Recovery of Recombinant Cutinase Using Detergent Foam

Fernandes, Sheryl; Mattìasson, Bo; Hatti‐Kaul, Rajni

doi:10.1021/bp010161m

Cited by 14 publications

(19 citation statements)

References 44 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We have recently reported the use of Triton X-114 foam for recombinant enzyme recovery from a culture broth (Fernandes et al, 2002). Hydrophobic interactions between the protein and the surfactant were important for good recovery of the enzyme.…”

Section: Resultsmentioning

confidence: 99%

“…The time for total collapse of the foam prepared from 0.005±1% native TX solution ranged from 0.5 to 7 h at 25°C (Fernandes et al, 2002). The collapse time of the foam from 0.1% surfactant increased with the concentration of the dye from about 4 h for TX to over 6 h for TX-CB:TX, and 8 h for TX-CB.…”

Section: Preparation and Characterization Of The Af®nity Surfactantmentioning

confidence: 99%

“…The solution was diluted with water to a desired detergent concentration, either directly or after mixing in a 1:1 ratio with 2% (w/w) solution of unmodi®ed Triton X-114 (henceforth referred to as TX-CB:TX). The diluted anity surfactant solution was used for the recovery of LDH according to the procedure described earlier (Fernandes et al, 2002), a¯ow scheme of which is shown in Figure 1. Fifty grams of the surfactant solution in a 250-mL beaker was stirred at 8000 rpm using a high speed impeller (Ultra Thurrax Antrieb-T25; IKA Laboratechnic, Staufen, Germany) for 15 min at 22°C, allowed to stand for about 3 min for the aphrons to cream and separate out as a foam from the aqueous phase.…”

Section: Recovery Of Ldh From Porcine Muscle Extract Using Tx-cb Foammentioning

confidence: 99%

“…Recently, colloidal gas aphrons (CGAs), i.e., the microbubbles generated by vigorous stirring of a surfactant solution, have attracted attention as yet another tool for separation of proteins (Fernandes et al, 2002;Jauregi and Varley, 1998;Noble et al, 1998;. The microbubbles (10±100 lm), consisting of a gaseous inner core surrounded by a thin, multilayered surfactant ®lm (Jauregi et al, 2000;Sebba, 1987), provide a high interfacial area for adsorption of molecules, and their high buoyancy leads to a quick and simple separation from the bulk solution .…”

Section: Introductionmentioning

confidence: 99%

“…Recombinant cutinase, a lipolytic enzyme was separated from a culture broth of Saccharomyces cerevisiae using foam prepared from a non-ionic surfactant, Triton X-114 (Fernandes et al, 2002). The enzyme with a hydrophobic fusion tag (Trp-Pro) 4 was recovered with a higher yield than the wild-type cutinase.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Selective recovery of lactate dehydrogenase using affinity foam

Fernandes

Hatti‐Kaul

Mattìasson

2002

Biotech & Bioengineering

View full text Add to dashboard Cite

Selective isolation of lactate dehydrogenase (LDH) from porcine muscle extract was studied using foam generated from the vigorous stirring of a non-ionic surfactant, Triton X-114 derivatized with Cibacron blue. The cloud point of the surfactant-dye conjugate was higher than that of the native Triton X-114, and also the foam prepared from the affinity surfactant was more rigid taking a longer time to collapse. The equilibrium dissociation constant between pure LDH and surfactant-dye conjugate was 5.0 microM as compared to the value of 2.2 microM for the enzyme and free dye as measured by differential spectroscopy. The isolation procedure involved mixing of the porcine muscle extract with the affinity foam, separating and collapsing the foam, and warming the solution formed to 37 degrees C to yield the surfactant-dye phase and an aqueous phase containing the enzyme. The effect of surfactant concentration and protein load on enzyme recovery and purification was investigated. Under optimal conditions, LDH was quantitatively recovered with high purification factor in a very short time. Both recovery and purification were higher when foam prepared from an equivalent mixture of surfactant-dye conjugate and unmodified surfactant was used. The selectivity of interaction between LDH and detergent-dye conjugate was confirmed by lowered recovery when NADH was included during the binding step.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Preparation and Characterization Of The Af®nity Surfactantmentioning

confidence: 99%

Section: Recovery Of Ldh From Porcine Muscle Extract Using Tx-cb Foammentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Selective recovery of lactate dehydrogenase using affinity foam

Fernandes

Hatti‐Kaul

Mattìasson

2002

Biotech & Bioengineering

View full text Add to dashboard Cite

show abstract

Selective separation of β‐lactoglobulin from sweet whey using CGAs generated from the cationic surfactant CTAB

Fuda

Bhatia²,

Pyle³

et al. 2005

Biotech & Bioengineering

View full text Add to dashboard Cite

The selective separation of whey proteins was studied using colloidal gas aphrons generated from the cationic surfactant cetyl trimethyl ammonium bromide (CTAB). From the titration curves obtained by zeta potential measurements of individual whey proteins, it was expected to selectively adsorb the major whey proteins, i.e., bovine serum albumin, alpha-lactalbumin, and beta-lactoglobulin to the aphrons and elute the remaining proteins (lactoferrin and lactoperoxidase) in the liquid phase. A number of process parameters including pH, ionic strength, and mass ratio of surfactant to protein (M(CTAB)/M(TP)) were varied in order to evaluate their effect on protein separation. Under optimum conditions (2 mmol/l CTAB, M(CTAB)/M(TP) = 0.26-0.35, pH 8, and ionic strength = 0.018 mol/l), 80-90% beta-lactoglobulin was removed from the liquid phase as a precipitate, while about 75% lactoferrin and lactoperoxidase, 80% bovine serum albumin, 95% immunoglobulin, and 65% alpha-lactalbumin were recovered in the liquid fraction. Mechanistic studies using zeta potential measurements and fluorescence spectroscopy proved that electrostatic interactions modulate only partially the selectivity of protein separation, as proteins with similar surface charges do not separate to the same extent between the two phases. The selectivity of recovery of beta-lactoglobulin probably occurs in two steps: the first being the selective interaction of the protein with opposite-charged surfactant molecules by means of electrostatic interactions, which leads to denaturation of the protein and subsequent formation and precipitation of the CTAB-beta-lactoglobulin complex. This is followed by the separation of CTAB-beta-lactoglobulin aggregates from the bulk liquid by flotation in the aphron phase. In this way, CGAs act as carriers which facilitate the removal of protein precipitate.

show abstract

Technology of streptomycin sulfate separation by two‐stage foam separation

2012

Biotechnology Progress

View full text Add to dashboard Cite

Industrial discharges from manufacturing streptomycin sulfate (SS) are inhibitory to biological wastewater treatment and need to be stripped of residual SS. For effective SS recovery from the wastewater, a two-stage foam separation technology was investigated using a column with a vertical ellipsoid-shaped channel (VEC) and a conventional one, and sodium dodecyl sulfate (SDS) served as the collector. The mechanism of enhancing foam drainage by VEC was theoretically analyzed. In the first stage, the column with VEC was used and under the optimal conditions of the liquid-loading volume 300 mL, volumetric airflow rate 100 mL/min, the initial pH 7.0 and the molar ratio of SDS to SS 8.0, an improved SS enrichment ratio of 16.7 was obtained. In the second stage, a conventional column was used and with a volumetric airflow rate of 450 mL/min, the foamate had a SS concentration of about 0.5 g/L, so it was used as the feed solution of the first stage. By the two-stage technology, the total SS recovery percentage reached as high as 99.7%. Thus, it was significantly effective for the two-stage foam separation technology to recover SS from the simulative wastewater.

show abstract

Recovery of Recombinant Cutinase Using Detergent Foam

Cited by 14 publications

References 44 publications

Selective recovery of lactate dehydrogenase using affinity foam

Selective recovery of lactate dehydrogenase using affinity foam

Selective separation of β‐lactoglobulin from sweet whey using CGAs generated from the cationic surfactant CTAB

Technology of streptomycin sulfate separation by two‐stage foam separation

Contact Info

Product

Resources

About