Purification of intracellular enzymes from whole bacterial cells using reversed micelles

Giovenco, S.; Verheggen, F.T.M.; Laane, C.

doi:10.1016/0141-0229(87)90099-8

Cited by 51 publications

(8 citation statements)

References 6 publications

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“…The selective release of the enzyme may be because of the selective permeation of the enzyme through the outer cell wall as a consequence of the action of the surfactant and solvent on the cell wall. Apparently the solvent and surfactant did not affect the inner cytoplasmic membrane unlike that reported by Giovenco et al 8 with CTAB±hexanol±isooctane solutions. The cytoplasmic proteins were, therefore, not mixed with the periplasmic proteins/enzymes, giving a better purity of the enzyme.…”

Section: à3contrasting

confidence: 77%

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Selective reverse micellar extraction of penicillin acylase from E coli

Gaikar

Kulkarni

2001

J of Chemical Tech & Biotech

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The recovery of penicillin acylase from E coli by a new reverse micellar treatment is described. The results are compared with the cell disruption by ultrasound followed by reverse micellar extraction. The process gave selective extraction of penicillin acylase directly from the periplasmic space of E coli without disrupting the cells. Unlike ultrasonication which breaks open the cells entirely, making subsequent processing dif®cult and expensive, reverse micellar treatment of cells gave a moderate recovery of 60% of enzyme activity in a highly pure form.

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Section: à3contrasting

confidence: 77%

“…1±7 In a modi®ed form of reverse micellar extraction, Giovenco et al 8 reported direct extraction of dehydrogenases from Azotobacter vinelandii. The whole cells were lysed by cetyl ammonium bromide (CTAB)±hexanol±isooctane solutions releasing the entire cell contents and the enzymes were solubilized into the reverse micelles.…”

Section: Introductionmentioning

confidence: 99%

Selective reverse micellar extraction of penicillin acylase from E coli

Gaikar

Kulkarni

2001

J of Chemical Tech & Biotech

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“…5 It seems that it is rather difficult to extract relatively large monomeric and oligomeric proteins into reversed micelles due to steric limitations 6. 7 Large molecular weight proteins have, however, been solubilized in reversed micelles by the injection method 8. 9…”

Section: Introductionmentioning

confidence: 99%

Extraction of yeast alcohol dehydrogenase using reversed micelles formed with CTAB

Zhang

Liu

Chen

2000

J. Chem. Technol. Biotechnol.

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A large molecular weight protein, alcohol dehydrogenase (ADH, 141 kDa), has been puri®ed from baker's yeast using reversed micelles formed with a cationic surfactant (cetyltrimethylammonium bromide, CTAB) using the phase transfer method. Various parameters, such as pH, ionic strength and contact time of the two phases in ADH forward and backward transfer were studied. The active ADH was successfully recovered after a full forward and backward extraction cycle. The recovery of ADH activity obtained was $90% and the puri®cation factor was 3.1 for the overall process.

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“…The injection mode takes a low time for extraction and ensures a high enzyme activity. However, a very few studies on extraction have been made using the injection mode for the solubilization and separation of proteins by reverse micelles [17,19]. In the present study, the effect of various process parameters on both forward and back extractions of β-galactosidase by the injection mode of RME was investigated to optimize the extraction efficiency of the AOT/iso-octane reverse micellar system.…”

Section: Introductionmentioning

confidence: 99%

Reverse Micellar Extraction of β-Galactosidase from Barley (Hordeum vulgare)

Hemavathi

Hebbar

Raghavarao

2008

Appl Biochem Biotechnol

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The reverse micellar system of sodium bis(2-ethylhexyl)sulfosuccinate (AOT)/isooctane was used for the extraction and primary purification of beta-galactosidase (EC 3.2.1.23) from the aqueous extract of barley (Hordeum vulgare) for the first time. The process parameters such as the concentration of the surfactant, the volume of the sample injected, and its protein concentration, pH, and ionic strength of the initial aqueous phase for forward extraction, buffer pH, and salt concentration for back extraction are varied to optimize the extraction efficiency. Studies carried out with both phase transfer and injection mode of reverse micellar extraction confirmed the injection mode to be more suitable for beta-galactosidase extraction. The extent of reverse micellar solubilization of proteins increased with an increase in protein concentration of the feed sample. However, back extraction efficiency remained almost constant (13-14.4%), which indicates the selectivity of AOT reverse micelles for a particular protein under given experimental conditions. beta-Galactosidase was extracted with an activity recovery of 98.74% and a degree of purification of 7.2-fold.

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Purification of intracellular enzymes from whole bacterial cells using reversed micelles

Cited by 51 publications

References 6 publications

Selective reverse micellar extraction of penicillin acylase from E coli

Selective reverse micellar extraction of penicillin acylase from E coli

Extraction of yeast alcohol dehydrogenase using reversed micelles formed with CTAB

Reverse Micellar Extraction of β-Galactosidase from Barley (Hordeum vulgare)

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