Application of Response Surface Methodology to Optimize Three Phase Partitioning for Purification of Laccase from<i>Pleurotus ostreatus</i>

Kumar, Vikas; Sathyaselvabala, Vasanthakumar; Kirupha, Selvaraj Dinesh; Murugesan, A.; Vidhyadevi, T.; Sivanesan, S.

doi:10.1080/01496395.2011.583306

Cited by 30 publications

(9 citation statements)

References 29 publications

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“…On the other hand, TPP makes it possible to isolate and concentrate proteins by means of a separate layer formed between aqueous and organic phases with the aid of a dissolved inorganic salt (generally ammonium sulfate) and tert‐butanol, so a back‐extraction step is not required. Therefore, TPP is a quick and easy way for enrichment of proteins that is easily scalable and that can be used directly with crude suspensions .…”

Section: Introductionmentioning

confidence: 99%

“…11 -14 Nevertheless, most of the studies regarding ILATPS are focused on the influence of different salts and ionic liquids on the phase formation and on the distribution coefficient of the solutes, 15 but very few references include the back-extraction step, which may be one of the main challenges that must be overcome. 5 On the other hand, TPP makes it possible to isolate and concentrate proteins by means of a separate layer formed between aqueous and organic phases with the aid of a dissolved inorganic salt (generally ammonium sulfate) and tert-butanol, 16,17 so www.soci.org E Alvarez-Guerra, A Irabien a back-extraction step is not required. Therefore, TPP is a quick and easy way for enrichment of proteins that is easily scalable and that can be used directly with crude suspensions.…”

Section: Introductionmentioning

confidence: 99%

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Ionic liquid‐based three phase partitioning (ILTPP) systems for whey protein recovery: ionic liquid selection

Alvarez‐Guerra

Irabien

2014

J of Chemical Tech & Biotech

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BACKGROUND The ionic liquid three phase partitioning (ILTPP) technique is a promising alternative to conventional processing technologies for protein separation and purification since it allows protein recovery at the liquid–liquid interface. In previous work, ILTPP has been developed with the BmimBF4/NaH2PO4 system for lactoferrin recovery, a bovine whey protein with important nutraceutical properties. However, BmimBF4 may suffer from hydrolysis when it is in contact with water, so the selection of an alternative ionic liquid with similar results regarding lactoferrin recovery should be sought. RESULTS The system based on BmimTfO and phosphate is selected as the most suitable since it combines relatively high LF recoveries at the liquid–liquid interface (83–99%) and chemical stability. The highest LF recoveries occur at moderate acidic pH (3.1–4.0), which may also be improved at relatively low protein concentrations. The process selectivity with respect to bovine serum albumin (another whey protein) varies from 1.1 to 3.0, depending on experimental conditions. CONCLUSION ILTPP is a promising technique to recover the protein fraction from bovine whey or to enrich specific proteins such as lactoferrin, so ILTPP could replace alternative techniques such as membranes. The risk of hydrolysis associated with previous ionic liquid ILTPP systems has been successfully overcome. © 2014 Society of Chemical Industry

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Ionic liquid‐based three phase partitioning (ILTPP) systems for whey protein recovery: ionic liquid selection

Alvarez‐Guerra

Irabien

2014

J of Chemical Tech & Biotech

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“…Therefore, in the light of the above basic demands, three-phase partitioning (TPP) seems to be a good and economical alternative where clarification, concentration and partial purification can be integrated in one step [18][19][20]. A protein-rich middle layer is formed in addition to the two liquid phases when such a system is applied to crude protein extracts [18][19][20][21][22][23][24][25][26][27][28][29]. A protein-rich middle layer is formed in addition to the two liquid phases when such a system is applied to crude protein extracts [18][19][20][21][22][23][24][25][26][27][28][29].…”

Section: Introductionmentioning

confidence: 99%

“…It involves the addition of a salt (generally ammonium sulfate) to the crude extract followed by the addition of an organic solvent (generally t-butanol). A protein-rich middle layer is formed in addition to the two liquid phases when such a system is applied to crude protein extracts [18][19][20][21][22][23][24][25][26][27][28][29].…”

Section: Introductionmentioning

confidence: 99%

“…TPP has been successfully used in the purification of other enzymes such as protease [19], cellulase [18,20], amylase [18], lipase [18], a-galactosidase [21], xylanase [20], laccase [22], peroxidase [25], alkaline phosphatase [23], phospholipase [24], polygalacturonase [26], aryl alcohol oxidase [27] and invertase [18,29]. The simplicity of the process and low cost of the phase forming materials make it feasible for large-scale protein purification using appropriate scale-up techniques [21].…”

Section: Introductionmentioning

confidence: 99%

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Aspergillus niger exo‐inulinase purification by three phase partitioning

Vaidyanathan

Periyaraman

Sathyaselvabala

et al. 2011

Engineering in Life Sciences

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Inulinase (2, 1‐β‐D‐fructan fructanohydrolase, EC 3.2.1.7) hydrolyses inulin into nearly pure fructose, which is an excellent alternative for the production of fructose syrup. Growing inulinase utilization in different industries encourages the search for high benefit/cost ratio purification techniques for such enzymes. Here, we adapted the three‐phase partitioning (TPP) technique for the downstream process of inulinase obtained from Aspergillus niger. TPP is a simple non‐chromatographic process used for purification and concentration of protein. The various conditions required for attaining efficient purification of inulinase were optimized. The optimum conditions for TPP were found to be 30% w/v ammonium sulfate saturation with 1.0 : 0.5 v/v ratio of t‐butanol to crude extract at pH 4.0 and temperature 25°C. The enzyme was purified by 10.2‐fold using two‐step TPP with an overall recovery of 88%. The enzyme's molecular mass was found to be 63.8 kDa by SDS‐PAGE analysis. Terminal hydrolysis fructose units from the inulin show that enzymes are exo‐inulinase. The recovery of purified exo‐inulinase achieved in this work shows the technical viability of enzyme purification by TPP.

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Ionic liquid recovery alternatives in ionic liquid‐based three‐phase partitioning (ILTPP)

et al. 2014

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in Wiley Online Library (wileyonlinelibrary.com) Ionic liquid-based three-phase partitioning (ILTPP) is a promising technique to recover high-added value proteins at the liquid-liquid interface. Its economic and environmental performance highly depends on the net ionic liquid consumption. Alternatives to maximize the fraction of ionic liquid that can be recycled are studied. It is demonstrated that the addition of extra salt, previously proposed in literature, has a very limited effect on ionic liquid recovery for relatively high protein concentrations in the feed stream, and that it may even lead to an increase of the ionic liquid losses under certain conditions. However, small additions of salt are shown to be effective and profitable from an economic point of view. Vacuum evaporation is shown to allow for the complete ionic liquid and salt recovery, reinforcing the sustainability and viability of ILTPP processes.

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Application of Response Surface Methodology to Optimize Three Phase Partitioning for Purification of Laccase fromPleurotus ostreatus

Cited by 30 publications

References 29 publications

Ionic liquid‐based three phase partitioning (ILTPP) systems for whey protein recovery: ionic liquid selection

Ionic liquid‐based three phase partitioning (ILTPP) systems for whey protein recovery: ionic liquid selection

Aspergillus niger exo‐inulinase purification by three phase partitioning

Ionic liquid recovery alternatives in ionic liquid‐based three‐phase partitioning (ILTPP)

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