Recovery of an Extracellular Alkaline Protease From Whole Fermentation Broth Using Reversed Micelles

Rahaman, R. S.; Chee, Jiun-Yee; Cabral, Joaquim M. S.; Hatton, T. Alan

doi:10.1002/btpr.5420040405

Cited by 117 publications

(23 citation statements)

References 4 publications

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“…fermentation broth (Rahaman et al 1988). Although these results are not yet those recommended for enzyme recovery, further efforts should be made to optimize the procedure by changing the other reversed micellar extraction parameters.…”

Section: Resultsmentioning

confidence: 89%

“…alkaline protease back-extraction (Rahaman et al 1988) and in the acid phosphatase from Aspergillus niger fermentation broth (Soni & Madamwar 2000), with high values of pH, a decrease of enzymatic activity took place while the total protein recovered increased. This was probably due to interaction of the surfactant with broth constituents (Soni & Madamwar 2000) or to protein instability in presence of organic solvent at high pH values (Rahaman et al 1988;Krieger et al 1997). The results were also analysed in terms of protein mass on the back-extraction and, apart from experimental errors, it can be concluded ( Table 2) that the protein mass balance can be considered closed.…”

Section: Resultsmentioning

confidence: 98%

“…fermentation broth (Rahaman et al 1988) when the same aqueous/organic volume ratios were used for enzyme purification.…”

Section: Resultsmentioning

confidence: 99%

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Liquid–liquid extraction of an extracellular alkaline protease from fermentation broth using aqueous two-phase and reversed micelles systems

Porto

Monteiro

Moreira

et al. 2005

World J Microbiol Biotechnol

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The study of recovery of an extracellular alkaline protease from fermentation broth produced by Norcadiopsis sp. was carried out with liquid-liquid extraction through sodium di-(2-ethylhexyl) sulphosuccinate/isooctane reversed micelles systems and aqueous two-phase systems (polyethylene glycol/potassium phosphate). The best conditions for extraction and back-extraction with the reversed micelles system was obtained at pH 9.0 and pH 5.0, respectively, showing a yield of protein of 6.16%, a specific activity of 4.10 U/ml and a purification factor of 1.80. The studies using aqueous two-phase systems of polyethylene glycol/potassium phosphate at pH 10.0 showed purification factors of 2 and 5, and protein yield of 11 and 4%, respectively, for polyethylene glycol 550/potassium phosphate and polyethylene glycol 8000/potassium phosphate. The results indicate that the aqueous two-phase systems are more attractive as a first step in the isolation and purification processes.

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Section: Resultsmentioning

confidence: 89%

Section: Resultsmentioning

confidence: 98%

See 1 more Smart Citation

Liquid–liquid extraction of an extracellular alkaline protease from fermentation broth using aqueous two-phase and reversed micelles systems

Porto

Monteiro

Moreira

et al. 2005

World J Microbiol Biotechnol

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“…Extraction in aqueous two-phase systems made of polymer/polymer and polymer/salt mixtures has become an established technique for achieving clari®cation, concentration, and partial puri®cation of proteins directly from crude feedstocks (Boland et al, 1989;Hustedt et al, 1985). Reverse micellar systems, which are water in oil microemulsions stabilized by surfactant molecules, have also been used to extract proteins from aqueous media (Rahaman et al 1988;Hatton, 1989). Protein separation by these techniques is determined by a combination of dierent interactions between the protein surface and the system components (Albertsson, 1986;Goklen and Hatton, 1985).…”

Section: Introductionmentioning

confidence: 99%

Selective recovery of lactate dehydrogenase using affinity foam

Fernandes

Hatti‐Kaul

Mattìasson

2002

Biotech & Bioengineering

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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.

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“…Electrostatic, steric, and hydrophobic interactions between protein and reverse micelles are considered to be the driving forces for the diffusion of solutes into the reverse micellar core [3]. Several reports on the application of reverse micellar extraction for separation and purification of biomolecules are available [4][5][6][7][8][9][10][11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%