Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase

Souza, Ranyere Lucena de; Lima, Rafaella A.; Coutinho, João A. P.; Soares, Cleide Mara Faria; Lima, Álvaro Silva

doi:10.1016/j.procbio.2015.05.015

Cited by 44 publications

(20 citation statements)

References 46 publications

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“…Table 2 reports the enzymatic activity (EA -U.mL -1 ), total protein concentration (C -mg.mL -1 ), specific activity (SA -U.mL -1 ) and purification factor (PF -fold) in the fermented broth and dialyzed. The purification factor of the dialysate was found to be 12.7 ± 0.2 fold, confirming the values previously reported by our group at the stage of pre-purification by dialysis [8,30,55]. The proposed application of this ATPS revealed a great performance in the purification of the lipolytic lipase produced from Bacillus sp.…”

Section: Production and Pre-purification Of Lipasesupporting

confidence: 87%

“…Showed lower purification factors than those here achieved (PF = 51 ± 2 fold) [30]. ATPS based in THF with the of potassium phosphate buffer (pH 7) were also tested and the results of purification (PF = 103.9 ± 0.9 fold) were again below to those described in this work [55]. All of these studies were reported for the lipase purification from Bacillus sp.…”

Section: Purification Of Lipase Using Atpsmentioning

confidence: 46%

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Aqueous two-phase systems based on cholinium salts and tetrahydrofuran and their use for lipase purification

Souza

Lima

Coutinho

et al. 2015

Separation and Purification Technology

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Section: Production and Pre-purification Of Lipasesupporting

confidence: 87%

Section: Purification Of Lipase Using Atpsmentioning

confidence: 46%

Aqueous two-phase systems based on cholinium salts and tetrahydrofuran and their use for lipase purification

Souza

Lima

Coutinho

et al. 2015

Separation and Purification Technology

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“…Moreover, the selection of the salting-out agent in IL-based ABS is limited by its effect on pH, and thus, often a phosphate buffer aiming at controlling the pH value of the coexisting aqueous phases is used. 19,51 Therefore, in order to examine the viability of selfbuffering GB-ILs as media for enzyme applications, the enzyme activity and stability 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 biocatalyst in biotechnological applications due to its multiplicity of catalytic applications. 41,[52][53] As depicted in Figure 3 …”

Section: Lipase Enzyme Activity In Gb-ilsmentioning

confidence: 99%

Evaluating Self-buffering Ionic Liquids for Biotechnological Applications

Lee

Vicente

Silva

et al. 2015

ACS Sustainable Chem. Eng.

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A new range of Good's buffer ionic liquids (GB-ILs), displaying simultaneously the properties of ionic liquids and Good's buffers, were synthesized by combination of Good's buffers anions (MOPSO, BES, TAPSO and CAPSO) and tetrabutylammonium, tetrabutylphosphonium and cholinium cations via an acid-base neutralization reaction. The activity and stability of a lipolytic enzyme from Pseudomonas cepacia in aqueous solutions of these buffers were evaluated and the results show their advantage as media for enzymatic reactions when compared to conventional phosphate buffers. Moreover aqueous biphasic systems (ABS) composed by these GB-ILs and potassium citrate were investigated and shown to be highly effective and selective for the partitioning of the lipolytic enzyme into the GB-IL-rich phase. The results allow the development of an efficient and biocompatible process combining the self-buffering and enzyme-stabilizing properties of the GB-ILs in the reaction step, with the advantages of GB-ILs as extraction solvents in ABS.

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“…where V R represents the volume ratio between top V T and bottom V B phases, while K E and K P are the enzyme and protein partition coefficients between two phases [5,26].…”

Section: Pf =mentioning

confidence: 99%

Purification of bacterial inulinase in aqueous two‐phase systems

Temkov¹,

Strinska

Dobrev

et al. 2018

Engineering in Life Sciences

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In this study, extracellular inulinase from Bacillus sp. 11/3 was partially purified and concentrated using aqueous two‐phase system (ATPS). Two different phase forming salts and four types of polyethylene glycol (PEG) were used. Binodal curves and tie‐length lines (TLLs) for eight ATPS were developed. For inulinase purification, concentrations of PEG and salt according to binodal curves (between 17 and 26%) were chosen. All ATPSs for inulinase purification were characterized. An ATPS consisted of 26% PEG1000 and 26% MgSO4 was found to be the most suitable for inulinase purification. This ATPS has 28.47% TLL, 1.03 of volume ratio, purification factor of 4.65 fold and recovery yield of 66.17%. On the SDS‐PAGE electrophoresis two protein bands with molecular weight of around 24 and 56 kDa were observed. The partially purified enzymes had optimal activity at pH 8.0 and 6.5, optimal temperature at 30 and 70°C and kinetic parameters Km = 26.32 mmol and Vmax = 526 mmol/min.

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Novel aqueous two-phase systems based on tetrahydrofuran and potassium phosphate buffer for purification of lipase

Cited by 44 publications

References 46 publications

Aqueous two-phase systems based on cholinium salts and tetrahydrofuran and their use for lipase purification

Aqueous two-phase systems based on cholinium salts and tetrahydrofuran and their use for lipase purification

Evaluating Self-buffering Ionic Liquids for Biotechnological Applications

Purification of bacterial inulinase in aqueous two‐phase systems

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