Studying the Influence of Salts on Partitioning of Proteins: Isoelectric Point Determination

Forciniti, Daniel

doi:10.1385/1-59259-028-4:201

Cited by 6 publications

(10 citation statements)

References 12 publications

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“…The highest selectivity (8.12) and purification fold of lipase (12.42) were achieved at the addition of 1% (w/w) NaCl. The addition of NaCl above 2% (w/w) will decrease the partition efficiency, as the chemical potential of the solute is affected due to the unequal partitioning of neutral salts between the two phases [39].…”

Section: Effect Of Nacl On Lipase Partitioning=mentioning

confidence: 99%

Direct purification of Burkholderia Pseudomallei lipase from fermentation broth using aqueous two-phase systems

Ooi

Tey

Hii

et al. 2009

Biotechnol Bioproc E

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An aqueous two-phase purification process was employed for the recovery of _ìêâÜçäÇÉêá~=éëÉìÇçã~ääÉá=lipase from fermentation broth. The partition behavior of _K=éëÉìÇçã~ääÉá=lipase was investigated with various parameters such as phase composition, tie-line length (TLL), volume ratio (V R ), sample loading, system pH, and addition of neutral salts. Optimum conditions for the purification of lipase were obtained in polyethylene glycol (PEG) 6000-potassium phosphate system using TLL of 42.2% (w/w), with V R of 2.70, and 1% (w/w) NaCl addition at pH 7 for 20% (w/w) crude load. Based on this system, the purification factor of lipase was enhanced to 12.42 fold, with a high yield of 93%. Hence, the simplicity and effectiveness of aqueous two-phase systems (ATPS) in the purification of lipase were proven in this study. © KSBB hÉóïçêÇëW=~èìÉçìë=íïçJéÜ~ëÉ=ëóëíÉãëI=äáé~ëÉI=éìêáÑáÅ~íáçåI=éçäóÉíÜóäÉåÉ=ÖäóÅçäI=êÉÅçîÉêó= = = = =

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Section: Effect Of Nacl On Lipase Partitioning=mentioning

confidence: 99%

Direct purification of Burkholderia Pseudomallei lipase from fermentation broth using aqueous two-phase systems

Ooi

Tey

Hii

et al. 2009

Biotechnol Bioproc E

View full text Add to dashboard Cite

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“…4b). This is due to the unequal partitioning of neutral salts between the two phases, which affects the chemical potential of the solute in each phase and thus its partition coefficient (Forciniti, 2000). With an increase in NaCl concentration the difference in free volume (V f ) observed was less (Fig.…”

Section: Effect Of Neutral Salt Concentrationmentioning

confidence: 99%

Aqueous two phase extraction for purification and concentration of betalains

Chethana¹,

Nayak²,

Raghavarao³

2007

Journal of Food Engineering

131

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“…The luminescence can be measured with standard plate readers widely available in biochemical, biological, and biotechnology laboratories, requiring no special expertise and instrumentation. The experimental procedures of the conventional pI determination methods are time-consuming and may contain several steps and laborious preparation of several solutions. ,,, The developed method is more sensitive compared to existing methods for the estimation of the pI as the required concentration of 210 μg/L is at least 100 times lower compared to those of all existing literature methods and more than 1000 times lower than those for the most commonly used methods such as IEF. ,,,, The low concentration can potentially allow the estimation of the pI for hydrophobic and weakly soluble proteins, which is impossible with IEF due to the required high concentration and the precipitation on a gel …”

Section: Discussionmentioning

confidence: 99%

“…A cross-partitioning − method utilizes the partition coefficient of proteins in the presence of salts. The partition coefficient versus pH is measured for alkali chloride and alkali sulfate systems.…”

mentioning

confidence: 99%

Method for Estimation of Protein Isoelectric Point

et al. 2012

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Adsorption of sample protein to Eu(3+) chelate-labeled nanoparticles is the basis of the developed noncompetitive and homogeneous method for the estimation of the protein isoelectric point (pI). The lanthanide ion of the nanoparticle surface-conjugated Eu(3+) chelate is dissociated at a low pH, therefore decreasing the luminescence signal. A nanoparticle-adsorbed sample protein prevents the dissociation of the chelate, leading to a high luminescence signal. The adsorption efficiency of the sample protein is reduced above the isoelectric point due to the decreased electrostatic attraction between the negatively charged protein and the negatively charged particle. Four proteins with isoelectric points ranging from ~5 to 9 were tested to show the performance of the method. These pI values measured with the developed method were close to the theoretical and experimental literature values. The method is sensitive and requires a low analyte concentration of submilligrams per liter, which is nearly 10000 times lower than the concentration required for the traditional isoelectric focusing. Moreover, the method is significantly faster and simpler than the existing methods, as a ready-to-go assay was prepared for the microtiter plate format. This mix-and-measure concept is a highly attractive alternative for routine laboratory work.

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Studying the Influence of Salts on Partitioning of Proteins: Isoelectric Point Determination

Cited by 6 publications

References 12 publications

Direct purification of Burkholderia Pseudomallei lipase from fermentation broth using aqueous two-phase systems

Direct purification of Burkholderia Pseudomallei lipase from fermentation broth using aqueous two-phase systems

Aqueous two phase extraction for purification and concentration of betalains

Method for Estimation of Protein Isoelectric Point

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