Three-phase partitioning for the separation of proteins, enzymes, biopolymers, oils and pigments: a review

Rajendran, Devi Sri; Chidambaram, Ashok; Kumar, Ponnusamy Senthil; Venkataraman, Swethaa; Muthusamy, Shanmugaprakash; Vo, Dai‐Viet N.; Rangasamy, Gayathri; Vaithyanathan, Vasanth Kumar; Vaidyanathan, Vinoth Kumar

doi:10.1007/s10311-022-01540-8

Cited by 7 publications

(7 citation statements)

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“…34 Much less investigated for this application, TPP systems can also be used for fractionation of biopolymer mixtures. 36,37 Some promising results for fractionation of biopolymers have been reported by Wang et al, 38 who demonstrated that separating microbial polysaccharide and protein can be achieved using (NH 4 ) 2 SO 4 / butanol TPP system. In this case, polysaccharide mostly migrated to the salt phase, with an extraction yield of 52% and less than 5% protein content.…”

Section: Introductionmentioning

confidence: 99%

Polymer and alcohol‐based three‐phase partitioning systems for separation of polysaccharide and protein

Antunes,

Temmink,

Schuur

2023

J of Chemical Tech & Biotech

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BackgroundNatural polymers are macromolecules produced by living organisms, with a wide range of applications and relevance for the development of a circular economy. However, large‐scale production is still hindered by several factors, such as downstream processing. In this work, three‐phase partitioning (TPP) systems were investigated for separation of model polysaccharide (dextran, alginate and gum arabic) from protein (Bovine serum albumin (BSA) and lysozyme). The recyclability of phase‐forming compounds used to form the extractive platform was assessed by ultrafiltration (UF). This study contributes to development of production processes for biopolymers from fermented waste by proposing an effective separation technique for fractionation of biopolymers. Previously such biopolymers were only collected as mixtures, and with the studied approaches, also fractionation may be applied of polysaccharides from proteins. With the chosen systems, the scope of TPP systems is expanded by using another class of phase‐forming compound (polymers), and in addition, UF was studied as a versatile regeneration approach.ResultsWithin the TPP approach, the best separation of dextran from BSA was achieved using TPP systems composed of 25 wt% PEG + 25 wt% K3C6H5O7 and 36 wt% EtOH + 10 wt% K3PO4, in which more than 95% of dextran and BSA were found as precipitate and partitioned to top phase (PEG or EtOH‐rich), respectively. By using other model compounds, it was found that the molecular weight and charge of biopolymer play a key role in the yield and selectivity of TPP systems. Finally, by using ultrafiltration/diafiltration, about 99% of ethanol and phosphate salt used to form the extractive platform could be retrieved in the permeate stream.ConclusionThe high extraction yields, selectivity and recyclability of phase‐forming compounds confirm the potential of polymer‐based and alcohol‐based TPP systems to fractionate biopolymer mixtures.This article is protected by copyright. All rights reserved.

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Section: Introductionmentioning

confidence: 99%

Polymer and alcohol‐based three‐phase partitioning systems for separation of polysaccharide and protein

Antunes,

Temmink,

Schuur

2023

J of Chemical Tech & Biotech

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“…In TPP, the partition of proteins is driven by multiple effects such as salting out, and kosmotropic, isoionic, osmolytic and co‐solvent precipitation 22 . Currently, there is a gap in the mechanisms of protein separation and the driving forces in TPP 21 . Besides, given the complexity of venoms, it would be hard to fully explain the partition of proteins and peptides.…”

Section: Resultsmentioning

confidence: 99%

“…22 Currently, there is a gap in the mechanisms of protein separation and the driving forces in TPP. 21 Besides, given the complexity of venoms, it would be hard to fully explain the partition of proteins and peptides. The top phase (tbutanol phase) favors the partition of nonpolar molecules; therefore, it was expected that most of the proteins were partitioned to the opposite phase.…”

Section: Partition Of Clarified Venomsmentioning

confidence: 99%

“…[17][18][19][20] On the other hand, TPP is a fast and easy separation method that allows concentration and partial purification of different molecules. 21 Three-phase separation occurs when the biological sample is mixed with ammonium sulfate, at a given saturation percentage, and t-butanol. 22 During TPP formation, a combination effect between salting out, kosmotropic precipitation, isoionic precipitation, protein hydration shifts, osmolytic, and cosolvent precipitation favors proteins and other molecules to precipitate at the interphase.…”

Section: Introductionmentioning

confidence: 99%

“…On the other hand, TPP is a fast and easy separation method that allows concentration and partial purification of different molecules 21 . Three‐phase separation occurs when the biological sample is mixed with ammonium sulfate, at a given saturation percentage, and t ‐butanol 22 .…”

Section: Introductionmentioning

confidence: 99%

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Primary recovery strategies of low‐molecular‐weight toxins from Crotalus molossus nigrescens and Crotalus atrox using aqueous two‐phase and three‐phase partition systems

Sánchez‐Trasviña,

Melendez‐Martínez,

González‐Castañón

et al. 2024

J of Chemical Tech & Biotech

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BACKGROUNDRattlesnake venoms are a complex source of different toxin families with biotechnological and biomedical applications. However, the complexity of the sample and the low abundance of some toxins can restrict their study and applications. Toxin isolation usually requires multiple chromatographic steps, implying elevated operative costs and processing times. This raises the need for recovery and enrichment of those proteins and peptides. For that reason, we tested two liquid–liquid separation methods – aqueous two‐phase systems (ATPS) and three‐phase partition (TPP) – since these two alternatives have shown great potential for the primary recovery of different toxin families from rattlesnake venoms.RESULTSPolyethylene glycol (PEG)–citrate and ionic liquid (IL)–potassium phosphate ATPS and TPP based on t‐butanol and ammonium sulfate were used to partition different toxin families from Crotalus molossus nigrescens and C. atrox venoms. PEG–citrate allowed a selective partition of proteins between both phases. The best enrichment case was observed with a myotoxin band (6.6 kDa) in the PEG‐rich phase from 2% to 41.3% of abundance. In IL–potassium phosphate systems, proteins showed a preference towards the IL‐rich phase with no selective partition. In TPP, proteins were partitioned mainly to the bottom phase and interphase, but serine proteases (29.1 kDa) were enriched from 3.7% to 56.6% in the top phase.CONCLUSIONThe present study demonstrated that PEG–citrate ATPS and TPP separation methods can generate a selective partition of some toxin families from rattlesnake venoms. These enrichment methods will facilitate the isolation of toxins, from animal venoms, that are relevant for biotechnological and biomedical applications. © 2024 Society of Chemical Industry (SCI).

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Extraction and optimization of biomolecules recovery from Chlorella sp. NITT 02: Unleashing potential through ultrasound and triphasic system of extraction

Anto,

Premalatha,

Mathimani

2024

J Appl Phycol

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Three-phase partitioning for the separation of proteins, enzymes, biopolymers, oils and pigments: a review

Cited by 7 publications

References 145 publications

Polymer and alcohol‐based three‐phase partitioning systems for separation of polysaccharide and protein

Polymer and alcohol‐based three‐phase partitioning systems for separation of polysaccharide and protein

Primary recovery strategies of low‐molecular‐weight toxins from Crotalus molossus nigrescens and Crotalus atrox using aqueous two‐phase and three‐phase partition systems

Extraction and optimization of biomolecules recovery from Chlorella sp. NITT 02: Unleashing potential through ultrasound and triphasic system of extraction

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