Purification of α‐toxin from <i>Clostridium perfringens</i> type A in PEG‐phosphate aqueous two‐phase systems: a factorial study

Cavalcanti, Maria Taciana Holanda; Neto, Benício de Barros; Lima-Filho, José Luiz de; Porto, Tatiana Souza; Pessoa, Adalberto

doi:10.1002/jctb.1798

Cited by 7 publications

(5 citation statements)

References 24 publications

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“…There are frequent reports in the literature regarding recoveries exceeding 100% for enzyme extraction using ATPS, as shown by Cavalcanti et al 25 and Cavalcanti et al. 31 These authors cited enzyme yields of around 230 and 168%, respectively, when the PEG/phosphate ATPS was used for phospholipase C extraction. According to Mayerhoff et al, 32 high values in the recovery of these biomolecules can be explained by the decrease in concentration of potential enzyme inhibitors and activation of enzyme interaction with components of the system, which can favor enzymatic activity.…”

Section: Figure 1 Effects Of Ph On Proteolytic Activitymentioning

confidence: 96%

Partition of proteases from Lentinus citrinus DPUA 1535 by the Peg/Phosphate Aqueous Two-Phase System

et al. 2012

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A full two-level factorial design was employed to study the influence of PEG molar mass (MM PEG), PEG concentration (C PEG) and phosphate concentration (C PHOSPH) on proteases partition by Lentinus citrinus DPUA 1535 in a PEG/phosphate aqueous two-phase system (ATPS). For all ATPS studied, proteases partitioned for the top phase and the best proteases extraction condition was obtained with MM PEG = 6000 g mol-1, C PEG = 17.5% (w/w) and C PHOSPH = 25% (w/w) with (1.1) purification factor and (151%) activity yield. Findings reported here demonstrate a practical strategy that serves as a first step for proteases purification from crude extract by L. citrinus

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Section: Figure 1 Effects Of Ph On Proteolytic Activitymentioning

confidence: 96%

Partition of proteases from Lentinus citrinus DPUA 1535 by the Peg/Phosphate Aqueous Two-Phase System

et al. 2012

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“…Moreover, the positive interaction among M PEG , C PEG and C PHOS also affected positively K, Y and PF, but had a negative effect on S (Table 3). Cavalcanti et al [56] and Kirsch et al [40] observed that C PEG and C PHOS influenced K, Y and PF. Moreover, pH was also considered to affect K directly [57].…”

Section: Partition Coefficient Activity Yield and Purification Factomentioning

confidence: 98%

“…In addition, the values obtained for the response variables PF and Y were 1.1 and 151%, respectively. In order to evaluate the purification of ␣toxin from Clostridium perfringens type A, Cavalcanti et al [56] used a PEG/phosphate ATPS and obtained the highest values for the PF (5.7) when using a C PEG and C PHOS of 17.5% and 15%, respectively.…”

Section: Partition Coefficient Activity Yield and Purification Factomentioning

confidence: 99%

Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system

Maciel

Ottoni

Herculano

et al. 2014

Fluid Phase Equilibria

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The partitioning and purification of polygalacturonases (PG) produced by Aspergillus niger URM 5162 were investigated in aqueous two-phase systems (ATPS), formed by polyethylene glycol and phosphate salts (PEG/phosphate). To evaluate the effect of the 4 independent variables-molar mass of polyethylene glycol (PEG) (400-8000 g/mol-M PEG), PEG concentration (12.5-17.5%, w/w-C PEG), phosphate concentration (15-25%, w/w, C PHOS) and pH (6.0-8.0)-on the 4 response variables: partition coefficient (K), activity yield (Y), purification factor (PF) and selectivity (S), a factorial design (2 4) was used. The endopolygalacturonases (endo-PG) and exo-polygalacturonase (exo-PG) were preferentially partitioned in the top phase. For endo-and exo-PG, the highest values for the response variables K (1.23 and 2.40), Y (74.04% and 17.97%), PF (8.18 and 1.98) and S (24.68 and 48.07), respectively, were obtained for a C PEG of 12.5% (w/w), M PEG of 8000 g/mol, and C PHOS of 25% (w/w) at pH 6.0. These conditions were considered the most suitable for the purification of PG produced by A. niger URM 5162. Furthermore, the most important independent variables for endo-and exo-PG were C PHOS and M PEG , respectively. All independent variables studied and their interactions significantly influenced the response variables. According to these results, the PEG/phosphate system is a useful cost-effective alternative for purification of PG produced by A. niger URM 5162.

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“…Hence, center-points characterize experimental runs at the center level of each factor range [6,19]. Furthermore, the pure error of the replicated center-point experiments is used to estimate the statistical significance of the calculated values of each factor, and thus evaluate the background variability of the process [6,19,20].…”

Section: Screening Of Significant Factorsmentioning

confidence: 99%

Optimization of PEG–salt aqueous two-phase systems by design of experiments

Glyk

Solle

Scheper

et al. 2015

Chemometrics and Intelligent Laboratory Systems

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Since the mechanism governing the partitioning behavior of biomolecules, such as proteins and enzymes, in polyethylene glycol (PEG)-salt aqueous two-phase systems (ATPS) is complex and not easily predictable, many laborious experiments have to be performed for an optimization of these systems, causing increased overall cost. However, the multivariate statistical design of experiments (DoE) methodology is representing a promising and efficient optimization technique which can overcome the limitations of traditional optimization methods. Therefore, DoE has emerged as a powerful and efficient optimization tool for PEGsalt ATPS, since it is faster, more efficient and cost-effective, allowing a simultaneous and rigorous evaluation of process/system parameters. In the present review, different DoE process steps are represented to highlight the feasibility of this approach to operate as a promising and efficient optimization tool, thus facilitating the evaluation of the partitioning behavior, recovery and purification of different proteins and enzymes in PEG-salt ATPS. In this context, several experimental designs, such as factorial and response surface designs, have been discussed and evaluated by statistical regression analysis and analysis of variance (ANOVA), as well as various applications of PEG-salt ATPS using DoE have been outlined which may further promote the optimization of these systems.

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Purification of α‐toxin from Clostridium perfringens type A in PEG‐phosphate aqueous two‐phase systems: a factorial study

Abstract: BACKGROUND: Purification of α-toxin produced by Clostridium perfringens type A in aqueous two-phase systems (ATPS) was studied with a full two-level factorial design on two factors (concentrations of 8000 g mol

Cited by 7 publications

References 24 publications

Partition of proteases from Lentinus citrinus DPUA 1535 by the Peg/Phosphate Aqueous Two-Phase System

Partition of proteases from Lentinus citrinus DPUA 1535 by the Peg/Phosphate Aqueous Two-Phase System

Purification of polygalacturonases produced by Aspergillus niger using an aqueous two-phase system

Optimization of PEG–salt aqueous two-phase systems by design of experiments

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