Partition behaviour and purification of a Mucor bacilliformis acid protease in aqueous two-phase systems

Fernández‐Lahore, Marcelo; Miranda, Magdalena; Fraile, Elda R.; Bonino, M.J. Biscoglio De Jimenez; Cascone, Osvaldo

doi:10.1016/0032-9592(94)00026-3

Cited by 31 publications

(15 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Aqueous two-phase systems (ATPS), mainly poly(ethylene glycol) (PEG)/salt systems, have been widely used for the bioseparation of enzymes and proteins because of their low cost and wide range of hydrophobicity differences between the two-phase systems [1][2][3][4]. Besides the type of polymer and salt, the molecular weight of the polymer, polymer and salt concentrations, ionic strength and pH of the medium, together with the target molecule characteristics (size, charge and hydrophobicity), are the main factors influencing the partition of the molecules [1,3,5].…”

Section: Introductionmentioning

confidence: 99%

ATPS applied to extraction of small molecules – polycetides – and simultaneous clarification of culture media with filamentous microorganisms

Esmanhoto

Kilikian

2004

Journal of Chromatography B

View full text Add to dashboard Cite

Section: Introductionmentioning

confidence: 99%

ATPS applied to extraction of small molecules – polycetides – and simultaneous clarification of culture media with filamentous microorganisms

Esmanhoto

Kilikian

2004

Journal of Chromatography B

View full text Add to dashboard Cite

“…1). Lahore et al (1995) and Reh et al (2007) also found that the acid protease partition increased toward the PEG-rich phase at increasing NaCl concentration. These authors attributed this effect to a decrease in the water activity in the bottom phase produced by the increase in the salt concentration and thereby hydrophobic protein is excluded from this phase.…”

Section: Resultsmentioning

confidence: 86%

“…This effect has been used to modify the partition coefficient value of the protease in favor of top phase, increasing the recovery of the enzyme by the liquid-liquid extraction method. The partitioning coefficient can be increased quite dramatically when higher concentrations of NaCl are added to ATPS (Lahore et al 1995;Reh et al 2007). This effect has been attributed to a possible hydrophobic interaction.…”

Section: Introductionmentioning

confidence: 98%

“…NaCl is non-toxic; therefore, it can be used to modify the partition process without affecting the uses of the enzyme in any biological process. Lahore et al (1995) found that concentration of NaCl had strong effect on the protease partitioning. They also found that the acid protease partition coefficient in absence and in the presence of NaCl depends strongly on the polyethylene glycol (PEG) molecular weight.…”

Section: Introductionmentioning

confidence: 98%

“…Polyethylene glycol/salt systems are suitable for partitioning and purification of proteases (Lahore et al 1995;Porto et al 2007;Reh et al 2007;Splenzini et al 2006). The partitioning in ATPS depends not only on the surface properties (such as the size, charge and hydrophobicity) of the enzyme but also on the physicochemical traits of the two phases.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Optimization of conditions for acid protease partitioning and purification in aqueous two-phase systems using response surface methodology

2008

View full text Add to dashboard Cite

Aspergillopepsin I, an acid protease, was purified using an aqueous two-phase system that comprised various combinations of polyethylene glycol (PEG), NaH2PO4 and NaCl. Partition of the enzyme depended upon the molecular mass of the PEG and the presence of NaCl. With PEG 1500, 4000 and 6000, the partition coefficients were increased by 1,500-, 1,800- and 560-fold compared to values without NaCl. The presence of NaCl (8.75%, w/w) increased purification by 3.8, 9.5 and 2.8 times into these respective PEGs. The optimal aqueous two-phase system for acid protease purification was developed using response surface methodology. This system contained 17.3% of PEG 4000 (w/w), 15% NaH2PO4 (w/w) and 8.75% NaCl (w/w) and provided the best partition coefficient (Ke > 1,100) and yield over 99% in the same phase. The optimal ATPS purification factor of acid protease was over 5.

show abstract

Purification of Pseudomonas sp. proteases through aqueous biphasic systems as an alternative source to obtain bioactive protein hydrolysates

Pillaca-Pullo

Intiquilla

Santos

et al. 2020

Biotechnology Progress

View full text Add to dashboard Cite

Aqueous biphasic systems (ABSs) are an interesting alternative for separating industrial enzymes due to easy scale-up and low operational cost. The proteases of Pseudomonas sp. M211 were purified through ABS platforms formed by polyethylene glycol (PEG) and citrate buffer salt. Two experimental designs 2 3 + 4 were performed to evaluate the following parameters: molar mass of PEG (M PEG ), concentration of PEG (C PEG ), concentration of citrate buffer (C Cit ), and pH. The partition coefficient (K), activity yield (Y), and purification factor (PF) were the responses analyzed. The best purification performance was obtained with the system composed of M PEG = 10,000 g/mol, C PEG = 22 wt%, C Cit = 12 wt%, pH = 8.0; the responses obtained were K = 4.9, Y = 84.5%, PF = 15.1, and tie-line length = 52.74%. The purified proteases of Pseudomonas sp. (PPP) were used to obtain hydrolysates of Lupinus mutabilis (Peruvian lupin cultivar) seed protein in comparison with the commercialprotease Alcalase ® 2.4L. A strong correlation between hydrolysis degree and radical scavenging activity was observed, and the highest antioxidant activity was obtained with Alcalase ® (1.40 and 3.47 μmol Trolox equivalent/mg protein, for 2,2 0 -azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) and oxygen radical absorbance capacity, respectively) compared with PPP (0.55 and 1.03 μmol Trolox/mg protein). Nevertheless, the IC 50 values were lower than those often observed for antioxidant hydrolysates from plant proteins. PEG/citrate buffer system is valuable to purify Pseudomonas proteases from the fermented broth, and the purified protease could be promising to produce antioxidant protein hydrolysates.

show abstract

Partition behaviour and purification of a Mucor bacilliformis acid protease in aqueous two-phase systems

Cited by 31 publications

References 16 publications

ATPS applied to extraction of small molecules – polycetides – and simultaneous clarification of culture media with filamentous microorganisms

ATPS applied to extraction of small molecules – polycetides – and simultaneous clarification of culture media with filamentous microorganisms

Optimization of conditions for acid protease partitioning and purification in aqueous two-phase systems using response surface methodology

Purification of Pseudomonas sp. proteases through aqueous biphasic systems as an alternative source to obtain bioactive protein hydrolysates

Contact Info

Product

Resources

About