Partitioning and purification of thaumatin in aqueous two-phase systems

Cascone, Osvaldo; Andrews, Bárbara A.; Asenjo, Juan A.

doi:10.1016/0141-0229(91)90076-m

Cited by 89 publications

(37 citation statements)

References 11 publications

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“…The elastase distribution pattern observed in these systems showed that increasing NaCl concentration beyond 2% could dramatically decrease the recovery. Although adding NaCl has been reported to be beneficial to targeted products (Silgia et al, 2000;Cascone et al, 1991;Marcos et al, 2002), in the current study addition of NaCl negatively influenced partitioning of elastase into the top phase. Others described similar results in other systems (Li and He, 1992;Isabel del-Val and Cristina, 2003).…”

Section: Effect Of Nacl Concentration On Elastase Partitioningmentioning

confidence: 66%

Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system

Li³

2005

J Zheijang Univ Sci B

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This paper presents the evaluation of an aqueous two-phase system (ATPS) for extracting elastase produced by Bacillus sp. EL31410. The elastase and cell partition behavior in polyethylene glycol (PEG)/salt systems was investigated. The suitable system for elastase extraction was PEG/KH 2 PO 4 -K 2 HPO 4 , in which elastase is mainly partitioned into the PEG-rich phase, while the cells remained in the other phase. The influence of defined system parameters (e.g. PEG molecular mass, pH, NaCl addition) on the partitioning behavior of elastase is described. The concentration of phase forming components, PEG and KH 2 PO 4 -K 2 HPO 4 , was optimized for elastase recovery by means of response surface methodology, and it was found that they greatly influenced extraction recovery. The optimal ATPS was 23.1% (w/w) PEG 2 000 and 11.7% (w/w) KH 2 PO 4 -K 2 HPO 4 . The predicted recovery was about 89.5%, so this process is suggested to be a rapid and convenient method for elastase extraction.

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Section: Effect Of Nacl Concentration On Elastase Partitioningmentioning

confidence: 66%

Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system

Li³

2005

J Zheijang Univ Sci B

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“…Given that the isoelectric point of ovalbumin is 4.5 [5], in all systems evaluated the protein is negatively charged. In PEG-salt ABS it has been reported that proteins prefer the top or the bottom phase depending on the nature of the phase formingcomponents and their concentration [41][42]. At pH 7.0, ovalbumin is negatively charged (isoelectric point = 4.5 [5] and still prefers the phase of lower ionic strength (polymer-rich) in all systems evaluated.…”

Section: Phase Diagrams and Tie-linesmentioning

confidence: 99%

Single-step purification of ovalbumin from egg white using aqueous biphasic systems

Pereira

Cruz

Almeida

et al. 2016

Process Biochemistry

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The ability of aqueous biphasic systems (ABS) composed of polyethylene glycols of different molecular weights (PEG 400, 600 and 1000) and buffered aqueous solutions of potassium citrate/ citric acid (pH = 5.0 -8.0) to selectively extract ovalbumin from egg white was here investigated. Phase diagrams, tie-lines and tie-line lengths were determined at 25ºC and the partitioning of ovalbumin in these systems was then evaluated. Aiming at optimizing the selective extraction of ovalbumin in the studied ABS, factors such as pH, PEG molecular weight and amount of the phase-forming components were initially investigated with pure commercial ovalbumin. In almost all ABS, it was observed a preferential partitioning of ovalbumin to the polymer-rich phase, with extraction efficiencies higher than 90%. The best ABS were then applied in the purification of ovalbumin from the real egg white matrix. In order to ascertain on the ovalbumin purity and yield, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion high performance liquid chromatography (SE-HPLC) analyses were conducted, confirming that the isolation/purification of ovalbumin from egg white was completely achieved in a single-step with a recovery yield of 65%. The results obtained show that polymer-salt-based ABS allow the selective extraction of ovalbumin from egg white with a simpler approach and better performance than previously reported. Finally, it is shown that ovalbumin can be completely recovered from the PEG-rich phase by an induced precipitation using an inexpensive and sustainable separation platform which can be easily applied on an industrial scale.

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“…Therefore, considerable reduction in PEG size would be unacceptable. [30,31] As seen from Table 2, the increase in the MW of PEG from 600 to 2000 Da for the same polymer composition led to about 24.9-fold decrease in K p , while K e increased slightly. This result showed that the selectiveness of the system increases with the increase in PEG MW.…”

Section: Selection Of Polymer Molecular Weight and System Compositionmentioning

confidence: 82%

Extraction of lipase fromRhizopus microsporusfermentation culture by aqueous two-phase partitioning

Anvari

2015

Biotechnology & Biotechnological Equipment

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The purification of a desired protein from the complex mixture of biomolecules in a fermentation broth is one of the main challenges in industrial-scale biotechnological processes. For partial purification of industrial enzymes, one possible approach is to use aqueous two-phase extraction (APTE) as a primary downstream processing step. In the present study, the feasibility of utilizing ATPE for the purification of lipase from Rhizopus microsporus fermentation culture was investigated. The effects of the phase composition and salt type, molecular weight of the polyethylene glycol (PEG), pH of the system, sample loading and addition of neutral salts on lipase partitioning were investigated at 24 C. In most of the examined aqueous two-phase systems (ATPSs), lipase showed affinity to the top phase. Optimum conditions for lipase purification were obtained in 20% PEG 2000/12% (NH 4 ) 2 SO 4 , with 5% Na 2 CO 3 addition at pH D 8 for 30% crude load. For the optimized ATPS, the recovery yield and purification factor in the top phase were determined to be 92.3% and 19.8, respectively.

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Partitioning and purification of thaumatin in aqueous two-phase systems

Cited by 89 publications

References 11 publications

Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system

Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system

Single-step purification of ovalbumin from egg white using aqueous biphasic systems

Extraction of lipase fromRhizopus microsporusfermentation culture by aqueous two-phase partitioning

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