Optimized extraction of a lectin from Crataeva tapia bark using AOT in isooctane reversed micelles

Nascimento, Cynthia O.; Costa, Romero Marcos Pedrosa Brandão; Araújo, Regina Maria Sousa de; Chaves, M. E. C.; Coelho, Luana Cassandra Breitenbach Barroso; Teixeira, J. A.; Correia, Maria T.S.; Carneiro-da-Cunha, Maria G.

doi:10.1016/j.procbio.2008.02.021

Cited by 24 publications

(21 citation statements)

References 23 publications

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“…High amount of salt promotes the interaction of salt ion with the surfactant, which in turn reduces the interaction between surfactant head and biomolecules leading to reduced debye length. This phenomenon results in the reduction of interaction between biomolecule and the surfactant head group and is termed as electrostatic screening effect 18, 29‐31…”

Section: Resultsmentioning

confidence: 99%

“…Further increase in surfactant concentration resulted in reduced activity recovery and extraction efficiency. The increase in extraction can be attributed to the fact that the increase in surfactant concentration up to a certain extent results in an increase in the number of reverse micelles 31, 32, 33. Beyond which extraction capacity decreased due to micellar clustering, which decreased the interfacial area available to the target biomolecule 32…”

Section: Resultsmentioning

confidence: 99%

“…Further increase in salt concentration up to 1.25 M resulted in decrease in these parameters due to the dominance of hydrophobic interactions, aggregation and precipitation of protein during backward extraction 36, 37. The electrostatic screening effect at optimum salt concentration (1 M) may be responsible for the decrease of the surfactant head group repulsions, leading to the smaller reverse micelles which in turn causes size exclusion effect and leads to higher extraction 31…”

Section: Resultsmentioning

confidence: 99%

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Single step purification of lactoperoxidase from whey involving reverse micelles‐assisted extraction and its comparison with reverse micellar extraction

Nandini

Rastogi

2010

Biotechnology Progress

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The extraction of lactoperoxidase (EC 1.11.1.7) from whey was studied using single step reverse micelles-assisted extraction and compared with reverse micellar extraction. The reverse micelles-assisted extraction resulted in extraction of contaminating proteins and recovery of lactoperoxidase in the aqueous phase leading to its purification. Reverse micellar extraction at the optimized condition after forward and backward steps resulted in activity recovery of lactoperoxidase and purification factor of the order of 86.60% and 3.25-fold, respectively. Whereas reverse micelles-assisted extraction resulted in higher activity recovery of lactoperoxidase (127.35%) and purification factor (3.39-fold). The sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profiles also evidenced that higher purification was obtained in reverse micelles-assisted extraction as compared of reverse micellar extracted lactoperoxidase.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Single step purification of lactoperoxidase from whey involving reverse micelles‐assisted extraction and its comparison with reverse micellar extraction

Nandini

Rastogi

2010

Biotechnology Progress

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“…A protein from the Crataeva tapia bark has been purified using reversed micelles in isooctane [16] and through chromatographic processes [17]. Named CrataBL ( Crata eva tapia B ark L ectin), this protein was shown to possess some specificity for binding glucose and galactose [17].…”

Section: Introductionmentioning

confidence: 99%

Crystal Structure of Crataeva tapia Bark Protein (CrataBL) and Its Effect in Human Prostate Cancer Cell Lines

et al. 2013

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A protein isolated from the bark of Crataeva tapia (CrataBL) is both a Kunitz-type plant protease inhibitor and a lectin. We have determined the amino acid sequence and three-dimensional structure of CrataBL, as well as characterized its selected biochemical and biological properties. We found two different isoforms of CrataBL isolated from the original source, differing in positions 31 (Pro/Leu); 92 (Ser/Leu); 93 (Ile/Thr); 95 (Arg/Gly) and 97 (Leu/Ser). CrataBL showed relatively weak inhibitory activity against trypsin (Kiapp = 43 µM) and was more potent against Factor Xa (Kiapp = 8.6 µM), but was not active against a number of other proteases. We have confirmed that CrataBL contains two glycosylation sites and forms a dimer at high concentration. The high-resolution crystal structures of two different crystal forms of isoform II verified the β-trefoil fold of CrataBL and have shown the presence of dimers consisting of two almost identical molecules making extensive contacts (∼645 Å2). The structure differs from those of the most closely related proteins by the lack of the N-terminal β-hairpin. In experiments aimed at investigating the biological properties of CrataBL, we have shown that addition of 40 µM of the protein for 48 h caused maximum growth inhibition in MTT assay (47% of DU145 cells and 43% of PC3 cells). The apoptosis of DU145 and PC3 cell lines was confirmed by flow cytometry using Annexin V/FITC and propidium iodide staining. Treatment with CrataBL resulted in the release of mitochondrial cytochrome c and in the activation of caspase-3 in DU145 and PC3 cells.

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“…As a rule, authors verified the following advantages in protein extraction by reverse micelles: surfactants and organic solvents may be repetitively used after recovering the same, with reduced extraction costs, and polar sites of reverse micelles maintain activities which are characteristics of dissolved proteins. Nascimento et al (2008) optimized extraction of lectin from the bark of Crataeva tapia from a crude extract using anionic surfactant sodium bis (2-ethylhexyl) sulfosuccinate (AOT) in isooctane reversed micelles. The back-extraction to a final aqueous phase was made by adding butanol.…”

Section: Introductionmentioning

confidence: 99%

<b>Optimization of protein extraction process from jackfruit seed flour by reverse micelle system

et al. 2016

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ABSTRACT. The extraction of protein from flour of jackfruit seeds by reverse micelles was evaluated. Reverse micelle system was composed of sodium dodecyl sulfate (SDS) as surfactant, butanol as solvent, and water. The effects of stirring time, temperature, molar ratio H 2 O SDS -1 , concentration of butanol (mass percentage) and flour mass were tested in batch systems. Based on the adjusted linear regression model, only butanol concentration provided optimum extraction conditions (41.16%). Based on the analysis of surface response, the best extraction yield could be obtained at 25°C, stirring time of 120 min, mass of flour of 100 mg, and a ratio H 2 O SDS -1 of 50. Experimental results showed that a 79.00% extraction yield could be obtained.

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Optimized extraction of a lectin from Crataeva tapia bark using AOT in isooctane reversed micelles

Abstract: 2.2. Preparation of extract C. tapia bark was collected in the region of Recife city (Pernambuco, Brazil) and the extract was obtained by pulverizing dried bark [10% (w/v) in 150 mM NaCl] followed by agitation overnight at 4 8C. Afterwards, the extract was filtered through

Cited by 24 publications

References 23 publications

Single step purification of lactoperoxidase from whey involving reverse micelles‐assisted extraction and its comparison with reverse micellar extraction

Single step purification of lactoperoxidase from whey involving reverse micelles‐assisted extraction and its comparison with reverse micellar extraction

Crystal Structure of Crataeva tapia Bark Protein (CrataBL) and Its Effect in Human Prostate Cancer Cell Lines

<b>Optimization of protein extraction process from jackfruit seed flour by reverse micelle system

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