radionuclide partitioning in sed...

  18. Optimization of DNA extraction for advancing coral microbiota investigations.

    Science.gov (United States)

    Weber, Laura; DeForce, Emelia; Apprill, Amy

    2017-02-08

    DNA-based sequencing approaches are commonly used to identify microorganisms and their genes and document trends in microbial community diversity in environmental samples. However, extraction of microbial DNA from complex environmental samples like corals can be technically challenging, and extraction methods may impart biases on microbial community structure. We designed a two-phase study in order to propose a comprehensive and efficient method for DNA extraction from microbial cells present in corals and investigate if extraction method influences microbial community composition. During phase I, total DNA was extracted from seven coral species in a replicated experimental design using four different MO BIO Laboratories, Inc., DNA Isolation kits: PowerSoil®, PowerPlant® Pro, PowerBiofilm®, and UltraClean® Tissue & Cells (with three homogenization permutations). Technical performance of the treatments was evaluated using DNA yield and amplification efficiency of small subunit ribosomal RNA (SSU ribosomal RNA (rRNA)) genes. During phase II, potential extraction biases were examined via microbial community analysis of SSU rRNA gene sequences amplified from the most successful DNA extraction treatments. In phase I of the study, the PowerSoil® and PowerPlant® Pro extracts contained low DNA concentrations, amplified poorly, and were not investigated further. Extracts from PowerBiofilm® and UltraClean® Tissue and Cells permutations were further investigated in phase II, and analysis of sequences demonstrated that overall microbial community composition was dictated by coral species and not extraction treatment. Finer pairwise comparisons of sequences obtained from Orbicella faveolata, Orbicella annularis, and Acropora humilis corals revealed subtle differences in community composition between the treatments; PowerBiofilm®-associated sequences generally had higher microbial richness and the highest coverage of dominant microbial groups in comparison to the Ultra

  19. A Method for Community Detection in Protein Networks Using Spectral Optimization

    Directory of Open Access Journals (Sweden)

    Sminu Izudheen

    2011-12-01

    Full Text Available Identification of community structures in complex networks has been a challenge in many domain and discipline. In protein networks these community interactions play a vital role in identifying the outcome of many cellular mechanisms. This paper reports the use of spectral optimization of triangular modularity as an effective method to identify these community structures. The algorithm has been carefully tested on real biological data and the results acknowledge that this is a powerful method for extracting community structures from protein networks.

  20. Automatically extracting functionally equivalent proteins from SwissProt

    Directory of Open Access Journals (Sweden)

    Martin Andrew CR

    2008-10-01

    Full Text Available Abstract Background There is a frequent need to obtain sets of functionally equivalent homologous proteins (FEPs from different species. While it is usually the case that orthology implies functional equivalence, this is not always true; therefore datasets of orthologous proteins are not appropriate. The information relevant to extracting FEPs is contained in databanks such as UniProtKB/Swiss-Prot and a manual analysis of these data allow FEPs to be extracted on a one-off basis. However there has been no resource allowing the easy, automatic extraction of groups of FEPs – for example, all instances of protein C. We have developed FOSTA, an automatically generated database of FEPs annotated as having the same function in UniProtKB/Swiss-Prot which can be used for large-scale analysis. The method builds a candidate list of homologues and filters out functionally diverged proteins on the basis of functional annotations using a simple text mining approach. Results Large scale evaluation of our FEP extraction method is difficult as there is no gold-standard dataset against which the method can be benchmarked. However, a manual analysis of five protein families confirmed a high level of performance. A more extensive comparison with two manually verified functional equivalence datasets also demonstrated very good performance. Conclusion In summary, FOSTA provides an automated analysis of annotations in UniProtKB/Swiss-Prot to enable groups of proteins already annotated as functionally equivalent, to be extracted. Our results demonstrate that the vast majority of UniProtKB/Swiss-Prot functional annotations are of high quality, and that FOSTA can interpret annotations successfully. Where FOSTA is not successful, we are able to highlight inconsistencies in UniProtKB/Swiss-Prot annotation. Most of these would have presented equal difficulties for manual interpretation of annotations. We discuss limitations and possible future extensions to FOSTA, and

  1. Optimization of carotenoids extraction from Penaeus semisulcatus shrimp wastes

    OpenAIRE

    Gholamreza jahed Khaniki; Parisa Sadighara; Ramin Nabizadeh Nodehi; Mahmood Alimohammadi; Naiema Vakili Saatloo

    2013-01-01

    Objective: To find effective method for carotenoids extraction from shrimp waste which is one of the important sources of natural carotenoids and produced in large quantities in Iran. Methods: Two methods of carotenoids extraction, enzymatic and alkaline (NaOH 1 normal) treatment, were assayed. About 5 g of gritted shrimp wastes were used at each stage. For alkaline treatment, sodium hydroxide were added to shrimp waste. After 48 h, the mixture was filtered and centrifuged. ...

  2. Response Surface Modeling and Optimization of Accelerated Solvent Extraction of Four Lignans from Fructus Schisandrae

    Directory of Open Access Journals (Sweden)

    Jian Liang

    2012-03-01

    Full Text Available A new method based on accelerated solvent extraction (ASE combined with response surface methodology (RSM modeling and optimization has been developed for the extraction of four lignans in Fructus Schisandrae (the fruits of Schisandra chinensis Baill. The RSM method, based on a three level and three variable Box-Behnken design (BBD, was employed to obtain the optimal combination of extraction condition. In brief, the lignans schizandrin, schisandrol B, deoxyschizandrin and schisandrin B were optimally extracted with 87% ethanol as extraction solvent, extraction temperature of 160 °C, static extraction time of 10 min, extraction pressure of 1,500 psi, flush volume of 60% and one extraction cycle. The 3D response surface plot and the contour plot derived from the mathematical models were applied to determine the optimal conditions. Under the above conditions, the experimental value of four lignans was 14.72 mg/g, which is in close agreement with the value predicted by the model.

  3. Operon Gene Order Is Optimized for Ordered Protein Complex Assembly.

    Science.gov (United States)

    Wells, Jonathan N; Bergendahl, L Therese; Marsh, Joseph A

    2016-02-02

    The assembly of heteromeric protein complexes is an inherently stochastic process in which multiple genes are expressed separately into proteins, which must then somehow find each other within the cell. Here, we considered one of the ways by which prokaryotic organisms have attempted to maximize the efficiency of protein complex assembly: the organization of subunit-encoding genes into operons. Using structure-based assembly predictions, we show that operon gene order has been optimized to match the order in which protein subunits assemble. Exceptions to this are almost entirely highly expressed proteins for which assembly is less stochastic and for which precisely ordered translation offers less benefit. Overall, these results show that ordered protein complex assembly pathways are of significant biological importance and represent a major evolutionary constraint on operon gene organization.

  4. Optimization of TRPO Process Parameters for Americium Extraction from High Level Waste

    Institute of Scientific and Technical Information of China (English)

    CHEN Jing; WANG Jianchen; SONG Chongli

    2001-01-01

    The numerical calculations for Am multistage fractional extraction by trialkyl phosphine oxide (TRPO) were verified by a hot test.1750 L/t-U high level waste (HLW) was used as the feed to the TRPO process.The analysis used the simple objective function to minimize the total waste content in the TRPO process streams.Some process parameters were optimized after other parameters were selected.The optimal process parameters for Am extraction by TRPO are:10 stages for extraction and 2 stages for scrubbing;a flow rate ratio of 0.931 for extraction and 4.42 for scrubbing;nitric acid concentration of 1.35 mol/L for the feed and 0.5 mol/L for the scrubbing solution.Finally,the nitric acid and Am concentration profiles in the optimal TRPO extraction process are given.

  5. Antioxidant Compound Extraction from Maqui (Aristotelia chilensis [Mol] Stuntz Berries: Optimization by Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Issis Quispe-Fuentes

    2017-02-01

    Full Text Available The optimum conditions for the antioxidant extraction from maqui berry were determined using a response surface methodology. A three level D-optimal design was used to investigate the effects of three independent variables namely, solvent type (methanol, acetone and ethanol, solvent concentration and extraction time over total antioxidant capacity by using the oxygen radical absorbance capacity (ORAC method. The D-optimal design considered 42 experiments including 10 central point replicates. A second-order polynomial model showed that more than 89% of the variation is explained with a satisfactory prediction (78%. ORAC values are higher when acetone was used as a solvent at lower concentrations, and the extraction time range studied showed no significant influence on ORAC values. The optimal conditions for antioxidant extraction obtained were 29% of acetone for 159 min under agitation. From the results obtained it can be concluded that the given predictive model describes an antioxidant extraction process from maqui berry.

  6. Determination of optimal parameters of basil supercritical fluid extraction by response surface methodology

    Directory of Open Access Journals (Sweden)

    Filip Snežana Đ.

    2016-01-01

    Full Text Available The supercritical fluid extraction of aroma compounds from basil (Ocimum basilicum L. was studied. Response surface methodology was used to optimize the parameters of the process. Full factorial design was applied to evaluate the effects of two independent variables (pressure and temperature on the extraction yield and linalool yield. From the response surface plots, pressure and temperature exhibited independent and interactive effect on the extraction yield. The optimal conditions to obtain the highest extraction yield (1.91% of O. basilicum were the pressure of 29.7 MPa and temperature of 59.2oC, whereas the highest yield of linalool (1.998 g•kg-1 was obtained at the pressure of 20 MPa and temperature of 40oC. The experimental values agreed with the predicted ones, indicating suitability of the response surface methodology for optimizing the extraction process. [Projekat Ministarstva nauke Republike Srbije, br. TR 31013

  7. Antioxidant Compound Extraction from Maqui (Aristotelia chilensis [Mol] Stuntz) Berries: Optimization by Response Surface Methodology

    Science.gov (United States)

    Quispe-Fuentes, Issis; Vega-Gálvez, Antonio; Campos-Requena, Víctor H.

    2017-01-01

    The optimum conditions for the antioxidant extraction from maqui berry were determined using a response surface methodology. A three level D-optimal design was used to investigate the effects of three independent variables namely, solvent type (methanol, acetone and ethanol), solvent concentration and extraction time over total antioxidant capacity by using the oxygen radical absorbance capacity (ORAC) method. The D-optimal design considered 42 experiments including 10 central point replicates. A second-order polynomial model showed that more than 89% of the variation is explained with a satisfactory prediction (78%). ORAC values are higher when acetone was used as a solvent at lower concentrations, and the extraction time range studied showed no significant influence on ORAC values. The optimal conditions for antioxidant extraction obtained were 29% of acetone for 159 min under agitation. From the results obtained it can be concluded that the given predictive model describes an antioxidant extraction process from maqui berry.

  8. Extraction and downstream processing of plant-derived recombinant proteins.

    Science.gov (United States)

    Buyel, J F; Twyman, R M; Fischer, R

    2015-11-01

    Plants offer the tantalizing prospect of low-cost automated manufacturing processes for biopharmaceutical proteins, but several challenges must be addressed before such goals are realized and the most significant hurdles are found during downstream processing (DSP). In contrast to the standardized microbial and mammalian cell platforms embraced by the biopharmaceutical industry, there are many different plant-based expression systems vying for attention, and those with the greatest potential to provide inexpensive biopharmaceuticals are also the ones with the most significant drawbacks in terms of DSP. This is because the most scalable plant systems are based on the expression of intracellular proteins in whole plants. The plant tissue must therefore be disrupted to extract the product, challenging the initial DSP steps with an unusually high load of both particulate and soluble contaminants. DSP platform technologies can accelerate and simplify process development, including centrifugation, filtration, flocculation, and integrated methods that combine solid-liquid separation, purification and concentration, such as aqueous two-phase separation systems. Protein tags can also facilitate these DSP steps, but they are difficult to transfer to a commercial environment and more generic, flexible and scalable strategies to separate target and host cell proteins are preferable, such as membrane technologies and heat/pH precipitation. In this context, clarified plant extracts behave similarly to the feed stream from microbes or mammalian cells and the corresponding purification methods can be applied, as long as they are adapted for plant-specific soluble contaminants such as the superabundant protein RuBisCO. Plant-derived pharmaceutical proteins cannot yet compete directly with established platforms but they are beginning to penetrate niche markets that allow the beneficial properties of plants to be exploited, such as the ability to produce 'biobetters' with tailored

  9. A global optimization algorithm for protein surface alignment

    Directory of Open Access Journals (Sweden)

    Guerra Concettina

    2010-09-01

    Full Text Available Abstract Background A relevant problem in drug design is the comparison and recognition of protein binding sites. Binding sites recognition is generally based on geometry often combined with physico-chemical properties of the site since the conformation, size and chemical composition of the protein surface are all relevant for the interaction with a specific ligand. Several matching strategies have been designed for the recognition of protein-ligand binding sites and of protein-protein interfaces but the problem cannot be considered solved. Results In this paper we propose a new method for local structural alignment of protein surfaces based on continuous global optimization techniques. Given the three-dimensional structures of two proteins, the method finds the isometric transformation (rotation plus translation that best superimposes active regions of two structures. We draw our inspiration from the well-known Iterative Closest Point (ICP method for three-dimensional (3D shapes registration. Our main contribution is in the adoption of a controlled random search as a more efficient global optimization approach along with a new dissimilarity measure. The reported computational experience and comparison show viability of the proposed approach. Conclusions Our method performs well to detect similarity in binding sites when this in fact exists. In the future we plan to do a more comprehensive evaluation of the method by considering large datasets of non-redundant proteins and applying a clustering technique to the results of all comparisons to classify binding sites.

  10. Optimal Coordinated Control of Power Extraction in LES of a Wind Farm with Entrance Effects

    Directory of Open Access Journals (Sweden)

    Jay P. Goit

    2016-01-01

    Full Text Available We investigate the use of optimal coordinated control techniques in large eddy simulations of wind farm boundary layer interaction with the aim of increasing the total energy extraction in wind farms. The individual wind turbines are considered as flow actuators, and their energy extraction is dynamically regulated in time, so as to optimally influence the flow field. We extend earlier work on wind farm optimal control in the fully-developed regime (Goit and Meyers 2015, J. Fluid Mech. 768, 5–50 to a ‘finite’ wind farm case, in which entrance effects play an important role. For the optimal control, a receding horizon framework is employed in which turbine thrust coefficients are optimized in time and per turbine. Optimization is performed with a conjugate gradient method, where gradients of the cost functional are obtained using adjoint large eddy simulations. Overall, the energy extraction is increased 7% by the optimal control. This increase in energy extraction is related to faster wake recovery throughout the farm. For the first row of turbines, the optimal control increases turbulence levels and Reynolds stresses in the wake, leading to better wake mixing and an inflow velocity for the second row that is significantly higher than in the uncontrolled case. For downstream rows, the optimal control mainly enhances the sideways mean transport of momentum. This is different from earlier observations by Goit and Meyers (2015 in the fully-developed regime, where mainly vertical transport was enhanced.

  11. Optimization of Extraction Technology of Ellagic Acid from Pomegranate Peels with Orthogonal Experiment

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    [Objective] This study aimed to investigate the optimal condition for extrac- tion of ellagic acid from pomegranate peels. [Method] With ellagic acid yield as an indicator, ultrasound extraction method was adopted to extract the ellagic acid from pomegranate peels, and the concentration of ellagic acid was measured by using ul- traviolet spectrophotometry; L9(34) orthogonal experiment was designed with four fac- tors at three levels, to investigate the effect of extraction temperature, extraction du- ration, solid-liquid ratio and ultrasound power on extraction rate of ellagic acid. [Re- sult] The optimal extraction condition for ellagic acid is extraction temperature of 30 ~C, solid-liquid ratio of 1:200, extraction duration of 20 min, and ultrasound power of 50 W. The relationship between primary and secondary factors affecting experimental indicators was solid-liquid ratio 〉 extraction duration 〉 ultrasound power 〉 extraction temperature. [Conclusion] Solid-liquid ratio has the maximum effect on extraction rate of ellagic acid. The ellagic acid extraction technology identified in this study is rea- sonable and feasible.

  12. Optimization of protein electroextraction from microalgae by a flow process.

    Science.gov (United States)

    Coustets, Mathilde; Joubert-Durigneux, Vanessa; Hérault, Josiane; Schoefs, Benoît; Blanckaert, Vincent; Garnier, Jean-Pierre; Teissié, Justin

    2015-06-01

    Classical methods, used for large scale treatments such as mechanical or chemical extractions, affect the integrity of extracted cytosolic protein by releasing proteases contained in vacuoles. Our previous experiments on flow processes electroextraction on yeasts proved that pulsed electric field technology allows preserving the integrity of released cytosolic proteins, by not affecting vacuole membranes. Furthermore, large cell culture volumes are easily treated by the flow technology. Based on this previous knowledge, we developed a new protocol in order to electro-extract total cytoplasmic proteins from microalgae (Nannochloropsis salina, Chlorella vulgaris and Haematococcus pluvialis). Given that induction of electropermeabilization is under the control of target cell size, as the mean diameter for N. salina is only 2.5 μm, we used repetitive 2 ms long pulses of alternating polarities with stronger field strengths than previously described for yeasts. The electric treatment was followed by a 24h incubation period in a salty buffer. The amount of total protein release was observed by a classical Bradford assay. A more accurate evaluation of protein release was obtained by SDS-PAGE. Similar results were obtained with C. vulgaris and H. pluvialis under milder electrical conditions as expected from their larger size.

  13. Optimization of ultrasound-assisted extraction of charantin from Momordica charantia fruits using response surface methodology

    OpenAIRE

    Javed Ahamad; Saima Amin; Showkat R. Mir

    2015-01-01

    Background: Momordica charantia Linn. (Cucurbitaceae) fruits are well known for their beneficial effects in diabetes that are often attributed to its bioactive component charantin. Objective: The aim of the present study is to develop and optimize an efficient protocol for the extraction of charantin from M. charantia fruits. Materials and Methods: Response surface methodology (RSM) was used for the optimization of ultrasound-assisted extraction (UAE) conditions. RSM was based on a three-leve...

  14. Extraction of flavonoids from Tagetes patula: process optimization and screening for biological activity

    Directory of Open Access Journals (Sweden)

    Vanessa M. Munhoz

    2014-10-01

    Full Text Available The flowers of Tagetes patula L., Asteraceae, commonly known as French marigold, are used in folk medicine as an antiseptic, diuretic, blood purifier and insect repellent. This study was conducted to optimize the extraction process through the biomonitoring of flavonoids, using a statistical mixture simplex-centroid design, to evaluate the effect of the solvents water, ethanol and acetone, as well as mixtures of these solvents, assessed by the total flavonoid content. The extracts were tested for dry residue, radical scavenging activity, chromatographic profile, and larvicidal activity. The acetone extract had the highest total flavonoid content, 25.13 ± 1.02% (4.07%; and the best radical scavenging activity, with IC50 of 15.74 μg/ml ± 1.09 (6.92%, but with lower dry residue, 6.62 ± 1.33% (20.10%. The water extracts showed higher levels of dry residue, but lower total flavonoid content and radical scavenging activity than the acetone extract. The positive correlation between the total flavonoid content and radical scavenging activity of the extracts showed that flavonoids contribute significantly to the antioxidant capacity. The statistical mixture design allowed us to optimize the extraction of flavonoids from flowers of T. patula, with acetone as the best extraction solvent. Preliminary studies on the biological activity of the optimized extracts demonstrated a larvicidal effect of the acetone extract on Aedes aegypti mosquitoes.

  15. Optimization of Method to Extract Collagen from "Emperor" Tissue of Soft-shelled Turtles.

    Science.gov (United States)

    Yamamoto, Tetsushi; Uemura, Kentaro; Sawashi, Yuki; Mitamura, Kuniko; Taga, Atsushi

    2016-01-01

    Soft-shelled turtles (Pelodiscus sinensis) are widely distributed in some Asian countries, and parts of this turtle contain abundant collagen. In this study, we optimized a method for extracting collagen from the soft-shelled turtle. We used three types of solvent and four extraction conditions to determine an effective collagen extraction method, which was extraction at 37°C with acetic acid after hydrochloric acid pretreatment. Next, we extracted collagen from three regions in the soft-shelled turtle: muscle, skin, and an area of soft tissue in the periphery of the turtle shell known in Japan and China as the "emperor." We determined that emperor tissue yielded the highest concentration and purity of collagen. We then optimized the pretreatment method for extraction from emperor tissue by using formic acid instead of hydrochloric acid, and the amount of extracted collagen increased by approximately 1.3-fold. Finally, we identified the optimal solvent out of four types of organic acid for collagen extraction from emperor tissue; the amount of extracted collagen from emperor tissue increased approximately 3-fold when citric acid was used as the extraction solvent instead of acetic acid. Emperor tissue can regenerate; thus, it is possible to obtain collagen from the emperor repeatedly without killing the turtle. Our findings suggest that the emperor tissue of softshelled turtles may be a good source of collagen for pharmaceutical and cosmetic applications.

  16. Optimized Baxter model of protein solutions: electrostatics versus adhesion

    NARCIS (Netherlands)

    Prinsen, P.; Odijk, T.

    2004-01-01

    A theory is set up of spherical proteins interacting by screened electrostatics and constant adhesion, in which the effective adhesion parameter is optimized by a variational principle for the free energy. An analytical approach to the second virial coefficient is first outlined by balancing the rep

  17. Optimization of cold neutron beam extraction at ESS

    DEFF Research Database (Denmark)

    Schönfeldt, Troels; Batkov, K.; Klinkby, Esben Bryndt

    The present study takes its origin in the baseline design of European Spallation Source where a cold and a thermal moderator are situated next to each other enabling bispectral extraction. The study aims at mapping the differences in various neutron distributions depending on the angle and position...... from which the moderator is viewed. This study does not only show changes in both cold and thermal neutron flux, depending on extraction position, but also shows that there are significant differences in the wavelength spectrum and origin of neutrons depending on the angel of view....

  18. Optimization and Technological Development Strategies of an Antimicrobial Extract from Achyrocline alata Assisted by Statistical Design

    Science.gov (United States)

    Demarque, Daniel P.; Fitts, Sonia Maria F.; Boaretto, Amanda G.; da Silva, Júlio César Leite; Vieira, Maria C.; Franco, Vanessa N. P.; Teixeira, Caroline B.; Toffoli-Kadri, Mônica C.; Carollo, Carlos A.

    2015-01-01

    Achyrocline alata, known as Jateí-ka-há, is traditionally used to treat several health problems, including inflammations and infections. This study aimed to optimize an active extract against Streptococcus mutans, the main bacteria that causes caries. The extract was developed using an accelerated solvent extraction and chemometric calculations. Factorial design and response surface methodologies were used to determine the most important variables, such as active compound selectivity. The standardized extraction recovered 99% of the four main compounds, gnaphaliin, helipyrone, obtusifolin and lepidissipyrone, which represent 44% of the extract. The optimized extract of A. alata has a MIC of 62.5 μg/mL against S. mutans and could be used in mouth care products. PMID:25710523

  19. Optimization and technological development strategies of an antimicrobial extract from Achyrocline alata assisted by statistical design.

    Science.gov (United States)

    Demarque, Daniel P; Fitts, Sonia Maria F; Boaretto, Amanda G; da Silva, Júlio César Leite; Vieira, Maria C; Franco, Vanessa N P; Teixeira, Caroline B; Toffoli-Kadri, Mônica C; Carollo, Carlos A

    2015-01-01

    Achyrocline alata, known as Jateí-ka-há, is traditionally used to treat several health problems, including inflammations and infections. This study aimed to optimize an active extract against Streptococcus mutans, the main bacteria that causes caries. The extract was developed using an accelerated solvent extraction and chemometric calculations. Factorial design and response surface methodologies were used to determine the most important variables, such as active compound selectivity. The standardized extraction recovered 99% of the four main compounds, gnaphaliin, helipyrone, obtusifolin and lepidissipyrone, which represent 44% of the extract. The optimized extract of A. alata has a MIC of 62.5 μg/mL against S. mutans and could be used in mouth care products.

  20. Optimization of Extraction Process for Polysaccharide in Salvia Miltiorrhiza Bunge Using Response Surface Methodology.

    Science.gov (United States)

    Yanhua, Wang; Fuhua, Wu; Zhaohan, Guo; Mingxing, Peng; Yanan, Zhang; Ling, Pang Zhen; Minhua, Du; Caiying, Zhang; Zian, Liang

    2015-01-01

    This study was aimed to optimize the extraction process for Salvia miltiorrhiza Bunge polysaccharide using response surface methodology The results showed that four operating parameters including microwave power, microwave time and the particle size had notable effects on the polysaccharide extraction of Salvia miltiorrhiza Bunge. The effects could be ranked in decreasing order of importance as follows:. Microwave power > microwave time > the comminution degree. The optimal extraction parameters were determined as 573.83W of Microwave power and 8.4min of microwave time and 67.51mesh of the comminution degree, resulting in the yield of Salvia miltiorrhiza Bunge polysaccharide of 101.161mg / g. The established regression model describing polysaccharide extraction from as a function of the three extraction parameters was highly significant (R 2 = 0.9953). The predicted and experimental results were found to be in good agreement. Thus, the model can be applicable for the prediction of polysaccharide extraction from Salvia miltiorrhiza Bunge.

  1. Optimization and technological development strategies of an antimicrobial extract from Achyrocline alata assisted by statistical design.

    Directory of Open Access Journals (Sweden)

    Daniel P Demarque

    Full Text Available Achyrocline alata, known as Jateí-ka-há, is traditionally used to treat several health problems, including inflammations and infections. This study aimed to optimize an active extract against Streptococcus mutans, the main bacteria that causes caries. The extract was developed using an accelerated solvent extraction and chemometric calculations. Factorial design and response surface methodologies were used to determine the most important variables, such as active compound selectivity. The standardized extraction recovered 99% of the four main compounds, gnaphaliin, helipyrone, obtusifolin and lepidissipyrone, which represent 44% of the extract. The optimized extract of A. alata has a MIC of 62.5 μg/mL against S. mutans and could be used in mouth care products.

  2. Extraction optimization of coumarins from radix angelicae pubescentis by HPLC-DAD coupled with uniform design

    Institute of Scientific and Technical Information of China (English)

    GUO Fang-qiu; HUANG Lan-fang; ZHOU Huo-fei; JIANG Qiu-ju

    2006-01-01

    Uniform design was used to optimize extraction condition of direct refluence extraction of coumarins from the Chinese traditional medicine of radix angelicae pubescentis(Duhuo); the sum peak area of coumarins separated with high performance liquid chromatography(HPLC) at detection wavelength of 320 nm was considered as detection index, two factors of solvent concentration and extraction time were mainly studied at extraction temperature of 85 ℃ and a volume ratio of solvent to sample of 10: 1. Optimal subclass, quadric polynomial step by step aggression and neural network method were applied to process the experimental results. The results show that the first and second methods give the same factors combination (concentration of ethanol: 95 %, extraction time: 3.6 h) and the second method is much better than the first one. The extraction model is consequently developed.

  3. Optimization of Smashing Tissue Extraction Technology of Schisandra chinensis Fruits by Orthogonal Test

    Institute of Scientific and Technical Information of China (English)

    TANG Yun; LIU Yan-ze; HAN Ling; ZHAO Yu-qing

    2012-01-01

    Objective To optimize the extract technology of active lignins from the fruits of Schisandra chinensis.Methods The content of schizandrin,gomisin A,and deoxyschizandrin were selected as standards to evaluate the efficiency of smashing tissue extraction (STE).Solid-liquid ratio,extracting times,ethanol concentration,and extracting time were investigated through orthogonal test.Results The optimized conditions for STE were ten times amount of 80% EtOH,extracting for three times,and 2 min for each time.Conclusion STE could obtain relatively higher yield,simplicity of operation,and benefit for environment protection.It could be better choice for the extraction ofS.chinensis.

  4. Mixture approach for optimizing lycopene extraction from tomato and tomato products.

    Science.gov (United States)

    Periago, María Jesús; Rincón, Francisco; Agüera, Maria Dolores; Ros, Gaspar

    2004-09-22

    A simple mixture process design based on the comparison of both quadratic and special cubic models and involving three mixture components (hexane/acetone/ethanol) as a solution for extracting lycopene from raw tomato, tomato sauce, and tomato paste was used to confirm the hypothesis that lycopene extraction rates are a function of the solvent used during the extraction process. Conventional criteria (p lycopene extraction was hexane, there was a positive secondary synergistic interaction of hexane with ethanol (all sample types) and with acetone (tomato paste samples); this suggests that a mixture including all three components is essential for optimizing the extraction process. The partial special cubic model yielded three stationary points, indicating the concentrations of hexane, acetone, and ethanol required to optimize lycopene extraction in raw tomato, tomato sauce, and paste.

  5. A twist on folding: Predicting optimal sequences and optimal folds of simple protein models with the hidden-force algorithm

    CERN Document Server

    Kolossváry, István

    2012-01-01

    We propose a new way of looking at global optimization of off-lattice protein models. We present a dual optimization concept of predicting optimal sequences as well as optimal folds. We validate the utility of the recently introduced hidden-force Monte Carlo optimization algorithm by finding significantly lower energy folds for minimalist protein models than previously reported. Further, we also find the protein sequence that yields the lowest energy fold amongst all sequences for a given chain length and residue mixture. In particular, for protein models with a binary sequence, we show that the sequence-optimized folds form more compact cores than the lowest energy folds of the historically fixed, Fibonacci-series sequences of chain lengths of 13, 21, 34, 55, and 89. We emphasize that while the protein model we used is minimalist, the methodology is applicable to detailed protein models, and sequence optimization may yield novel folds and aid de novo protein design.

  6. Optimization of ultrasound-assisted extraction of bioactive compounds from wild garlic (Allium ursinum L.).

    Science.gov (United States)

    Tomšik, Alena; Pavlić, Branimir; Vladić, Jelena; Ramić, Milica; Brindza, Ján; Vidović, Senka

    2016-03-01

    Ultrasound-assisted extraction was used for extraction of bioactive compounds and for production of Allium ursinum liquid extract. The experiments were carried out according to tree level, four variables, face-centered cubic experimental design (FDC) combined with response surface methodology (RSM). Temperature (from 40 to 80 °C), ethanol concentration (from 30% to 70%), extraction time (from 40 to 80 min) and ultrasonic power (from 19.2 to 38.4 W/L) were investigated as independent variables in order to obtain the optimal conditions for extraction and to maximize the yield of total phenols (TP), flavonoids (TF) and antioxidant activity of obtained extracts. Experimental results were fitted to the second order polynomial model where multiple regression and analysis of variance were used to determine the fitness of the model and optimal condition for investigated responses. The predicted values of the TP (1.60 g GAE/100 g DW), TF (0.35 g CE/100 g DW), antioxidant activity, IC50 (0.71 mg/ml) and EY (38.1%) were determined at the optimal conditions for ultrasound assisted extraction: 80 °C temperature, 70% ethanol, 79.8 min and 20.06 W/L ultrasonic power. The predicted results matched well with the experimental results obtained using optimal extraction conditions which validated the RSM model with a good correlation.

  7. Optimization of pressurized liquid extraction of carotenoids and chlorophylls from Chlorella vulgaris.

    Science.gov (United States)

    Cha, Kwang Hyun; Lee, Hee Ju; Koo, Song Yi; Song, Dae-Geun; Lee, Dong-Un; Pan, Cheol-Ho

    2010-01-27

    Pressurized liquid extraction (PLE) was applied to the extraction of carotenoids and chlorophylls from the green microalga Chlorella vulgaris. Four extraction techniques such as maceration (MAC), Soxhlet extraction (SOX), ultrasound assisted extraction (UAE), and PLE were compared, and both the extraction temperature (50, 105, and 160 degrees C) and the extraction time (8, 19, and 30 min), which are the two main factors for PLE, were optimized with a central composite design to obtain the highest extraction efficiency. The extraction solvent (90% ethanol/water) could adequately extract the functional components from C. vulgaris. PLE showed higher extraction efficiencies than MAC, SOX, and UAE. Temperature was the key parameter having the strongest influence on the extraction of carotenoids and chlorophylls from chlorella. In addition, high heat treatment (>110 degrees C) by PLE minimized the formation of pheophorbide a, a harmful chlorophyll derivative. These results indicate that PLE may be a useful extraction method for the simultaneous extraction of carotenoids and chlorophylls from C. vulgaris.

  8. Extraction and characterisation of protein fractions from five insect species.

    Science.gov (United States)

    Yi, Liya; Lakemond, Catriona M M; Sagis, Leonard M C; Eisner-Schadler, Verena; van Huis, Arnold; van Boekel, Martinus A J S

    2013-12-15

    Tenebrio molitor, Zophobas morio, Alphitobius diaperinus, Acheta domesticus and Blaptica dubia were evaluated for their potential as a future protein source. Crude protein content ranged from 19% to 22% (Dumas analysis). Essential amino acid levels in all insect species were comparable with soybean proteins, but lower than for casein. After aqueous extraction, next to a fat fraction, a supernatant, pellet, and residue were obtained, containing 17-23%, 33-39%, 31-47% of total protein, respectively. At 3% (w/v), supernatant fractions did not form stable foams and gels at pH 3, 5, 7, and 10, except for gelation for A. domesticus at pH 7. At 30% w/v, gels at pH 7 and pH 10 were formed, but not at pH 3 and pH 5. In conclusion, the insect species studied have potential to be used in foods due to: (1) absolute protein levels; (2) protein quality; (3) ability to form gels.

  9. Optimization of the extraction of carrageenan from Kappaphycus alvarezii using response surface methodology

    OpenAIRE

    Vanessa Webber; Sabrina Matos de Carvalho; Paulo José Ogliari; Leila Hayashi; Pedro Luiz Manique Barreto

    2012-01-01

    This study aims to optimize an alternative method of extraction of carrageenan without previous alkaline treatment and ethanol precipitation using Response Surface Methodology (RSM). In order to introduce an innovation in the isolation step, atomization drying was used reducing the time for obtaining dry carrageenan powder. The effects of extraction time and temperature on yield, gel strength, and viscosity were evaluated. Furthermore, the extracted material was submitted to structural analys...

  10. Reversals and collisions optimize protein exchange in bacterial swarms

    Energy Technology Data Exchange (ETDEWEB)

    Amiri, Aboutaleb; Harvey, Cameron; Buchmann, Amy; Christley, Scott; Shrout, Joshua D.; Aranson, Igor S.; Alber, Mark

    2017-03-01

    Swarming groups of bacteria coordinate their behavior by self-organizing as a population to move over surfaces in search of nutrients and optimal niches for colonization. Many open questions remain about the cues used by swarming bacteria to achieve this self-organization. While chemical cue signaling known as quorum sensing is well-described, swarming bacteria often act and coordinate on time scales that could not be achieved via these extracellular quorum sensing cues. Here, cell-cell contact-dependent protein exchange is explored as amechanism of intercellular signaling for the bacterium Myxococcus xanthus. A detailed biologically calibrated computational model is used to study how M. xanthus optimizes the connection rate between cells and maximizes the spread of an extracellular protein within the population. The maximum rate of protein spreading is observed for cells that reverse direction optimally for swarming. Cells that reverse too slowly or too fast fail to spread extracellular protein efficiently. In particular, a specific range of cell reversal frequencies was observed to maximize the cell-cell connection rate and minimize the time of protein spreading. Furthermore, our findings suggest that predesigned motion reversal can be employed to enhance the collective behavior of biological synthetic active systems.

  11. Reversals and collisions optimize protein exchange in bacterial swarms

    Science.gov (United States)

    Amiri, Aboutaleb; Harvey, Cameron; Buchmann, Amy; Christley, Scott; Shrout, Joshua D.; Aranson, Igor S.; Alber, Mark

    2017-03-01

    Swarming groups of bacteria coordinate their behavior by self-organizing as a population to move over surfaces in search of nutrients and optimal niches for colonization. Many open questions remain about the cues used by swarming bacteria to achieve this self-organization. While chemical cue signaling known as quorum sensing is well-described, swarming bacteria often act and coordinate on time scales that could not be achieved via these extracellular quorum sensing cues. Here, cell-cell contact-dependent protein exchange is explored as a mechanism of intercellular signaling for the bacterium Myxococcus xanthus. A detailed biologically calibrated computational model is used to study how M. xanthus optimizes the connection rate between cells and maximizes the spread of an extracellular protein within the population. The maximum rate of protein spreading is observed for cells that reverse direction optimally for swarming. Cells that reverse too slowly or too fast fail to spread extracellular protein efficiently. In particular, a specific range of cell reversal frequencies was observed to maximize the cell-cell connection rate and minimize the time of protein spreading. Furthermore, our findings suggest that predesigned motion reversal can be employed to enhance the collective behavior of biological synthetic active systems.

  12. Optimized liquid-liquid extractive rerefining of spent lubricants.

    Science.gov (United States)

    Kamal, Muhammad Ashraf; Naqvi, Syed Mumtaz Danish; Khan, Fasihullah

    2014-01-01

    Central composite design methodology has been employed to model the sludge yield data obtained during liquid-liquid extractive rerefining of spent lubricants using an alcohol (1-butanol) and a ketone (methyl ethyl ketone) as prospective solvents. The study has resulted in two reasonably accurate multivariate process models that relate the sludge yield (R (2) = 0.9065 and 0.9072 for alcohol and ketone, resp.) to process variables (settling time t, operating temperature T, and oil to solvent ratio r). Construction of such models has allowed the maximization of the sludge yield (more than 8% and 3% in case of alcohol and ketone, resp.) so that the extraction of useable oil components from spent lubricants can economically be performed under extremely mild conditions (t = 16.7 h, T = 10°C, and r = 2) and fairly moderate conditions (t = 26.6 h, T = 10°C, and r = 5) established for the alcohol and ketone correspondingly. Based on these performance parameters alcohol appears to be superior over ketone for this extraction process. Additionally extractive treatment results in oil stocks with lesser quantity of environmentally hazardous polyaromatic hydrocarbons that are largely left in the separated sludge.

  13. Optimized Liquid-Liquid Extractive Rerefining of Spent Lubricants

    Directory of Open Access Journals (Sweden)

    Muhammad Ashraf Kamal

    2014-01-01

    Full Text Available Central composite design methodology has been employed to model the sludge yield data obtained during liquid-liquid extractive rerefining of spent lubricants using an alcohol (1-butanol and a ketone (methyl ethyl ketone as prospective solvents. The study has resulted in two reasonably accurate multivariate process models that relate the sludge yield (R2=0.9065 and 0.9072 for alcohol and ketone, resp. to process variables (settling time t, operating temperature T, and oil to solvent ratio r. Construction of such models has allowed the maximization of the sludge yield (more than 8% and 3% in case of alcohol and ketone, resp. so that the extraction of useable oil components from spent lubricants can economically be performed under extremely mild conditions (t = 16.7 h, T=10°C, and r=2 and fairly moderate conditions (t = 26.6 h, T=10°C, and r=5 established for the alcohol and ketone correspondingly. Based on these performance parameters alcohol appears to be superior over ketone for this extraction process. Additionally extractive treatment results in oil stocks with lesser quantity of environmentally hazardous polyaromatic hydrocarbons that are largely left in the separated sludge.

  14. An Optimized Approach for Extracting Approximate Functional Dependencies in XML Documents

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    In this paper, the definition of approximate XFDs based on value equality is proposed. Two metrics, support and strength, are presented for measuring the degree of approximate XFD. A basic algorithm is designed for extracting minimal set of approximate XFDs, and then two optimized strategies are proposed to improve the performance. Finally, the experimental results show that the optimized algorithms are correct and effective.

  15. Extraction of Fennel (Foeniculum vulgare) Seeds: Process Optimization and Antioxidant Capacity of the Extracts

    OpenAIRE

    2016-01-01

    This paper presents the study on the extraction of bioactive substances from fennel seeds. The impact of the main process variables (solvent composition, liquid-to-solid ratio, temperature, contact time) on the concentration of the target substances (polyphenols and flavonoids) in the extracts is studied resulting in the selection of a set of operating parameters, at which their content is maximized. Extracts with higher concentration of target compounds demonstrate higher antioxidant capacit...

  16. Optimization of Antifungal Extracts from Ficus hirta Fruits Using Response Surface Methodology and Antifungal Activity Tests

    Directory of Open Access Journals (Sweden)

    Chuying Chen

    2015-10-01

    Full Text Available The fruits of Ficus hirta (FH display strong antifungal activity against Penicillium italicum and Penicillium digitatum. In order to optimize the extraction conditions of antifungal extracts from FH fruit, various extraction parameters, such as ethanol concentration, extraction time, solvent to solid ratio and temperature, were chosen to identify their effects on the diameters of inhibition zones (DIZs against these two Penicillium molds. Response surface methodology (RSM was applied to obtain the optimal combination of these parameters. Results showed that the optimal extraction parameters for maximum antifungal activity were: 90% (v/v ethanol concentration, 65 min extraction time, 31 mL/g solvent to solid ratio and 51 °C temperature. Under the abovementioned extraction conditions, the experimental DIZs values obtained experimentally were 57.17 ± 0.75 and 39.33 ± 0.82 mm, which were very close to the values of 57.26 and 39.29 mm predicted by the model. Further, nine kinds of phytopathogens were tested in vitro to explore the antifungal activity of the FH extracts. It was found for the first time that the FH extracts showed significant inhibition on the growth of P. italicum, A. citri, P. vexans, P. cytosporella and P. digitatum.

  17. Effect of pH and postmortem aging on protein extraction from broiler breast muscle.

    Science.gov (United States)

    Eady, M; Samuel, D; Bowker, B

    2014-07-01

    This study determined the effects of extraction buffer pH and postmortem aging on the extraction of salt-soluble and water-soluble proteins from broiler pectoralis muscle. Deboned broiler breast fillets were collected at 4 h postmortem, packaged, and then stored at 4°C until 1, 5, or 8 d postmortem. After the designated aging period, salt-soluble and water-soluble protein extractions were performed using buffers at 7 different pH levels (pH 5.4, 6.4, 6.9, 7.2, 7.5, 8.0, 9.0). Protein concentrations of the extracts were measured and SDS-PAGE analysis was performed. Salt-soluble protein concentration increased (P protein concentration increased (P extraction buffer pH by aging treatment interaction for the total protein concentration of either the salt-soluble or water-soluble protein extracts. The protein concentrations of salt-soluble extracts were similar at both 1 and 8 d postmortem but lower (P protein concentrations of water-soluble extracts were similar at both 1 and 5 d postmortem, but higher (P extraction buffer pH and postmortem aging influenced the SDS-PAGE protein profiles of salt-soluble and water-soluble protein extracts from breast muscles. Data demonstrate that postmortem aging and extraction buffer pH influence both the total amount and the composition of the myofibrillar and sarcoplasmic proteins that can be extracted from broiler breast fillets.

  18. Protein extraction and enzymatic hydrolysis of ammonia-treated cassava leaves (Manihot esculenta Crantz).

    Science.gov (United States)

    Urribarrí, Lauris; Chacón, David; González, Orlaidy; Ferrer, Alexis

    2009-05-01

    In the present work, cassava leaves were treated with 0.5 kg ammonia/kg dry matter at 78 degrees C and 30% moisture content in a 2-kg reactor. Protein extraction was carried out with a calcium hydroxide solution (pH 10) for 30 min at several temperatures (30 degrees C, 45 degrees C, 60 degrees C, 75 degrees C, and 90 degrees C) and solid/liquid ratios (1:10 and 1:15) in a thermostatized bath. Soluble protein content of the extracts was determined by Lowry's method. Dry substrate concentrations of 5%, 7.5%, and 10% and enzyme doses of 2 and 5 IU/g dry matter were used for the enzymatic hydrolysis in an orbital incubator at 50 degrees C and 100 rpm. Both cellulase and xylanase were used. Reducing sugars produced were determined with the dinitrosalicylic acid method. The highest protein extraction yield for the ammonia-treated leaves was 29.10%, which was 50% higher than with the untreated leaves (20%), and was obtained at 90 degrees C with a 1:10 solid/liquid ratio. The concentrate had a protein content of 36.35% and the amino acid profile was suitable for swine and poultry. The highest sugar yield was 54.72% with respect to theoretical and was obtained with 5% solids and an enzyme dose of 5 IU/g dry matter. This yield was 3.4 times higher than the yield of the untreated leaves (16.13%). These results indicate that cassava leaves have a great potential for animal feeding and ethanol production. Both protein extraction and sugar yields may be enhanced by optimizing the ammonia treatment.

  19. Understanding leaf membrane protein extraction to develop a food-grade process.

    Science.gov (United States)

    Tamayo Tenorio, Angelica; Boom, Remko M; van der Goot, Atze Jan

    2017-02-15

    Leaf membrane proteins are an underutilised protein fraction for food applications. Proteins from leaves can contribute to a more complete use of resources and help to meet the increasing protein demand. Leaf protein extraction and purification is applied by other disciplines, such as proteomics. Therefore, this study analysed proteomic extraction methods for membrane proteins as an inspiration for a food-grade alternative process. Sugar beet leaves were extracted with two proteomic protocols: solvent extraction and Triton X-114 phase partitioning method. Extraction steps contributed to protein purity and/or to selective fractionation, enabling the purification of specific proteins. It was observed that membrane proteins distributed among different solvents, buffers and solutions used due to their physicochemical heterogeneity. This heterogeneity does not allow a total membrane protein extraction by a unique method or even combinations of processing steps, but it enables the creation of different fractions with different physicochemical properties useful for food applications.

  20. Optimization of microwave-assisted extraction of polyphenols from Myrtus communis L. leaves.

    Science.gov (United States)

    Dahmoune, Farid; Nayak, Balunkeswar; Moussi, Kamal; Remini, Hocine; Madani, Khodir

    2015-01-01

    Phytochemicals, such as phenolic compounds, are of great interest due to their health-benefitting antioxidant properties and possible protection against inflammation, cardiovascular diseases and certain types of cancer. Maximum retention of these phytochemicals during extraction requires optimised process parameter conditions. A microwave-assisted extraction (MAE) method was investigated for extraction of total phenolics from Myrtus communis leaves. The total phenolic capacity (TPC) of leaf extracts at optimised MAE conditions was compared with ultrasound-assisted extraction (UAE) and conventional solvent extraction (CSE). The influence of extraction parameters including ethanol concentration, microwave power, irradiation time and solvent-to-solid ratio on the extraction of TPC was modeled by using a second-order regression equation. The optimal MAE conditions were 42% ethanol concentration, 500 W microwave power, 62 s irradiation time and 32 mL/g solvent to material ratio. Ethanol concentration and liquid-to-solid ratio were the significant parameters for the extraction process (pMyrtus leaves using MAE compared with UAE and CSE, it was also observed that tannins (32.65 ± 0.01 mg/g), total flavonoids (5.02 ± 0.05 mg QE/g) and antioxidant activities (38.20 ± 1.08 μg GAE/mL) in MAE extracts were higher than the other two extracts. These findings further illustrate that extraction of bioactive phytochemicals from plant materials using MAE method consumes less extraction solvent and saves time.

  1. Optimization and comparative evaluation of nucleic acids extraction protocols

    Directory of Open Access Journals (Sweden)

    Lucian Negura

    2011-12-01

    Full Text Available Modern molecular applications have grown the need of biobanks, which contain DNA and RNA of high purety, quality, and quantity. Nucleic acid extraction methods have widely variate and evolve in time, from methods using toxic reagents to enzymatic protocols, and furthermore to DNA or RNA-binding polymers, separating membranes or highly eulogized commercially kits. In order to establish a constant, reproducible and ergonomic system in generating biobanks, we compared different available methods for the extraction of genomic DNA and total RNA, from peripheram blood or solid tumoral tissues. We evaluated the cost/effectiveness and time consumption of each method, tracking RNA/DNA quantity, quality and integrity. We imagined a “E-ratio” value to define these parameters, and a “NA- estimation” to integrate “E-ratio” with quality and integrity data.

  2. A Bayesian Sampler for Optimization of Protein Domain Hierarchies

    Science.gov (United States)

    2014-01-01

    Abstract The process of identifying and modeling functionally divergent subgroups for a specific protein domain class and arranging these subgroups hierarchically has, thus far, largely been done via manual curation. How to accomplish this automatically and optimally is an unsolved statistical and algorithmic problem that is addressed here via Markov chain Monte Carlo sampling. Taking as input a (typically very large) multiple-sequence alignment, the sampler creates and optimizes a hierarchy by adding and deleting leaf nodes, by moving nodes and subtrees up and down the hierarchy, by inserting or deleting internal nodes, and by redefining the sequences and conserved patterns associated with each node. All such operations are based on a probability distribution that models the conserved and divergent patterns defining each subgroup. When we view these patterns as sequence determinants of protein function, each node or subtree in such a hierarchy corresponds to a subgroup of sequences with similar biological properties. The sampler can be applied either de novo or to an existing hierarchy. When applied to 60 protein domains from multiple starting points in this way, it converged on similar solutions with nearly identical log-likelihood ratio scores, suggesting that it typically finds the optimal peak in the posterior probability distribution. Similarities and differences between independently generated, nearly optimal hierarchies for a given domain help distinguish robust from statistically uncertain features. Thus, a future application of the sampler is to provide confidence measures for various features of a domain hierarchy. PMID:24494927

  3. A Pareto-optimal refinement method for protein design scaffolds.

    Science.gov (United States)

    Nivón, Lucas Gregorio; Moretti, Rocco; Baker, David

    2013-01-01

    Computational design of protein function involves a search for amino acids with the lowest energy subject to a set of constraints specifying function. In many cases a set of natural protein backbone structures, or "scaffolds", are searched to find regions where functional sites (an enzyme active site, ligand binding pocket, protein-protein interaction region, etc.) can be placed, and the identities of the surrounding amino acids are optimized to satisfy functional constraints. Input native protein structures almost invariably have regions that score very poorly with the design force field, and any design based on these unmodified structures may result in mutations away from the native sequence solely as a result of the energetic strain. Because the input structure is already a stable protein, it is desirable to keep the total number of mutations to a minimum and to avoid mutations resulting from poorly-scoring input structures. Here we describe a protocol using cycles of minimization with combined backbone/sidechain restraints that is Pareto-optimal with respect to RMSD to the native structure and energetic strain reduction. The protocol should be broadly useful in the preparation of scaffold libraries for functional site design.

  4. Selection of an Appropriate Protein Extraction Method to Study the Phosphoproteome of Maize Photosynthetic Tissue

    OpenAIRE

    2016-01-01

    Often plant tissues are recalcitrant and, due to that, methods relying on protein precipitation, such as TCA/acetone precipitation and phenol extraction, are usually the methods of choice for protein extraction in plant proteomic studies. However, the addition of precipitation steps to protein extraction methods may negatively impact protein recovery, due to problems associated with protein re-solubilization. Moreover, we show that when working with non-recalcitrant plant tissues, such as you...

  5. Extraction of lipids from microalgae by ultrasound application: prospection of the optimal extraction method.

    Science.gov (United States)

    Araujo, Glacio S; Matos, Leonardo J B L; Fernandes, Jader O; Cartaxo, Samuel J M; Gonçalves, Luciana R B; Fernandes, Fabiano A N; Farias, Wladimir R L

    2013-01-01

    Microalgae have the ability to grow rapidly, synthesize and accumulate large amounts (approximately 20-50% of dry weight) of lipids. A successful and economically viable algae based oil industry will depend on the selection of appropriate microalgal strains and the selection of the most suitable lipid extraction method. In this paper, five extraction methods were evaluated regarding the extraction of lipids from Chlorella vulgaris: Bligh and Dyer, Chen, Folch, Hara and Radin, and Soxhlet. Furthermore, the addition of silica powder was studied to evaluate the introduction of more shear stress to the system as to increase the disruption of cell walls. Among the studied methods, the Bligh and Dyer method assisted by ultrasound resulted in the highest extraction of oil from C. vulgaris (52.5% w/w). Addition of powder silica did not improve the extraction of oil. Copyright © 2012 Elsevier B.V. All rights reserved.

  6. Optimization of the extraction conditions of the volatile compounds from chili peppers by headspace solid phase micro-extraction.

    Science.gov (United States)

    Junior, Stanislau Bogusz; de Marchi Tavares de Melo, Arlete; Zini, Cláudia Alcaraz; Godoy, Helena Teixeira

    2011-05-27

    A method involving headspace-solid phase micro-extraction (HS-SPME), gas chromatography with flame ionization detection (GC-FID) and gas chromatography with mass spectrometry (GC-MS) was developed and optimized to investigate the volatile composition of Capsicum chili peppers. Five SPME fibers were tested for extraction: carboxen/polydimethylsiloxane (CAR/PDMS-75μm), polydimethylsiloxane (PDMS-100μm), divinylbenzene/polydimethylsiloxane (DVB/PDMS-65μm), carbowax/divinylbenzene (CW/DVB-70μm), and divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS-50/30μm), the last of which was shown to be the most efficient fiber to trap the volatile compounds. Optimization of the extraction conditions was carried out using multivariate strategies such as factorial design and response surface methodology. Eighty three compounds were identified by GC-MS when using the optimized extraction conditions, the majority of which were esters. Copyright © 2011 Elsevier B.V. All rights reserved.

  7. An effective placental cotyledons proteins extraction method for 2D gel electrophoresis.

    Science.gov (United States)

    Tan, Niu J; Daim, Leona D J; Jamil, Amilia A M; Mohtarrudin, Norhafizah; Thilakavathy, Karuppiah

    2017-03-01

    Effective protein extraction is essential especially in producing a well-resolved proteome on 2D gels. A well-resolved placental cotyledon proteome, with good reproducibility, have allowed researchers to study the proteins underlying the physiology and pathophysiology of pregnancy. The aim of this study is to determine the best protein extraction protocol for the extraction of protein from placental cotyledons tissues for a two-dimensional gel electrophoresis (2D-GE). Based on widely used protein extraction strategies, 12 different extraction methodologies were carefully selected, which included one chemical extraction, two mechanical extraction coupled protein precipitations, and nine chemical extraction coupled protein precipitations. Extracted proteins were resolved in a one-dimensional gel electrophoresis and 2D-GE; then, it was compared with set criteria: extraction efficacy, protein resolution, reproducibility, and recovery efficiency. Our results revealed that a better profile was obtained by chemical extraction in comparison to mechanical extraction. We further compared chemical extraction coupled protein precipitation methodologies, where the DNase/lithium chloride-dense sucrose homogenization coupled dichloromethane-methanol precipitation (DNase/LiCl-DSH-D/MPE) method showed good protein extraction efficiency. This, however, was carried out with the best protein resolution and proteome reproducibility on 2D-gels. DNase/LiCl-DSH-D/MPE was efficient in the extraction of proteins from placental cotyledons tissues. In addition, this methodology could hypothetically allow the protein extraction of any tissue that contains highly abundant lipid and glycogen. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Multiresponse Optimization and Prediction of Antioxidant Properties of Aqueous Ginger Extract

    Science.gov (United States)

    Makanjuola, Solomon Akinremi; Enujiugha, Victor Ndigwe; Omoba, Olufunmilayo Sade

    2016-01-01

    The influence of extraction temperature, powder concentration, and extraction time on the antioxidant properties of aqueous ginger extract was investigated. The possibility of estimating the antioxidant properties of the extract from its absorbance and colour properties was also investigated. Results indicated that powder concentration was the most significant factor to consider in optimizing antioxidant extraction. However, temperature and time still influenced the 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activity while extraction temperature influenced the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of the extract. Using the total phenol content, total flavonoid content, ABTS radical scavenging activity, and DPPH radical scavenging activity of the extract, the multiresponse optimization condition for extraction of antioxidant based on the experimental range studied is 96°C, 2.10 g/100 mL, and 90 min. The absorbance of the ginger extract at 610 nm could be exploited for rapid estimation of its total flavonoid and polyphenol with a R2 of 0.713 and 0.753, respectively. PMID:28078258

  9. Process optimization and analysis of microwave assisted extraction of pectin from dragon fruit peel.

    Science.gov (United States)

    Thirugnanasambandham, K; Sivakumar, V; Prakash Maran, J

    2014-11-04

    Microwave assisted extraction (MAE) technique was employed for the extraction of pectin from dragon fruit peel. The extracting parameters were optimized by using four-variable-three-level Box-Behnken design (BBD) coupled with response surface methodology (RSM). RSM analysis indicated good correspondence between experimental and predicted values. 3D response surface plots were used to study the interactive effects of process variables on extraction of pectin. The optimum extraction conditions for the maximum yield of pectin were power of 400 W, temperature of 45 °C, extracting time of 20 min and solid-liquid ratio of 24 g/mL. Under these conditions, 7.5% of pectin was extracted. Copyright © 2014 Elsevier Ltd. All rights reserved.

  10. DNA extraction of birch leaves by improved CTAB method and optimization of its ISSR system

    Institute of Scientific and Technical Information of China (English)

    PAN hua; YANG Chuan-ping; WEI Zhi-gang; JIANG Jing

    2006-01-01

    The basic method of DNA extraction (CTAB) was improved as the multi-times STE-CTAB extraction method and used to extract the DNA of birch leaved in this experiment. Results showed that the improved method is suitable not only for genomic DNA extraction of birch but also for that of other plants. The purity of genomic DNA extracted by the multi-times STE-CTAB extraction method is higher than that by one time STE-CTAB method, and it does not need the process of RNase. The factors of influencing ISSR system were explored based on the genomic DNA of birch extracted by the two methods. The optimal conditions for ISSR system were determined as follows: cycles of denaturation for 30 s at 94℃, annealing for 30 s at 51 ℃, extension for 30 s at 72℃, and a final 7 min extension at 72 ℃.

  11. Optimization of supercritical fluid consecutive extractions of fatty acids and polyphenols from Vitis vinifera grape wastes.

    Science.gov (United States)

    Aizpurua-Olaizola, Oier; Ormazabal, Markel; Vallejo, Asier; Olivares, Maitane; Navarro, Patricia; Etxebarria, Nestor; Usobiaga, Aresatz

    2015-01-01

    In this study, supercritical fluid extraction has been successfully applied to a sequential fractionation of fatty acids and polyphenols from wine wastes (2 different vitis vinifera grapes). To this aim, in a 1st step just fatty acids were extracted and in a 2nd one the polyphenols. The variables that affected to the extraction efficiency were separately optimized in both steps following an experimental design approach. The effect of extraction temperature flow, pressure, and time were thoroughly evaluated for the extraction of fatty acids, whereas the addition of methanol was also considered in the case of the polyphenols extraction. A quantitative extraction with high efficiency was achieved at a very short time and low temperatures. Concerning quantification, fatty acids were determined by means of gas chromatography coupled to mass spectrometry after a derivatization step, whereas the polyphenols were analyzed by means of high performance liquid chromatography coupled to tandem mass spectrometry and the Folin-Ciocalteu method.

  12. Optimized Ultrasonic-assisted Extraction of Flavonoids from Pyracantha fortuneana Fruit and Evaluation of Antibacterial Activities

    Directory of Open Access Journals (Sweden)

    Yun Zhang

    2014-12-01

    Full Text Available Ultrasonic-Assisted Extraction (UAE method was developed for extraction of total flavonoids from Pyracantha fortuneana fruit. The UAE parameters and the antioxidant activities of flavonoids extract were investigated. Important variables and their levels were obtained using single factor analysis method and central composite design. Through Response Surface Methodology (RSM design experiments, the processing conditions were optimized as follows: ethanol concentration, 81.15%; liquid-solid ratio, 30.00 mL/g; extraction time, 3.14 h; and temperature, 69.55°C. Under the optimum condition, the extraction yield was 1.261%, which was well matched with the predicted values of 1.263%. The antibacterial activities of extract against four bacterial strains were determined by agar well diffusion method and the results indicated that the extract showed potent antibacterial activities, which concludes its application as natural antibacterial agent.

  13. Determination of extractability of pine bark using supercritical CO(2) extraction and different solvents: optimization and prediction.

    Science.gov (United States)

    Yesil-Celiktas, Ozlem; Otto, Frank; Gruener, Sabine; Parlar, Harun

    2009-01-28

    Bark from Pinus brutia was extracted with supercritical fluid extraction (SFE), using CO(2), at various extraction conditions both at laboratory and at pilot scale. Optimized parameters were 200 bar, 60 degrees C, and 3% ethanol at a solvent/feed ratio of 30. Additionally, the pine bark was sonicated (1 h at 50 degrees C) by different solvents (n-hexane, dichloromethane, ethyl acetate, and ethanol) to investigate the correlation between the different extraction setups and to obtain information on SFE up-scaling possibilities. Analyzed by HPLC, 7.2% of (-)-catechin was extractable at laboratory scale, and 58.4% (800 bar) and 47.8% (200 bar), both with modifiers, at pilot scale. By sonication with ethanol, 46.8% of (-)-catechin and almost 100% of (-)-epicatechin and (-)-catechin gallate were extracted. Ethyl acetate extract revealed high correlations with the laboratory scale SFE (r = 0.98) and also pilot scale SFE runs at 200 (r = 0.99) and 800 bar (r = 0.98) without modifiers.

  14. 正交试验法优化马兰头叶蛋白的提取工艺研究%Study on Optimization of the Extraction Technology of Protein from the Kalimeris indica(L.) Sch. -Bip.by Orthogonal Test

    Institute of Scientific and Technical Information of China (English)

    扶庆权; 华春; 周峰; 周泉澄

    2012-01-01

    采用碱提酸沉法从马兰头中提取蛋白质。在单因素试验的基础上,采用L9(34)正交试验设计,研究pH、温度、水解时间和料液比对马兰头蛋白提取率的影响。结果表明,在各影响因素中,影响程度依次为:pH〉温度〉水解时间〉料液比,碱法提取马兰头蛋白的最佳工艺条件为:pH 9.0、提取温度50℃、水解时间60min、料液比1︰25(g/mL),在此条件下测得马兰头蛋白提取率为75.86%。%The protein was extracted from Kalimeris indica(L.) Sch.-Bip.by alkali extraction.Based on single factor test,the effects of pH,extraction temperature,extraction time,extraction solid/liquid ratio(g/mL) on the extraction rate of protein were studied according to L9(34) orthogonal test.The results showed that factors influencing the extraction of protein from the Kalimeris indica(L.) Sch.-Bip.by alkali-extracting method were in the order as follows: pHextraction temperatureextraction solid/liquid ratio(g/mL)extraction time,the optimum extracting conditions of protein from the Kalimeris indica(L.) Sch.-Bip.for orthogonal test were adding 25 times amount water solutions of pH 9.0 and extracting for 60 min at 50℃,and the extraction rate of protein was 75.86%.

  15. Optimal Fixation Conditions and DNA Extraction Methods for MLPA Analysis on FFPE Tissue-Derived DNA.

    Science.gov (United States)

    Atanesyan, Lilit; Steenkamer, Maryvonne J; Horstman, Anja; Moelans, Cathy B; Schouten, Jan P; Savola, Suvi P

    2017-01-01

    Molecular genetic analysis of formalin-fixed, paraffin-embedded (FFPE) tissues is of great importance both for research and diagnostics. Multiplex ligation-dependent probe amplification (MLPA) is a widely used technique for gene copy number determination, and it has been successfully used for FFPE tissue-extracted DNA analysis. However, there have been no studies addressing the effect of tissue fixation procedures and DNA extraction methods on MLPA. This study therefore focuses on selecting optimal preanalytic conditions such as FFPE tissue preparation conditions and DNA extraction methods. Healthy tissues were fixed in buffered or nonbuffered formalin for 1 hour, 12 to 24 hours, or 48 to 60 hours at 4 °C or at room temperature. DNA extracted from differently fixed and subsequently paraffin-embedded tissues was used for MLPA. Four commercial DNA extraction kits and one in-house method were compared. Tissues fixed for 12 to 24 hours in buffered formalin at room temperature produced DNA with the most optimal quality for MLPA. The in-house FFPE DNA extraction method was shown to perform as efficient as or even superior to other methods in terms of suitability for MLPA, time and cost-efficiency, and ease of performance. FFPE-extracted DNA is well suitable for MLPA analysis, given that optimal tissue fixation and DNA extraction methods are chosen.

  16. Optimization of PEG-based extraction of polysaccharides from Dendrobium nobile Lindl. and bioactivity study.

    Science.gov (United States)

    Zhang, Yi; Wang, Hongxin; Wang, Peng; Ma, ChaoYang; He, GuoHua; Rahman, Md Ramim Tanver

    2016-11-01

    Polyethylene glycol (PEG) as a green solvent was employed to extract polysaccharide. The optimal conditions for PEG-based ultrasonic extraction of Dendrobium nobile Lindl. polysaccharide (JCP) were determined by response surface methodology. Under the optimal conditions: extraction temperature of 58.5°C; ultrasound power of 193W, and the concentration of polyethylene glycol-200 (PEG-200) solution of 45%, the highest JCP yield was obtained as 15.23±0.57%, which was close to the predicted yield, 15.57%. UV and FT-IR analysis revealed the general characteristic absorption peaks of both JCP with water extraction (JCPw) and PEG-200 solvent extraction (JCPp). Thermal analysis of both JCPs was performed with Thermal Gravimetric Analyzer (TGA) and Differential Scanning Calorimeter (DSC). Antioxidant activities of two polysaccharides were also compared and no significant difference in vitro was obtained. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Optimization of the Ocimum basilicum L. extraction process regarding the antioxidant activity

    Directory of Open Access Journals (Sweden)

    Vidović Senka S.

    2012-01-01

    Full Text Available The levels of input variables (temperature and extraction solvent that optimize a particular response (total phenols content, total flavonoids content and antioxidant activity of the Ocimum basilicum L. extraction process were determined by the response surface methodology (RSM. The influence of theextraction temperature on extraction process was investigated in the range from 33.8ºC to 76.2ºC, as well as of extraction solvent ethanol, in the range of concentrations from 21.7% to 78.3%. For the preparation of basil dry extract, characterized with minimal IC50 value, the calculated optimal values of temperature and ethanol concentration were: 75.33ºC and 73.66% (w/w. [Projekat Ministarstva nauke Republike Srbije, br. TR 31013

  18. Tree kernel-based protein-protein interaction extraction from biomedical literature.

    Science.gov (United States)

    Qian, Longhua; Zhou, Guodong

    2012-06-01

    There is a surge of research interest in protein-protein interaction (PPI) extraction from biomedical literature. While most of the state-of-the-art PPI extraction systems focus on dependency-based structured information, the rich structured information inherent in constituent parse trees has not been extensively explored for PPI extraction. In this paper, we propose a novel approach to tree kernel-based PPI extraction, where the tree representation generated from a constituent syntactic parser is further refined using the shortest dependency path between two proteins derived from a dependency parser. Specifically, all the constituent tree nodes associated with the nodes on the shortest dependency path are kept intact, while other nodes are removed safely to make the constituent tree concise and precise for PPI extraction. Compared with previously used constituent tree setups, our dependency-motivated constituent tree setup achieves the best results across five commonly used PPI corpora. Moreover, our tree kernel-based method outperforms other single kernel-based ones and performs comparably with some multiple kernel ones on the most commonly tested AIMed corpus.

  19. Comparing extraction buffers to identify optimal method to extract somatic coliphages from sewage sludges.

    Science.gov (United States)

    Murthi, Poornima; Praveen, Chandni; Jesudhasan, Palmy R; Pillai, Suresh D

    2012-08-01

    Somatic coliphages are present in high numbers in sewage sludge. Since they are conservative indicators of viruses during wastewater treatment processes, they are being used to evaluate the effectiveness of sludge treatment processes. However, efficient methods to extract them from sludge are lacking. The objective was to compare different virus extraction procedures and develop a method to extract coliphages from sewage sludge. Twelve different extraction buffers and procedures varying in composition, pH, and sonication were compared in their ability to recover indigenous phages from sludges. The 3% buffered beef extract (BBE) (pH 9.0), the 10% BBE (pH 9.0), and the 10% BBE (pH 7.0) with sonication were short-listed and their recovery efficiency was determined using coliphage-spiked samples. The highest recovery was 16% for the extraction that involved 10% BBE at pH 9.0. There is a need to develop methods to extract somatic phages from sludges for monitoring sludge treatment processes.

  20. Optimization of olive leaf extract obtained by ultrasound-assisted extraction with response surface methodology.

    Science.gov (United States)

    Şahin, Selin; Samlı, Rüya

    2013-01-01

    In the present article, ultrasound-assisted extraction (UAE) of polyphenols from agricultural and industrial waste of olive oil and table oil productions, olive tree (Olea europaea) leaves were investigated. The aim of the study is to examine the extraction parameters such as solvent concentration (0-100% ethanol (EtOH), v/v), the ratio of solid to solvent (25-50mg/mL) and extraction time (20-60 min), and to obtain the best possible combinations of these parameters through response surface methodology (RSM). The extract yield was stated as mg extract per g of dried leaf (DL). Total phenolic content was expressed in gallic acid equivalent (GAE) per g of dried leaf. Free radical scavenging activity for the antioxidant capacity was tested by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical. The second order polynomial model gave a satisfactory description of the experimental data. 201.2158 mg extract/g DL, 25.0626 mg GAE/g DL, and 95.5610% in respect to inhibition of DPPH radical were predicted at the optimum operating conditions (500 mg solid to 10 mL solvent ratio, 60 min of extraction time and 50% EtOH composition), respectively.

  1. Optimized protein extraction methods for proteomic analysis of Rhizoctonia solani

    Science.gov (United States)

    Rhizoctonia solani (Teleomorph: Thanatephorus cucumeris) is a basidiomycetous fungus which includes important plant pathogens, saprophytes and mycorrhizae. R. solani displays several hyphal anastomosis groups (AGs) with distinct host plant specializations. In order to facilitate studies on its biol...

  2. Optimization of Ultrasonic-Assisted Extraction of Cordycepin from Cordyceps militaris Using Orthogonal Experimental Design

    Directory of Open Access Journals (Sweden)

    Hsiu-Ju Wang

    2014-12-01

    Full Text Available This study reports on the optimization of the extraction conditions of cordycepin from Cordyceps militaris by using ultrasonication. For this purpose, the orthogonal experimental design was used to investigate the effects of factors on the ultrasonic-assisted extraction (UAE. Four factors: extraction time (min, ethanol concentration (%, extraction temperature (°C and extraction frequency (kHz, were studied. The results showed that the highest cordycepin yield of 7.04 mg/g (86.98% ± 0.23% was obtained with an extraction time of 60 min, ethanol concentration of 50%, extraction temperature of 65 °C and extraction frequency of 56 kHz. It was found that the cordycepin extraction yield increased with the effect of ultrasonication during the extraction process. Therefore, UAE can be used as an alternative to conventional immersion extraction with respect to the recovery of cordycepin from C. militaris, with the advantages of shorter extraction time and reduced solvent consumption.

  3. Optimization of microwave-assisted enzymatic extraction of polysaccharides from the fruit of Schisandra chinensis Baill.

    Science.gov (United States)

    Cheng, Zhenyu; Song, Haiyan; Yang, Yingjie; Liu, Yan; Liu, Zhigang; Hu, Haobin; Zhang, Yang

    2015-05-01

    A microwave-assisted enzymatic extraction (MAEE) method had been developed, which was optimized by response surface methodology (RSM) and orthogonal test design, to enhance the extraction of crude polysaccharides (CPS) from the fruit of Schisandra chinensis Baill. The optimum conditions were as follows: microwave irradiation time of 10 min, extraction pH of 4.21, extraction temperature of 47.58°C, extraction time of 3h and enzyme concentration of 1.5% (wt% of S. chinensis powder) for cellulase, papain and pectinase, respectively. Under these conditions, the extraction yield of CPS was 7.38 ± 0.21%, which was well in close agreement with the value predicted by the model. The three methods including heat-refluxing extraction (HRE), ultrasonic-assisted extraction (UAE) and enzyme-assisted extraction (EAE) for extracting CPS by RSM were further compared. Results indicated MAEE method had the highest extraction yields of CPS at lower temperature. It was indicated that the proposed approach in this study was a simple and efficient technique for extraction of CPS in S. chinensis Baill.

  4. Pressure-assisted protein extraction: a novel method for recovering proteins from archival tissue for proteomic analysis.

    Science.gov (United States)

    Fowler, Carol B; Waybright, Timothy J; Veenstra, Timothy D; O'Leary, Timothy J; Mason, Jeffrey T

    2012-04-06

    Formaldehyde-fixed, paraffin-embedded (FFPE) tissue repositories represent a valuable resource for the retrospective study of disease progression and response to therapy. However, the proteomic analysis of FFPE tissues has been hampered by formaldehyde-induced protein modifications, which reduce protein extraction efficiency and may lead to protein misidentification. Here, we demonstrate the use of heat augmented with high hydrostatic pressure (40,000 psi) as a novel method for the recovery of intact proteins from FFPE mouse liver. When FFPE mouse liver was extracted using heat and elevated pressure, there was a 4-fold increase in protein extraction efficiency, a 3-fold increase in the extraction of intact proteins, and up to a 30-fold increase in the number of nonredundant proteins identified by mass spectrometry, compared to matched tissue extracted with heat alone. More importantly, the number of nonredundant proteins identified in the FFPE tissue was nearly identical to that of matched fresh-frozen tissue.

  5. Integrating semantic information into multiple kernels for protein-protein interaction extraction from biomedical literatures.

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    Lishuang Li

    Full Text Available Protein-Protein Interaction (PPI extraction is an important task in the biomedical information extraction. Presently, many machine learning methods for PPI extraction have achieved promising results. However, the performance is still not satisfactory. One reason is that the semantic resources were basically ignored. In this paper, we propose a multiple-kernel learning-based approach to extract PPIs, combining the feature-based kernel, tree kernel and semantic kernel. Particularly, we extend the shortest path-enclosed tree kernel (SPT by a dynamic extended strategy to retrieve the richer syntactic information. Our semantic kernel calculates the protein-protein pair similarity and the context similarity based on two semantic resources: WordNet and Medical Subject Heading (MeSH. We evaluate our method with Support Vector Machine (SVM and achieve an F-score of 69.40% and an AUC of 92.00%, which show that our method outperforms most of the state-of-the-art systems by integrating semantic information.

  6. A Shortest Dependency Path Based Convolutional Neural Network for Protein-Protein Relation Extraction

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    Lei Hua

    2016-01-01

    Full Text Available The state-of-the-art methods for protein-protein interaction (PPI extraction are primarily based on kernel methods, and their performances strongly depend on the handcraft features. In this paper, we tackle PPI extraction by using convolutional neural networks (CNN and propose a shortest dependency path based CNN (sdpCNN model. The proposed method (1 only takes the sdp and word embedding as input and (2 could avoid bias from feature selection by using CNN. We performed experiments on standard Aimed and BioInfer datasets, and the experimental results demonstrated that our approach outperformed state-of-the-art kernel based methods. In particular, by tracking the sdpCNN model, we find that sdpCNN could extract key features automatically and it is verified that pretrained word embedding is crucial in PPI task.

  7. A Shortest Dependency Path Based Convolutional Neural Network for Protein-Protein Relation Extraction.

    Science.gov (United States)

    Hua, Lei; Quan, Chanqin

    2016-01-01

    The state-of-the-art methods for protein-protein interaction (PPI) extraction are primarily based on kernel methods, and their performances strongly depend on the handcraft features. In this paper, we tackle PPI extraction by using convolutional neural networks (CNN) and propose a shortest dependency path based CNN (sdpCNN) model. The proposed method (1) only takes the sdp and word embedding as input and (2) could avoid bias from feature selection by using CNN. We performed experiments on standard Aimed and BioInfer datasets, and the experimental results demonstrated that our approach outperformed state-of-the-art kernel based methods. In particular, by tracking the sdpCNN model, we find that sdpCNN could extract key features automatically and it is verified that pretrained word embedding is crucial in PPI task.

  8. Utilization of baker's yeast (Saccharomyces cerevisiae for the production of yeast extract: effects of different enzymatic treatments on solid, protein and carbohydrate recovery

    Directory of Open Access Journals (Sweden)

    TATJANA VUKASINOVIC MILIC

    2007-05-01

    Full Text Available Yeast extract (YE was produced from commercial pressed baker's yeast (active and inactivated using two enzymes: papain and lyticase. The effects of enzyme concentration and hydrolysis time on the recovery of solid, protein and carbohydrate were investigated. Autolysis, as a basic method for cell lysis was also used and the results compared. The optimal extraction conditions were investigated. The optimal concentrations of papain and lyticase were found to be 2.5 % and 0.025 %, respectively.

  9. Optimization of DNA Extractions from Iron-rich Microbial Mats

    Science.gov (United States)

    Fullerton, H.; Hilton, T. S.; Moyer, C. L.

    2013-12-01

    Iron is the fourth most abundant element in the Earth's crust and is potentially one of the most abundant energy sources on the earth as an electron donor for chemolithoautotrophicgrowth coupled to Fe(II) oxidation. Many microbes have adapted to this energy source. One such bacterial class are the Zetaproteobacteria, which dominate Iron-rich microbial mats at Loihi seamount. Although cell counts are very high (up to 5.3x108 cells/ml), efficient DNA yields are low in comparison. In this study we compared extraction efficiency across different methods and with the addition of various buffers. Regardless of protocol (i.e., kit), the addition of sodium citrate drastically increased the DNA yield. The addition of sodium citrate did not alter community structure as determined by T-RFLP and qPCR. Citrate is a well-known ferric iron chelator and will bind ferrous as well. The chelated iron is then unable to participate in the Fenton reaction and this stops the generation of hydroxyl radicals which in turn can react and degrade the extracted DNA. We have utilized this relationship to allow us to obtain nearly an order of magnitude more microbial community DNA per sample, which should also have implications when processing low biomass samples, e.g., from the deep subsurface.

  10. Simultaneous extraction, optimization, and analysis of flavonoids and polyphenols from peach and pumpkin extracts using a TLC-densitometric method.

    Science.gov (United States)

    Altemimi, Ammar; Watson, Dennis G; Kinsel, Mary; Lightfoot, David A

    2015-01-01

    The use of medicinal plants has been reported throughout human history. In the fight against illnesses, medicinal plants represent the primary health care system for 60 % of the world's population. Flavonoids are polyphenolic compounds with active anti-microbial properties; they are produced in plants as pigments. Quercetin, myricetin, and rutin are among the most well-known and prevalent flavonoids in plants, with an antioxidant activity capable of decreasing the oxidation of low density lipoproteins [LDLs]. To date, this research is the first of its kind to employ a coupled thin-layer chromatography (TLC) and a densitometric quantification method with a Box-Behnken design (BBD) response surface methodology (RSM) for optimization of ultrasonic-assisted extraction and determination of rutin and quercetin from peach and ellagic acid and myricetin from pumpkin fruits. The effect of process variables (extraction temperature (°C), extraction power (%) and extraction time (min)) on ultrasound-assisted extraction (UAE) were examined by using BBD and RSM. TLC followed by Quantity-One™ (BioRad) image analysis as a simple and rapid method was used for identification and quantification of the compounds in complex mixtures. The results were consistent under optimal conditions among the experimental values and their predicted values. A mass spectrometry (MALDI-TOF MS) technique was also used to confirm the identity of the natural products in the TLC spots resolved. The results show that the coupled TLC-densitometric methods & BBD can be a very powerful approach to qualitative and quantitative analysis of; rutin and quercetin from peach extracts; and ellagic acid and myricetin contents from pumpkin extracts.

  11. Selection of an Appropriate Protein Extraction Method to Study the Phosphoproteome of Maize Photosynthetic Tissue

    Science.gov (United States)

    Luís, Inês M.; Alexandre, Bruno M.; Oliveira, M. Margarida

    2016-01-01

    Often plant tissues are recalcitrant and, due to that, methods relying on protein precipitation, such as TCA/acetone precipitation and phenol extraction, are usually the methods of choice for protein extraction in plant proteomic studies. However, the addition of precipitation steps to protein extraction methods may negatively impact protein recovery, due to problems associated with protein re-solubilization. Moreover, we show that when working with non-recalcitrant plant tissues, such as young maize leaves, protein extraction methods with precipitation steps compromise the maintenance of some labile post-translational modifications (PTMs), such as phosphorylation. Therefore, a critical issue when studying PTMs in plant proteins is to ensure that the protein extraction method is the most appropriate, both at qualitative and quantitative levels. In this work, we compared five methods for protein extraction of the C4-photosynthesis related proteins, in the tip of fully expanded third-leaves. These included: TCA/Acetone Precipitation; Phenol Extraction; TCA/Acetone Precipitation followed by Phenol Extraction; direct extraction in Lysis Buffer (a urea-based buffer); and direct extraction in Lysis Buffer followed by Cleanup with a commercial kit. Protein extraction in Lysis Buffer performed better in comparison to the other methods. It gave one of the highest protein yields, good coverage of the extracted proteome and phosphoproteome, high reproducibility, and little protein degradation. This was also the easiest and fastest method, warranting minimal sample handling. We also show that this method is adequate for the successful extraction of key enzymes of the C4-photosynthetic metabolism, such as PEPC, PPDK, PEPCK, and NADP-ME. This was confirmed by MALDI-TOF/TOF MS analysis of excised spots of 2DE analyses of the extracted protein pools. Staining for phosphorylated proteins in 2DE revealed the presence of several phosphorylated isoforms of PEPC, PPDK, and PEPCK. PMID

  12. DNA, RNA, and Protein Extraction: The Past and The Present

    Directory of Open Access Journals (Sweden)

    Beow Chin Yiap

    2009-01-01

    Full Text Available Extraction of DNA, RNA, and protein is the basic method used in molecular biology. These biomolecules can be isolated from any biological material for subsequent downstream processes, analytical, or preparative purposes. In the past, the process of extraction and purification of nucleic acids used to be complicated, time-consuming, labor-intensive, and limited in terms of overall throughput. Currently, there are many specialized methods that can be used to extract pure biomolecules, such as solution-based and column-based protocols. Manual method has certainly come a long way over time with various commercial offerings which included complete kits containing most of the components needed to isolate nucleic acid, but most of them require repeated centrifugation steps, followed by removal of supernatants depending on the type of specimen and additional mechanical treatment. Automated systems designed for medium-to-large laboratories have grown in demand over recent years. It is an alternative to labor-intensive manual methods. The technology should allow a high throughput of samples; the yield, purity, reproducibility, and scalability of the biomolecules as well as the speed, accuracy, and reliability of the assay should be maximal, while minimizing the risk of cross-contamination.

  13. Optimization of the extraction of carrageenan from Kappaphycus alvarezii using response surface methodology

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    Vanessa Webber

    2012-12-01

    Full Text Available This study aims to optimize an alternative method of extraction of carrageenan without previous alkaline treatment and ethanol precipitation using Response Surface Methodology (RSM. In order to introduce an innovation in the isolation step, atomization drying was used reducing the time for obtaining dry carrageenan powder. The effects of extraction time and temperature on yield, gel strength, and viscosity were evaluated. Furthermore, the extracted material was submitted to structural analysis, by infrared spectroscopy and nuclear magnetic resonance spectroscopy (¹H-NMR, and chemical composition analysis. Results showed that the generated regression models adequately explained the data variation. Carrageenan yield and gel viscosity were influenced only by the extraction temperature. However, gel strength was influenced by both, extraction time and extraction temperature. Optimal extraction conditions were 74 ºC and 4 hours. In these conditions, the carrageenan extract properties determined by the polynomial model were 31.17%, 158.27 g.cm-2, and 29.5 cP for yield, gel strength, and viscosity, respectively, while under the experimental conditions they were 35.8 ± 4.68%, 112.50 ± 4.96 g.cm-2, and 16.01 ± 1.03 cP, respectively. The chemical composition, nuclear magnetic resonance spectroscopy, and infrared spectroscopy analyses showed that the crude carrageenan extracted is composed mainly of κ-carrageenan.

  14. Optimization of liquid-liquid extraction of biosurfactants from corn steep liquor.

    Science.gov (United States)

    Vecino, X; Barbosa-Pereira, L; Devesa-Rey, R; Cruz, J M; Moldes, A B

    2015-09-01

    In this work, the optimization of the operational conditions for the chloroform-based extraction of surface-active compounds from corn steep liquor (CSL) was carried out and the nutritional properties of the remnant aqueous phase (CSL-less biosurfactant) was evaluated as microbial fermentation medium. The optimal conditions to obtain biosurfactants from CSL were as follows: chloroform/CSL ratio 2 (v/v), 56 °C at extraction times >30 min. At the optima conditions, 100 % of biosurfactant extract can be obtained from CSL, obtaining 12.0 ± 0.5 g of biosurfactant extract/Kg of CSL. The critical micelle concentration (CMC) of the biosurfactant extract was 399.4 mg L(-1). This value is similar to the CMC of cetrimonium bromide (CTAB), a cationic surfactant used in the formulation of nanoparticles. The extraction of biosurfactant can be also carried out at room temperature although in this case, the extraction yield decreased about 15 %. The extraction of surface-active compounds from agroindustrial streams can suppose important advances for the bio-based surfactants industry. Biosurfactants obtained in this work are not only more eco-friendly than chemical detergents but also can be cost competitive with its chemical counterparts. Furthermore, after the extraction of surface-active compounds, CSL-less biosurfactant was found to be suitable as nutritional supplement for lactic acid bacteria, maintaining its nutritional properties in comparison with regular CSL.

  15. Information Extraction of High Resolution Remote Sensing Images Based on the Calculation of Optimal Segmentation Parameters.

    Science.gov (United States)

    Zhu, Hongchun; Cai, Lijie; Liu, Haiying; Huang, Wei

    2016-01-01

    Multi-scale image segmentation and the selection of optimal segmentation parameters are the key processes in the object-oriented information extraction of high-resolution remote sensing images. The accuracy of remote sensing special subject information depends on this extraction. On the basis of WorldView-2 high-resolution data, the optimal segmentation parameters methodof object-oriented image segmentation and high-resolution image information extraction, the following processes were conducted in this study. Firstly, the best combination of the bands and weights was determined for the information extraction of high-resolution remote sensing image. An improved weighted mean-variance method was proposed andused to calculatethe optimal segmentation scale. Thereafter, the best shape factor parameter and compact factor parameters were computed with the use of the control variables and the combination of the heterogeneity and homogeneity indexes. Different types of image segmentation parameters were obtained according to the surface features. The high-resolution remote sensing images were multi-scale segmented with the optimal segmentation parameters. Ahierarchical network structure was established by setting the information extraction rules to achieve object-oriented information extraction. This study presents an effective and practical method that can explain expert input judgment by reproducible quantitative measurements. Furthermore the results of this procedure may be incorporated into a classification scheme.

  16. Information Extraction of High Resolution Remote Sensing Images Based on the Calculation of Optimal Segmentation Parameters.

    Directory of Open Access Journals (Sweden)

    Hongchun Zhu

    Full Text Available Multi-scale image segmentation and the selection of optimal segmentation parameters are the key processes in the object-oriented information extraction of high-resolution remote sensing images. The accuracy of remote sensing special subject information depends on this extraction. On the basis of WorldView-2 high-resolution data, the optimal segmentation parameters methodof object-oriented image segmentation and high-resolution image information extraction, the following processes were conducted in this study. Firstly, the best combination of the bands and weights was determined for the information extraction of high-resolution remote sensing image. An improved weighted mean-variance method was proposed andused to calculatethe optimal segmentation scale. Thereafter, the best shape factor parameter and compact factor parameters were computed with the use of the control variables and the combination of the heterogeneity and homogeneity indexes. Different types of image segmentation parameters were obtained according to the surface features. The high-resolution remote sensing images were multi-scale segmented with the optimal segmentation parameters. Ahierarchical network structure was established by setting the information extraction rules to achieve object-oriented information extraction. This study presents an effective and practical method that can explain expert input judgment by reproducible quantitative measurements. Furthermore the results of this procedure may be incorporated into a classification scheme.

  17. Optimization of polysaccharides extraction from Tricholoma mongolicum Imai and their antioxidant and antiproliferative activities.

    Science.gov (United States)

    Wang, Jin; Zhao, Yongming; Li, Wei; Wang, Zhibao; Shen, Lixia

    2015-10-20

    Response surface methodology was used to optimize the enzyme-assisted extraction parameters for polysaccharides from Tricholoma mongolicum Imai (TMIPs). The orthogonal test design was employed to determine the optimal concentration of three kinds of enzyme (trypsin, pectinase and papain) and the optimal concentrations of trypsin, pectinase and papain were 1.5%, 2.5%, and 2.0% (dry weight % of T. mongolicum Imai powder), respectively. In addition, three variables that remarkably affected the yield of polysaccharides such as extraction temperature, pH and extraction time were studied based on a Box-Behnken design. The results demonstrated that extraction time was the most remarkable factor affecting the TMIPs yield, followed by pH and temperature. Optimal extraction was obtained at 48.4°C, pH 5.4, and extraction time of 132min. Under these optimum conditions, the yield was 24.01%, which is consistent with the predicted value. Furthermore, crude polysaccharides were purified to obtain four fractions. In vitro antioxidant and antiproliferative activities results showed that TMIP-4 had stronger antioxidant and antiproliferative capacity than other fractions.

  18. Development and extraction optimization of baicalein and pinostrobin from Scutellaria violacea through response surface methodology

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    Shankar Subramaniam

    2015-01-01

    Full Text Available Objective: To develop a process that involves optimization of the amount of baicalein and pinostrobin from the hydro-methanolic extract of the leaves of Scutellaria violacea by response surface methodology (RSM. Materials and Methods: The combinatorial influence of various extraction parameters on the extraction yield was investigated by adopting Box-Behnken experimental design. Preliminary experiments carried out based on the traditional one variable at a time optimization revealed four such operational parameters to play a crucial role by influencing the yield. These four process parameters at three levels were considered to obtain the Box-Behnken experimental design. Results: RSM based model fitted to the resulting experimental data suggested that 52.3% methanol/water, 12.46:1 solvent-solid ratio, 285 rpm agitation and 6.07 h of extraction time are the optimal conditions which yielded a maximized amount of baicalein and pinostrobin of 2.9 and 4.05 mg/g DM. Analysis of variance revealed a high correlation coefficient (R2 = 0.999 for baicalein and 0.994 for pinostrobin, signifying a good fit between the regression model (second order and the experimental observations. Conclusion: The present study signifies that both the metabolites have been extracted from S. violacea for the first time. Further, this study developed an optimized extraction procedure to obtain maximum yield of the metabolites, which is unique and better than conventional extraction methodology. The operational parameters under optimized conditions accounts for the lowest cost in extraction process thus, providing an efficient, rapid and cost-effective method for isolation and scale up of these commercially vital flavonoids.

  19. Optimization and evaluation of wheat germ oil extracted by supercritical CO2

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    Niu, LiYa

    2011-06-01

    Full Text Available Box-Behnken design combined with response surface methodology (RSM was used to optimize the parameters of supercritical CO2 extraction (SFE of wheat germ oil. The quality of the oil and residual meal obtained by SFE and solvent extraction (SE were evaluated from proximate analysis, fatty acid composition and antioxidant activity. A maximum oil yield of 10.46% was achieved under the optimal conditions of wheat germ particle size 60-80 mesh; water content 4.37%; pressure 30MPa; temperature 40°C extraction time 1.7h. The oil obtained by SFE showed stronger DPPH radical scavenging ability than SE oil at the same concentration. The fatty acid composition of SFE oil was similar to SE oil. Higher contents of protein (34.3% and lysine (2.47g/100g were found in the residual meal obtained by SFE. The results show that oil and defatted meal obtained by SFE can be promising nutritional sources for food.Un diseño Box-Behnken combinado con metodología de superficie de respuesta (RSM fue usado para optimizar los parámetros de extracción con fluido supercrítico (SFE del aceite de germen de trigo. La calidad del aceite y de la harina residual obtenida por SFE y por extracción con solvente (SE fue evaluada mediante su análisis porcentual, composición de ácidos grasos y actividad antioxidante. Un máximo rendimiento de aceite del 10.46% fue obtenido con las condiciones óptimas de 60-80 mesh de tamaño de partícula del germen de trigo; 4.37% contenido de agua; 30MPa de presión; 40°C de temperatura y 1.7 h de tiempo de extracción. El aceite obtenido por SFE mostró una capacidad atrapadora de radicales libres mucho mayor que el aceite obtenido por SE a la misma concentración. La composición de ácidos grasos del aceite SFE fue similar al aceite SE. El mayor contenido de proteínas (34% y de lisina (2.47g/100g fue encontrado en las harinas residuales obtenidas por SFE. Los resultados muestran que el aceite y la harina desengrasada obtenidas por SFE

  20. Optimization extraction, preliminary characterization and antioxidant activity in vitro of polysaccharides from Stachys sieboldii Miq. tubers.

    Science.gov (United States)

    Feng, Kai; Chen, Wei; Sun, Liwei; Liu, Jianzeng; Zhao, Yangxin; Li, Luxi; Wang, Yuxing; Zhang, Wenjing

    2015-07-10

    Response surface methodology was used to optimize the extraction conditions of water-soluble polysaccharides from Stachys sieboldii Miq. tubers. A central composite design was used to optimize the extraction processing parameters. The optimum extraction conditions are as follows: extraction temperature, 95°C; extraction time, 2.5h; water to raw material ratio, 16; and extraction frequency, 3. Under the optimized conditions, an experimental yield of 9.21 ± 0.18%, which is in good agreement with the predicted yield, was obtained. Purified polysaccharide SSP II-a was successfully obtained using diethylaminoethanol-Sepharose and Sepharose CL-6B column chromatography. SSP II-a was found to be an acidic polysaccharide fraction with an average molecular weight of 168kDa and composed of rhamnose, glucuronic acid, galacturonic acid, glucose, galactose and arabinose. In vitro antioxidant activity assays suggested that SSP II-a presents high scavenging activity toward superoxide anion, hydroxyl, and 2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) radicals but relatively lower scavenging activity toward 1,1-diphenyl-2-picrylhydrazyl radicals. The results indicated that response surface methodology is an effective method for the extraction of polysaccharides from S. sieboldii Miq. tubers and the polysaccharides could be explored as a potential antioxidant agent for use in medicine or functional food.

  1. Palatability of water-soluble extracts of protein sources and replacement of fishmeal by a selected mixture of protein sources for juvenile turbot ( Scophthalmus maximus)

    Science.gov (United States)

    Dong, Chun; He, Gen; Mai, Kangsen; Zhou, Huihui; Xu, Wei

    2016-06-01

    Poor palatability is a limiting factor for replacing fishmeal with other protein sources in aquaculture. The water-soluble molecules with low molecular weights are the major determinants of the palatability of diets. The present study was conducted to investigate the palatability of water-soluble extracts from single protein source (single extract pellets) and the mixture of these extracts with different proportions (blended extract pellets) in juvenile turbot ( Scophthalmus maximus). Then according to the palatability of blended extract pellets, an optimal mixture proportion was selected, and a new protein source made from raw protein materials with the selected proportion was formulated to replace fishmeal. Summarily, the palatability of single extract pellets for turbot was descendent from fishmeal to pet-food grade poultry by-product meal, wheat gluten meal, soybean meal, peanut meal, meat and bone meal, and corn gluten meal. Subsequently, according to the palatability of single extract pellets, 52 kinds of blended extract pellets were designed to test their palatability. The results showed that the pellets presented remarkably different palatability, and the optimal one was diet 52 (wheat gluten meal: pet-food grade poultry by-product meal: meat and bone meal: corn gluten meal = 1:6:1:2). The highest ingestion ratio (the number of pellets ingested/the number of pellets fed) was 0.73 ± 0.03, which was observed in Diet 52. Then five isonitrogenous (52% crude protein) and isocaloric (20 kJ g-1 gross energy) diets were formulated by replacing 0 (control), 35%, 50%, 65% and 80% of fishmeal with No.52 blending proportion. After a 10-weeks feeding trial, a consistent feed intake was found among all replacement treatments. Replacement level of fishmeal up to 35% did not significantly influence final body weight, specific growth rate, feed efficiency ratio, and protein efficiency ratio of turbot. Therefore, the water-soluble extracts of protein sources play an

  2. Optimized sample preparation for two-dimensional gel electrophoresis of soluble proteins from chicken bursa of Fabricius

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    Zheng Xiaojuan

    2009-10-01

    Full Text Available Abstract Background Two-dimensional gel electrophoresis (2-DE is a powerful method to study protein expression and function in living organisms and diseases. This technique, however, has not been applied to avian bursa of Fabricius (BF, a central immune organ. Here, optimized 2-DE sample preparation methodologies were constructed for the chicken BF tissue. Using the optimized protocol, we performed further 2-DE analysis on a soluble protein extract from the BF of chickens infected with virulent avibirnavirus. To demonstrate the quality of the extracted proteins, several differentially expressed protein spots selected were cut from 2-DE gels and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS. Results An extraction buffer containing 7 M urea, 2 M thiourea, 2% (w/v 3-[(3-cholamidopropyl-dimethylammonio]-1-propanesulfonate (CHAPS, 50 mM dithiothreitol (DTT, 0.2% Bio-Lyte 3/10, 1 mM phenylmethylsulfonyl fluoride (PMSF, 20 U/ml Deoxyribonuclease I (DNase I, and 0.25 mg/ml Ribonuclease A (RNase A, combined with sonication and vortex, yielded the best 2-DE data. Relative to non-frozen immobilized pH gradient (IPG strips, frozen IPG strips did not result in significant changes in the 2-DE patterns after isoelectric focusing (IEF. When the optimized protocol was used to analyze the spleen and thymus, as well as avibirnavirus-infected bursa, high quality 2-DE protein expression profiles were obtained. 2-DE maps of BF of chickens infected with virulent avibirnavirus were visibly different and many differentially expressed proteins were found. Conclusion These results showed that method C, in concert extraction buffer IV, was the most favorable for preparing samples for IEF and subsequent protein separation and yielded the best quality 2-DE patterns. The optimized protocol is a useful sample preparation method for comparative proteomics analysis of chicken BF tissues.

  3. 响应面法优化秘鲁鱿鱼(Dosidicus gigas)肌肉盐溶蛋白的提取和凝胶形成条件%Response Surface Optimization of Conditions for Extraction and Gel Preparation of Salt-soluble Proteins from Dosidicus gigas Muscle

    Institute of Scientific and Technical Information of China (English)

    金淼; 周逸; 徐亦及; 唐剑波; 杨文鸽; 张进杰

    2013-01-01

    The effect of extraction and gel preparation conditions of salt-soluble protein from Dosidicus gigas on its gel characteristics were discussed.Using respond surface methodology,the effects of KCl concentration,pH and lowtemperature heating time on the water holding capacity(WHC) and texture characteristics of salt-soluble protein gel from squid meat were studied.Result:(1) The effect of KCl concentration on the WHC of gel was extremely significant; the effect of KCl concentration and low-temperature heating time on the hardness of gel was significant ; both the effect of pH on the WHC and low-temperature heating time on the stickiness of gel were significant.(2) Response surface and contour were graphed with the WHC,hardness and stickiness of gel as the response value,the optimal extraction and gel preparation conditions of salt-soluble proteins from Dosidicus gigas muscle were 0.16mol· L-1 KCl,pH7.10 and 55min of low-temperature heating time,respectively.Experimentally,these conditions produced a salt-soluble protein gel with a WHC of 88.00%,hardness of 510.00 g,stickiness of-11.00 g,confirming the model prediction.%探讨了秘鲁鱿鱼肉盐溶蛋白的提取及凝胶形成条件对其凝胶特性的影响.采用响应面分析法,研究KCl浓度、pH值和低温加热时间对鱿鱼肉盐溶蛋白凝胶保水性和质构特性的影响.结果显示:(1)提取液KCl浓度对保水性影响极其显著,对硬度影响显著;提取液pH值对保水性影响显著;低温加热时间对硬度和粘性影响显著;(2)在分析各因素显著性及其交互作用的基础上,确定鱿鱼肉盐溶蛋白提取和凝胶形成的适宜条件为:KCl浓度0.16 mol·L-1、pH 7.10、40℃加热55min,此时凝胶保水性、硬度、粘性分别达到88.00%、510.00g、-ll.00g,与模型预测值相符.

  4. Extraction of proteins with low fluoride level from Antarctic krill (Euphausia superba) and their composition analysis.

    Science.gov (United States)

    Wang, Lingzhao; Xue, Changhu; Wang, Yuming; Yang, Bao

    2011-06-08

    The extraction of proteins with low fluoride level (LFP) from Antarctic krill (Euphausia superba) was investigated in this work. The optimal conditions for protein solubilization were determined to be pH 11.5 and 4 °C. The proteins were solubilized two times; a water/krill ratio (mL/g) of 6 and a time of 30 min were used for the first step, whereas the second used a water/krill residue ratio (mL/g) of 3 and a time of 30 min. The optimum pH for protein precipitation was 4.6. A LFP with fluoride content of 9.86 mg/kg (dry weight) was finally obtained through a fluoride removal program. The protein yield of LFP was 52.68%. Composition analysis of LFP indicated it was composed of 66.96% of crude proteins (dry weight) and 33.01% of total lipids (dry weight),, and all nine essential amino acids were in sufficient amounts to meet FAO/WHO/UNU requirements for adults and infants. In addition, LFP could be taken as a good source of EPA and DHA for consideration of use as a food item for human consumption.

  5. OPTIMIZATION OF FRUCTANS EXTRACTION FROM in vitro CULTIVATED CHICORY ‘HAIRY’ ROOTS

    Directory of Open Access Journals (Sweden)

    K. S. Maznik

    2013-06-01

    Full Text Available Dependence of efficiency of fructans extraction on soaking time, temperature and time of high temperature extraction was investigated. Dried and powdered chicory Cichorium intybus L. cv Pala rossa «hairy» roots obtained by Agrobacterium rhizogenes-mediated transformation with pCB161 vector were used for study. There were used low- and high-temperature extractions without heating at +22 °C during 0.5; 1 and 24 hours and with heating at +70 °C, +80 °C and +90 °C during 10, 20 and 30 minutes. Fructans fractionation was conducted by two ways: separation of high molecular weight fraction by crystallization at +4 °C and low molecular weight separation by extraction with 95% ethanol. To determine fructans concentration in the extracts McRary and Slattery method was used. Based on the experimental data, a mathematical model of fructan extraction process was created. Its adequacy was tested with the Fisher criterion and coefficient of determination. Optimal parameters of the extraction process chosen using the methods of linear programming were determined. Extraction for 30 minutes at 90 °C without soaking identified as the most tech nological one. It allowed to extract fructans general amount from transgenic roots (146 ±8,77 mg/g of root dry weight. Optimal regime of fructan obtaining from chicory «hairy» roots is extraction at +90 °C for 30 min. Preliminary soaking time does not affect any effectiveness for such extraction. The most effective mode of obtaining of low- and high molecular fractions of fructans from transgenic chicory roots is twostage extraction with 95% ethanol at +80 °C and water at +90 °C with the duration of each stage of 30 minutes.

  6. Response surface optimization of enzyme-assisted extraction polysaccharides from Dictyophora indusiata.

    Science.gov (United States)

    Wu, Songhai; Gong, Guili; Wang, Yanyan; Li, Feng; Jia, Shaoyi; Qin, Fengxiang; Ren, Haitao; Liu, Yong

    2013-10-01

    An enzyme-assisted procedure for the extraction of the water-soluble polysaccharides from the stipe of Dictyophora indusiata was investigated using response surface methodology. The orthogonal array design was employed to optimize the concentration of three kinds of enzyme (cellulase, papain and pectolyase) and the optimal cellulose, papain and pectolyase concentration were 2.0% (wt.% of D. indusiata powder), 2.0% and 1.5%, respectively. And then the effect such as temperature, time and pH was studied based on a three-level three-factor Box-Behnken design. The optimized conditions were as follows: extraction temperature 52.5 °C, extraction time 105 min and pH 5.25. Under these conditions, the experimental yield of polysaccharides was 9.77±0.18%, which was well matched with the predictive yield of 9.87%. As it turned out, enzyme-assisted procedure was an effective method.

  7. Optimization of Multistage Extraction of Olive Leaves for Recovery of Phenolic Compounds at Moderated Temperatures and Short Extraction Times

    Directory of Open Access Journals (Sweden)

    Konstantinos Stamatopoulos

    2013-12-01

    Full Text Available The aim of the present study was to improve the recovery of polyphenols from olive leaves (OL by optimizing a multistage extraction scheme; provided that the olive leaves have been previously steam blanched. The maximum total phenol content expressed in ppm caffeic acid equivalents was obtained at pH 2, particle size 0.315 mm, solid-liquid ratio 1:7 and aqueous ethanol concentration 70% (v/v. The optimum duration time of each extraction stage and the operation temperature, were chosen based on qualitative and quantitative analysis of oleuropein (OLE, verbascoside, luteolin-7-O-glucoside and apigenin-7-O-glucoside performed by high performance liquid chromatography with diode array detector (HPLC-DAD. The optimum conditions for multistage extraction were 30 min total extraction time (10 min × 3 stages at 85 °C. The 80% of the total yield of polyphenols was obtained at the 1st stage of the extraction. The total extraction yield of oleuropein was found 23 times higher (103.1 mg OLE/g dry weight (d.w. OL compared to the yield (4.6 mg OLE/g d.w. OL obtained by the conventional extraction method (40 °C, 48 h. However, from an energetic and hence from an economical point of view it is preferable to work at 40 °C, since the total extraction yield of polyphenolic compounds was only 17% higher for a double increase in the operating temperature (i.e., 85 °C.

  8. Beiwei ZHU%Optimization of Enzyme Extraction Conditions of Cordycepin Polysaccharide Using Response Surface Methodology

    Institute of Scientific and Technical Information of China (English)

    Dong AN; Beiwei ZHU

    2012-01-01

    [Objective] This study aimed to optimize extraction from the leftover of Cordyceps militaris the conditions for polysaccharide culture medium. [Method] Firstly the compositions of C. militaris culture medium were detected, before the cordycepin polysaccharide in medium was extracted using enzymatic hydrolysis. The optimum hydrolytic enzyme was selected through single factor test. Then, the extraction tem- perature, pH, enzyme content and solid-liquid ratio were optimized by response sur- face methodology, and confirmed by mathematical simulation. [Result] Acid hydrolytic enzyme was the optimum for extracting polysaccharides from the leftover of C. mili- taris culture medium. The optimum extraction conditions were as follows: extraction temperature 39.89 ℃, solid-liquid ratio 1:75.78, enzyme content 2.39% and pH 3.12. Under these conditions, the extraction rate of polysaccharides reached 9.96%. [Con- clusion] The study could provide a certain theoretical direction for extracting polysac- charities from the leftover of C. mliltaris culture medium on a large scale.

  9. Optimization of Variables for Aqueous Extraction of Gum from Grewia mollis Powder

    Directory of Open Access Journals (Sweden)

    Emmanuel Panyoo Akdowa

    2014-01-01

    Full Text Available Grewia gum is a polysaccharide derived from the inner stem bark of the edible plant Grewia mollis. Juss (family Tiliaceae. It is a savanna shrub that grows wildly but is usually cultivated in Nigeria and Northern part of Cameroon. The main goal of the present study was to investigate the effect of aqueous extraction conditions on the extraction yield and physicochemical properties of the Grewia mollis. The studied aqueous extraction variables were water/powder (W/P ratio (10 : 1–80 : 1 w/p, temperature (25.0–85.0∘C, time (1–3 h, and pH (4.0–10.0. The results indicated that the aqueous extraction variables exhibited the least significant (P<0.05 effect on the yield and the viscosity of the gum. The result shows that the ratio of extraction is the main factor affecting the extraction of gum. The optimized extraction condition for higher viscosity was at the powder/water ratio of 1 : 55.4, pH of 7, time of 1 h, and temperature of 50∘C. However, the optimized extraction condition for higher yield was at the powder/water ratio of 1 : 80, pH of 4, time of 3 h, and temperature of 73∘C.

  10. Optimization of ultrasound-assisted extraction of camptothecin from Camptotheca acuminata seeds

    Institute of Scientific and Technical Information of China (English)

    JING Li-jia; LI Si-yang; CHANG Zui; WANG Yang; YAN Xiu-feng

    2011-01-01

    Naturally occurring camptothecin (CPT) is an important source of chemotherapeutic agents. The extraction from Camptotheca acuminata is still the main approach to obtain CPT compared with total synthesis. In the present study, ultrasound-assisted extractions (UAE) of CPT from C. acuminata seeds with alkaline solutions were investigated and CPT yield were determined by High Performance Liquid Chromatography. The conditions of alkaline species and concentrations, extraction time, extraction temperature and ultrasonic power were optimized.Results show that both Na3PO4 and Na2CO3 solutions gain good extraction yields, whereas Na3PO4 solution has stronger basicity and need higher concentration than Na2CO3 solution does, thus aqueous Na2CO3 is more beneficial for the extraction. The optimal condition was ultrasonically extracted with 0.5% aqueous Na2CO3 at 50℃ and ultrasonic power of 400 W for 60 min. Comparing with UAE with ethanol, the extraction with 0.5% Na2CO3 solution achieves higher yield. Moreover, aqueous Na2CO3 as a solvent has various advantages including non-toxicity, inflammable, non-corrosive and low cost, which ensure this UAE method is a superior method with high utilizing prospect.

  11. Optimization of an effective extraction procedure for the analysis of microcystins in soils and lake sediments

    Energy Technology Data Exchange (ETDEWEB)

    Chen Wei [State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China); Graduate School of Chinese Academy of Sciences, Beijing 100049 (China); Li Lin [State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China); Gan Nanqin [State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China); Song Lirong [State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072 (China)]. E-mail: lrsong@ihb.ac.cn

    2006-09-15

    Microcystin analysis in sediments and soils is considered very difficult due to low recovery for extraction. This is the primary limiting factor for understanding the fate of toxins in the interface between water and sediment in both the aquatic ecosystem as well as in soils. In the present study, a wide range of extraction solvents were evaluated over a wide range of pH, extraction approaches and equilibration time to optimize an effective extraction procedure for the analysis of microcystins in soils and lake sediments. The number of extractions required and acids in extraction solutions were also studied. In this procedure, EDTA-sodium pyrophosphate solution was selected as an extraction solvent based on the adsorption mechanism study. The optimized procedure proved to be highly efficient and achieved over 90% recovery. Finally, the developed procedure was applied to field soil and sediment sample collected from Chinese lakes during bloom seasons and microcystins were determined in six of ten samples. - Efficiency of extraction of microcystins from soil and sediment was greatly increased.

  12. Optimization of ultrasonic circulating extraction of polysaccharides from Asparagus officinalis using response surface methodology.

    Science.gov (United States)

    Zhao, Qingsheng; Kennedy, John F; Wang, Xiaodong; Yuan, Xiaofan; Zhao, Bing; Peng, Youshun; Huang, Yunxiang

    2011-08-01

    Polysaccharides were extracted from Asparagus officinalis. A novel ultrasonic circulating extraction (UCE) technology was applied for the polysaccharide extraction. Three-factor-three-level Box-Behnken design was employed to optimize ultrasonic power, extraction time and the liquid-solid ratio to obtain a high polysaccharide yield. The optimal extraction conditions were as follows: ultrasonic power was 600 W, extraction time was 46 min, the liquid-solid ratio was 35 mL/g. Under these conditions, the experimental yield of polysaccharides was 3.134%, which was agreed closely to the predicted value. The average molecular weight of A. officinalis polysaccharide was about 6.18×10(4) Da. The polysaccharides were composed of glucose, fucose, arabinose, galactose and rhamnose in a ratio of 2.18:1.86:1.50:0.98:1.53. Compared with hot water extraction (HWE), UCE showed time-saving, higher yield and no influence on the structure of asparagus polysaccharides. The results indicated that ultrasonic circulating extraction technology could be an effective and advisable technique for the large scale production of plant polysaccharides. Copyright © 2011 Elsevier B.V. All rights reserved.

  13. [Fetal electrocardiogram extraction based on independent component analysis and quantum particle swarm optimizer algorithm].

    Science.gov (United States)

    Du, Yanqin; Huang, Hua

    2011-10-01

    Fetal electrocardiogram (FECG) is an objective index of the activities of fetal cardiac electrophysiology. The acquired FECG is interfered by maternal electrocardiogram (MECG). How to extract the fetus ECG quickly and effectively has become an important research topic. During the non-invasive FECG extraction algorithms, independent component analysis(ICA) algorithm is considered as the best method, but the existing algorithms of obtaining the decomposition of the convergence properties of the matrix do not work effectively. Quantum particle swarm optimization (QPSO) is an intelligent optimization algorithm converging in the global. In order to extract the FECG signal effectively and quickly, we propose a method combining ICA and QPSO. The results show that this approach can extract the useful signal more clearly and accurately than other non-invasive methods.

  14. Optimization of ultrasonic-assisted extraction and in vitro antioxidant activities of polysaccharides from Trametes orientalis.

    Science.gov (United States)

    Zheng, Yi; Li, Yong; Wang, Wei-dong

    2014-10-13

    A Box-Behnken design was employed to optimize ultrasonic-assisted extraction of Trametes orientalis polysaccharides (TOP). The crude polysaccharides were purified by DEAE cellulose-52 chromatography, giving a main fraction named as PTOP. The antioxidant properties of PTOP were evaluated by different in vitro antioxidant assays, such as 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities, reducing power, superoxide radical scavenging activities, and chelating ability of ferrous ions. The results showed that optimal extraction parameters were as follows: ratio of water to raw material 30.6 mL/g, ultrasonic power 109.8 W, extraction temperature 40.2 °C, and extraction time 42.2 min. Under these conditions, the experimental yield of polysaccharides was 7.49 ± 0.14%, which agreed closely with the predicted value (7.47%). Furthermore, PTOP exhibited antioxidant capacity in a concentration-dependent manner in all assays.

  15. [Optimization of supercritical fluid extraction of bioactive components in Ligusticum chuanxiong by orthogonal array design].

    Science.gov (United States)

    Hu, Li-Cui; Wu, Xun; Yang, Xue-Dong

    2013-10-01

    With the yields of ferulic acid, coniferylferulate, Z-ligustilide, senkyunolide A, butylidenephthalide, butylphthalide, senkyunolide I, senkyunolide H, riligustilide, levistolide A, and total pharmacologically active ingredient as evaluation indexes, the extraction of Ligusticum chuanxiong by supercritical fluid technology was investigated through an orthogonal experiment L9 (3(4)). Four factors, namely temperature, pressure, flow rate of carbon dioxide, co-solvent concentration of the supercritical fluid, were investigated and optimized. Under the optimized conditions, namely 65 degrees C of temperature, 35 MPa of pressure, 1 L x min(-1) of CO2 flow rate, 8% of co-solvent concetration, supercritical fluid extraction could achieve a better yield than the conventional reflux extraction using methanol. And the supercritical fluid extraction process was validated to be stable and reliable.

  16. Lead Generation and Optimization Based on Protein-Ligand Complementarity

    Directory of Open Access Journals (Sweden)

    Koji Ogata

    2010-06-01

    Full Text Available This work proposes a computational procedure for structure-based lead generation and optimization, which relies on the complementarity of the protein-ligand interactions. This procedure takes as input the known structure of a protein-ligand complex. Retaining the positions of the ligand heavy atoms in the protein binding site it designs structurally similar compounds considering all possible combinations of atomic species (N, C, O, CH3, NH,etc. Compounds are ranked based on a score which incorporates energetic contributions evaluated using molecular mechanics force fields. This procedure was used to design new inhibitor molecules for three serine/threonine protein kinases (p38 MAP kinase, p42 MAP kinase (ERK2, and c-Jun N-terminal kinase 3 (JNK3. For each enzyme, the calculations produce a set of potential inhibitors whose scores are in agreement with IC50 data and Ki values. Furthermore, the native ligands for each protein target, scored within the five top-ranking compounds predicted by our method, one of the top-ranking compounds predicted to inhibit JNK3 was synthesized and his inhibitory activity confirmed against ATP hydrolysis. Our computational procedure is therefore deemed to be a useful tool for generating chemically diverse molecules active against known target proteins.

  17. Enzymatic aqueous technology for simultaneous coconut protein and oil extraction

    Directory of Open Access Journals (Sweden)

    Coelho, Maria A. Z.

    2003-03-01

    Full Text Available In this study, the following commercial enzymes were evaluated in the enzymatic extraction of oil and protein from coconut: Celluclast, Termamyl, Viscozyme, Neutrase y Protease. Preliminary experiments were carried out for selection of enzymes, enzyme concentration and incubation time. In order to verify the effects of pH and water/substrate ratio, a response surface methodology was applied based in a 32 full factorial experimental design. The pH is the most meaningful parameter on oil and protein extraction yields, with a significance level higher than 90%. The maximal extraction yields of oil and protein emulsion (83% was reached using Viscozyme L and subsequently Neutrase 1.5 MG at concentrations of 0.6% (w/w and 0.3% (w/w, respectively, total incubation time of 60 minutes, substrate/water ratio 1:6 and pH around 7 (no adjustment.En este trabajo diferentes enzimas hidroliticos fueron usados para la extracción de aceite y proteína de coco: Celluclast, Termamyl, Viscozyme, Neutrase y Protease producidos por Novo Nordisk. Experimientos preliminares fueron encaminados a la selección de los extractos enzimaticos, de las concentraciones de las enzimas y de los tiempos de incubación. La técnica de análisis de varianza, mediante un delineamiento factorial en tres niveles, fue usada para la optimización de los procesos con relación a los parámetros pH y razón sustrato/agua. Comparando con la extracción acuosa todos los enzimas hidroliticos incrementan la extracción de aceite y proteína vegetal de coco. Los coeficientes de correlación muestran que el pH fué el parámetro mas significativo (pViscozyme y Neutrase 0.6 % (w/w y 0.3 % (w/w, respectivamente, tiempo de incubación 30 minutos, razón sustrato/agua 1:6 y pH 7.

  18. Road centerline extraction from airborne LiDAR point cloud based on hierarchical fusion and optimization

    Science.gov (United States)

    Hui, Zhenyang; Hu, Youjian; Jin, Shuanggen; Yevenyo, Yao Ziggah

    2016-08-01

    Road information acquisition is an important part of city informatization construction. Airborne LiDAR provides a new means of acquiring road information. However, the existing road extraction methods using LiDAR point clouds always decide the road intensity threshold based on experience, which cannot obtain the optimal threshold to extract a road point cloud. Moreover, these existing methods are deficient in removing the interference of narrow roads and several attached areas (e.g., parking lot and bare ground) to main roads extraction, thereby imparting low completeness and correctness to the city road network extraction result. Aiming at resolving the key technical issues of road extraction from airborne LiDAR point clouds, this paper proposes a novel method to extract road centerlines from airborne LiDAR point clouds. The proposed approach is mainly composed of three key algorithms, namely, Skewness balancing, Rotating neighborhood, and Hierarchical fusion and optimization (SRH). The skewness balancing algorithm used for the filtering was adopted as a new method for obtaining an optimal intensity threshold such that the "pure" road point cloud can be obtained. The rotating neighborhood algorithm on the other hand was developed to remove narrow roads (corridors leading to parking lots or sidewalks), which are not the main roads to be extracted. The proposed hierarchical fusion and optimization algorithm caused the road centerlines to be unaffected by certain attached areas and ensured the road integrity as much as possible. The proposed method was tested using the Vaihingen dataset. The results demonstrated that the proposed method can effectively extract road centerlines in a complex urban environment with 91.4% correctness and 80.4% completeness.

  19. Extraction of resinoids from St. John's wort (Hypericum perforatum L: I. Efficiency and optimization of extraction

    Directory of Open Access Journals (Sweden)

    Milenović Dragan M.

    2002-01-01

    Full Text Available The extraction of resinoids from St. John's wort (Hypericum perforatum L was studied in a series of two papers. In the first part, the effects of the operating conditions on the yield of resinoids (total extract were analyzed, while the mathematical models of extraction kinetics were compared in the second one. The extraction was carried out using an aqueous solution of ethanol (70 and 95 % v/v at a hydromodulus (plant material to solvent ratio, w/v of 1:5 or 1:10. The plant material was disintegrated and divided into three fractions (mean particle size: 0.23, 0.57 and 1.05 mm. The temperature was 25, 50 or about 80°C (boiling temperature. A higher yield of resinoids was obtained when the plant material of greater disintegration degree (0.23 mm was treated with 70% v/v aqueous ethanol solution at higher hydromoduli (1:10 and temperatures (80°C. The effects of the operating factors on the yield of resinoids were estimated by using both the full factorial experimental plan 24 and artificial neuronic networks (ANN of 3-4-1 topology. Of the two methods, the ANN one was found to be advantageous because of its capability of estimating the yield of resinoids in the whole range of the applied operating conditions.

  20. Response surface optimization of ultrasound assisted extraction of pectin from pomegranate peel.

    Science.gov (United States)

    Moorthy, I Ganesh; Maran, J Prakash; Surya, S Muneeswari; Naganyashree, S; Shivamathi, C S

    2015-01-01

    Ultrasound assisted extraction of pectin from waste pomegranate peel was investigated and optimized using Box-Behnken response surface design coupled with numerical optimization technique. The individual and interactive effect of process variables (solid-liquid ratio, pH, extraction time and temperature) on the pectin yield was studied. The experimental data obtained were analyzed by Pareto analysis of variance (ANOVA) and second-order polynomial models were developed using multiple regression analysis. The models developed from the experimental design were predictive and good fit with the experimental data with high coefficient of determination (R(2)) value. The optimal extraction condition was found to be 1:17.52 g/ml of solid-liquid ratio, 1.27 of pH, 28.31 min of extraction time and 61.90 °C of extraction temperature respectively. Under the optimal conditions, experimental yield was very close to the predicted values. Copyright © 2014 Elsevier B.V. All rights reserved.

  1. Optimization and evaluation of wheat germ oil extracted by supercritical CO2

    OpenAIRE

    2011-01-01

    Box-Behnken design combined with response surface methodology (RSM) was used to optimize the parameters of supercritical CO2 extraction (SFE) of wheat germ oil. The quality of the oil and residual meal obtained by SFE and solvent extraction (SE) were evaluated from proximate analysis, fatty acid composition and antioxidant activity. A maximum oil yield of 10.46% was achieved under the optimal conditions of wheat germ particle size 60-80 mesh; water content 4.37%; pressure 30MPa; temperature 4...

  2. Optimization of extraction conditions for secondary biomolecules from various plant species

    Directory of Open Access Journals (Sweden)

    Šibul Filip S.

    2016-01-01

    Full Text Available Extraction of plant secondary metabolites is an essential step in isolation of natural products. Non-optimized extraction conditions can lead to losses, degradation and modification of the biomolecules. In this paper, the influence of different solvent mixtures, solvent amounts, temperature, extraction time, and procedures for defatting on yield and profile of various classes of secondary metabolites was investigated. Rumex alpinus was used for the extraction of anthraquinones, Glycine max for isoflavonoids, Chaerophyllum bulbosum for flavonoids and phenolic acids, Anthriscus sylvestris for lignans and coumarins, alkaloids were extracted from Lupinus albus and sesquiterpene lactones from Artemisia absinthium. Extraction efficiency was evaluated by use of LC-DAD-ESI-MS/MS. The compromise extraction solvent for all of the examined compounds is 80 % methanol, mixed in ratio 13 : 1 with plant material. Maceration should last for six hours, repeated four times with fresh solvent. Defatting of the extracts does not lead to significant losses of the compounds of interest. It is acceptable to use extraction and evaporation temperature of 60ºC, while the extracts should be stored in the dark, on -20ºC. [Projekat Ministarstva nauke Republike Srbije, br. 172058

  3. An Improved Method of Optimizing the Extraction of Polyphenol Oxidase from Potato (Solanum tuberosum L. Peel

    Directory of Open Access Journals (Sweden)

    Suprabhat MUKHERJEE

    2012-02-01

    Full Text Available The present study has an objective to optimize the extraction of Polyphenol Oxidase (PPO from potato (Solanum tuberosum L. peel. Response surface methodology (RSM was used to design experiments and study the effect of six influential extraction parameters: extraction buffer concentration (100-500 mM, pH of extraction buffer (4.5-8.5, time (1-12 hours, temperature (4-40�C, concentration of PMSF (1-5 mM and volume of extraction buffer (200-1000 ml on the extraction of PPO. The dependent variable was considered as response function which was specific activity (SA of extracted PPO. ANOVA was performed to obtain the regression equation that could predict the responses within given range. From RSM generated model, the optimum conditions for the maximum extraction of PPO were phosphate buffer concentration of 100 mm, buffer pH of 4.5, extraction time of 1 hour, 40�C temperature, PMSF concentration of 5 mM and buffer volume of 200 ml. Finally, this study illustrates a cost effective and less time consuming method to maximize the extraction of PPO from a vegetable waste.

  4. An optimal control method for maximizing the efficiency of direct drive ocean wave energy extraction system.

    Science.gov (United States)

    Chen, Zhongxian; Yu, Haitao; Wen, Cheng

    2014-01-01

    The goal of direct drive ocean wave energy extraction system is to convert ocean wave energy into electricity. The problem explored in this paper is the design and optimal control for the direct drive ocean wave energy extraction system. An optimal control method based on internal model proportion integration differentiation (IM-PID) is proposed in this paper though most of ocean wave energy extraction systems are optimized by the structure, weight, and material. With this control method, the heavy speed of outer heavy buoy of the energy extraction system is in resonance with incident wave, and the system efficiency is largely improved. Validity of the proposed optimal control method is verified in both regular and irregular ocean waves, and it is shown that IM-PID control method is optimal in that it maximizes the energy conversion efficiency. In addition, the anti-interference ability of IM-PID control method has been assessed, and the results show that the IM-PID control method has good robustness, high precision, and strong anti-interference ability.

  5. Ternary diagram of extract proteins / solvent systems: Sesame, soybean and lupine proteins

    Directory of Open Access Journals (Sweden)

    Mohamed, S. S.

    2004-09-01

    Full Text Available Solvent extraction as a method of extracting protein from oilseed meals offers the advantage of higher efficiency. Unfortunately, the published literature points to the gap in the work concerned with the necessary equilibrium diagram to design due process equipment for such extracts. Initiated by this lack of basic knowledge, the present study has been undertaken to provide the equilibrium data for three different ternary systems, namely: sesame protein / sodium hydroxide solution system, soybean protein / sodium hydroxide solution system and lupine protein / sodium hydroxide solution system. These oilseed meals were selected because of their high protein content (53.4 %, 46.2 % and 42.3 % protein, respectively. The study also concentrated on the evaluation of the major parameters affecting the extraction process, i.e. the normality of the sodium hydroxide solution used as extracting solvent and the initial oilseed solvent to meal feeding ratio. The results obtained indicate that the best normality of sodium hydroxide solution used for extracting soybean and lupine protein is 0.02N, while 0.04N solution is required for extracting sesame protein. Also, operating at a liquid to solid feed ratio of 30:1 and 50:1 for soybean, sesame and lupine, respectively, is enough to reach a high protein extract. Correlations were presented for each locus of under flow compositions, graphically acquired, and the data are compared with those calculated by analytical solutions.La extracción con disolventes es un método de extracción de proteínas de las harinas de semillas oleaginosas que ofrece la ventaja de su elevada eficacia. Desafortunadamente, la bibliografía coincide en el vacío existente con respecto a los diagramas de equilibrio necesarios para el diseño de los equipos adecuados. Debido a esta falta de conocimientos, el presente estudio se ha llevado a cabo para obtener datos de tres sistemas ternarios: sistema proteína de sésamo / disolución de

  6. Extraction design and low energy beam transport optimization of space charge dominated multispecies ion beam sources

    Science.gov (United States)

    Delferrière, O.; De Menezes, D.

    2004-05-01

    In all accelerator projects, the low energy part of the accelerator has to be carefully optimized to match the beam characteristic requirements of the higher energy parts. Since 1994 with the beginning of the Injector of Protons for High Intensity (IPHI) project and Source of Light Ions with High Intensities (SILHI) electron cyclotron resonance (ECR) ion source development at CEA/Saclay, we are using a set of two-dimensional (2D) codes for extraction system optimization (AXCEL, OPERA-2D) and beam transport (MULTIPART). The 95 keV SILHI extraction system optimization has largely increased the extracted current, and improved the beam line transmission. From these good results, a 130 mA D+ extraction system for the International Fusion Material Irradiation Facility project has been designed in the same way as SILHI one. We are also now involved in the SPIRAL 2 project for the building of a 40 keV D+ ECR ion source, continuously tunable from 0.1 to 5 mA, for which a special four-electrode extraction system has been studied. In this article we will describe the 2D design process and present the different extraction geometries and beam characteristics. Simulation results of SILHI H+ beam emittance will be compared with experimental measurements.

  7. Optimization of subcritical fluid extraction of bioactive compounds using Hansen solubility parameters.

    Science.gov (United States)

    Srinivas, K; King, J W; Monrad, J K; Howard, L R; Hansen, C M

    2009-08-01

    Process engineering operations in food and nutraceutical industries pertaining to the design of extraction of value-added products from biomass using pressurized liquids involve a careful selection of the solvent and optimal temperature conditions to achieve maximum yield. Complex molecular structure and limited physical property data in the literature of biological solutes extracted from biomass compounds have necessitated the process modeling of such operations. In this study, we have applied the Hansen 3-dimensional solubility parameter concept to optimize the extraction of molecularly complex solutes using subcritical fluid solvents. Hansen solubility spheres characterized by the relative energy differences (RED) have been used to characterize and quantify the solute-subcritical solvent interactions as a function of temperature. The solvent power of subcritical water and compressed hydroethanolic mixtures above their boiling points has been characterized using the above-mentioned method. The use of group contribution methods in collaboration with computerized algorithms to plot the Hansen spheres provides a quantitative prediction tool for optimizing the design of extraction conditions. The method can be used to estimate conditions for solute-solvent miscibility, an optimum temperature range for conducting extractions under pressurized conditions, and approximate extraction conditions of solutes from natural matrices.

  8. Optimization of ultrasonic-assisted extraction of natural antioxidants from rice bran using response surface methodology.

    Science.gov (United States)

    Tabaraki, Reza; Nateghi, Ashraf

    2011-11-01

    Ultrasonic technology was applied for extraction of polyphenols and antioxidants from the rice bran using ethanol as a food grade solvent. Response surface methodology (RSM) was used to optimize experimental conditions for extraction of polyphenols and antioxidants. Three independent variables such as solvent percentage (%), temperature (°C) and time (min) were studied. Effect of ethanol concentration was found to be significant on all responses. Total phenolic content (TPC) varied from 2.37 to 6.35mg gallic acid equivalent/g of dry sample. Antioxidant activity of the extracts was determined by the ferric reducing antioxidant power (FRAP) assay and scavenging activity of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. FRAP and DPPH values varied from 31.74 to 57.23μmol Fe(2+)/g of dry sample and 16.88% to 55.61% inhibition, respectively. Extraction yields ranged from 11 to 20.2%. Optimal ultrasonic-assisted extraction (UAE) conditions were identified as 65-67% ethanol, 51-54°C, 40-45min. The experimental values agreed with those predicted by SRM models, thus indicating suitability of the model employed and the success of RSM in optimizing the extraction conditions.

  9. Encapsulation of Beetroot Pomace Extract: RSM Optimization, Storage and Gastrointestinal Stability

    Directory of Open Access Journals (Sweden)

    Vesna Tumbas Šaponjac

    2016-04-01

    Full Text Available One of the great problems in food production are surplus by-products, usually utilized for feeding animals and for preparation of dietary fibre or biofuel. These products represent potential sources of bioactive antioxidants and colour-giving compounds which could be used in the pharmaceutical industry and as food additives. In the present study beetroot pomace extract was encapsulated in soy protein by a freeze drying method. Process parameters (core: wall ratio, extract concentration and mixing time were optimized using response surface methodology (RSM in order to obtain the optimum encapsulate (OE with the highest polyphenol encapsulation efficiency (EE and radical scavenging activity on DPPH radicals (SA. Using the calculated optimum conditions, the EE (86.14% and SA (1668.37 μmol Trolox equivalents/100 g of OE did not differ significantly (p < 0.05 from the predicted ones. The contents of total polyphenols (326.51 mg GAE/100 g, flavonoids (10.23 mg RE/100 g, and betalains (60.52 mg betanin/100 g and 61.33 mg vulgaxanthin-I/100 g, individual content of phenolic compounds and betalains by HPLC, and the ability to reduce Fe3+ ions, i.e., reducing power (394.95 μmol Trolox equivalents/100 g of OE were determined as well. During three months of storage at room temperature, polyphenol retention was much higher (76.67% than for betalain pigments, betacyanins (17.77% and betaxanthins (17.72%. In vitro digestion and release of phenolics from OE showed higher release rate in simulated intestinal fluid than in gastric fluid. These results suggest encapsulation as a contemporary method for valorisation of sensitive bioactive compounds from food industry by-products.

  10. Bio-inspired optimization algorithms for optical parameter extraction of dielectric materials: A comparative study

    Science.gov (United States)

    Ghulam Saber, Md; Arif Shahriar, Kh; Ahmed, Ashik; Hasan Sagor, Rakibul

    2016-10-01

    Particle swarm optimization (PSO) and invasive weed optimization (IWO) algorithms are used for extracting the modeling parameters of materials useful for optics and photonics research community. These two bio-inspired algorithms are used here for the first time in this particular field to the best of our knowledge. The algorithms are used for modeling graphene oxide and the performances of the two are compared. Two objective functions are used for different boundary values. Root mean square (RMS) deviation is determined and compared.

  11. An AIL/IL-based liquid/liquid extraction system for the purification of His-tagged proteins.

    Science.gov (United States)

    Xu, Weiyuan; Cao, Huazhen; Ren, Guangwei; Xie, Hujun; Huang, Jianying; Li, Shijun

    2014-06-01

    A sorbent based on affinity ionic liquid (AIL), triazacyclononane-ionic liquid, was synthesized, characterized, and applied to the extraction of histidine (His)-tagged proteins from aqueous buffer to ionic liquid (IL) phase. The adsorbed His-tagged proteins could be back-extracted from the IL phase to the aqueous buffer with an imidazole solution. The specific binding of His-tagged proteins with AIL/IL could be affected by a few factors including the ionic strength and coordinated metal ions. In the case of His-tagged enhanced green fluorescent protein (EGFP), the maximum binding capacity of Cu(2+)-AIL/IL reached 2.58 μg/μmol under the optimized adsorption conditions. The eluted His-tagged EGFP kept fluorescent and remained active through the purification process. Moreover, a tandem extraction process successively using Cu(2+)-AIL/IL and Zn(2+)-AIL/IL systems was developed, which was proven very efficient to obtain the ultimate protein with a purity of about 90 %. An effective reclamation method for the AIL/IL extraction system was further established. The sorbent could be easily regenerated by removing metal ions with EDTA and the followed reimmobilization of metal ions. Easy handling of the presented M(2+)-AIL/IL system and highly specific ability to absorb His-tagged proteins make it attractive and potentially applicable in biomolecular separation.

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

    Science.gov (United States)

    Morrison, Kerrie A; Akram, Aneel; Mathews, Ashlyn; Khan, Zoeya A; Patel, Jaimin H; Zhou, Chumin; Hardy, David J; Moore-Kelly, Charles; Patel, Roshani; Odiba, Victor; Knowles, Tim J; Javed, Masood-Ul-Hassan; Chmel, Nikola P; Dafforn, Timothy R; Rothnie, Alice J

    2016-12-01

    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. © 2016 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

  13. Optimization of Solid-Liquid Extraction of Antioxidants from Black Mulberry Leaves by Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Zoran Zeković

    2012-01-01

    Full Text Available The extraction of active components from natural sources depends on different factors. The knowledge of the effects of different extraction parameters is useful for the optimization of the process, as well for the ability to predict the extraction yield. The aim of this study is to examine the influence of solvent concentration (ethanol/water 40–80 %, by volume, temperature (40–80 °C and solvent/raw material ratio (10–30 mL/g on the extraction yield of phenolic compounds, flavonoids and antioxidant activity from black mulberry (Morus nigra L. leaves. Experimental values of total phenolic content were in the range from 18.6 to 48.7 mg of chlorogenic acid equivalents per g of dried leaves and total flavonoids in the range from 6.0 to 21.4 mg of rutin eqivalents per g of dried leaves. Antioxidant activity expressed as the inhibition concentration at 50 % (IC50 value was in the range from 0.019 to 0.078 mg of mulberry extract per mL. Response surface methodology (RSM was used to determine the optimum extraction conditions and to investigate the effect of different variables on the observed properties of mulberry leaf extracts. The results show a good fit to the proposed model (R˄2>0.90. The optimal conditions for obtaining the highest extraction yield of phenolics and flavonoids were within the experimental range. The experimental values agreed with those predicted, thus indicating suitability of the used model and the success of RSM in optimizing the investigated extraction conditions.

  14. Calorimetric Quantification of Cyclodextrin-Mediated Detergent Extraction for Membrane-Protein Reconstitution.

    Science.gov (United States)

    Textor, Martin; Keller, Sandro

    2016-01-01

    For many in vitro studies, purified membrane proteins need to be reconstituted from detergent micelles into lipid bilayers to regain their native structures and functions. Stoichiometric complexation of detergent by cyclodextrin provides a tightly controllable strategy for detergent extraction. Here, we describe a practical approach making use of isothermal titration calorimetry to obtain a complete set of thermodynamic parameters that allows for quantitative prediction of the transition from micelles to bilayer membranes during reconstitution. These parameters include the dissociation constant of the cyclodextrin/detergent inclusion complex, the critical micellar concentration of the detergent, and the phase boundaries of the lipid/detergent phase diagram. The underlying theoretical framework involves linked equilibria among all pseudophases, as described previously (Textor, Vargas, & Keller, 2015). This chapter focuses on practical aspects of the approach and discusses caveats and calorimetry-specific details of data analysis. With the entire parameter set at hand, exploration of different reconstitution trajectories within the lipid/detergent phase diagram is possible. Together with the straightforward control over the rate of detergent extraction offered by cyclodextrin complexation, this opens the possibility of systematically tuning and optimizing the reconstitution process of membrane proteins. Provided some particular precautions are taken, the approach can be adapted to many other combinations of proteins, lipids, detergents, and cyclodextrins.

  15. A Novel Feature Extraction Method with Feature Selection to Identify Golgi-Resident Protein Types from Imbalanced Data.

    Science.gov (United States)

    Yang, Runtao; Zhang, Chengjin; Gao, Rui; Zhang, Lina

    2016-02-06

    The Golgi Apparatus (GA) is a major collection and dispatch station for numerous proteins destined for secretion, plasma membranes and lysosomes. The dysfunction of GA proteins can result in neurodegenerative diseases. Therefore, accurate identification of protein subGolgi localizations may assist in drug development and understanding the mechanisms of the GA involved in various cellular processes. In this paper, a new computational method is proposed for identifying cis-Golgi proteins from trans-Golgi proteins. Based on the concept of Common Spatial Patterns (CSP), a novel feature extraction technique is developed to extract evolutionary information from protein sequences. To deal with the imbalanced benchmark dataset, the Synthetic Minority Over-sampling Technique (SMOTE) is adopted. A feature selection method called Random Forest-Recursive Feature Elimination (RF-RFE) is employed to search the optimal features from the CSP based features and g-gap dipeptide composition. Based on the optimal features, a Random Forest (RF) module is used to distinguish cis-Golgi proteins from trans-Golgi proteins. Through the jackknife cross-validation, the proposed method achieves a promising performance with a sensitivity of 0.889, a specificity of 0.880, an accuracy of 0.885, and a Matthew's Correlation Coefficient (MCC) of 0.765, which remarkably outperforms previous methods. Moreover, when tested on a common independent dataset, our method also achieves a significantly improved performance. These results highlight the promising performance of the proposed method to identify Golgi-resident protein types. Furthermore, the CSP based feature extraction method may provide guidelines for protein function predictions.

  16. A Novel Feature Extraction Method with Feature Selection to Identify Golgi-Resident Protein Types from Imbalanced Data

    Directory of Open Access Journals (Sweden)

    Runtao Yang

    2016-02-01

    Full Text Available The Golgi Apparatus (GA is a major collection and dispatch station for numerous proteins destined for secretion, plasma membranes and lysosomes. The dysfunction of GA proteins can result in neurodegenerative diseases. Therefore, accurate identification of protein subGolgi localizations may assist in drug development and understanding the mechanisms of the GA involved in various cellular processes. In this paper, a new computational method is proposed for identifying cis-Golgi proteins from trans-Golgi proteins. Based on the concept of Common Spatial Patterns (CSP, a novel feature extraction technique is developed to extract evolutionary information from protein sequences. To deal with the imbalanced benchmark dataset, the Synthetic Minority Over-sampling Technique (SMOTE is adopted. A feature selection method called Random Forest-Recursive Feature Elimination (RF-RFE is employed to search the optimal features from the CSP based features and g-gap dipeptide composition. Based on the optimal features, a Random Forest (RF module is used to distinguish cis-Golgi proteins from trans-Golgi proteins. Through the jackknife cross-validation, the proposed method achieves a promising performance with a sensitivity of 0.889, a specificity of 0.880, an accuracy of 0.885, and a Matthew’s Correlation Coefficient (MCC of 0.765, which remarkably outperforms previous methods. Moreover, when tested on a common independent dataset, our method also achieves a significantly improved performance. These results highlight the promising performance of the proposed method to identify Golgi-resident protein types. Furthermore, the CSP based feature extraction method may provide guidelines for protein function predictions.

  17. Optimization of Extraction of Novel Pectinase Enzyme Discovered in Red Pitaya (Hylocereus polyrhizus) Peel

    OpenAIRE

    Nor Khanani Zohdi; Mehrnoush Amid

    2013-01-01

    Plant peels could be a potential source of novel pectinases for use in various industrial applications due to their broad substrate specificity with high stability under extreme conditions. Therefore, the extraction conditions of a novel pectinase enzyme from pitaya peel was optimized in this study. The effect of extraction variables, namely buffer to sample ratio (2:1 to 8:1, X1), extraction temperature (−15 to +25 °C, X2) and buffer pH (4.0 to 12.0, X3) on specific activity, temperature st...

  18. Motion Key-Frame Extraction by Using Optimized t-Stochastic Neighbor Embedding

    Directory of Open Access Journals (Sweden)

    Qiang Zhang

    2015-04-01

    Full Text Available Key-frame extracting technology has been widely used in the field of human motion synthesis. Efficient and accurate key frames extraction methods can improve the accuracy of motion synthesis. In this paper, we use an optimized t-Stochastic Neighbor Embedding (t-SNE for short algorithm to reduce the data and on this basis extract the key frames. The experimental results show that the validity of this method is better than the existing methods under the same experimental data.

  19. Preventing protein adsorption from a range of surfaces using an aqueous fish protein extract

    DEFF Research Database (Denmark)

    Pillai, Saju; Arpanaei, Ayyoob; Meyer, Rikke L.;

    2009-01-01

    We utilize an aqueous extract of fish proteins (FPs) as a coating for minimizing the adsorption of fibrinogen (Fg) and human serum albumin (HSA). The surfaces include stainless steel (SS), gold (Au), silicon dioxide (SiO2), and poly(styrene) (PS). The adsorption processes (kinetics and adsorbed...... Fg- or HSA-coated surfaces are exposed to the FPs, a significant increase in adsorbed mass occurs because the FPs are highly surface-active displacing Fg. Additionally, fluorescence microscopy confirms that very little Fg adsorbs to the FP-coated surfaces. We propose that FP coatings prevent protein...

  20. Protein structure prediction using bee colony optimization metaheuristic

    DEFF Research Database (Denmark)

    Fonseca, Rasmus; Paluszewski, Martin; Winter, Pawel

    2010-01-01

    Predicting the native structure of proteins is one of the most challenging problems in molecular biology. The goal is to determine the three-dimensional struc- ture from the one-dimensional amino acid sequence. De novo prediction algorithms seek to do this by developing a representation of the pr......Predicting the native structure of proteins is one of the most challenging problems in molecular biology. The goal is to determine the three-dimensional struc- ture from the one-dimensional amino acid sequence. De novo prediction algorithms seek to do this by developing a representation...... of the proteins structure, an energy potential and some optimization algorithm that ¿nds the structure with minimal energy. Bee Colony Optimization (BCO) is a relatively new approach to solving opti- mization problems based on the foraging behaviour of bees. Several variants of BCO have been suggested...... in the literature. We have devised a new variant that uni¿es the existing and is much more ¿exible with respect to replacing the various elements of the BCO. In particular this applies to the choice of the local search as well as the method for generating scout locations and performing the waggle dance. We apply...

  1. Evolutionary optimization of kernel weights improves protein complex comembership prediction.

    Science.gov (United States)

    Hulsman, Marc; Reinders, Marcel J T; de Ridder, Dick

    2009-01-01

    In recent years, more and more high-throughput data sources useful for protein complex prediction have become available (e.g., gene sequence, mRNA expression, and interactions). The integration of these different data sources can be challenging. Recently, it has been recognized that kernel-based classifiers are well suited for this task. However, the different kernels (data sources) are often combined using equal weights. Although several methods have been developed to optimize kernel weights, no large-scale example of an improvement in classifier performance has been shown yet. In this work, we employ an evolutionary algorithm to determine weights for a larger set of kernels by optimizing a criterion based on the area under the ROC curve. We show that setting the right kernel weights can indeed improve performance. We compare this to the existing kernel weight optimization methods (i.e., (regularized) optimization of the SVM criterion or aligning the kernel with an ideal kernel) and find that these do not result in a significant performance improvement and can even cause a decrease in performance. Results also show that an expert approach of assigning high weights to features with high individual performance is not necessarily the best strategy.

  2. TCR/pMHC Optimized Protein crystallization Screen

    Science.gov (United States)

    Bulek, Anna M.; Madura, Florian; Fuller, Anna; Holland, Christopher J.; Schauenburg, Andrea J.A.; Sewell, Andrew K.; Rizkallah, Pierre J.; Cole, David K.

    2012-01-01

    The interaction between the clonotypic αβ T cell receptor (TCR), expressed on the T cell surface, and peptide-major histocompatibility complex (pMHC) molecules, expressed on the target cell surface, governs T cell mediated autoimmunity and immunity against pathogens and cancer. Structural investigations of this interaction have been limited because of the challenges inherent in the production of good quality TCR/pMHC protein crystals. Here, we report the development of an ‘intelligently designed’ crystallization screen that reproducibly generates high quality TCR/pMHC complex crystals suitable for X-ray crystallographic studies, thereby reducing protein consumption. Over the last 2 years, we have implemented this screen to produce 32 T cell related protein structures at high resolution, substantially contributing to the current immune protein database. Protein crystallography, used to study this interaction, has already extended our understanding of the molecular rules that govern T cell immunity. Subsequently, these data may help to guide the intelligent design of T cell based therapies that target human diseases, underlining the importance of developing optimized approaches for crystallizing novel TCR/pMHC complexes. PMID:22705983

  3. Evaluation of protein extraction protocols for 2DE in marine ecotoxicoproteomics.

    Science.gov (United States)

    Wu, Huifeng; Ji, Chenglong; Wei, Lei; Zhao, Jianmin

    2013-11-01

    In ecotoxicoproteomics, an accurate and reproducible extraction of proteins is a critical step for 2DE analysis and further protein identification using MS. The criteria for the assessment of protein extraction quality include protein yield, protein spots resolved in a 2DE gel, matched protein spots in replicate gels, reproducibility, and compatibility with MS. In this work, we evaluated three protein extraction systems, straightforward lysis buffer, trichloroacetic acid-acetone, and TRIzol reagent with some modifications, for the protein extraction from three animal species including mussel Mytilus galloprovincialis, flounder Paralichthys olivaceus, and polychaete Nereis diversicolor used in marine ecotoxicology. Our results indicated that these methods could extract significantly different protein profiles. The method using TRIzol reagent resulted in the most matched protein spots resolved in four replicate 2DE gels and highest reproducibilities for the gill of M. galloprovincialis and liver of P. olivaceus. However, a modified trichloroacetic acid-acetone solvent system was best for the whole soft tissue of N. diversicolor. This work provides the fundamental information of the extraction quality of protein extraction protocols from different marine animals, which may facilitate the selection of a suitable protein extraction protocol for ecotoxicoproteomics.

  4. Extraction, composition and functional properties of pennycress (Thlaspi arvense L.) press cake protein

    Science.gov (United States)

    This study compared two methods for extracting the protein in pennycress (Thlaspi arvense L.) press cake and determined the composition and functional properties of the protein products. Proteins in pennycress press cake were extracted by using the conventional alkali solubilization-acid precipitati...

  5. Optimal Extraction of Gallic Acid from Suaeda glauca Bge. Leaves and Enhanced Efficiency by Ionic Liquids

    Directory of Open Access Journals (Sweden)

    Xin-Hong Wang

    2016-01-01

    Full Text Available The ultrasound-assisted extraction (UAE was initially applied to extract gallic acid from Suaeda glauca Bge. using 70% ethanol as extraction solvent. Temperature, liquid-solid ratio, and extraction time were optimized by response surface methodology (RSM, obtaining maximum levels of gallic acid (6.30 mg·g−1 at 51°C, 19.52 mL·g−1, and 42.68 min, respectively. The obtained model was statistically significant (p<0.0001. The verification experiments at the optimum conditions yielded gallic acid for 6.21 mg·g−1. Subsequently, under optimal conditions, four ionic liquids were used to extract gallic acid from Suaeda glauca Bge. The results indicated that the presence of 1-hexyl-3-methylimidazolium chloride allowed increasing the EE of gallic acid up to 8.90 mg·g−1. This might be interpreted in terms of the molecular interaction between ionic liquid and gallic acid. The use of ionic liquids involves a stronger gallic acid extraction capacity than conventional organic volatile solvents. A promising alternative process is proposed for the extraction of gallic acid of Suaeda glauca Bge.

  6. Optimization of solvent extraction of shea butter (Vitellaria paradoxa) using response surface methodology and its characterization.

    Science.gov (United States)

    Ajala, E O; Aberuagba, F; Olaniyan, A M; Onifade, K R

    2016-01-01

    Shea butter (SB) was extracted from its kernel by using n-hexane as solvent in an optimization study. This was to determine the optima operating variables that would give optimum yield of SB and to study the effect of solvent on the physico-chemical properties and chemical composition of SB extracted using n-hexane. A Box-behnken response surface methodology (RSM) was used for the optimization study while statistical analysis using ANOVA was used to test the significance of the variables for the process. The variables considered for this study were: sample weight (g), solvent volume (ml) and extraction time (min). The physico-chemical properties of SB extracted were determined using standard methods and Fourier Transform Infrared Spectroscopy (FTIR) for the chemical composition. The results of RSM analysis showed that the three variables investigated have significant effect (p butter extracted using traditional method (SBT) showed that it is a more suitable raw material for food, biodiesel production, cosmetics, medicinal and pharmaceutical purposes than shea butter extracted using solvent extraction method (SBS). Fourier Transform Infrared Spectroscopy (FTIR) results obtained for the two samples were similar to what was obtainable from other vegetable oil.

  7. Flat-relative optimal extraction. A quick and efficient algorithm for stabilised spectrographs

    CERN Document Server

    Zechmeister, M; Reiners, A

    2013-01-01

    Optimal extraction is a key step in processing the raw images of spectra as registered by two-dimensional detector arrays to a one-dimensional format. Previously reported algorithms reconstruct models for a mean one-dimensional spatial profile to assist a properly weighted extraction. We outline a simple optimal extraction algorithm including error propagation, which is very suitable for stabilised, fibre-fed spectrographs and does not model the spatial profile shape. A high signal-to-noise, master-flat image serves as reference image and is directly used as an extraction profile mask. Each extracted spectral value is the scaling factor relative to the cross-section of the unnormalised master-flat which contains all information about the spatial profile as well as pixel-to-pixel variations, fringing, and blaze. The extracted spectrum is measured relative to the flat spectrum. Using echelle spectra of the HARPS spectrograph we demonstrate a competitive extraction performance in terms of signal-to-noise and sho...

  8. Extraction optimization, characterization and antioxidant activity of polysaccharide from Gentiana scabra bge.

    Science.gov (United States)

    Cheng, Zhenyu; Zhang, Yuewei; Song, Haiyan; Zhou, Hongli; Zhong, Fangli; Hu, Haobin; Feng, Yu

    2016-12-01

    In this study, optimization of smashing tissue extraction (STE), preliminary chemical characterization and antioxidant activity in vitro of crude polysaccharides (CPS) from Gentiana scabra bge (G. scabra) were investigated. The optimal extraction conditions were determined as follows: sample particle size of 80mesh, solid/liquid ratio of 1:34, extraction voltage of 157.09V and extraction time of 130.38s. Under these conditions, the extraction yield of CPS had reached 15.03±0.14% (n=3). Chemical composition analysis indicated CPS was mainly composed of mannose, rhamnose, galacturonic acid, glcose, galactose, arabinose and fucose in a molar ratio of 1.00:9.89:51.59:35.37:38.06:99.13:21.34, respectively. The average molecular weight of CPS was estimated to be 3.8×10(4)Da. In addition, the potential antioxidant activity of CPS extracted by STE were demonstrated by DPPH radical scavenging assay, superoxide anion radical scavenging assay, hydroxyl radical scavenging assay and ferric reducing power assay. Overall, this study provided an effective extraction technique for G. scabra polysaccharides which would be explored as a promising natural antioxidant agent applied in functional foods or medicines.

  9. Optimization of microwave-assisted extraction of polysaccharide from Psidium guajava L. fruits.

    Science.gov (United States)

    Amutha Gnana Arasi, Michael Antony Samy; Gopal Rao, Manchineela; Bagyalakshmi, Janardanan

    2016-10-01

    This study deals with the optimization of microwave assisted extraction of polysaccharide from Psidium guajava L. fruit using Response surface methodology. To evaluate the effect of three independent variables, Water to plant material ratio, microwave power used for extraction and Irradiation time, central composite design has been employed. The yield is considered as dependent variable. The design model estimated the optimum yield of 6.81677% at 200W microwave power level, 3:1 water to plant material ratio and 20min of irradiation time. Three factors three levels Central composite design coupled with RSM was used to model the extraction process. ANOVA was performed to find the significance of the model. The polysaccharide extracted using microwave assisted extraction process was analyzed using FTIR Spectroscopy.

  10. Challenges of protein extraction from recalcitrant plant tissues for proteomics

    Science.gov (United States)

    Proteins play an important role in several biological processes. Proteomics encompasses basically four principal applications, namely protein mining, protein expression profiling, protein-network mapping and mapping of protein modifications. The results in these applications depend mostly on the c...

  11. Optimal cuts to extract the third-order elastic constants of langasite single crystals.

    Science.gov (United States)

    Zhang, Haifeng

    2011-06-01

    Optimal cuts to determine the third-order elastic constants of langasite single crystals by the resonator method are proposed. By designing a small number of langasite resonators with optimal cut angles and measuring their force-frequency effects, the third-order elastic constants of langasite single crystals may be extracted separately. The numerical method to search for these optimal cut angles is presented. All 14 third-order elastic constants may be determined through a series of experimental measurements. This method will simplify traditional methods used to determine the third-order elastic constants and could potentially produce more accurate results.

  12. Biorefinery process for protein extraction from oriental mustard (Brassica juncea (L.) Czern.) using ethanol stillage.

    Science.gov (United States)

    Ratanapariyanuch, Kornsulee; Tyler, Robert T; Shim, Youn Young; Reaney, Martin Jt

    2012-01-12

    Large volumes of treated process water are required for protein extraction. Evaporation of this water contributes greatly to the energy consumed in enriching protein products. Thin stillage remaining from ethanol production is available in large volumes and may be suitable for extracting protein rich materials. In this work protein was extracted from ground defatted oriental mustard (Brassica juncea (L.) Czern.) meal using thin stillage. Protein extraction efficiency was studied at pHs between 7.6 and 10.4 and salt concentrations between 3.4 × 10-2 and 1.2 M. The optimum extraction efficiency was pH 10.0 and 1.0 M NaCl. Napin and cruciferin were the most prevalent proteins in the isolate. The isolate exhibited high in vitro digestibility (74.9 ± 0.80%) and lysine content (5.2 ± 0.2 g/100 g of protein). No differences in the efficiency of extraction, SDS-PAGE profile, digestibility, lysine availability, or amino acid composition were observed between protein extracted with thin stillage and that extracted with NaCl solution. The use of thin stillage, in lieu of water, for protein extraction would decrease the energy requirements and waste disposal costs of the protein isolation and biofuel production processes.

  13. Improving Jatropha curcas seed protein recovery by using counter current multistage extraction

    NARCIS (Netherlands)

    Lestari, D.; Mulder, W.J.; Sanders, J.P.M.

    2010-01-01

    Jatropha curcas seed press cake contains 23 wt% proteins (dry basis). Due to the toxic compounds in Jatropha, we will use the proteins for non-food applications. Related to non-food applications, an efficient protein extraction to obtain a high protein recovery and high protein concentration with go

  14. Understanding leaf membrane protein extraction to develop a food-grade process

    NARCIS (Netherlands)

    Tamayo Tenorio, Angelica; Boom, Remko M.; Goot, van der Atze Jan

    2017-01-01

    Leaf membrane proteins are an underutilised protein fraction for food applications. Proteins from leaves can contribute to a more complete use of resources and help to meet the increasing protein demand. Leaf protein extraction and purification is applied by other disciplines, such as proteomics.

  15. Optimization of antioxidant phenolic compounds extraction from quinoa (Chenopodium quinoa) seeds.

    Science.gov (United States)

    Carciochi, Ramiro Ariel; Manrique, Guillermo Daniel; Dimitrov, Krasimir

    2015-07-01

    The objective of this study was to optimize the extraction conditions of phenolic and flavonoids compounds from quinoa (Chenopodium quinoa) seeds using ultrasound assistance technology. A randomized central composite face-centered design was used to evaluate the effect of extraction temperature, ethanol concentration in the solvent, and ultrasound power on the total phenolic content (TPC), total flavonoid content (TFC) and antioxidant activity by response surface analysis. Predicted model equations were obtained to describe the experimental data regarding TPC, TFC and antioxidant activity, with significant variation in the linear, quadratic, and interaction effects of the independent variables. Regression analysis showed that more than 88 % of the variability was explained by the models. The best extraction conditions obtained by simultaneous maximization of the responses were: extraction temperature of 60 °C, 80 % ethanol as solvent and non-application of ultrasounds. Under the optimal conditions, the corresponding predicted response values were 103.6 mg GAE/100 g dry weight (dw), 25.0 mg quercetin equiv./100 g dw and 28.6 % DPPH radical scavenging, for TPC, TFC and antioxidant activity, respectively. The experimental values agreed with those predicted within a 95 % confidence level, indicating the suitability of the employed model. HPLC analysis of the obtained extracts confirmed the highest phenolic compound yield in the extract obtained under optimal extraction conditions. Considering the characteristics of the antioxidant-rich extracts obtained, they could be consider for potential application in the food industry, as nutraceutical and functional foods ingredient or well as replacement of synthetic antioxidants.

  16. Assessing the impact of protein extraction methods for human gut metaproteomics.

    Science.gov (United States)

    Zhang, Xu; Li, Leyuan; Mayne, Janice; Ning, Zhibin; Stintzi, Alain; Figeys, Daniel

    2017-07-10

    Metaproteomics is a promising methodology for the functional characterizations of the gut microbiome. However, the performance of metaproteomic analysis is affected by protein extraction protocols in terms of the amount of protein recovered and the relative abundance of different bacteria observed in microbiome. Currently, there is a lack of consistency on protein extraction methods in published metaproteomics studies. Here we evaluated the effects of different protein extraction methods on human fecal metaproteome characterizations. We found that sodium dodecyl sulfate (SDS)-based lysis buffer obtained higher protein yields and peptide/protein group identifications compared to urea and the non-ionic detergent-based B-Per buffer. The addition of bead beating to any of the extraction buffers increased both protein yields and protein identifications. As well, bead beating led to a significant increase of the relative abundances of Firmicutes and Actinobacteria. We also demonstrated that ultrasonication, another commonly used mechanical disruption approach, performed even better than bead beating for gut microbial protein extractions. Importantly, proteins of the basic metabolic pathways showed significantly higher relative abundances when using ultrasonication. Overall, these results demonstrate that protein extraction protocols markedly impact the metaproteomic results and recommend a protein extraction protocol with both SDS and ultrasonication for metaproteomic studies. The gut microbiome is emerging as an important factor influencing human health. Metaproteomics is promising for advancing the understanding of the functional roles of the microbiome in disease. However, metaproteomics suffers from a lack of consistent sample preparation procedures. In the present study, protein extraction protocols for fecal microbiome samples were evaluated for their effects on protein yields, peptide identifications, protein group identifications, taxonomic compositions and

  17. Optimization of a wet microalgal lipid extraction procedure for improved lipid recovery for biofuel and bioproduct production.

    Science.gov (United States)

    Sathish, Ashik; Marlar, Tyler; Sims, Ronald C

    2015-10-01

    Methods to convert microalgal biomass to bio based fuels and chemicals are limited by several processing and economic hurdles. Research conducted in this study modified/optimized a previously published procedure capable of extracting transesterifiable lipids from wet algal biomass. This optimization resulted in the extraction of 77% of the total transesterifiable lipids, while reducing the amount of materials and temperature required in the procedure. In addition, characterization of side streams generated demonstrated that: (1) the C/N ratio of the residual biomass or lipid extracted (LE) biomass increased to 54.6 versus 10.1 for the original biomass, (2) the aqueous phase generated contains nitrogen, phosphorous, and carbon, and (3) the solid precipitate phase was composed of up to 11.2 wt% nitrogen (70% protein). The ability to isolate algal lipids and the possibility of utilizing generated side streams as products and/or feedstock material for downstream processes helps promote the algal biorefinery concept. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Real-time implementation of optimized maximum noise fraction transform for feature extraction of hyperspectral images

    Science.gov (United States)

    Wu, Yuanfeng; Gao, Lianru; Zhang, Bing; Zhao, Haina; Li, Jun

    2014-01-01

    We present a parallel implementation of the optimized maximum noise fraction (G-OMNF) transform algorithm for feature extraction of hyperspectral images on commodity graphics processing units (GPUs). The proposed approach explored the algorithm data-level concurrency and optimized the computing flow. We first defined a three-dimensional grid, in which each thread calculates a sub-block data to easily facilitate the spatial and spectral neighborhood data searches in noise estimation, which is one of the most important steps involved in OMNF. Then, we optimized the processing flow and computed the noise covariance matrix before computing the image covariance matrix to reduce the original hyperspectral image data transmission. These optimization strategies can greatly improve the computing efficiency and can be applied to other feature extraction algorithms. The proposed parallel feature extraction algorithm was implemented on an Nvidia Tesla GPU using the compute unified device architecture and basic linear algebra subroutines library. Through the experiments on several real hyperspectral images, our GPU parallel implementation provides a significant speedup of the algorithm compared with the CPU implementation, especially for highly data parallelizable and arithmetically intensive algorithm parts, such as noise estimation. In order to further evaluate the effectiveness of G-OMNF, we used two different applications: spectral unmixing and classification for evaluation. Considering the sensor scanning rate and the data acquisition time, the proposed parallel implementation met the on-board real-time feature extraction.

  19. Optimization of enzyme-assisted extraction and characterization of polysaccharides from Hericium erinaceus.

    Science.gov (United States)

    Zhu, Yang; Li, Qian; Mao, Guanghua; Zou, Ye; Feng, Weiwei; Zheng, Daheng; Wang, Wei; Zhou, Lulu; Zhang, Tianxiu; Yang, Jun; Yang, Liuqing; Wu, Xiangyang

    2014-01-30

    The enzyme-assisted extraction (EAE) of polysaccharides from the fruits of Hericium erinaceus was studied. In this study, response surface methodology and the Box-Behnken design based on single-factor and orthogonal experiments were applied to optimize the EAE conditions. The optimal extraction conditions were as follows: a pH of 5.71, a temperature of 52.03°C and a time of 33.79 min. The optimal extraction conditions resulted in the highest H. erinaceus polysaccharides (HEP) yield, with a value 13.46 ± 0.37%, which represented an increase of 67.72% compared to hot water extraction (HWE). The polysaccharides were characterized by FT-IR, SEM, CD, AFM, and GC. The results showed that HEP was composed of mannose, glucose, xylose, and galactose in a molar ratio of 15.16:5.55:4.21:1. The functional groups of the H. erinaceus polysaccharides extracted by HWE and EAE were fundamentally identical but had apparent conformational changes.

  20. Response surface methodology to optimize enzyme-assisted aqueous extraction of maize germ oil.

    Science.gov (United States)

    Shende, Deepika; Sidhu, Gagandeep Kaur

    2016-08-01

    The present study was aimed at optimizing the enzyme assisted aqueous extraction (EAAE) process for extraction of maize germ oil. The different EAAE process parameter viz., pH of the slurry, seed to water ratio, the temperature of incubation and time of hydrolysis were optimized for improving oil recovery and oil quality. The combined effect of independent variables on recovery of oil, time taken for oil extraction and various quality parameters were studied using response surface methodology. The designed experimental runs were conducted to obtain the optimal conditions as 5.85 pH of slurry, 1:6.92 seed to water ratio, 45.12 °C temperature of incubation and 1 h time of hydrolysis. Oil extracted under these conditions was light yellowish in color and had a pleasant nutty taste, with maximum oil recovery of 70 %. The extraction variables viz., pH of slurry, seed to water ratio, the temperature of incubation had a significant effect on recovery of oil and various quality characteristics, however the time of hydrolysis had a non significant effect.

  1. Extraction optimization, characterization and immunity activity of polysaccharides from Fructus Jujubae.

    Science.gov (United States)

    Cui, Guoting; Zhang, Wuxia; Wang, Qingjie; Zhang, Amin; Mu, Haibo; Bai, Hongjin; Duan, Jinyou

    2014-10-13

    The versatile Fructus Jujubae is widely used in Chinese and Korean traditional medicine. In this study, the extraction optimization, characterization and immunostimulatory activities of polysaccharides from Fructus Jujubae were investigated. Based on a four-variable-three-level Box-Behnken statistical design, the optimal extraction parameters were optimized as follows: extraction temperature 90 °C, extraction time 3.23 h, water to raw material ratio 33:1 and extraction 3 times. Under these conditions, the experimental yield of polysaccharides was 6.47 ± 0.26%, which was close to the predicted yield value (6.54%). The crude Fructus Jujubae polysaccharide was further purified by DEAE-cellulose chromatography repeatedly, and two homogenous fractions, designated as RQP1d and RQP2d with molecular weight of 83.8 and 123.0 kDa respectively, were obtained. Their structures were determined by chemical analysis, Fourier transform infrared (FT-IR) spectroscopy and scanning electron microscopy (SEM). Finally, preliminary immunological tests indicated that both RQP1d and RQP2d significantly stimulated NO production in RAW264.7 macrophages, and promoted LPS-induced splenocyte proliferation. These data implied Fructus Jujubae polysaccharides had the potential to be explored as novel natural immunostimulant for using in functional foods or medicine.

  2. Optimization of cloud point extraction and solid phase extraction methods for speciation of arsenic in natural water using multivariate technique

    Energy Technology Data Exchange (ETDEWEB)

    Baig, Jameel A., E-mail: jab_mughal@yahoo.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080, Sindh (Pakistan); Kazi, Tasneem G., E-mail: tgkazi@yahoo.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080, Sindh (Pakistan); Shah, Abdul Q., E-mail: aqshah07@yahoo.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080, Sindh (Pakistan); Arain, Mohammad B. [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080, Sindh (Pakistan); Afridi, Hassan I., E-mail: hassanimranafridi@yahoo.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080, Sindh (Pakistan); Kandhro, Ghulam A., E-mail: gakandhro@yahoo.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080, Sindh (Pakistan); Khan, Sumaira, E-mail: skhanzai@gmail.com [National Center of Excellence in Analytical Chemistry, University of Sindh, Jamshoro 76080, Sindh (Pakistan)

    2009-09-28

    The simple and rapid pre-concentration techniques viz. cloud point extraction (CPE) and solid phase extraction (SPE) were applied for the determination of As{sup 3+} and total inorganic arsenic (iAs) in surface and ground water samples. The As{sup 3+} was formed complex with ammonium pyrrolidinedithiocarbamate (APDC) and extracted by surfactant-rich phases in the non-ionic surfactant Triton X-114, after centrifugation the surfactant-rich phase was diluted with 0.1 mol L{sup -1} HNO{sub 3} in methanol. While total iAs in water samples was adsorbed on titanium dioxide (TiO{sub 2}); after centrifugation, the solid phase was prepared to be slurry for determination. The extracted As species were determined by electrothermal atomic absorption spectrometry. The multivariate strategy was applied to estimate the optimum values of experimental factors for the recovery of As{sup 3+} and total iAs by CPE and SPE. The standard addition method was used to validate the optimized methods. The obtained result showed sufficient recoveries for As{sup 3+} and iAs (>98.0%). The concentration factor in both cases was found to be 40.

  3. fast_protein_cluster: parallel and optimized clustering of large-scale protein modeling data.

    Science.gov (United States)

    Hung, Ling-Hong; Samudrala, Ram

    2014-06-15

    fast_protein_cluster is a fast, parallel and memory efficient package used to cluster 60 000 sets of protein models (with up to 550 000 models per set) generated by the Nutritious Rice for the World project. fast_protein_cluster is an optimized and extensible toolkit that supports Root Mean Square Deviation after optimal superposition (RMSD) and Template Modeling score (TM-score) as metrics. RMSD calculations using a laptop CPU are 60× faster than qcprot and 3× faster than current graphics processing unit (GPU) implementations. New GPU code further increases the speed of RMSD and TM-score calculations. fast_protein_cluster provides novel k-means and hierarchical clustering methods that are up to 250× and 2000× faster, respectively, than Clusco, and identify significantly more accurate models than Spicker and Clusco. fast_protein_cluster is written in C++ using OpenMP for multi-threading support. Custom streaming Single Instruction Multiple Data (SIMD) extensions and advanced vector extension intrinsics code accelerate CPU calculations, and OpenCL kernels support AMD and Nvidia GPUs. fast_protein_cluster is available under the M.I.T. license. (http://software.compbio.washington.edu/fast_protein_cluster) © The Author 2014. Published by Oxford University Press.

  4. Inactivation disinfection property of Moringa Oleifera seed extract: optimization and kinetic studies

    Science.gov (United States)

    Idris, M. A.; Jami, M. S.; Hammed, A. M.

    2017-05-01

    This paper presents the statistical optimization study of disinfection inactivation parameters of defatted Moringa oleifera seed extract on Pseudomonas aeruginosa bacterial cells. Three level factorial design was used to estimate the optimum range and the kinetics of the inactivation process was also carried. The inactivation process involved comparing different disinfection models of Chicks-Watson, Collins-Selleck and Homs models. The results from analysis of variance (ANOVA) of the statistical optimization process revealed that only contact time was significant. The optimum disinfection range of the seed extract was 125 mg/L, 30 minutes and 120rpm agitation. At the optimum dose, the inactivation kinetics followed the Collin-Selleck model with coefficient of determination (R2) of 0.6320. This study is the first of its kind in determining the inactivation kinetics of pseudomonas aeruginosa using the defatted seed extract.

  5. Optimization of ethanol production from hot-water extracts of sugar maple chips

    Energy Technology Data Exchange (ETDEWEB)

    Xu, Jian; Liu, Shijie [Department of Paper and Bioprocess Engineering, College of Environmental Science and Forestry, State University of New York, 1 Forestry Drive, Syracuse, NY 13210 (United States)

    2009-11-15

    Hot-water extracts from sugar maple chips prior to papermaking was employed in this study to produce ethanol by Pichia stipitis 58784. The effects of several factors, seed culture age, fermentation time, inoculum quantity, agitation rate, percent extract, concentration of inorganic nitrogen source (NH{sub 4}){sub 2}SO{sub 4} and pH value, on ethanol production were investigated by orthogonal experiments. Orthogonal analysis shows that the optimal fermentation was obtained in the condition of 48-h seed culture, 120-h fermentation, 16% inoculum, 180 rpm, containing 30% extracts, 8% ammonium sulphate supplement and pH 5. This optimal condition was verified at 800-mL level in a 1.3 L fermentor. The ethanol yield reached 82.27% of the theoretical (20.57 g/L) after 120 h. (author)

  6. Application of D-optimal design to extract the pectin from lime bagasse using microwave green irradiation.

    Science.gov (United States)

    Thirugnanasambandham, K; Sivakumar, V

    2015-01-01

    The main objective of the present study is to extract the pectin from lime bagasse under various extraction conditions such as microwave power, extraction time, temperature, pH and mass of the sample. RSM coupled with D-optimal experimental design was used to optimize and investigate the extraction conditions on the pectin yield. Second order polynomial equation was developed and its adequacy was analyzed by analysis of variance (ANOVA). The optimal extraction was found to be as follows; microwave power of 400 W, extraction time of 500 s, temperature of 30 °C, pH of 1 and mass of the sample of 6g. Under these optimal scheme, 7.8 g/100g of pectin was extracted. Molecular weight of the pectin polymer and the distribution of the pectin compounds were determined by gel filtration chromatography.

  7. Optimization Extraction, Preliminary Characterization and Antioxidant Activities of Polysaccharides from Semen Juglandis

    Directory of Open Access Journals (Sweden)

    Xueyong Ren

    2016-10-01

    Full Text Available The optimization extraction process, preliminary characterization and antioxidant activities of polysaccharides from Semen Juglandis (SJP were studied in this paper. Based on the Box-Behnken experimental design and response surface methodology, the optimal extraction conditions for the SJP extraction were obtained as follows: temperature 88 °C, extraction time 125 min and ratio of liquid to solid 31 mL/g. Under these conditions, experimental extraction yield of SJP was (5.73 ± 0.014% (n = 5, similar to the predicted value of 5.78%. Furtherly, the purified SJP obtained from SJP extract by DEAE-52 and Sephacryl S-100 chromatography was analyzed to be rhamnose, galacturonic acid, galactose, arabinose and fucose in the molar ratio of 1:6.34:1.38:3.21:1.56. And the weight-average molecular weight and radius of gyration of the purified SJP in 0.1 M NaCl were determined to be 2.76 × 104 g/mol and 122 nm by SEC-MALLS, respectively. More importantly, it exhibited appreciable antioxidant activities compared to the standard Vc, such as DPPH radical scavenging activity (IC50 0.21 mg/mL, strong reducing power, ABTS radical scavenging activity (IC50 0.29 mg/mL, and hydroxyl radical scavenging activity (IC50 0.38 mg/mL. These results indicate that SJP may be useful for developing functional health products or natural antioxidant.

  8. Optimization of enzyme-assisted extraction of polysaccharides from alfalfa and its antioxidant activity.

    Science.gov (United States)

    Wang, Shaopu; Dong, Xiaofang; Tong, Jianming

    2013-11-01

    In this present study, an efficient complex enzyme-assisted extraction technology was developed and optimized to extract polysaccharides from alfalfa using four factors at five levels central composite rotatable response surface design (CCRD). The experimental data was fitted to a second order polynomial equation with high coefficient of determination values (R(2)>0.95). The results of statistical analysis showed that the linear and quadratic terms of these four variables had significant effects (Penzyme concentration of 2.5%, 2.0%, 3.0% (weight of alfalfa) of cellulase, papain and pectase, extraction temperature 52.7 °C, extraction pH 3.87, ratio of water to raw material 78.92 mL/g and extraction time 2.73 h. Under the optimal conditions, the experimental extraction yield of alfalfa polysaccharides was 5.05 ± 0.02%, which was well matched with the value (5.09%) predicted by the CCRD model. Moreover, evaluation of the antioxidant activity of polysaccharides from alfalfa in vitro suggested that the polysaccharides had good antioxidant effect, especially scavenging activity for hydroxyl radical and DPPH radical, which indicated that the polysaccharides from alfalfa may be explored as a novel natural antioxidant.

  9. Extraction optimization, characterization and antioxidant activity in vitro of polysaccharides from mulberry (Morus alba L.) leaves.

    Science.gov (United States)

    Yuan, Qingxia; Xie, Yufeng; Wang, Wei; Yan, Yuhua; Ye, Hong; Jabbar, Saqib; Zeng, Xiaoxiong

    2015-09-05

    Extraction optimization, characterization and antioxidant activity in vitro of polysaccharides from mulberry leaves (MLP) were investigated in the present study. The optimal extraction conditions with an extraction yield of 10.0 ± 0.5% for MLP were determined as follows: extraction temperature 92 °C, extraction time 3.5h and ratio (v/w, mL/g) of extraction solvent (water) to raw material 34. Two purified fractions, MLP-3a and MLP-3b with molecular weights of 80.99 and 3.64 kDa, respectively, were obtained from crude MLP by chromatography of DEAE-Cellulose 52 and Sephadex G-100. Fourier transform-infrared spectroscopy revealed that crude MLP, MLP-3a and MLP-3b were acidic polysaccharides. Furthermore, crude MLP and MLP-3a had more complicated monosaccharide compositions, while MLP-3b had a relatively higher content of uronic acid. Crude MLP, MLP-3a and MLP-3b exhibited potent Fe(2+) chelating power and scavenging activities on 1,1-diphenyl-2-picrylhydrazyl, hydroxyl, superoxide and 2,2'-azinobis-(3-ethyl-benzothiazolin-6-sulfonic acid) radicals. The results suggested that MLP could be explored as natural antioxidant.

  10. Optimization of extraction, characterization and antioxidant activity of polysaccharides from Brassica rapa L.

    Science.gov (United States)

    Wang, Wei; Wang, Xiaoqing; Ye, Hong; Hu, Bing; Zhou, Li; Jabbar, Saqib; Zeng, Xiaoxiong; Shen, Wenbiao

    2016-01-01

    The root of Brassica rapa L. has been traditionally used as a Uyghur folk medicine to cure cough and asthma by Uyghur nationality in Xinjiang Uygur Autonomous Region of China. In the present study, therefore, extraction optimization, characterization and antioxidant activity in vitro of polysaccharides from the root of B. rapa L. (BRP) were investigated. The optimal extraction conditions with an extraction yield of 21.48 ± 0.41% for crude BRP were obtained as follows: extraction temperature 93°C, extraction time 4.3h and ratio of extraction solvent (water) to raw material 75 mL/g. The crude BRP was purified by chromatographic columns of DEAE-52 cellulose and Sephadex G-100, affording three purified fractions of BRP-1-1, BRP-2-1 and BRP-2-2 with average molecular weight of 1510, 1110 and 838 kDa, respectively. Monosaccharide composition analysis indicated that BRP-1-1 was composed of mannose, rhamnose, glucose, galactose and arabinose, BRP-2-1 was composed of rhamnose, galacturonic acid, galactose and arabinose, and BRP-2-2 was composed of rhamnose and galacturonic acid in a molar ratio of 1.27: 54.92. Furthermore, the crude BRP exhibited relatively higher antioxidant activity in vitro than purified fractions; hence, it could be used as a natural antioxidant in functional foods or medicines.

  11. Optimization of Ficus deltoidea Using Ultrasound-Assisted Extraction by Box-Behnken Statistical Design

    Directory of Open Access Journals (Sweden)

    L. J. Ong

    2016-09-01

    Full Text Available In this study, the effect of extraction parameters (ethanol concentration, sonication time, and solvent-to-sample ratio on Ficus deltoidea leaves was investigated using ultrasound-assisted extraction by response surface methodology (RSM. Total phenolic content (TPC of F. deltoidea extracts was identified using Folin-Ciocalteu method and expressed in gallic acid equivalent (GAE per g. Box-Behnken statistical design (BBD was the tool used to find the optimal conditions for maximum TPC. Besides, the extraction yield was measured and stated in percentage. The optimized TPC attained was 455.78 mg GAE/g at 64% ethanol concentration, 10 minutes sonication time, and 20 mL/g solvent-to-sample ratio whereas the greatest extraction yield was 33% with ethanol concentration of 70%, sonication time of 40 minutes, and solvent-to-material ratio at 40 mL/g. The determination coefficient, R2, for TPC indicates that 99.5% capriciousness in the response could be clarified by the ANOVA model and the value of 0.9681 of predicted R2 is in equitable agreement with the 0.9890 of adjusted R2. The present study shows that ethanol water as solvent, a short time of 10 minutes, and adequate solvent-to-sample ratio (20 mL/g are the best conditions for extraction.

  12. Oil extraction from Scenedesmus obliquus using a continuous microwave system--design, optimization, and quality characterization.

    Science.gov (United States)

    Balasubramanian, Sundar; Allen, James D; Kanitkar, Akanksha; Boldor, Dorin

    2011-02-01

    A 1.2 kW, 2450 MHz resonant continuous microwave processing system was designed and optimized for oil extraction from green algae (Scenedesmus obliquus). Algae-water suspension (1:1 w/w) was heated to 80 and 95°C, and subjected to extraction for up to 30 min. Maximum oil yield was achieved at 95°C and 30 min. The microwave system extracted 76-77% of total recoverable oil at 20-30 min and 95°C, compared to only 43-47% for water bath control. Extraction time and temperature had significant influence (pextraction yield. Oil analysis indicated that microwaves extracted oil containing higher percentages of unsaturated and essential fatty acids (indicating higher quality). This study validates for the first time the efficiency of a continuous microwave system for extraction of lipids from algae. Higher oil yields, faster extraction rates and superior oil quality demonstrate this system's feasibility for oil extraction from a variety of feedstock.

  13. An Extraction Process for Optimal Utilization of Naphtha Based on Molecule Management

    Institute of Scientific and Technical Information of China (English)

    Wang Tianxiao; Shen Benxian; Sun Hui

    2016-01-01

    In this paper, the separation of aromatics from light naphtha by using extraction process was investigated for im-proving the utilization efficiency of naphtha. It is indicated that, using a mixture of propylene carbonate-diethylene glycol as the solvent, the optimal extraction conditions cover: a volume fraction of propylene carbonate in the mixed solvent of 0.3, a solvent to feed ratio of 8, and an extraction temperature of 308 K. Through the extraction process, the aromatics mass frac-tion increases from 10.05% in naphtha to 27.74% in extract oil. It is found that the aromatics yield of extract oil,RA, reaches 92.11%. As a result, in comparison with naphtha, the potential aromatics content of extract oil increases impressively by 18.03%. Meanwhile, the aromatics content of raffinate oil decreases to 1.33%, and the normal paraffin yield of raffinate oil, Rp, is 76.61%. Accordingly, higher total olefins yields can be obtained when using raffinate oil as the raw material for steam cracking. The present results show that the utilization efficiency of naphtha is improved through extraction process.

  14. Effect of chickpea aqueous extracts, organic extracts, and protein concentrates on cell proliferation.

    Science.gov (United States)

    Girón-Calle, Julio; Vioque, Javier; del Mar Yust, María; Pedroche, Justo; Alaiz, Manuel; Millán, Francisco

    2004-01-01

    Pulses should be part of a healthy diet, and it is also becoming clear that they have health-promoting effects. Nevertheless, most studies on the bioactive or health-promoting properties of pulses have been carried out using soybeans. We have studied cell growth-regulating properties, which may be responsible for anti-cancer properties, in chickpea seeds. Chickpea seeds are a staple in the traditional diet of many Mediterranean, Asian, and South and Central American countries. In addition, chickpea seeds have industrial applications since they can be used for the preparation of protein concentrates and isolates. The cell lines Caco-2 (epithelial intestinal) and J774 (macrophages) have been exposed to chickpea seed extracts and protein preparations in order to screen the different chickpea fractions for effects on cell growth. Both cell growth-promoting and cell growth-inhibiting effects were found. Most interestingly, a fraction soluble in ethanol and acetone specifically and almost completely inhibited the growth of Caco-2 cells exhibiting a cancerous phenotype. It is concluded that chickpea seeds are a source of bioactive components and deserve further study for their possible anti-cancer effect.

  15. Protein biomarkers of external root resorption: A new protein extraction protocol. Are we going in the right direction?

    OpenAIRE

    2014-01-01

    OBJECTIVE: The aim of this study is to determine a protocol of gingival crevicular fluid protein extraction used for the first dimension of 2-DE gels. It also aims at conducting a review on the current candidates for protein markers of this pathology, all of which may be used to prevent the disease. METHODS: Gingival crevicular fluid was collected from two groups of 60 patients each, with and without external root resorption. Samples were extracted by means of various methods of protein extra...

  16. Extraction and characterization of protein from Irish brown seaweed Ascophyllum nodosum.

    Science.gov (United States)

    Kadam, Shekhar U; Álvarez, Carlos; Tiwari, Brijesh K; O'Donnell, Colm P

    2017-09-01

    This study investigates traditional and non-conventional methods of extraction of protein from Irish brown seaweed A. nodosum. Acid, alkali, combined acid-alkali with and without ultrasound pretreatment were investigated for extraction of protein from A. nodosum. Molecular weight of protein was determined using high performance size exclusion chromatography and amino acid profiling was carried out using an amino acid analyzer. Combination of first acid and then alkali extraction was found to be the most efficient method of extraction among all methods investigated (59% of recovery); followed by single step of alkali extraction assisted with ultrasound (68.4μm) which was able to extract 57% of total protein. Alkaline extraction was shown to yield the best protein/algae liquefaction ratio (1.28). This can be attributed to the release of polysaccharide complexes first by acid and then solubilization of proteins by alkali solvent. The molecular weight of extracted protein was found to be relatively low, in the range of 2-4kDa average MW. The alkali method of extraction was found to be optimum for extraction of amino acids from A. nodosum. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Optimising the Use of TRIzol-extracted Proteins in Surface Enhanced Laser Desorption/ Ionization (SELDI Analysis

    Directory of Open Access Journals (Sweden)

    Perlaky Laszlo

    2006-03-01

    Full Text Available Abstract Background Research with clinical specimens is always hampered by the limited availability of relevant samples, necessitating the use of a single sample for multiple assays. TRIzol is a common reagent for RNA extraction, but DNA and protein fractions can also be used for other studies. However, little is known about using TRIzol-extracted proteins in proteomic research, partly because proteins extracted from TRIzol are very resistant to solubilization. Results To facilitate the use of TRIzol-extracted proteins, we first compared the ability of four different common solubilizing reagents to solubilize the TRIzol-extracted proteins from an osteosarcoma cell line, U2-OS. Then we analyzed the solubilized proteins by Surface Enhanced Laser Desorption/ Ionization technique (SELDI. The results showed that solubilization of TRIzol-extracted proteins with 9.5 M Urea and 2% CHAPS ([3-[(3-cholamidopropyl-dimethylammonio]propanesulfonate] (UREA-CHAPS was significantly better than the standard 1% SDS in terms of solubilization efficiency and the number of detectable ion peaks. Using three different types of SELDI arrays (CM10, H50, and IMAC-Cu, we demonstrated that peak detection with proteins solubilized by UREA-CHAPS was reproducible (r > 0.9. Further SELDI analysis indicated that the number of ion peaks detected in TRIzol-extracted proteins was comparable to a direct extraction method, suggesting many proteins still remain in the TRIzol protein fraction. Conclusion Our results suggest that UREA-CHAPS performed very well in solubilizing TRIzol-extracted proteins for SELDI applications. Protein fractions left over after TRIzol RNA extraction could be a valuable but neglected source for proteomic or biochemical analysis when additional samples are not available.

  18. Protein extraction and gel-based separation methods to analyze responses to pathogens in carnation (Dianthus caryophyllus L).

    Science.gov (United States)

    Ardila, Harold Duban; Fernández, Raquel González; Higuera, Blanca Ligia; Redondo, Inmaculada; Martínez, Sixta Tulia

    2014-01-01

    We are currently using a 2-DE-based proteomics approach to study plant responses to pathogenic fungi by using the carnation (Dianthus caryophyllus L)-Fusarium oxysporum f. sp. dianthi pathosystem. It is clear that the protocols for the first stages of a standard proteomics workflow must be optimized to each biological system and objectives of the research. The optimization procedure for the extraction and separation of proteins by 1-DE and 2-DE in the indicated system is reported. This strategy can be extrapolated to other plant-pathogen interaction systems in order to perform an evaluation of the changes in the host protein profile caused by the pathogen and to identify proteins which, at early stages, are involved or implicated in the plant defense response.

  19. Anti-Cancer Effects of Protein Extracts from Calvatia lilacina, Pleurotus ostreatus and Volvariella volvacea

    Directory of Open Access Journals (Sweden)

    Jin-Yi Wu

    2011-01-01

    Full Text Available Calvatia lilacina (CL, Pleurotus ostreatus (PO and Volvariella volvacea (VV are widely distributed worldwide and commonly eaten as mushrooms. In this study, cell viabilities were evaluated for a human colorectal adenocarcinoma cell line (SW480 cells and a human monocytic leukemia cell line (THP-1 cells. Apoptotic mechanisms induced by the protein extracts of PO and VV were evaluated for SW480 cells. The viabilities of THP-1 and SW480 cells decreased in a concentration-dependent manner after 24 h of treatment with the protein extracts of CL, PO or VV. Apoptosis analysis revealed that the percentage of SW480 cells in the SubG1 phase (a marker of apoptosis was increased upon PO and VV protein-extract treatments, indicating that oligonucleosomal DNA fragmentation existed concomitantly with cellular death. The PO and VV protein extracts induced reactive oxygen species (ROS production, glutathione (GSH depletion and mitochondrial transmembrane potential (ΔΨm loss in SW480 cells. Pretreatment with N-acetylcysteine, GSH or cyclosporine A partially prevented the apoptosis induced by PO protein extracts, but not that induced by VV extracts, in SW480 cells. The protein extracts of CL, PO and VV exhibited therapeutic efficacy against human colorectal adenocarcinoma cells and human monocytic leukemia cells. The PO protein extracts induced apoptosis in SW480 cells partially through ROS production, GSH depletion and mitochondrial dysfunction. Therefore, the protein extracts of these mushrooms could be considered an important source of new anti-cancer drugs.

  20. Characterization of the immune response of domestic fowl following immunization with proteins extracted from Dermanyssus gallinae.

    Science.gov (United States)

    Harrington, David; Din, Hatem Mohi El; Guy, Jonathan; Robinson, Karen; Sparagano, Olivier

    2009-03-23

    Dermanyssus gallinae is the most significant ectoparasite of European poultry egg laying production systems due to high costs of control and associated production losses as well as adverse effects on bird welfare. In this study, soluble proteins were extracted from unfed D. gallinae (DGE) using a urea-based detergent and ultra-filtration, passed through a 0.22 microm filter and blended aseptically with adjuvant. One group of laying hens was immunized with DGE and adjuvant (Montanide ISA 50 V) whilst another group (Control) received physiological saline and adjuvant. All birds were immunized on two occasions, 21 days apart. Antibody response to immunization was determined by ELISA and western blotting using immunoglobulins (Igs) extracted from egg yolk. DGE immunization of hens resulted in a significant (Pgallinae can be used to stimulate a protective immune response in laying hens. Further work is needed to identify other proteins of interest that could confer higher protection against D. gallinae, as well as optimization of the vaccination and in vitro testing protocol.

  1. Process Optimization of Ultrasonic-Assisted Extraction of Arabinogalactan from Dihydroquercetin Extracted Residues by Response Surface Methodology and Evaluation of Its Antioxidant Activity

    Directory of Open Access Journals (Sweden)

    Zaizhi Liu

    2013-01-01

    Full Text Available Ultrasound was used for the extraction of larch arabinogalactan from Larix gmelinii. The optimal conditions for ultrasound extraction were determined by response surface methodology. Specifically, the Box-Behnken design was employed to evaluate the effects of three independent variables: ultrasound time, temperature, and liquid-solid ratio. The highest arabinogalactan yield (11.18% was obtained under the optimal extraction condition (extraction temperature 41.5°C, extraction time 24.3 min, and liquid-solid ratio 40 mL/g. In addition, the antioxidant activity of arabinogalactan that was extracted from dihydroquercetin extraction residues exhibited a moderate and concentration-dependent hydroxyl radical-scavenging capacity, ferric-reducing power, and reducing power. The wood material was characterized before and after processing by scanning electron microscopy and Fourier-transform infrared spectroscopy.

  2. Optimization of supercritical carbon dioxide extraction of Piper Betel Linn leaves oil and total phenolic content

    Science.gov (United States)

    Aziz, A. H. A.; Yunus, M. A. C.; Arsad, N. H.; Lee, N. Y.; Idham, Z.; Razak, A. Q. A.

    2016-11-01

    Supercritical Carbon Dioxide (SC-CO2) Extraction was applied to extract piper betel linn leaves. The piper betel leaves oil was used antioxidant, anti-diabetic, anticancer and antistroke. The aim of this study was to optimize the conditions of pressure, temperature and flowrate for oil yield and total phenolic content. The operational conditions of SC-CO2 studied were pressure (10, 20, 30 MPa), temperature (40, 60, 80 °C) and flowrate carbon dioxide (4, 6, 8 mL/min). The constant parameters were average particle size and extraction regime, 355pm and 3.5 hours respectively. First order polynomial expression was used to express the extracted oil while second order polynomial expression was used to express the total phenolic content and the both results were satisfactory. The best conditions to maximize the total extraction oil yields and total phenolic content were 30 MPa, 80 °C and 4.42 mL/min leading to 7.32% of oil and 29.72 MPa, 67.53 °C and 7.98 mL/min leading to 845.085 mg GAE/g sample. In terms of optimum condition with high extraction yield and high total phenolic content in the extracts, the best operating conditions were 30 MPa, 78 °C and 8 mL/min with 7.05% yield and 791.709 mg gallic acid equivalent (GAE)/g sample. The most dominant condition for extraction of oil yield and phenolic content were pressure and CO2 flowrate. The results show a good fit to the proposed model and the optimal conditions obtained were within the experimental range with the value of R2 was 96.13% for percentage yield and 98.52% for total phenolic content.

  3. Aqueous Two-Phase Extraction of Polyphenols Using a Microchannel System – Process Optimization and Intensification

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    Ivana Rukavina

    2011-01-01

    Full Text Available Polyphenols are one of the most numerous and widespread groups of compounds in the plant world. Nowadays, organic solvents such as methanol, ethanol, acetone, dimethylformamide, ethyl acetate and diethylether are mainly used for the extraction of polyphenols. These solvents require special process conditions and special care in the disposal of the used solvents. In this paper, the extraction of polyphenols from the model solution was performed using the aqueous two-phase system which contains 80.90 % water and represents low burden on the environment. The aqueous solution of gallic acid (GA was used as a model solution of polyphenols. The extraction was performed in the aqueous two-phase system containing PEG6000/H2O/(NH42SO4 in a macroextractor (V=10 mL and microextractor (V=14 ƒμL. The influence of the process parameters, the concentration of gallic acid, pH and composition of the aqueous two-phase system was investigated in order to maximize the partition coefficient. The method of multifactor experimental planning was used to optimize the extraction process and the results were statistically analysed using the evolutionary operation method (EVOP. Optimal operating conditions of the extraction process were pH=6.50, γGA=4.50 g/L, the mass fraction of polyethylene glycol (PEG wPEG=0.1037 g/g and the mass fraction of ammonium sulphate (AMS wAMS=0.0925 g/g. Under these conditions the maximal partition coefficient of K=5.54 and the extraction efficiency of E=89.11 % were achieved and successfully applied for total phenol extraction from white wine in the macro- and microextractor. Approximately the same partition coefficients and extraction efficiency were achieved in the microextractor within a 60-fold shorter residence time.

  4. Optimization of Supercritical CO2 Extraction of Fish Oil from Viscera of African Catfish (Clarias gariepinus

    Directory of Open Access Journals (Sweden)

    Mohamed Zaidul Islam Sarker

    2012-09-01

    Full Text Available Fish oil was extracted from the viscera of African Catfish using supercritical carbon dioxide (SC-CO2. A Central Composite Design of Response Surface methodology (RSM was employed to optimize the SC-CO2 extraction parameters. The oil yield (Y as response variable was executed against the four independent variables, namely pressure, temperature, flow rate and soaking time. The oil yield varied with the linear, quadratic and interaction of pressure, temperature, flow rate and soaking time. Optimum points were observed within the variables of temperature from 35 °C to 80 °C, pressure from 10 MPa to 40 MPa, flow rate from 1 mL/min to 3 mL/min and soaking time from 1 h to 4 h. However, the extraction parameters were found to be optimized at temperature 57.5 °C, pressure 40 MPa, flow rate 2.0 mL/min and soaking time 2.5 h. At this optimized condition, the highest oil yields were found to be 67.0% (g oil/100 g sample on dry basis in the viscera of catfish which was reasonable to the yields of 78.0% extracted using the Soxhlet method.

  5. Optimization of lycopene extraction from tomato cell suspension culture by response surface methodology.

    Science.gov (United States)

    Lu, Chi-Hua; Engelmann, Nancy J; Lila, Mary Ann; Erdman, John W

    2008-09-10

    Radioisotope-labeled lycopene is an important tool for biomedical research but currently is not commercially available. A tomato cell suspension culture system for the production of radioisotope-labeled lycopene was previously developed in our laboratory. In the current study, the goal was to optimize the lycopene extraction efficiency from tomato cell cultures for preparatory high-performance liquid chromatography (HPLC) separation. We employed response surface methodology (RSM), which combines fractional factorial design and a second-degree polynomial model. Tomato cells were homogenized with ethanol, saponified by KOH, and extracted with hexane, and the lycopene content was analyzed by HPLC-PDA. We varied five factors at five levels: ethanol volume (1.33-4 mL/g); homogenization period (0-40 s/g); saturated KOH solution volume (0-0.67 mL/g); hexane volume (1.67-3 mL/g); and vortex period (5-25 s/g). Ridge analysis by SAS suggested that the optimal extraction procedure to extract 1 g of tomato cells was at 1.56 mL of ethanol, 28 s homogenization, 0.29 mL of KOH, 2.49 mL of hexane, and 17.5 s vortex. These optimal conditions predicted by RSM were confirmed to enhance lycopene yield from standardized tomato cell cultures by more than 3-fold.

  6. Optimization of the Aqueous Extraction of Phenolic Compounds from Olive Leaves.

    Science.gov (United States)

    Goldsmith, Chloe D; Vuong, Quan V; Stathopoulos, Costas E; Roach, Paul D; Scarlett, Christopher J

    2014-10-23

    Olive leaves are an agricultural waste of the olive-oil industry representing up to 10% of the dry weight arriving at olive mills. Disposal of this waste adds additional expense to farmers. Olive leaves have been shown to have a high concentration of phenolic compounds. In an attempt to utilize this waste product for phenolic compounds, we optimized their extraction using water-a "green" extraction solvent that has not yet been investigated for this purpose. Experiments were carried out according to a Box Behnken design, and the best possible combination of temperature, extraction time and sample-to-solvent ratio for the extraction of phenolic compounds with a high antioxidant activity was obtained using RSM; the optimal conditions for the highest yield of phenolic compounds was 90 °C for 70 min at a sample-to-solvent ratio of 1:100 g/mL; however, at 1:60 g/mL, we retained 80% of the total phenolic compounds and maximized antioxidant capacity. Therefore the sample-to-solvent ratio of 1:60 was chosen as optimal and used for further validation. The validation test fell inside the confidence range indicated by the RSM output; hence, the statistical model was trusted. The proposed method is inexpensive, easily up-scaled to industry and shows potential as an additional source of income for olive growers.

  7. Glycosylation of therapeutic proteins: an effective strategy to optimize efficacy.

    Science.gov (United States)

    Solá, Ricardo J; Griebenow, Kai

    2010-02-01

    During their development and administration, protein-based drugs routinely display suboptimal therapeutic efficacies due to their poor physicochemical and pharmacological properties. These innate liabilities have driven the development of molecular strategies to improve the therapeutic behavior of protein drugs. Among the currently developed approaches, glycoengineering is one of the most promising, because it has been shown to simultaneously afford improvements in most of the parameters necessary for optimization of in vivo efficacy while allowing for targeting to the desired site of action. These include increased in vitro and in vivo molecular stability (due to reduced oxidation, cross-linking, pH-, chemical-, heating-, and freezing-induced unfolding/denaturation, precipitation, kinetic inactivation, and aggregation), as well as modulated pharmacodynamic responses (due to altered potencies from diminished in vitro enzymatic activities and altered receptor binding affinities) and improved pharmacokinetic profiles (due to altered absorption and distribution behaviors, longer circulation lifetimes, and decreased clearance rates). This article provides an account of the effects that glycosylation has on the therapeutic efficacy of protein drugs and describes the current understanding of the mechanisms by which glycosylation leads to such effects.

  8. Spray Drying of Rhodomyrtus tomentosa (Ait. Hassk. Flavonoids Extract: Optimization and Physicochemical, Morphological, and Antioxidant Properties

    Directory of Open Access Journals (Sweden)

    Pingping Wu

    2014-01-01

    Full Text Available The optimal condition of spray drying purified flavonoids extract from R. tomentosa berries was studied by response surface methodology. The optimized condition for microencapsulation was of maltodextrin to gum Arabic ratio 1 : 1.3, total solid content 27.4%, glycerol monostearate content 0.25%, and core to coating material ratio 3 : 7, resulting in EE 91.75%. Prepared at the optimized condition, the flavonoids extract microcapsules (FEMs were irregularly spherical particles with low moisture content (3.27%, high solubility (92.35%, and high bulk density (0.346 g/cm3. DPPH radical scavenging activity of FEMs was not decreased after spray drying (P>0.05 and higher than those in citric acid and rutin at the same concentration. Moreover, FEMs effectively retarded the oxidation of fresh lard during the 10-day storage period compared with vitamin C, nonencapsulated flavonoids extract, and rutin. Therefore, FEMs produced at the optimized condition could be used as powder ingredients with antioxidant capacities.

  9. Spray Drying of Rhodomyrtus tomentosa (Ait.) Hassk. Flavonoids Extract: Optimization and Physicochemical, Morphological, and Antioxidant Properties.

    Science.gov (United States)

    Wu, Pingping; Deng, Qian; Ma, Guangzhi; Li, Nianghui; Yin, Yanyan; Zhu, Baojun; Chen, Meiling; Huang, Ruqiang

    2014-01-01

    The optimal condition of spray drying purified flavonoids extract from R. tomentosa berries was studied by response surface methodology. The optimized condition for microencapsulation was of maltodextrin to gum Arabic ratio 1 : 1.3, total solid content 27.4%, glycerol monostearate content 0.25%, and core to coating material ratio 3 : 7, resulting in EE 91.75%. Prepared at the optimized condition, the flavonoids extract microcapsules (FEMs) were irregularly spherical particles with low moisture content (3.27%), high solubility (92.35%), and high bulk density (0.346 g/cm(3)). DPPH radical scavenging activity of FEMs was not decreased after spray drying (P > 0.05) and higher than those in citric acid and rutin at the same concentration. Moreover, FEMs effectively retarded the oxidation of fresh lard during the 10-day storage period compared with vitamin C, nonencapsulated flavonoids extract, and rutin. Therefore, FEMs produced at the optimized condition could be used as powder ingredients with antioxidant capacities.

  10. Parametric optimization of rice bran oil extraction using response surface methodology

    Directory of Open Access Journals (Sweden)

    Ahmad Syed W.

    2016-09-01

    Full Text Available Use of bran oil in various edible and nonedible industries is very common. In this research work, efficient and optimized methodology for the recovery of rice bran oil has been investigated. The present statistical study includes parametric optimization, based on experimental results of rice bran oil extraction. In this study, three solvents, acetone, ethanol and solvent mixture (SM [acetone: ethanol (1:1 v/v] were employed in extraction investigations. Response surface methodology (RSM, an optimization technique, was exploited for this purpose. A five level central composite design (CCD consisting four operating parameter, like temperature, stirring rate, solvent-bran ratio and contact time were examined to optimize rice bran oil extraction. Experimental results showed that oil recovery can be enhanced from 71% to 82% when temperature, solvent-bran ratio, stirring rate and contact time were kept at 55°C, 6:1, 180 rpm and 45 minutes, respectively while fixing the pH of the mixture at 7.1.

  11. Optimization of ultrasound-assisted extraction to obtain mycosterols from Agaricus bisporus L. by response surface methodology and comparison with conventional Soxhlet extraction.

    Science.gov (United States)

    Heleno, Sandrina A; Diz, Patrícia; Prieto, M A; Barros, Lillian; Rodrigues, Alírio; Barreiro, Maria Filomena; Ferreira, Isabel C F R

    2016-04-15

    Ergosterol, a molecule with high commercial value, is the most abundant mycosterol in Agaricus bisporus L. To replace common conventional extraction techniques (e.g. Soxhlet), the present study reports the optimal ultrasound-assisted extraction conditions for ergosterol. After preliminary tests, the results showed that solvents, time and ultrasound power altered the extraction efficiency. Using response surface methodology, models were developed to investigate the favourable experimental conditions that maximize the extraction efficiency. All statistical criteria demonstrated the validity of the proposed models. Overall, ultrasound-assisted extraction with ethanol at 375 W during 15 min proved to be as efficient as the Soxhlet extraction, yielding 671.5 ± 0.5mg ergosterol/100 g dw. However, with n-hexane extracts with higher purity (mg ergosterol/g extract) were obtained. Finally, it was proposed for the removal of the saponification step, which simplifies the extraction process and makes it more feasible for its industrial transference.

  12. Supercritical Carbon Dioxide Extraction of Bioactive Compounds from Ampelopsis grossedentata Stems: Process Optimization and Antioxidant Activity

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    Da Sun

    2011-10-01

    Full Text Available Supercritical carbon dioxide (SC-CO2 extraction of bioactive compounds including flavonoids and phenolics from Ampelopsis grossedentata stems was carried out. Extraction parameters such as pressure, temperature, dynamic time and modifier, were optimized using an orthogonal array design of L9 (34, and antioxidant activities of the extracts were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH free radical scavenging assay and ferrous ion chelating (FIC assay. The best conditions obtained for SC-CO2 extraction of flavonoids was 250 bar, 40 °C, 50 min, and with a modifier of methanol/ethanol (1:3, v/v, and that for phenolics extraction was 250 bar, 40 °C, 50 min, and with a modifier of methanol/ethanol (1:1, v/v. Meantime, flavonoids and phenolics were found to be mainly responsible for the DPPH scavenging activity of the extracts, but not for the chelating activity on ferrous ion according to Pearson correlation analysis. Furthermore, several unreported flavonoids such as apigenin, vitexin, luteolin, etc., have been detected in the extracts from A. grossedentata stems.

  13. Optimization of Ultrasound-Assisted Extraction of Natural Antioxidants from the Osmanthus fragrans Flower.

    Science.gov (United States)

    Li, An-Na; Li, Sha; Li, Ya; Xu, Dong-Ping; Li, Hua-Bin

    2016-02-18

    An ultrasound-assisted extraction (UAE) method was developed to extract natural antioxidants from the Osmanthus fragrans flower. The effect of UAE on antioxidant activity of the extract from the Osmanthus fragrans flower was studied using a Trolox equivalent antioxidant capacity (TEAC) assay. Optimization conditions were firstly determined using a single-factor experiment, and response surface methodology was then used to evaluate interaction of several experimental parameters. Analysis of the coefficient of determination showed that second-order polynomial models produced a highly satisfactory fitting of the experimental data with regard to TEAC values (R² = 0.9829, p extraction for 35.2 min at 59.4 °C. Under these conditions, the maximum TEAC value was 584.9 ± 6.0 μmol Trolox/g DW, which was higher than those obtained by the conventional extracting method (486.4 ± 12.6 μmol Trolox/g DW) and the Soxhlet extraction method (339.1 ± 16.2 μmol Trolox/g DW). The crude extract obtained could be used either as a food additive or in pharmaceuticals for the prevention and treatment of diseases caused by oxidative stress.

  14. A predictive control framework for optimal energy extraction of wind farms

    Science.gov (United States)

    Vali, M.; van Wingerden, J. W.; Boersma, S.; Petrović, V.; Kühn, M.

    2016-09-01

    This paper proposes an adjoint-based model predictive control for optimal energy extraction of wind farms. It employs the axial induction factor of wind turbines to influence their aerodynamic interactions through the wake. The performance index is defined here as the total power production of the wind farm over a finite prediction horizon. A medium-fidelity wind farm model is utilized to predict the inflow propagation in advance. The adjoint method is employed to solve the formulated optimization problem in a cost effective way and the first part of the optimal solution is implemented over the control horizon. This procedure is repeated at the next controller sample time providing the feedback into the optimization. The effectiveness and some key features of the proposed approach are studied for a two turbine test case through simulations.

  15. Optimizing gas extraction at landfills in Denmark; Optimering af gasindvinding pae deponeringsanlaeg i Danmark

    Energy Technology Data Exchange (ETDEWEB)

    Willumsen, H.C. [LFG Consult (Denmark)

    2005-07-01

    In landfills which contain organic material and anaerobic decomposition takes place, and landfill gas (LFG) is produced. The LFG contains approximately 50% methane, which contributes to the greenhouse effect when emitting from the landfill. Approximately 19% of the greenhouse gases in the atmosphere are methane, and around 8% of this is emission from landfills. This means that roughly 1.5% of the global warming is related to emission from landfills. Extraction of LFG for energy purposes was started 30 years ago in USA. In Denmark 26 LFG plants have been established since 1985. The gas is utilized for CHP or pure power production in gas engine/generator units. In some cases the LFG is used in gas burners in connection with boilers for district heating systems. 24 million m{sup 3} was recovered in 2004, which is equivalent to a reduction of CO{sub 2} of 160.000 tons CO{sub 2}/year. (BA)

  16. Footprints of Optimal Protein Assembly Strategies in the Operonic Structure of Prokaryotes

    Directory of Open Access Journals (Sweden)

    Jan Ewald

    2015-04-01

    Full Text Available In this work, we investigate optimality principles behind synthesis strategies for protein complexes using a dynamic optimization approach. We show that the cellular capacity of protein synthesis has a strong influence on optimal synthesis strategies reaching from a simultaneous to a sequential synthesis of the subunits of a protein complex. Sequential synthesis is preferred if protein synthesis is strongly limited, whereas a simultaneous synthesis is optimal in situations with a high protein synthesis capacity. We confirm the predictions of our optimization approach through the analysis of the operonic organization of protein complexes in several hundred prokaryotes. Thereby, we are able to show that cellular protein synthesis capacity is a driving force in the dissolution of operons comprising the subunits of a protein complex. Thus, we also provide a tested hypothesis explaining why the subunits of many prokaryotic protein complexes are distributed across several operons despite the presumably less precise co-regulation.

  17. An Effective Vacuum Assisted Extraction Method for the Optimization of Labdane Diterpenoids from Andrographis paniculata by Response Surface Methodology

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    Ya-Qi Wang

    2014-12-01

    Full Text Available An effective vacuum assisted extraction (VAE technique was proposed for the first time and applied to extract bioactive components from Andrographis paniculata. The process was carefully optimized by response surface methodology (RSM. Under the optimized experimental conditions, the best results were obtained using a boiling temperature of 65 °C, 50% ethanol concentration, 16 min of extraction time, one extraction cycles and a 12:1 liquid-solid ratio. Compared with conventional ultrasonic assisted extraction and heat reflux extraction, the VAE technique gave shorter extraction times and remarkable higher extraction efficiency, which indicated that a certain degree of vacuum gave the solvent a better penetration of the solvent into the pores and between the matrix particles, and enhanced the process of mass transfer. The present results demonstrated that VAE is an efficient, simple and fast method for extracting bioactive components from A. paniculata, which shows great potential for becoming an alternative technique for industrial scale-up applications.

  18. Protein purification using chromatography: selection of type, modelling and optimization of operating conditions.

    Science.gov (United States)

    Asenjo, J A; Andrews, B A

    2009-01-01

    To achieve a high level of purity in the purification of recombinant proteins for therapeutic or analytical application, it is necessary to use several chromatographic steps. There is a range of techniques available including anion and cation exchange, which can be carried out at different pHs, hydrophobic interaction chromatography, gel filtration and affinity chromatography. In the case of a complex mixture of partially unknown proteins or a clarified cell extract, there are many different routes one can take in order to choose the minimum and most efficient number of purification steps to achieve a desired level of purity (e.g. 98%, 99.5% or 99.9%). This review shows how an initial 'proteomic' characterization of the complex mixture of target protein and protein contaminants can be used to select the most efficient chromatographic separation steps in order to achieve a specific level of purity with a minimum number of steps. The chosen methodology was implemented in a computer- based Expert System. Two algorithms were developed, the first algorithm was used to select the most efficient purification method to separate a protein from its contaminants based on the physicochemical properties of the protein product and the protein contaminants and the second algorithm was used to predict the number and concentration of contaminants after each separation as well as protein product purity. The application of the Expert System approach was experimentally tested and validated with a mixture of four proteins and the experimental validation was also carried out with a supernatant of Bacillus subtilis producing a recombinant beta-1,3-glucanase. Once the type of chromatography is chosen, optimization of the operating conditions is essential. Chromatographic elution curves for a three-protein mixture (alpha-lactoalbumin, ovalbumin and beta-lactoglobulin), carried out under different flow rates and ionic strength conditions, were simulated using two different mathematical

  19. Process optimization and characterization of fragrant oil from red pepper (Capsicum annuum L.) seed extracted by subcritical butane extraction.

    Science.gov (United States)

    Gu, Ling-Biao; Pang, Hui-Li; Lu, Ke-Ke; Liu, Hua-Min; Wang, Xue-De; Qin, Guang-Yong

    2017-04-01

    Red pepper seeds account for 450-500 g kg(-1) of the total pepper weight and are often discarded as waste. In this study, process optimization and characterization of fragrant oil from roasted red pepper seed extracted by subcritical butane extraction were carried out. The optimal conditions of extraction were a temperature of 74.61 °C, a time of 68.65 min and a liquid/solid ratio of 30.24:1. The oil had a refractive index (25 °C) of 1.471, a relative density of 0.900, an acid value of 1.421 mg g(-1) oil, an iodine value of 127.035 g per 100 g, a saponification value of 184.060 mg KOH g(-1) , an unsaponifiable matter content of 12.400 g kg(-1) , a peroxide value of 2.465 meq. O2 kg(-1) and a viscosity of 52.094 cP. The main fatty acids in the oil were linoleic acid (72.95%) followed by palmitic acid (11.43%) and oleic acid (10.00%). The oil showed desirable thermal and oxidative stability. A total of 19 volatile compounds, mostly aldehydes and alkenes, were identified from the oil. The results indicated that the method is appropriate for the preparation of fragrant red pepper seed oil, and the oil is suitable for used as edible oil. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  20. Optimization of HPLC method for the isolation of Hypericum perforatum L. methanol extract

    Directory of Open Access Journals (Sweden)

    Stamenković, J.

    2013-12-01

    Full Text Available St. John's Wort (Hypericum perforatum L. is one of the most studied plant species in the family Hypericaceae. The aim of this study was the identification of the constituents of methanol extract of H. perforatum and optimization of conditions for their isolation. The main components of the methanol extract were isolated on preparative ZORBAX Eclipse XDB C18 column with solvent system consisting of methanol and 1x10-2 M ammonium acetate in water. Constituents of the extract were identified by comparing their retention times with the retention times of the standards, with the literature data and the UV spectra. By varying the conditions of chromatography, the optimal conditions for isolation of the methanol extract constituents were determined: mobile phase consisting of methanol and 1x10-2 M ammonium acetate in water in ratio 1 : 1, sample concentration 100 mg/mL, sample volume 30 µL, flow 2 mL/min. Under these conditions 7 components of the methanol extract were isolated.

  1. Extraction optimization of polysaccharide from Zanthoxylum bungeanum using RSM and its antioxidant activity.

    Science.gov (United States)

    Li, Peiqin; Zhou, Ligang; Mou, Yan; Mao, Ziling

    2015-01-01

    Polysaccharide prepared from pericarp of Zanthoxylum bungeanum was proved to possess excellent antioxidant activities in vitro by using reducing ferric iron power, DPPH radical scavenging activity, chelating ferrous iron capacity, and hydroxyl radical scavenging activity assays in the present study. In those four antioxidant assay models, Z. bungeanum polysaccharide (ZBP) displayed prominent antioxidant activities with low EC50 values of 0.011, 0.021, 0.056 and 0.008 mg/mL, respectively. Moreover, the extraction process of ZBP was further optimized by response surface methodology combined with Box-Behnken design. The highest polysaccharide yield 13.96%, which agreed closely with the predicted yield 13.20%, was obtained under the optimal extraction conditions as follows: extraction temperature 89 °C, extraction time 3h, ratio of water volume (mL) to raw material weight (g) 29 (v/w), and extraction number two times. The present research not only provide theoretical basis for exploitation of natural polysaccharide antioxidants, but also establish the foundation of large-scale production and further system utilization of ZBP.

  2. Optimization of extraction of high purity all-trans-lycopene from tomato pulp waste.

    Science.gov (United States)

    Poojary, Mahesha M; Passamonti, Paolo

    2015-12-01

    The aim of this work was to optimize the extraction of pure all-trans-lycopene from the pulp fractions of tomato processing waste. A full factorial design (FFD) consisting of four independent variables including extraction temperature (30-50 °C), time (1-60 min), percentage of acetone in n-hexane (25-75%, v/v) and solvent volume (10-30 ml) was used to investigate the effects of process variables on the extraction. The absolute amount of lycopene present in the pulp waste was found to be 0.038 mg/g. The optimal conditions for extraction were as follows: extraction temperature 20 °C, time 40 min, a solvent composition of 25% acetone in n-hexane (v/v) and solvent volume 40 ml. Under these conditions, the maximal recovery of lycopene was 94.7%. The HPLC-DAD analysis demonstrated that, lycopene was obtained in the all-trans-configuration at a very high purity grade of 98.3% while the amount of cis-isomers and other carotenoids were limited.

  3. Optimization of the extraction process of polyphenols from cashew apple agro-industrial residues

    Directory of Open Access Journals (Sweden)

    Renata Araújo Milanez de Sena Andrade

    2015-06-01

    Full Text Available The goal of this study was to determine the chemical composition of cashew apples agro-industrial residue and optimize the process of polyphenols extraction in this residue. The extraction process conditions were defined using a 24-1 fractional factorial experimental design using acetone and methanol as solvents. The independent variables were: time (30 to 90 min, temperature (30 to 50 °C, solvent concentrations (50% to 90%, agitation speed (100 to 300 rpm; the dependent variables were: total phenolic content and DPPH scavenging capacity. The optimized process was carried out by applying the Central Composite Rotational Design (CCRD considering the results obtained with the 24-1 fractional factorial experimental design. The residue presented bioactive compounds in its composition, with emphasis on the content of total phenolic compounds (1975.64 mg/ 100 g. The extraction process was not affected by methanol; however, acetone affected the amounts of extracted phytochemicals. Extracts with high levels of polyphenols and strong DPPH scavenging capacity (> 80% were obtained using 55% acetone, 30 minutes, 30 °C, and 150 rpm. The results showed that cashew apple residue is a potential natural source of bioactive compounds with strong antioxidant capacity. These compounds could be used partially or totally to replace synthetic antioxidants.

  4. Optimization of Vacuum-Microwave Radiation Pretreatment on Extraction of Ganoderma Polysaccharides

    Directory of Open Access Journals (Sweden)

    C. F. Song

    2015-01-01

    Full Text Available A new process of vacuum-microwave (VM radiation pretreatment for extracting polysaccharides from the Ganoderma lucidum was proposed, and the parameters were optimized by response surface methodology (RSM. The orthogonal-central composite design scheme was used and the responsive surfaces methodology of three factors and five levels was adopted, and the factors influencing the technological parameters and its interaction terms were analyzed and regressed. The optimal parameters were obtained as follows: the infiltration time of 70 min, microwave power density of 11.2 W/g, and VM irradiation time of 180 s. In consequence, the extraction yield was up to 1.775% when VM radiation was conducted in advance. Compared to the traditional hot-water extraction method, VM pretreatment can shorten the extraction time by more than a half, and the polysaccharide extraction yield was increased by 48.1%. It holds significant potential for further investigation, development, and application.

  5. Extraction Optimization, Purification and Physicochemical Properties of Polysaccharides from Gynura medica

    Directory of Open Access Journals (Sweden)

    Fengwei Li

    2016-03-01

    Full Text Available Extraction of polysaccharides from Gynura medica (GMPs was optimized by response surface methodology (RSM. A central composition design including three parameters, namely extraction temperature (X1, ratio of water to raw material (X2 and extraction time (X3, was used. The best conditions were extraction temperature of 91.7 °C, extraction time of 4.06 h and ratio of water to raw material of 29.1 mL/g. Under the optimized conditions, the yield of GMPs was 5.56%, which was similar to the predicted polysaccharides yield of 5.66%. A fraction named GMP-1 was obtained after isolation and purification by DEAE-52 and Sephadex G-100 gel chromatography, respectively. GMP-1, with a molecular weight of 401 kDa, mainly consisted of galacturonic acid (GalA, xylose (Xyl, glucose (Glu. Infrared spectroscopy was used to characterize the major functional groups of GMP-1 and the results indicated that it was an acidic polysaccharide. The antioxidant and α-glucosidase inhibitory activities of GMPs and GMP-1 were determined in vitro. The results indicated that GMPs and GMP-1 show potential for use in functional foods or medicines.

  6. Biochemical Characterization of Tunisian Cichorium Intybus L. Roots and Optimization of Ultrasonic Inulin Extraction

    Directory of Open Access Journals (Sweden)

    Youkabed Ouederni Zarroug

    2016-11-01

    Full Text Available In this study Cichorium intybus L. roots were tested for its chemical composition, antioxidant activity, and phenolic profile. Optimization of ultrasonic inulin extraction using response surface methodology (RSM was further investigated. Chicory roots were found to have high value of total carbohydrates (70.43%, soluble fiber (66.93, Neutral detergent fiber (NDF (33.07%, potassium (380 mg/100g, calcium (540 mg/100g and sodium (140 mg/100g. Chicory roots exhibit a high content of flavonoids, polyphenols, and tannins. Antioxidant activity measurement reveals the capacity of Chicory roots to scavenge diphenylpicrylhydrazyl (DPPH radicals. Phenolic acids profile shows the abundance of vanillic acid (19.64% followed by protocatechuic acid (15.67%. The effect of three independent variables namely extraction time, the ratio of water to raw material and temperature on inulin extraction was studied. Optimum deciding responses were Inulin content, Total Soluble Solids (TSS content and Water produced inulin yield. The optimal ultrasonic extraction conditions were: extraction time 87 min, liquid to solid ratio 38 (ml/g and ultrasonic temperature 61 ̊ C. Under these conditions, the inulin content, TSS content and produced inulin yield were 35.92%, 24.72%, and 32.53%, respectively. The produced inulin was characterized by the Fourier infrared transformation (FTIR and observed by means of scanning electron microscopy (SEM.  

  7. Extracting Tenebrio molitor protein while preventing browning: effect of pH and NaCl on protein yield

    NARCIS (Netherlands)

    Yi, L.; Boekel, van T.; Lakemond, C.M.M.

    2017-01-01

    The potential of insects as an alternative protein source for food applications was investigated by studying the effect of pH and NaCl on extraction yield of water-soluble proteins from Tenebrio molitor, while preventing browning due to polyphenol oxidation. Minimum protein solubility (29.6%) was at

  8. Extracting Tenebrio molitor protein while preventing browning: effect of pH and NaCl on protein yield

    NARCIS (Netherlands)

    Yi, L.; Boekel, van T.; Lakemond, C.M.M.

    2016-01-01

    The potential of insects as an alternative protein source for food applications was investigated by studying the effect of pH and NaCl on extraction yield of water-soluble proteins from Tenebrio molitor, while preventing browning due to polyphenol oxidation. Minimum protein solubility (29.6%) was at

  9. Characterization of rice starch and protein obtained by a fast alkaline extraction method.

    Science.gov (United States)

    Souza, Daiana de; Sbardelotto, Arthur Francisco; Ziegler, Denize Righetto; Marczak, Ligia Damasceno Ferreira; Tessaro, Isabel Cristina

    2016-01-15

    This study evaluated the characteristics of rice starch and protein obtained by a fast alkaline extraction method on rice flour (RF) derived from broken rice. The extraction was conducted using 0.18% NaOH at 30°C for 30min followed by centrifugation to separate the starch rich and the protein rich fractions. This fast extraction method allowed to obtain an isoelectric precipitation protein concentrate (IPPC) with 79% protein and a starchy product with low protein content. The amino acid content of IPPC was practically unchanged compared to the protein in RF. The proteins of the IPPC underwent denaturation during extraction and some of the starch suffered the cold gelatinization phenomenon, due to the alkaline treatment. With some modifications, the fast method can be interesting in a technological point of view as it enables process cost reduction and useful ingredients obtention to the food and chemical industries.

  10. Class Dependent LDA Optimization Using Genetic Algorithm for Robust MFCC Extraction

    Science.gov (United States)

    Abbasian, Houman; Nasersharif, Babak; Akbari, Ahmad

    Linear Discrimination analysis (LDA) finds transformations that maximizes the between-class scatter and minimizes within-class scatter. In this paper, we propose a method to use class-dependent LDA for speech recognition and MFCC extraction. To this end, we first use logarithm of clean speech Mel filter bank energies (LMFE) of each class then we obtain class-dependent LDA transformation matrix using multidimensional genetic algorithm (MGA) and use this matrix in place of DCT in MFCC feature extraction. The experimental results show that proposed speech recognition and optimization methods using class-dependent LDA, achieves to a significant isolated word recognition rate on Aurora2 database.

  11. Comparative evaluation of extraction methods for apoplastic proteins from maize leaves

    Directory of Open Access Journals (Sweden)

    Witzel Katja

    2011-12-01

    Full Text Available Abstract Proteins in the plant apoplast are essential for many physiological processes. We have analysed and compared six different infiltration solutions for proteins contained in the apoplast to recognize the most suitable method for leaves and to establish proteome maps for each extraction. The efficiency of protocols was evaluated by comparing the protein patterns resolved by 1-DE and 2-DE, and revealed distinct characteristics for each infiltration solution. Nano-LC-ESI-Q-TOF MS analysis of all fractions was applied to cover all proteins differentially extracted by infiltration solutions and led to the identification of 328 proteins in total in apoplast preparations. The predicted subcellular protein localisation distinguished the examined infiltration solutions in those with high or low amounts of intracellular protein contaminations, and with high or low quantities of secreted proteins. All tested infiltration solution extracted different subsets of proteins, and those implications on apoplast-specific studies are discussed.

  12. Potato tuber proteomics: Comparison of two complementary extraction methods designed for 2-DE of acidic proteins

    NARCIS (Netherlands)

    Delaplace, P.; Wal, van der F.; Dierick, J.F.; Cordewener, J.H.G.; Fauconnier, M.L.; Jardin, du P.; America, A.H.P.

    2006-01-01

    Two protein extraction procedures were tested in order to remove interfering compounds prior to 2-DE of potato tubers. These methods using SDS lysis buffer and phenol-phase extraction were compared regarding the quality of the resulting 2-D gel. While the resolution of SDS extracts on semipreparativ

  13. Application of optimized chemiluminescence assay for determination of the antioxidant capacity of herbal extracts.

    Science.gov (United States)

    Pogačnik, Lea; Ulrih, Nataša Poklar

    2012-01-01

    A chemiluminescence (CL) assay for the determination of antioxidant capacity (AOC) has been optimized and applied to analyses of herbal extracts in the present study. The optimal concentrations of reagents (luminol, H2O2, horseradish peroxidase) have been determined, as well as the optimal reaction conditions (wavelength, pH, temperature, sample volume). All of the measurements were performed at the emission maximum of the oxidized form of luminol (425 nm). The optimal concentrations of the reagents were determined as follows: 1.6 mmol/L luminol, 7.5 mmol/L H2O2 and 0.14 U/mL horseradish peroxidase activity in the reaction mixture. Analyses were carried out in phosphate buffer, pH 7.4, at room temperature. With the optimized CL assay, the AOCs of various water and methanol herbal extracts were determined (dog rose hips, plantain leaves and coltsfoot and thyme flowers) and the results were compared to those obtained by other classical methods for the evaluation of antioxidants. Strong correlations (r > 0.9) with the Folin-Ciocalteau assay and the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH)(●) assay are confirmed, although there is no correlation between AOC and the concentration of ascorbic acid in the samples analysed. This optimized CL assay is simple, rapid and reliable, and it represents a good alternative to classical methods (Folin-Ciocalteau, DPPH(●)) for the determination of AOC of herbal extracts and other food samples.

  14. Weak transient fault feature extraction based on an optimized Morlet wavelet and kurtosis

    Science.gov (United States)

    Qin, Yi; Xing, Jianfeng; Mao, Yongfang

    2016-08-01

    Aimed at solving the key problem in weak transient detection, the present study proposes a new transient feature extraction approach using the optimized Morlet wavelet transform, kurtosis index and soft-thresholding. Firstly, a fast optimization algorithm based on the Shannon entropy is developed to obtain the optimized Morlet wavelet parameter. Compared to the existing Morlet wavelet parameter optimization algorithm, this algorithm has lower computation complexity. After performing the optimized Morlet wavelet transform on the analyzed signal, the kurtosis index is used to select the characteristic scales and obtain the corresponding wavelet coefficients. From the time-frequency distribution of the periodic impulsive signal, it is found that the transient signal can be reconstructed by the wavelet coefficients at several characteristic scales, rather than the wavelet coefficients at just one characteristic scale, so as to improve the accuracy of transient detection. Due to the noise influence on the characteristic wavelet coefficients, the adaptive soft-thresholding method is applied to denoise these coefficients. With the denoised wavelet coefficients, the transient signal can be reconstructed. The proposed method was applied to the analysis of two simulated signals, and the diagnosis of a rolling bearing fault and a gearbox fault. The superiority of the method over the fast kurtogram method was verified by the results of simulation analysis and real experiments. It is concluded that the proposed method is extremely suitable for extracting the periodic impulsive feature from strong background noise.

  15. Sorghum Protein Extraction by Sonication and Its Relationship to Ethanol Fermentation

    Science.gov (United States)

    The objectives of this research were to develop a rapid method for extracting proteins from mashed and non-mashed sorghum meal using sonication (ultrasound), and to determine the relationships between the levels of extractable proteins and ethanol fermentation. Nine grain sorghum samples with a bro...

  16. Chaperone potential of Pulicaria undulata extract in preventing aggregation of stressed proteins.

    Science.gov (United States)

    Ghahghaei, Arezou; Valizadeh, Jafar; Nazari, Shahrzad; Ravandeh, Mehdi

    2014-06-01

    This study examined the effect of an aqueous extract of Pulicaria undulata on the 1,4-dithiothreitol (DTT)-induced aggregation of proteins. The effects of the chaperone properties of P. undulata extract on protein aggregation were determined by measuring light scattering absorption, fluorescence, and circular dichroism (CD) spectroscopy. The aqueous extract of P. undulata possesses good chaperone properties but the protection effect was varied in different protein. The extract showed a higher level of protection in high molecular weight proteins than in those of low molecular weight. Using a fluorescence study, the present study provides information on the hydrophobic area of proteins interacting with the P. undulata extract. In fact, by increasing the concentration of the P. undulata extract, the hydrophic area of the protein decreased. CD spectroscopy also revealed that DTT caused changes in both the tertiary and the secondary structure of the proteins, while in the presence of P. undulata extract, there was little change. Our finding suggests the possibility of using P. undulata extract for the inhibition of aggregation and the deposition of protein in disease.

  17. Critical parameters in cost-effective alkaline extraction for high protein yield from leaves

    NARCIS (Netherlands)

    Zhang, C.; Sanders, J.P.M.; Bruins, M.E.

    2014-01-01

    Leaves are potential resources for feed or food, but their applications are limited due to a high proportion of insoluble protein and inefficient processing. To overcome these problems, parameters of alkaline extraction were evaluated using green tea residue (GTR). Protein extraction could be maximi

  18. Advance chromatin extraction improves capture performance of protein A affinity chromatography.

    Science.gov (United States)

    Nian, Rui; Zhang, Wei; Tan, Lihan; Lee, Jeremy; Bi, Xeuzhi; Yang, Yuansheng; Gan, Hui Theng; Gagnon, Pete

    2016-01-29

    Practical effects of advance chromatin removal on performance of protein A affinity chromatography were evaluated using a caprylic acid-allantoin-based extraction method. Lacking this treatment, the practice of increasing loading residence time to increase capacity was shown to increase host protein contamination of the eluted IgG. Advance chromatin extraction suspended that compromise. Protein A ligand leakage from columns loaded with chromatin-extracted harvest was half the level observed on protein A columns loaded with non-extracted harvest. Columns loaded with chromatin-extracted harvest were cleaned more effectively by 50-100mM NaOH than columns loaded with non-extracted harvest that were cleaned with 250-500mM NaOH. Two protein A media with IgG capacities in excess of 50g/L were loaded with chromatin-extracted harvest, washed with 2.0M NaCl before elution, and the eluted IgG fraction titrated to pH 5.5 before microfiltration. Host protein contamination in the filtrate was reduced to protein A leakage to 0.5ppm, and aggregates to 1.0%. Caprylic acid and allantoin were both reduced below 5ppm. Step recovery of IgG was 99.4%. Addition of a single polishing step reduced residual protein A beneath the level of detection and aggregates to <0.1%. Overall process recovery including chromatin extraction was 90%.

  19. Adhesive performance of sorghum protein extracted from sorghum DDGS and flour

    Science.gov (United States)

    Distillers dried grains with solubles (DDGS) is the main co-product from grain-based ethanol production. The objective of this research was to compare the adhesive performance of three types of sorghum proteins: acetic acid-extracted sorghum protein from DDGS (PI), aqueous ethanol-extracted sorghum ...

  20. Characterisation of kiwifruit and asparagus enzyme extracts, and their activities toward meat proteins.

    Science.gov (United States)

    Ha, Minh; Bekhit, Alaa El-Din; Carne, Alan; Hopkins, David L

    2013-01-15

    Two plant enzyme extracts from kiwifruit and asparagus were evaluated for their ability to hydrolyse commercially available substrates and proteins present in both beef connective tissue and topside myofibrillar extracts. The results show significant differences in protease activity depending on the assay used. Protease assays with connective tissue and meat myofibrillar extracts provide a more realistic evaluation of the potential of the enzymes for application in meat tenderization. Overall, the kiwifruit protease extract was found to be more effective at hydrolysing myofibrillar and collagen proteins than the asparagus protease extract. The two protease extracts appeared to target meat myofibrillar and collagen proteins differently, suggesting the potential of a synergistic effect of these proteases in improving the tenderness of specific cuts of meat, based on their intrinsic protein composition.

  1. Comparison of Three Methods of Protein Extraction from Dermatophagoides Pteronyssinus for Two-dimensional Electrophoresis

    Institute of Scientific and Technical Information of China (English)

    Jin-lu Sun; Hong-yu Zhang; Zhi-yi Guo; Wan-tao Ying; Xiao-hong Qian; Jing-lan Wang

    2009-01-01

    Objective To explore an effective method of Dermatophagoides pteronyssinus protein extraction suitable for two-dimensional electrophoresis (2-DE) analysis. Methods The extracts of Dermatophagoides pteronyssinus were prepared with Coca's solution, lysis buffer of 2-DE, and Trizol reagent, respectively. Bicinchoninic acid (BCA) assay was used to determine the total protein concentration of the samples. The efficiency of different protein extraction methods were evaluated with 2-DE analysis. Results The concentrations of extracted protein by methods of Coca's solution, lysis buffer, and Trizol reagent were 0.63 g/L, 0.90 g/L, and 0.80 g/L, respectively. The 2-DE analysis results showed that some protein spots in low molecular weight (LMW) range could be detected with the Coca's solution method. With the lysis buffer of 2-DE method, more protein spots in LMW range could be detected, while the medium molecular weight (MMW) protein spots were absent. Several MMW protein spots (174-178 kD and 133 kD) and more LMW protein spots were detected with Trizol reagent method. Conclusions Among Coca's solution, lysis buffer of 2-DE, and Trizol reagent, the concentration of extracted protein of Dermatophagoides pteronyssinus by lysis buffer of 2-DE is the highest. However, most protein components of Dermatophagoides pteronyssinus purified mite bodies can be extracted by Trizol reagent, which may generally reflect the whole profile of Dermatophagoides pteronyssinus allergens.

  2. Optimization of Parameters for Biosynthesis of Silver Nanoparticles Using Leaf Extract of Aegle marmelos

    Directory of Open Access Journals (Sweden)

    JohnSamuel Godwin Christopher

    2015-10-01

    Full Text Available ABSTRACTThe aim of this study was to optimize the biosynthesis of silver nanoparticles using leaves ofAegle marmelos as the primary source. The optimal reaction medium comprised 2:1 concentration of leaf extract and 6mM concentration of silver nitrate solution (pH 7. The biosynthesized silver nanoparticles were confirmed by UV-Vis spectroscopy at 420 nm, XRD and FTIR analysis. The antimicrobial properties of silver nanoparticles were confirmed withBacillus subtilis andPseudomonas aeruginosa.

  3. Starch extraction process coupled to protein recovery from leguminous tuberous roots (Pachyrhizus ahipa).

    Science.gov (United States)

    Díaz, Andrea; Dini, Cecilia; Viña, Sonia Z; García, María A

    2016-11-05

    The objective of this work was to fit together the starch extraction from Pachyrhizus ahipa roots and the recovery of the proteins present in these storage organs, making an improved use of this novel raw material. The replacement of water by buffer PO4(-3)/NaCl as solvent in the first extraction steps improved protein extraction without lowering the starch yield. The starches obtained from the traditional and the proposed methods exhibited some differences in appearance and technological and thermal properties, which were endorsed to the adjustment in the methodology of extraction rather than to the use of buffer as solvent. Thus, P. ahipa starch obtaining procedure could be coupled to protein extraction with a minimum change in the methodology. This innovation did not significantly shift the characteristics of the starch obtained and allowed to obtain a protein yield of 135.7mg BSA equivalent protein/100g of fresh roots.

  4. Current Physical and SDS Extraction Methods Do Not Efficiently Remove Exosporium Proteins from Bacillus anthracis spores

    OpenAIRE

    Thompson, Brian M.; Binkley, Jana M; Stewart, George C.

    2011-01-01

    Biochemical studies of the outermost spore layers of the Bacillus cereus family are hindered by difficulties in efficient dispersal of the external spore layers and difficulties in dissociating protein complexes that comprise the exosporium layer. Detergent and physical methods have been utilized to disrupt the exosporium layer. Herein we compare commonly used SDS extraction buffers used to extract spore proteins and demonstrate the incomplete extractability of the exosporium layer by these m...

  5. Optimization of Ultrasound-assisted Extraction Procedure to Determine Astaxanthin in Xanthophyllomyces dendrorhous by Box-Behnken Designn

    Directory of Open Access Journals (Sweden)

    Wei Wu

    2013-11-01

    Full Text Available An ultrasonic-assisted extraction method has been developed for the effective extraction of astaxanthin from Xanthophyllomyces dendrorhous. Single-factor experiment design was employed to optimize the disruption temperature, disruption time, ethanol concentration, extraction time. The ethanol concentration was 60% ethanol in ethyl lactate and temperature for disrupting (40-60°C, time for disrupting extraction time (15-35 min, extraction time (20-40 min were used for further optimization of extraction conditions. The optimal conditions for the ultrasound-assisted extraction of astaxanthin were determined using Response Surface Methodology (RSM by Box-Behnken design. The optimal extraction conditions were that the yeast was disrupted at 49.08°C for 26.09 min under ultrasound irradiation of 200 W and then extracted for 32.43 min. Under optimal conditions, the astaxanthin content was 1472.85±43.64 &mug/g DW. This study introduces a simple, green and highly efficient method for extraction of astaxanthin fromX. dendrorhous

  6. Back to Osborne. Sequential protein extraction and LC-MS analysis for the characterization of the Holm oak seed proteome.

    Science.gov (United States)

    Romero-Rodríguez, M Cristina; Maldonado-Alconada, Ana M; Valledor, Luis; Jorrin-Novo, Jesus V

    2014-01-01

    It is impossible to capture in just one experiment all or most of the total set of protein species that constitute the cell's proteome. Thus, according to our results, and even considering that they depend on the experimental system carried out (plant, yeast, fungi, or bacteria), the best protein extraction protocol yielded less than 20 % of the total amount of proteins, as determined by the Kjeldahl method. For this reason, protein cataloguing and the whole proteome characterization require the use of firstly, fractionation techniques at the cellular, subcellular, protein, or peptide level, and secondly, the use of complementary approaches.Within our current research on Holm oak (Quercus ilex subsp. ballota), we aim to characterize its seed proteome. For that we have optimized an experimental workflow in which the Osborne sequential protein extraction (Osborne, Science 28:417-427, 1908) is combined with downstream electrophoretic protein separation or shotgun MS analysis. In general, it can be used to study any plant seed, as well as to investigate on seed maturation and germination, genotype characterization, allergens identification, food traceability, and substantial equivalence, among others.

  7. Evaluation of extraction solutions for biochemical analyses of the proteins in rice grains.

    Science.gov (United States)

    Lang, Gang-hua; Kagiya, Yukari; Ohnishi-Kameyama, Mayumi; Kitta, Kazumi

    2013-01-01

    The influence of different extraction solutions on the proteins extracted from rice grains was investigated. The largest amounts of salt-soluble proteins were extracted with solutions supplemented with Tris-HCl at pH 8.0. Rice allergens were analyzed by multiplex immunodetection. Except for α-globulin extracted with the solutions at pH 8.0, which showed a low-molecular-weight band besides the main band, no significant solution-dependent difference among the allergens was found. Total proteins were extracted with four kinds of solution. The extraction of the basic subunit of glutelin was found to be SDS-dependent, and more protein was obtained with extraction solutions supplemented with SDS. The contents of α-globulin and α-amylase/trypsin inhibitors were higher in the extracts without SDS than with SDS. We conclude from the present data that, in order to obtain comparable data from rice grain salt-soluble and total protein analyses, differences in the protein extraction efficiency of solutions used should be taken into consideration.

  8. Modeling and optimization of ultrasound-assisted extraction of polysaccharide from Cucurbita moschata.

    Science.gov (United States)

    Prakash Maran, J; Mekala, V; Manikandan, S

    2013-02-15

    Polysaccharides from pumpkin were extracted by ultrasound-assisted extraction technology using four factors at five levels central composite rotatable response surface design (CCRD). On using single factor analysis, process variables such as extraction temperature (50-70 °C), power of ultrasound (50-70 W), time (15-25 min) and solid-liquid ratio (1:10-1:20 g/ml) were selected. Experiments were conducted to evaluate the effects of four independent variables on the maximum extraction yield of polysaccharides. From the experimental data, second order polynomial mathematical model were developed with high coefficient of determination values (R(2)>0.96). From response surface plots, temperature and ultrasound power exhibited independent and interactive effects on the extraction yields. Extraction temperature of 70 °C, ultrasound power of 70 W, time of 23 min and solid-liquid ratio of 1:10 g/ml were determined as optimal conditions with a maximum polysaccharides yield of 16.21%, which was confirmed through the validation of the experiments.

  9. Optimization of Extraction of Novel Pectinase Enzyme Discovered in Red Pitaya (Hylocereus polyrhizus Peel

    Directory of Open Access Journals (Sweden)

    Nor Khanani Zohdi

    2013-11-01

    Full Text Available Plant peels could be a potential source of novel pectinases for use in various industrial applications due to their broad substrate specificity with high stability under extreme conditions. Therefore, the extraction conditions of a novel pectinase enzyme from pitaya peel was optimized in this study. The effect of extraction variables, namely buffer to sample ratio (2:1 to 8:1, X1, extraction temperature (−15 to +25 °C, X2 and buffer pH (4.0 to 12.0, X3 on specific activity, temperature stability, storage stability and surfactant agent stability of pectinase from pitaya peel was investigated. The study demonstrated that the optimum conditions for the extraction of pectinase from pitaya sources could improve the enzymatic characteristics of the enzyme and protect its activity and stability during the extraction procedure. The optimum extraction conditions cause the pectinase to achieve high specific activity (15.31 U/mg, temperature stability (78%, storage stability (88% and surfactant agent stability (83%. The most desirable conditions to achieve the highest activity and stability of pectinase enzyme from pitaya peel were the use of 5:1 buffer to sample ratio at 5 °C and pH 8.0.

  10. Optimization of polyphenols extraction from dried chokeberry using maceration as traditional technique.

    Science.gov (United States)

    Ćujić, Nada; Šavikin, Katarina; Janković, Teodora; Pljevljakušić, Dejan; Zdunić, Gordana; Ibrić, Svetlana

    2016-03-01

    Traditional maceration method was used for the extraction of polyphenols from chokeberry (Aronia melanocarpa) dried fruit, and the effects of several extraction parameters on the total phenolics and anthocyanins contents were studied. Various solvents, particle size, solid-solvent ratio and extraction time have been investigated as independent variables in two level factorial design. Among examined variables, time was not statistically important factor for the extraction of polyphenols. The optimal extraction conditions were maceration of 0.75mm size berries by 50% ethanol, with solid-solvent ratio of 1:20, and predicted values were 27.7mgGAE/g for total phenolics and 0.27% for total anthocyanins. Under selected conditions, the experimental total phenolics were 27.8mgGAE/g, and total anthocyanins were 0.27%, which is in agreement with the predicted values. In addition, a complementary quantitative analysis of individual phenolic compounds was performed using HPLC method. The study indicated that maceration was effective and simple technique for the extraction of bioactive compounds from chokeberry fruit.

  11. Optimizing conditions for the extraction of catechins from green tea using hot water.

    Science.gov (United States)

    Vuong, Quan V; Golding, John B; Stathopoulos, Costas E; Nguyen, Minh H; Roach, Paul D

    2011-11-01

    Six different factors involved in the extraction of catechins from green tea using water were examined for their impact on the yield of catechins and on the efficiency of water use. The best temperature and time combination for catechin extraction was at 80°C for 30 min. The yield of catechins was also optimal with a tea particle size of 1 mm, a brewing solution pH tea-to-water ratio at 50:1 (mL/g). In terms of efficient use of water in a single extraction, a water-to-tea ratio of 20:1 (mL/g) gave the best results; 2.5 times less water was used per gram of green tea. At the water-to-tea ratio of 20:1 mL/g, the highest yield of catechins per gram of green tea was achieved by extracting the same sample of green tea twice. However, for the most efficient use of water, the best extraction was found to be once at a water-to-tea ratio of 12:1 (mL/g) and once at a water-to-tea ratio of 8:1 (mL/g). Therefore, all six of the factors investigated had an impact on the yield of catechins extracted from green tea using water and two had an impact on the efficiency of water use.

  12. Optimization of extraction of novel pectinase enzyme discovered in red pitaya (Hylocereus polyrhizus) peel.

    Science.gov (United States)

    Zohdi, Nor Khanani; Amid, Mehrnoush

    2013-11-20

    Plant peels could be a potential source of novel pectinases for use in various industrial applications due to their broad substrate specificity with high stability under extreme conditions. Therefore, the extraction conditions of a novel pectinase enzyme from pitaya peel was optimized in this study. The effect of extraction variables, namely buffer to sample ratio (2:1 to 8:1, X₁), extraction temperature (-15 to +25 °C, X₂) and buffer pH (4.0 to 12.0, X₃) on specific activity, temperature stability, storage stability and surfactant agent stability of pectinase from pitaya peel was investigated. The study demonstrated that the optimum conditions for the extraction of pectinase from pitaya sources could improve the enzymatic characteristics of the enzyme and protect its activity and stability during the extraction procedure. The optimum extraction conditions cause the pectinase to achieve high specific activity (15.31 U/mg), temperature stability (78%), storage stability (88%) and surfactant agent stability (83%). The most desirable conditions to achieve the highest activity and stability of pectinase enzyme from pitaya peel were the use of 5:1 buffer to sample ratio at 5 °C and pH 8.0.

  13. Optimization and stabilization of gold nanoparticles by using herbal plant extract with microwave heating

    OpenAIRE

    Yasmin, Akbar; Ramesh, Kumaraswamy; Rajeshkumar, Shanmugam

    2014-01-01

    In this study, we have synthesized the gold nanoparticles by using Hibiscus rosa-sinensis, a medicinal plant. The gold nanoparticles were synthesized rapidly by the involvement of microwave heating. By changing of plant extract concentration, gold solution concentration, microwave heating time and power of microwave heating the optimized condition was identified. The surface Plasmon resonance found at 520 nm confirmed the gold nanoparticles synthesis. The spherical sized nanoparticles in the ...

  14. Optimization of Lycopene Extraction from Tomato Cell Suspension Culture by Response Surface Methodology

    OpenAIRE

    Lu, Chi-Hua; Engelmann, Nancy J.; Lila, Mary Ann; Erdman, John W

    2008-01-01

    Radioisotope-labeled lycopene is an important tool for biomedical research but currently is not commercially available. A tomato cell suspension culture system for the production of radioisotope-labeled lycopene was previously developed in our laboratory. In the current study, the goal was to optimize the lycopene extraction efficiency from tomato cell cultures for preparatory high-performance liquid chromatography (HPLC) separation. We employed response surface methodology (RSM), which combi...

  15. Improved proteomic analysis following trichloroacetic acid extraction of Bacillus anthracis spore proteins.

    Science.gov (United States)

    Deatherage Kaiser, Brooke L; Wunschel, David S; Sydor, Michael A; Warner, Marvin G; Wahl, Karen L; Hutchison, Janine R

    2015-11-01

    Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Analysis of cellular proteins is dependent upon efficient extraction from bacterial samples, which can be challenging with increasing complexity and refractory characteristics. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrichment for certain classes of proteins. The method presented here is technically simple, does not require specialized equipment such as a mechanical disrupter, and is effective for protein extraction of the particularly challenging sample type of Bacillus anthracis Sterne spores. The ability of Trichloroacetic acid (TCA) extraction to isolate proteins from spores and enrich for spore-specific proteins was compared to the traditional mechanical disruption method of bead beating. TCA extraction improved the total average number of proteins identified within a sample as compared to bead beating (547 vs 495, respectively). Further, TCA extraction enriched for 270 spore proteins, including those typically identified by first isolating the spore coat and exosporium layers. Bead beating enriched for 156 spore proteins more typically identified from whole spore proteome analyses. The total average number of proteins identified was equal using TCA or bead beating for easily lysed samples, such as B. anthracis vegetative cells. As with all assays, supplemental methods such as implementation of an alternative preparation method may simplify sample preparation and provide additional insight to the protein biology of the organism being studied.

  16. Artificial neural network modeling and optimization of ultrahigh pressure extraction of green tea polyphenols.

    Science.gov (United States)

    Xi, Jun; Xue, Yujing; Xu, Yinxiang; Shen, Yuhong

    2013-11-01

    In this study, the ultrahigh pressure extraction of green tea polyphenols was modeled and optimized by a three-layer artificial neural network. A feed-forward neural network trained with an error back-propagation algorithm was used to evaluate the effects of pressure, liquid/solid ratio and ethanol concentration on the total phenolic content of green tea extracts. The neural network coupled with genetic algorithms was also used to optimize the conditions needed to obtain the highest yield of tea polyphenols. The obtained optimal architecture of artificial neural network model involved a feed-forward neural network with three input neurons, one hidden layer with eight neurons and one output layer including single neuron. The trained network gave the minimum value in the MSE of 0.03 and the maximum value in the R(2) of 0.9571, which implied a good agreement between the predicted value and the actual value, and confirmed a good generalization of the network. Based on the combination of neural network and genetic algorithms, the optimum extraction conditions for the highest yield of green tea polyphenols were determined as follows: 498.8 MPa for pressure, 20.8 mL/g for liquid/solid ratio and 53.6% for ethanol concentration. The total phenolic content of the actual measurement under the optimum predicated extraction conditions was 582.4 ± 0.63 mg/g DW, which was well matched with the predicted value (597.2mg/g DW). This suggests that the artificial neural network model described in this work is an efficient quantitative tool to predict the extraction efficiency of green tea polyphenols. Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.

  17. Optimization of continuous and intermittent microwave extraction of pectin from banana peels.

    Science.gov (United States)

    Swamy, Gabriela John; Muthukumarappan, Kasiviswanathan

    2017-04-01

    Continuous and intermittent microwave-assisted extractions were used to extract pectin from banana peels. Extraction parameters which were employed in the continuous process were microwave power (300-900W), time (100-300s), pH (1-3) and in the intermittent process were microwave power (300-900W), pulse ratio (0.5-1), pH (1-3). The independent factors were optimized with the Box-Behnken response surface design (BBD) (three factor three level) with the desirability function methodology. Results indicate that the independent factors have substantial effect on the pectin yield. Optimized solutions for highest pectin yield (2.18%) from banana peels were obtained with microwave power of 900W, time 100s and pH 3.00 in the continuous method while the intermittent process yielded the highest pectin content (2.58%) at microwave power of 900W, pulse ratio of 0.5 and pH of 3.00. The optimized conditions were validated and close agreement was observed with the validation experiment and predicted value.

  18. Integrating kinetics with thermodynamics to study the alkaline extraction of protein from Caragana korshinskii Kom.

    Science.gov (United States)

    Zhong, Cheng; Zhou, Zhao; Zhang, Yu-Ming; Jia, Shi-Ru; Sun, Zhuo; Dale, Bruce E

    2014-09-01

    Extraction and recovery of protein from abundant plant biomass is one potential way to improve the economic feasibility of biorefineries. However, valorization of the protein fraction is challenging due to its low yield (kg protein extraction/kg biomass). In order to reveal the limiting operation parameters, the alkaline extraction process of protein from Caragana korshinskii Kom. was investigated by an integrative analysis of kinetics and thermodynamics. Both a two-site kinetic extraction model and a second-order model indicated that particle size is the most pivotal factor affecting protein extraction yield. In a two-site model, most proteins are extracted quickly from broken cells, while protein removal from the intact cells takes much longer; these are the faster and slower processes, respectively. A decrease of particle size from 20-40 to 60-80 mesh resulted in a decrease of C2 (protein yield in the slower process) from 14.02 to 7.32 mg g(-1), but a great increase of C1 (protein yield in the faster process) from 20.61 to 59.07 mg g(-1) . However, the protein yield was dominated by the faster process when the average particle size is under 80 mesh. The maximum initial extraction rate was 72.20 mg g(-1) min(-1) with the particle size of 60-80 mesh, almost ninefold of that with 20-40 mesh. Thermodynamic analysis revealed that the enthalpy change (ΔH) and entropy change (ΔS) in the protein extraction process were calculated as 21.08 kJ mol(-1) and 84.76 J K(-1), respectively. The standard free energy (ΔG) had a magnitude from -3.77 to -5.46, suggesting that the extraction process was spontaneous and physically feasible.

  19. Optimization of a greener method for removal phenol species by cloud point extraction and spectrophotometry.

    Science.gov (United States)

    Zain, N N M; Abu Bakar, N K; Mohamad, S; Saleh, N Md

    2014-01-24

    A greener method based on cloud point extraction was developed for removing phenol species including 2,4-dichlorophenol (2,4-DCP), 2,4,6-trichlorophenol (2,4,6-TCP) and 4-nitrophenol (4-NP) in water samples by using the UV-Vis spectrophotometric method. The non-ionic surfactant DC193C was chosen as an extraction solvent due to its low water content in a surfactant rich phase and it is well-known as an environmentally-friendly solvent. The parameters affecting the extraction efficiency such as pH, temperature and incubation time, concentration of surfactant and salt, amount of surfactant and water content were evaluated and optimized. The proposed method was successfully applied for removing phenol species in real water samples.

  20. Feature extraction of induction motor stator fault based on particle swarm optimization and wavelet packet

    Institute of Scientific and Technical Information of China (English)

    WANG Pan-pan; SHI Li-ping; HU Yong-jun; MIAO Chang-xin

    2012-01-01

    To effectively extract the interturn short circuit fault features of induction motor from stator current signal,a novel feature extraction method based on the bare-bones particle swarm optimization (BBPSO) algorithm and wavelet packet was proposed.First,according to the maximum inner product between the current signal and the cosine basis functions,this method could precisely estimate the waveform parameters of the fundamental component using the powerful global search capability of the BBPSO,which can eliminate the fundamental component and not affect other harmonic components.Then,the harmonic components of residual current signal were decomposed to a series of frequency bands by wavelet packet to extract the interturn circuit fault features of the induction motor.Finally,the results of simulation and laboratory tests demonstrated the effectiveness of the proposed method.

  1. Optimization of a greener method for removal phenol species by cloud point extraction and spectrophotometry

    Science.gov (United States)

    Zain, N. N. M.; Abu Bakar, N. K.; Mohamad, S.; Saleh, N. Md.

    2014-01-01

    A greener method based on cloud point extraction was developed for removing phenol species including 2,4-dichlorophenol (2,4-DCP), 2,4,6-trichlorophenol (2,4,6-TCP) and 4-nitrophenol (4-NP) in water samples by using the UV-Vis spectrophotometric method. The non-ionic surfactant DC193C was chosen as an extraction solvent due to its low water content in a surfactant rich phase and it is well-known as an environmentally-friendly solvent. The parameters affecting the extraction efficiency such as pH, temperature and incubation time, concentration of surfactant and salt, amount of surfactant and water content were evaluated and optimized. The proposed method was successfully applied for removing phenol species in real water samples.

  2. Optimization of protease extraction from horse mango (Mangifera foetida Lour) kernels by a response surface methodology.

    Science.gov (United States)

    Ahmad, Mohammad Norazmi; Liew, Siew Ling; Yarmo, Mohd Ambar; Said, Mamot

    2012-01-01

    Protease is one of the most important industrial enzymes with a multitude of applications in both food and non-food sectors. Although most commercial proteases are microbial proteases, the potential of non-conventional protease sources, especially plants, should not be overlooked. In this study, horse mango (Mangifera foetida Lour) fruit, known to produce latex with a blistering effect upon contact with human skin, was chosen as a source of protease, and the effect of the extraction process on its protease activity evaluated. The crude enzyme was extracted from the kernels and extraction was optimized by a response surface methodology (RSM) using a central composite rotatable design (CCRD). The variables studied were pH (x(1)), CaCl(2) (x(2)), Triton X-100 (x(3)), and 1,4-dithryeitol (x(4)). The results obtained indicate that the quadratic model is significant for all the variables tested. Based on the RSM model generated, optimal extraction conditions were obtained at pH 6.0, 8.16 mM CaCl(2), 5.0% Triton X-100, and 10.0 mM DTT, and the estimated response was 95.5% (w/w). Verification test results showed that the difference between the calculated and the experimental protease activity value was only 2%. Based on the t-value, the effects of the variables arranged in ascending order of strength were CaCl(2) < pH < DTT < Triton X-100.

  3. Less is More--Optimization of DNA Extraction from Canine Feces.

    Science.gov (United States)

    Lindquist, Christina D; Wictum, Elizabeth J

    2016-01-01

    Although most DNA crime laboratories may not encounter fecal samples often, they are a familiar sample type in non-human forensic laboratories due to their prevalence in the environment. Fecal matter can be challenging due to low numbers of nucleated cells and the presence of inhibitors that impede amplification success. Sampling location (internal vs. external), sampling quantity (10-200 mg), and various extraction protocols (silica matrix, bead beating, and clean-up column) were evaluated to maximize DNA yield. The greatest yield of intact DNA was obtained using a modified silica matrix extraction protocol (VGL-Fecal) on 30-50 mg of fecal matter collected from the external surface of a stool that had been dried for 24 h. This optimized sampling and extraction protocol was applied to a pilot study where DNA yield and genotyping success were evaluated. By optimizing our collection, sampling, and extraction procedures, a reliable method for maximizing the yield of canine fecal DNA was developed. © 2015 American Academy of Forensic Sciences.

  4. HIGH HYDROSTATIC PRESSURE EXTRACTION OF ANTIOXIDANTS FROM MORINDA CITRIFOLIA FRUIT – PROCESS PARAMETERS OPTIMIZATION

    Directory of Open Access Journals (Sweden)

    PRAVEEN KUMAR

    2006-06-01

    Full Text Available A modified version of high hydrostatic pressure extraction has been performed for extraction of antioxidants from M. citrifolia fruit at 5, 15, 25 bar and temperature 30° to 70°C for time duration 1, 2, 4 and 6 hours. The antioxidant activity of the extracts was determined by di-phenylpicrylhydrazyl radical scavenging method. The process parameters were optimized for antioxidant activity by central composite design method of response surface methodology using the statistical package, design expert. The results are expressed as 3D surface graphs. The optimum antioxidant activity was achieved at 58°C and 5 hours for 25bar. The optimal result achieved was within the region of response surface methodology. The statistical results were compared with the experimental result at 25bar, 2hour and 30° to 70°C and were found to be in proximate. The antioxidant activities of the extracts were found to increase with increase in pressure. It was also found that the response surface methodology works effectively for shorter range of parameters considered.

  5. Responsive Surface Methodology Optimizes Extraction Conditions of Industrial by-products, Camellia japonica Seed Cake

    Science.gov (United States)

    Kim, Jae Kyeom; Lim, Ho-Jeong; Kim, Mi-So; Choi, Soo Jung; Kim, Mi-Jeong; Kim, Cho Rong; Shin, Dong-Hoon; Shin, Eui-Cheol

    2016-01-01

    Background: The central nervous system is easily damaged by oxidative stress due to high oxygen consumption and poor defensive capacity. Hence, multiple studies have demonstrated that inhibiting oxidative stress-induced damage, through an antioxidant-rich diet, might be a reasonable approach to prevent neurodegenerative disease. Objective: In the present study, response surface methodology was utilized to optimize the extraction for neuro-protective constituents of Camellia japonica byproducts. Materials and Methods: Rat pheochromocytoma cells were used to evaluate protective potential of Camellia japonica byproducts. Results: Optimum conditions were 33.84 min, 75.24%, and 75.82°C for time, ethanol concentration and temperature. Further, we demonstrated that major organic acid contents were significantly impacted by the extraction conditions, which may explain varying magnitude of protective potential between fractions. Conclusions: Given the paucity of information in regards to defatted C. japonica seed cake and their health promoting potential, our results herein provide interesting preliminary data for utilization of this byproduct from oil processing in both academic and industrial applications. SUMMARY Neuro-protective potential of C. japonica seed cake on cell viability was affected by extraction conditionsExtraction conditions effectively influenced on active constituents of C. japonica seed cakeBiological activity of C. japonica seed cake was optimized by the responsive surface methodology. Abbreviations used: GC-MS: Gas chromatography-mass spectrometer, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, PC12 cells: Pheochromocytoma, RSM: Response surface methodology. PMID:27601847

  6. Application of 2k Full Factorial Design in Optimization of Solvent-Free Microwave Extraction of Ginger Essential Oil

    Directory of Open Access Journals (Sweden)

    Mumtaj Shah

    2014-01-01

    Full Text Available The solvent-free microwave extraction of essential oil from ginger was optimized using a 23 full factorial design in terms of oil yield to determine the optimum extraction conditions. Sixteen experiments were carried out with three varying parameters, extraction time, microwave power, and type of sample for two levels of each. A first order regression equation best fits the experimental data. The predicted values calculated by the regression model were in good agreement with the experimental values. The results showed that the extraction time is the most prominent factor followed by microwave power level and sample type for extraction process. An average of 0.25% of ginger oil can be extracted using current setup. The optimum conditions for the ginger oil extraction using SFME were the extraction time 30 minutes, microwave power level 640 watts, and sample type, crushed sample. Solvent-free microwave extraction proves a green and promising technique for essential oil extraction.

  7. [Optimization of extraction technology for salidroside, tyrosol, crenulatin and gallic acid in Rhodiolae Crenulatae Radix et Rhizoma with orthogonal test].

    Science.gov (United States)

    Luo, Xin; Wang, Xue-jing; Zhao, Yi-wu; Huang, Wen-zhe; Wang, Zhen-zhong; Xiao, Wei

    2015-09-01

    The extracting technology of salidroside, tyrosol, crenulatin and gallic acid from Rhodiolae Crenulatae Radix et Rhizoma was optimized. With extraction rate of salidroside, tyrosol, crenulatin and gallic acid as indexes, orthogonal test was used to evaluate effect of 4 factors on extracting technology, including concentration of solvent, the dosage of solvent, duration of extraction, and frequency of extraction. The results showed that, the best extracting technology was to extract in 70% alcohol with 8 times the weight of herbal medicine for 2 times, with 3 hours once. High extraction rate of salidroside, tyrosol, crenulatin and gallic acid were obtained with the present technology. The extracting technology was stable and feasible with high extraction rate of four compounds from Rhodiolae Crenulatae Radix et Rhizoma, it was suitable for industrial production.

  8. Optimization of the ethanol recycling reflux extraction process for saponins using a design space approach.

    Science.gov (United States)

    Gong, Xingchu; Zhang, Ying; Pan, Jianyang; Qu, Haibin

    2014-01-01

    A solvent recycling reflux extraction process for Panax notoginseng was optimized using a design space approach to improve the batch-to-batch consistency of the extract. Saponin yields, total saponin purity, and pigment yield were defined as the process critical quality attributes (CQAs). Ethanol content, extraction time, and the ratio of the recycling ethanol flow rate and initial solvent volume in the extraction tank (RES) were identified as the critical process parameters (CPPs) via quantitative risk assessment. Box-Behnken design experiments were performed. Quadratic models between CPPs and process CQAs were developed, with determination coefficients higher than 0.88. As the ethanol concentration decreases, saponin yields first increase and then decrease. A longer extraction time leads to higher yields of the ginsenosides Rb1 and Rd. The total saponin purity increases as the ethanol concentration increases. The pigment yield increases as the ethanol concentration decreases or extraction time increases. The design space was calculated using a Monte-Carlo simulation method with an acceptable probability of 0.90. Normal operation ranges to attain process CQA criteria with a probability of more than 0.914 are recommended as follows: ethanol content of 79-82%, extraction time of 6.1-7.1 h, and RES of 0.039-0.040 min-1. Most of the results of the verification experiments agreed well with the predictions. The verification experiment results showed that the selection of proper operating ethanol content, extraction time, and RES within the design space can ensure that the CQA criteria are met.

  9. Optimization of the ethanol recycling reflux extraction process for saponins using a design space approach.

    Directory of Open Access Journals (Sweden)

    Xingchu Gong

    Full Text Available A solvent recycling reflux extraction process for Panax notoginseng was optimized using a design space approach to improve the batch-to-batch consistency of the extract. Saponin yields, total saponin purity, and pigment yield were defined as the process critical quality attributes (CQAs. Ethanol content, extraction time, and the ratio of the recycling ethanol flow rate and initial solvent volume in the extraction tank (RES were identified as the critical process parameters (CPPs via quantitative risk assessment. Box-Behnken design experiments were performed. Quadratic models between CPPs and process CQAs were developed, with determination coefficients higher than 0.88. As the ethanol concentration decreases, saponin yields first increase and then decrease. A longer extraction time leads to higher yields of the ginsenosides Rb1 and Rd. The total saponin purity increases as the ethanol concentration increases. The pigment yield increases as the ethanol concentration decreases or extraction time increases. The design space was calculated using a Monte-Carlo simulation method with an acceptable probability of 0.90. Normal operation ranges to attain process CQA criteria with a probability of more than 0.914 are recommended as follows: ethanol content of 79-82%, extraction time of 6.1-7.1 h, and RES of 0.039-0.040 min-1. Most of the results of the verification experiments agreed well with the predictions. The verification experiment results showed that the selection of proper operating ethanol content, extraction time, and RES within the design space can ensure that the CQA criteria are met.

  10. Optimal extraction of small-scale surface water storage in Asia

    Science.gov (United States)

    Mahendrarajah, S.; Warr, P. G.; Jakeman, A. J.

    1992-05-01

    This paper analyzes the optimization of water storage of small-scale dams (tanks) found in the semiarid regions of Asia. The focus is on monsoonal water storage in small tank systems in Sri Lanka, and in particular on optimal extraction for supplementary irrigation under double cropping, also allowing for nonirrigation uses of storage. The problem of intraseasonal allocation of storage for irrigation is solved by deterministic dynamic programming (DP) using simulated crop response functions. An approach to solving the optimal interseasonal allocation problem is demonstrated by operating in sequence the DP models for the two seasons. An important feature is the generation and use of seasonal water response functions with respect to each cropping season ensuring optimality in both crop area and intraseasonal distribution of irrigation. The determination of the scarcity value of water in this framework is also illustrated. Our results show that in most years both the optimal conservation of storage from the wet season and the optimal area of irrigation in the dry season are much higher than the current practice.

  11. Optimal Elbow Angle for Extracting sEMG Signals During Fatiguing Dynamic Contraction

    Directory of Open Access Journals (Sweden)

    Mohamed R. Al-Mulla

    2015-09-01

    Full Text Available Surface electromyographic (sEMG activity of the biceps muscle was recorded from 13 subjects. Data was recorded while subjects performed dynamic contraction until fatigue and the signals were segmented into two parts (Non-Fatigue and Fatigue. An evolutionary algorithm was used to determine the elbow angles that best separate (using Davies-Bouldin Index, DBI both Non-Fatigue and Fatigue segments of the sEMG signal. Establishing the optimal elbow angle for feature extraction used in the evolutionary process was based on 70% of the conducted sEMG trials. After completing 26 independent evolution runs, the best run containing the optimal elbow angles for separation (Non-Fatigue and Fatigue was selected and then tested on the remaining 30% of the data to measure the classification performance. Testing the performance of the optimal angle was undertaken on nine features extracted from each of the two classes (Non-Fatigue and Fatigue to quantify the performance. Results showed that the optimal elbow angles can be used for fatigue classification, showing 87.90% highest correct classification for one of the features and on average of all eight features (including worst performing features giving 78.45%.

  12. Comparison of protein-extraction methods for gills of the shore crab, Carcinus maenas (L.), and application to 2DE.

    Science.gov (United States)

    Panchout, François; Letendre, Julie; Bultelle, Florence; Denier, Xavier; Rocher, Béatrice; Chan, Philippe; Vaudry, David; Durand, Fabrice

    2013-12-01

    As it is well-established that protein extraction constitutes a crucial step for two-dimensional electrophoresis (2DE), this work was done as a prerequisite to further the study of alterations in the proteome in gills of the shore crab Carcinus maenas under contrasted environmental conditions. Because of the presence of a chitin layer, shore crab gills have an unusual structure. Consequently, they are considered as a hard tissue and represent a challenge for optimal protein extraction. In this study, we compared three published extraction procedures for subsequent applications to 2DE: the first one uses homogenization process, the second one included an additional TCA-acetone precipitation step, and finally, the third one associated grinding in liquid nitrogen (N2) and TCA-acetone precipitation. Extracted proteins were then resolved using 1DE and 2DE. Although interesting patterns were obtained using 1DE with the three methods, only the one involving grinding in liquid N2 and TCA-acetone precipitation led to proper resolution after 2DE, showing a good level of reproducibility at technical (85%) and biological (84%) levels. This last method is therefore proposed for analysis of gill proteomes in the shore crab.

  13. Design and optimization of the lattice of the superconducting synchrotron SIS300 for slow extraction

    Energy Technology Data Exchange (ETDEWEB)

    Saa Hernandez, Angela

    2011-10-15

    The superconducting synchrotron SIS300 is planned to be built at the new Facility for Antiproton and Ion Research (FAIR), at GSI-Darmstadt. SIS300 will be a versatile machine, which by means of a low-energy stretcher-mode or a high-energy ramped-mode will provide slowly extracted heavy ion beams towards the experimental areas. To reach the required maximum field of 4.5 T, cos({theta}) magnets are necessary. Thus, SIS300 will become the first superconducting synchrotron worldwide with cos({theta}) magnets providing resonant slow extraction. Since SIS300 will be installed in the same tunnel as the SIS100 synchrotron, the dipole layout of SIS300 cannot be freely chosen. Thus, a standard lattice cannot be applied. A redesign of the SIS300 lattice accepting compromises concerning the positions and phase advances between the optical elements has been proposed. Using the analytical model of the slow extraction, firstly proposed by Kobayashi, and the analytical description of the resonance driving modes, a multiobjective optimization algorithm has been developed for the optimization of the lattice under the given boundary conditions. The final goal of the lattice optimization is a higher efficiency of the slow extraction. The results are evaluated by means of tracking simulations performed with the code Elegant. The field quality in superconducting cos({theta}) magnets is determined by the positions of the superconducting cable and the static and time-dependent effects of the current in the cable. Furthermore, the fast ramp rates of 1 T/s in the dipoles, which are fifty times faster than in any other superconducting cos({theta}) magnet, together with the fact that the aperture is smaller than in conventional accelerator magnets, makes it extremely difficult to obtain a high-quality magnetic field. The unavoidable field errors affect the beam dynamics and worsen the slow extraction efficiency. Therefore, the field errors in the SIS300 dipoles have been estimated, and their

  14. Optimized DNA extraction from neonatal dried blood spots: application in methylome profiling.

    Science.gov (United States)

    Ghantous, Akram; Saffery, Richard; Cros, Marie-Pierre; Ponsonby, Anne-Louise; Hirschfeld, Steven; Kasten, Carol; Dwyer, Terence; Herceg, Zdenko; Hernandez-Vargas, Hector

    2014-07-01

    Neonatal dried blood spots (DBS) represent an inexpensive method for long-term biobanking worldwide and are considered gold mines for research for several human diseases, including those of metabolic, infectious, genetic and epigenetic origin. However, the utility of DBS is restricted by the limited amount and quality of extractable biomolecules (including DNA), especially for genome wide profiling. Degradation of DNA in DBS often occurs during storage and extraction. Moreover, amplifying small quantities of DNA often leads to a bias in subsequent data, particularly in methylome profiles. Thus it is important to develop methodologies that maximize both the yield and quality of DNA from DBS for downstream analyses. Using combinations of in-house-derived and modified commercial extraction kits, we developed a robust and efficient protocol, compatible with methylome studies, many of which require stringent bisulfite conversion steps. Several parameters were tested in a step-wise manner, including blood extraction, cell lysis, protein digestion, and DNA precipitation, purification and elution. DNA quality was assessed based on spectrophotometric measurements, DNA detectability by PCR, and DNA integrity by gel electrophoresis and bioanalyzer analyses. Genome scale Infinium HumanMethylation450 and locus-specific pyrosequencing data generated using the refined DBS extraction protocol were of high quality, reproducible and consistent. This study may prove useful to meet the increased demand for research on prenatal, particularly epigenetic, origins of human diseases and for newborn screening programs, all of which are often based on DNA extracted from DBS.

  15. Extraction of Protein-Protein Interaction from Scientific Articles by Predicting Dominant Keywords

    Directory of Open Access Journals (Sweden)

    Shun Koyabu

    2015-01-01

    Full Text Available For the automatic extraction of protein-protein interaction information from scientific articles, a machine learning approach is useful. The classifier is generated from training data represented using several features to decide whether a protein pair in each sentence has an interaction. Such a specific keyword that is directly related to interaction as “bind” or “interact” plays an important role for training classifiers. We call it a dominant keyword that affects the capability of the classifier. Although it is important to identify the dominant keywords, whether a keyword is dominant depends on the context in which it occurs. Therefore, we propose a method for predicting whether a keyword is dominant for each instance. In this method, a keyword that derives imbalanced classification results is tentatively assumed to be a dominant keyword initially. Then the classifiers are separately trained from the instance with and without the assumed dominant keywords. The validity of the assumed dominant keyword is evaluated based on the classification results of the generated classifiers. The assumption is updated by the evaluation result. Repeating this process increases the prediction accuracy of the dominant keyword. Our experimental results using five corpora show the effectiveness of our proposed method with dominant keyword prediction.

  16. Extraction of Protein-Protein Interaction from Scientific Articles by Predicting Dominant Keywords.

    Science.gov (United States)

    Koyabu, Shun; Phan, Thi Thanh Thuy; Ohkawa, Takenao

    2015-01-01

    For the automatic extraction of protein-protein interaction information from scientific articles, a machine learning approach is useful. The classifier is generated from training data represented using several features to decide whether a protein pair in each sentence has an interaction. Such a specific keyword that is directly related to interaction as "bind" or "interact" plays an important role for training classifiers. We call it a dominant keyword that affects the capability of the classifier. Although it is important to identify the dominant keywords, whether a keyword is dominant depends on the context in which it occurs. Therefore, we propose a method for predicting whether a keyword is dominant for each instance. In this method, a keyword that derives imbalanced classification results is tentatively assumed to be a dominant keyword initially. Then the classifiers are separately trained from the instance with and without the assumed dominant keywords. The validity of the assumed dominant keyword is evaluated based on the classification results of the generated classifiers. The assumption is updated by the evaluation result. Repeating this process increases the prediction accuracy of the dominant keyword. Our experimental results using five corpora show the effectiveness of our proposed method with dominant keyword prediction.

  17. Which metaproteome? The impact of protein extraction bias on metaproteomic analyses.

    Science.gov (United States)

    Leary, Dagmar Hajkova; Hervey, W Judson; Deschamps, Jeffrey R; Kusterbeck, Anne W; Vora, Gary J

    2013-01-01

    Culture-independent techniques such as LC-MS/MS-based metaproteomic analyses are being increasingly utilized for the study of microbial composition and function in complex environmental samples. Although several studies have documented the many challenges and sources of bias that must be considered in these types of analyses, none have systematically characterized the effect of protein extraction bias on the biological interpretation of true environmental biofilm metaproteomes. In this study, we compared three protein extraction methods commonly used in the analyses of environmental samples [guanidine hydrochloride (GuHCl), B-PER, sequential citrate-phenol (SCP)] using nano-LC-MS/MS and an environmental marine biofilm to determine the unique biases introduced by each method and their effect on the interpretation of the derived metaproteomes. While the protein extraction efficiencies of the three methods ranged from 2.0 to 4.3%, there was little overlap in the sequence (1.9%), function (8.3% of total assigned protein families) and origin of the identified proteins from each extract. Each extraction method enriched for different protein families (GuHCl - photosynthesis, carbohydrate metabolism; B-PER - membrane transport, oxidative stress; SCP - calcium binding, structural) while 23.7-45.4% of the identified proteins lacked SwissProt annotations. Taken together, the results demonstrated that even the most basic interpretations of this complex microbial assemblage (species composition, ratio of prokaryotic to eukaryotic proteins, predominant functions) varied with little overlap based on the protein extraction method employed. These findings demonstrate the heavy influence of protein extraction on biofilm metaproteomics and provide caveats for the interpretation of such data sets when utilizing single protein extraction methods for the description of complex microbial assemblages.

  18. Reprint of "Which metaproteome? The impact of protein extraction bias on metaproteomic analyses".

    Science.gov (United States)

    Leary, Dagmar Hajkova; Hervey, W Judson; Deschamps, Jeffrey R; Kusterbeck, Anne W; Vora, Gary J

    2014-01-01

    Culture-independent techniques such as LC-MS/MS-based metaproteomic analyses are being increasingly utilized for the study of microbial composition and function in complex environmental samples. Although several studies have documented the many challenges and sources of bias that must be considered in these types of analyses, none have systematically characterized the effect of protein extraction bias on the biological interpretation of true environmental biofilm metaproteomes. In this study, we compared three protein extraction methods commonly used in the analyses of environmental samples [guanidine hydrochloride (GuHCl), B-PER, sequential citrate-phenol (SCP)] using nano-LC-MS/MS and an environmental marine biofilm to determine the unique biases introduced by each method and their effect on the interpretation of the derived metaproteomes. While the protein extraction efficiencies of the three methods ranged from 2.0 to 4.3%, there was little overlap in the sequence (1.9%), function (8.3% of total assigned protein families) and origin of the identified proteins from each extract. Each extraction method enriched for different protein families (GuHCl--photosynthesis, carbohydrate metabolism; B-PER--membrane transport, oxidative stress; SCP--calcium binding, structural) while 23.7-45.4% of the identified proteins lacked SwissProt annotations. Taken together, the results demonstrated that even the most basic interpretations of this complex microbial assemblage (species composition, ratio of prokaryotic to eukaryotic proteins, predominant functions) varied with little overlap based on the protein extraction method employed. These findings demonstrate the heavy influence of protein extraction on biofilm metaproteomics and provide caveats for the interpretation of such data sets when utilizing single protein extraction methods for the description of complex microbial assemblages.

  19. Anticariogenic and Hemolytic Activity of Selected Seed Protein Extracts In vitro conditions.

    Directory of Open Access Journals (Sweden)

    Kalpesh B Ishnava

    2014-10-01

    Full Text Available This study aimed to assess the anticariogenic and hemolytic activity of crude plant seed protein extracts against tooth decaying bacteria.The proteins from seeds of 12 different plants were extracted and used for antimicrobial assay against six different organisms. The extraction was carried out in 10mM of sodium phosphate buffer (pH 7.0. Protein concentrations were determined as described by Bradford method. Anticariogenic activity was studied by agar well diffusion method and Minimum Inhibitory Concentration (MIC was evaluated by the two-fold serial broth dilution method. Hemolytic activity, treatment of proteinase K and Kinetic study in Mimusops elengi crude seed protein extract.The anticariogenic assay demonstrated the activity of Mimusops elengi against Staphylococcus aureus and Streptococcus pyogenes. A minor activity of Glycine wightii against Streptococcus mutans was also found. The protein content of Mimusops elengi seed protein extract was 5.84mg/ml. The MIC values for Staphylococcus aureus and Streptococcus pyogenes against Mimusops elengi seed protein extract were 364.36μg/ml and 182.19μg/ml, respectively. Kinetic study further elucidated the mode of inhibition in the presence of the Mimusops elengi plant seed protein with respect to time. The concentration of crude extract which gave 50% hemolysis compared to Triton X-100 treatment (HC50 value was 1.58 mg/ml; which is more than five times larger than that of the MIC. Treatment with proteinase K of the Mimusops elengi seed protein resulted in absence of the inhibition zone; which clearly indicates that the activity was only due to protein.Our results showed the prominence of Mimusops elengi plant seed protein extract as an effective herbal medication against tooth decaying bacteria.

  20. GRIFFIN: A versatile methodology for optimization of protein-lipid interfaces for membrane protein simulations.

    Science.gov (United States)

    Staritzbichler, René; Anselmi, Claudio; Forrest, Lucy R; Faraldo-Gómez, José D

    2011-04-12

    As new atomic structures of membrane proteins are resolved, they reveal increasingly complex transmembrane topologies, and highly irregular surfaces with crevices and pores. In many cases, specific interactions formed with the lipid membrane are functionally crucial, as is the overall lipid composition. Compounded with increasing protein size, these characteristics pose a challenge for the construction of simulation models of membrane proteins in lipid environments; clearly, that these models are sufficiently realistic bears upon the reliability of simulation-based studies of these systems. Here, we introduce GRIFFIN, which uses a versatile framework to automate and improve a widely-used membrane-embedding protocol. Initially, GRIFFIN carves out lipid and water molecules from a volume equivalent to that of the protein, so as to conserve the system density. In the subsequent optimization phase GRIFFIN adds an implicit grid-based protein force-field to a molecular dynamics simulation of the pre-carved membrane. In this force-field, atoms inside the implicit protein volume experience an outward force that will expel them from that volume, whereas those outside are subject to electrostatic and van-der-Waals interactions with the implicit protein. At each step of the simulation, these forces are updated by GRIFFIN and combined with the intermolecular forces of the explicit lipid-water system. This procedure enables the construction of realistic and reproducible starting configurations of the protein-membrane interface within a reasonable timeframe and with minimal intervention. GRIFFIN is a standalone tool designed to work alongside any existing molecular dynamics package, such as NAMD or GROMACS.

  1. Investigation of antibacterial mechanism and identification of bacterial protein targets mediated by antibacterial medicinal plant extracts.

    Science.gov (United States)

    Yong, Ann-Li; Ooh, Keng-Fei; Ong, Hean-Chooi; Chai, Tsun-Thai; Wong, Fai-Chu

    2015-11-01

    In this paper, we investigated the antibacterial mechanism and potential therapeutic targets of three antibacterial medicinal plants. Upon treatment with the plant extracts, bacterial proteins were extracted and resolved using denaturing gel electrophoresis. Differentially-expressed bacterial proteins were excised from the gels and subjected to sequence analysis by MALDI TOF-TOF mass spectrometry. From our study, seven differentially expressed bacterial proteins (triacylglycerol lipase, N-acetylmuramoyl-L-alanine amidase, flagellin, outer membrane protein A, stringent starvation protein A, 30S ribosomal protein s1 and 60 kDa chaperonin) were identified. Additionally, scanning electron microscope study indicated morphological damages induced on bacterial cell surfaces. To the best of our knowledge, this represents the first time these bacterial proteins are being reported, following treatments with the antibacterial plant extracts. Further studies in this direction could lead to the detailed understanding of their inhibition mechanism and discovery of target-specific antibacterial agents. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. Optimization and numerical simulation for the extraction system of the H- multicusp ion source

    Science.gov (United States)

    Hosseinzadeh, M.; Afarideh, H.

    2014-05-01

    A new ion source has been designed and manufactured for the CYCLONE30 accelerator, which has a much advanced performance compared with the original. It is expected that the newly designed ion source extraction system will transport a very large percentage of the beam without deteriorating the beam optics, which is designed to deliver an H- beam at 30 keV. The accelerator assembly consists of three circular aperture electrodes made of copper. The simulation study was focused on finding parameter sets that raise the beam perveance as large as possible and which reduce the beam divergence as low as possible. Ion beams of the highest quality are extracted whenever the half-angular divergence is minimum, for which the perveance current intensity and the extraction gap have optimum values. The triode extraction system is designed and optimized by using CST software (for Particle Beam Simulations). The physical design of the extraction system is given in this paper. From the simulation results, it is concluded that it is possible to achieve this goal by decreasing the thickness of the plasma electrode, shortening the first gap, and adjusting the acceleration electrode voltage.

  3. Optimization of ultrasonic extraction of total flavonoids from Tussilago farfara L. using response surface methodology.

    Science.gov (United States)

    Liu, Caihong; Qin, Kun; Qi, Yongxiu; Li, Ke; Li, Yuqin; Jia, Baoxiu

    2014-04-01

    In this work, ultrasound technology was used for the extraction of total flavonoids from Tussilago farfara L. Response surface methodology (RSM), based on a Box-Behnken design (BBD), was used to optimize the effects of processing parameters on total flavonoids yields. The parameters were ultrasonic frequency (X1), extraction time (X2) and ratio of liquid to solid (X3). The statistical analysis indicated that the parameters of ultrasonic power and ratio of liquid to solid, and the square effects among them had significant effects on the yield of flavonoids from Tussilago farfara L. The interaction between ultrasonic power and extraction time, and ultrasonic power and ratio of liquid to solid also caused significant effects on the yields. The optimum extraction conditions were determined as follows: ultrasonic power 420 W, extraction time 30 min, ratio of liquid to solid 25 mL/g. Under these conditions, the experimental yield 6.59 +/- 0.061% agreed closely with the predicted yield (6.64%).

  4. Optimization of Insoluble and Soluble Fibres Extraction from Agave americana L. Using Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Amine Bouaziz

    2014-01-01

    Full Text Available Experimental design methodology was used to determine significant factors affecting the extraction yield of soluble and insoluble fibres from Agave americana L. and in second time to find optimum conditions leading to the highest yield. Results clearly indicated that the temperature, the powder to water (P/W ratio, and the agitation speed were the most important factors influencing fibres extraction yield which increased with temperature, P/W ratio, and agitation speed. Ionic strength affected significantly soluble fibre extraction yield and was the most important factor among nonsignificant ones influencing insoluble fibres extraction yield. Then, a Box-Behnken design was carried out to maximise fibres extraction. Selected optimal conditions were temperature: 90°C; P/W ratio: 0.1625; agitation speed: 400 rpm; and ionic strength: 1.5 g/L. These conditions yielded 93.02% and 80.46% of insoluble and soluble fibres, respectively. Concentrates showed high fibres purity and good functional properties.

  5. Improved Proteomic Analysis Following Trichloroacetic Acid Extraction of Bacillus anthracis Spore Proteins

    Energy Technology Data Exchange (ETDEWEB)

    Kaiser, Brooke LD; Wunschel, David S.; Sydor, Michael A.; Warner, Marvin G.; Wahl, Karen L.; Hutchison, Janine R.

    2015-08-07

    Proteomic analysis of bacterial samples provides valuable information about cellular responses and functions under different environmental pressures. Proteomic analysis is dependent upon efficient extraction of proteins from bacterial samples without introducing bias toward extraction of particular protein classes. While no single method can recover 100% of the bacterial proteins, selected protocols can improve overall protein isolation, peptide recovery, or enrich for certain classes of proteins. The method presented here is technically simple and does not require specialized equipment such as a mechanical disrupter. Our data reveal that for particularly challenging samples, such as B. anthracis Sterne spores, trichloroacetic acid extraction improved the number of proteins identified within a sample compared to bead beating (714 vs 660, respectively). Further, TCA extraction enriched for 103 known spore specific proteins whereas bead beating resulted in 49 unique proteins. Analysis of C. botulinum samples grown to 5 days, composed of vegetative biomass and spores, showed a similar trend with improved protein yields and identification using our method compared to bead beating. Interestingly, easily lysed samples, such as B. anthracis vegetative cells, were equally as effectively processed via TCA and bead beating, but TCA extraction remains the easiest and most cost effective option. As with all assays, supplemental methods such as implementation of an alternative preparation method may provide additional insight to the protein biology of the bacteria being studied.

  6. Independent Research Projects Using Protein Extraction: Affordable Ways to Inquire, Discover & Publish for Undergraduate Students

    Science.gov (United States)

    Pu, Rongsun

    2010-01-01

    This article describes how to use protein extraction, quantification, and analysis in the undergraduate teaching laboratory to engage students in inquiry-based, discovery-driven learning. Detailed instructions for obtaining proteins from animal tissues, using BCA assay to quantify the proteins, and data analysis are provided. The experimental…

  7. How Does Alkali Aid Protein Extraction in Green Tea Leaf Residue: A Basis for Integrated Biorefinery of Leaves

    OpenAIRE

    Chen Zhang; Sanders, Johan P. M.; Xiao, Ting T.; Bruins, Marieke E.

    2015-01-01

    Leaf protein can be obtained cost-efficiently by alkaline extraction, but overuse of chemicals and low quality of (denatured) protein limits its application. The research objective was to investigate how alkali aids protein extraction of green tea leaf residue, and use these results for further improvements in alkaline protein biorefinery. Protein extraction yield was studied for correlation to morphology of leaf tissue structure, protein solubility and hydrolysis degree, and yields of non-pr...

  8. Optimization of the Extraction of Antioxidants and Caffeine from Maté (Ilex paraguariensis Leaves by Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Vanessa Graciela Hartwig

    2013-04-01

    Full Text Available Optimal conditions for the industrial extraction of total polyphenols from maté (Ilex paraguariensis were determined using response surface methodology, with two independent variables: ethanol percentage of the extraction solution and liquid to solid ratio. Response variables were total polyphenol content, antioxidant capacity, concentration of total polyphenols and caffeine content. The optimal conditions found were a liquid to solid ratio from 8 - 9 w w-1 and ethanol percentage of the extraction solution from 30 -50 % w w-1. Under these conditions the main predicted values corresponding to leaf extracts were 40 μg chlorogenic acid equivalents mL-1 of original extract, 13 g chlorogenic acid equivalents per 100 g dry matter for total polyphenol content, 22 g Trolox equivalents and 15.5 g ascorbic acid equivalents per 100 g dry matter for antioxidant capacity. The total polyphenol content of twig extracts was 36% lower than that in the leaf extracts.

  9. Optimization of ultrasonic assisted extraction of antioxidants from black soybean (Glycine max var) sprouts using response surface methodology.

    Science.gov (United States)

    Lai, Jixiang; Xin, Can; Zhao, Ya; Feng, Bing; He, Congfen; Dong, Yinmao; Fang, Yun; Wei, Shaomin

    2013-01-16

    Response surface methodology (RSM) using a central composite