Reverse Micellar Extraction for Downstream Processing of Proteins/Enzymes

Krishna, S. Hari; Srinivas, N. D.; Raghavarao, K.S.M.S.; Karanth, N. G.

doi:10.1007/3-540-44604-4_5

Cited by 77 publications

(91 citation statements)

References 282 publications

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“…As observed for lipases in organic waterfree solvents (Sharma et al, 2001), this hydrophobic environment could favor the transesterification process, thereby leading to the synthesis of high molecular mass polyesters. These lipase-catalyzed esterifications are well documented for both the synthesis of small esters and the synthesis of high-M r polyesters (Wescott and Klibanov, 1994;Krishna et al, 2002;Kobayashi, 2010). These studies have clearly shown that the lipase-catalyzed esterification can involve both primary and secondary alcohol groups, depending on the source of lipase and environmental conditions (e.g.…”

Section: Discussion Slcus1 Catalysis Impacts the Ester Cross-linking mentioning

confidence: 96%

Ester Cross-Link Profiling of the Cutin Polymer of Wild-Type and Cutin Synthase Tomato Mutants Highlights Different Mechanisms of Polymerization

et al. 2015

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Cuticle function is closely related to the structure of the cutin polymer. However, the structure and formation of this hydrophobic polyester of glycerol and hydroxy/epoxy fatty acids has not been fully resolved. An apoplastic GDSL-lipase known as CUTIN SYNTHASE1 (CUS1) is required for cutin deposition in tomato (Solanum lycopersicum) fruit exocarp. In vitro, CUS1 catalyzes the self-transesterification of 2-monoacylglycerol of 9(10),16-dihydroxyhexadecanoic acid, the major tomato cutin monomer. This reaction releases glycerol and leads to the formation of oligomers with the secondary hydroxyl group remaining nonesterified. To check this mechanism in planta, a benzyl etherification of nonesterified hydroxyl groups of glycerol and hydroxy fatty acids was performed within cutin. Remarkably, in addition to a significant decrease in cutin deposition, mid-chain hydroxyl esterification of the dihydroxyhexadecanoic acid was affected in tomato RNA interference and ethyl methanesulfonate-cus1 mutants. Furthermore, in these mutants, the esterification of both sn-1,3 and sn-2 positions of glycerol was impacted, and their cutin contained a higher molar glycerol-to-dihydroxyhexadecanoic acid ratio. Therefore, in planta, CUS1 can catalyze the esterification of both primary and secondary alcohol groups of cutin monomers, and another enzymatic or nonenzymatic mechanism of polymerization may coexist with CUS1-catalyzed polymerization. This mechanism is poorly efficient with secondary alcohol groups and produces polyesters with lower molecular size. Confocal Raman imaging of benzyl etherified cutins showed that the polymerization is heterogenous at the fruit surface. Finally, by comparing tomato mutants either affected or not in cutin polymerization, we concluded that the level of cutin cross-linking had no significant impact on water permeance.Cuticles are ubiquitous hydrophobic barriers at the surfaces of aerial plant organs. These complex hydrophobic assemblies consist of a biopolymer, cutin, coated and filled with waxes and can also comprise embedded cell wall polysaccharides. Waxes comprise solventsoluble aliphatic molecules with long hydrocarbon chains, terpenes, and steroids (Kunst and Samuels,

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Section: Discussion Slcus1 Catalysis Impacts the Ester Cross-linking mentioning

confidence: 96%

Ester Cross-Link Profiling of the Cutin Polymer of Wild-Type and Cutin Synthase Tomato Mutants Highlights Different Mechanisms of Polymerization

et al. 2015

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“…However, increasing the AOT concentration from 80.0-200.0 g/L has significantly lowered the percentage of solubilized erythromycin from 95.87 % to 41.65 % with a slight precipitation observed at the interface of the solution. As clarified by Harikrishna et al, 15 at high surfactant concentration, the extraction yield would become lower because of intermicellar collision and hindrance to the diffusion of solute by surfactant aggregates. Meanwhile, Juang et al 16 appointed that the biomolecule solubilization will increase in conjunction to higher surfactant concentration because of the increment in the size of the micelle.…”

Section: Effect Of the Aot Concentrationmentioning

confidence: 96%

“…Considering the evidence from this study, it has been confirmed that reverse micelles extraction is controlled by electrostatic, steric and hydrophobic interactions between the bio-molecules and micelles. 15 The reduction of interactions in the repulsive head group occurring at the surface resulted in larger sized micelles than in either of the pure component micelles, hence the CMC value of the mixture is also lesser. The resulted aqueous solution after the solubilization of erythromycin was observed to be a clear solutionwith a transparent interface when the concentration of AOT varied from 20-140 g/L, thereby indicative of the successful solubilization of erythromycin in the reverse micelle phase.…”

Section: Effect Of the Aot Concentrationmentioning

confidence: 99%

“…Central to the entire discipline of reverse micelle extraction is the ability of biochemicals including amino acids, proteins, enzymes, and nucleic acids to solubilize in the micelle components, which then could be recovered from such solutions without losing its microbial utility. 4 Reverse micelles system consists of several components including surfactant, co-surfactant, oil, and water. As a whole, they make up a group of surfactant molecules with the hydrophilic part pointing inwards and dissolve in a continuous organic solvent medium.…”

Section: Introductionmentioning

confidence: 99%

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The solubilization and extraction parameters of erythromycin from aqueous phase into mixed AOT/SB3-18 reverse micelle phase

Aziz

Sakinah

2017

Chem. Eng. Res. Bull.

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A liquid anionic-zwitterionic surfactant-based aqueous two-phase extraction was developed and applied for the extraction of erythromycin. Erythromycin solubilization from aqueous to reverse micelle phase was studied. Zwitterion SB3-18 surfactant was added to ionic solution of bis(2-ethylhexyl) sulfosuccinate sodium salt (AOT) to form an organic mixed micellar phase. Erythromycin was then added to the mixtures and two clear phases were formed. The parameters affecting erythromycin solubilization including AOT concentration (20.0 ̶ 120.0 g/L), zwitterion concentration (6.0 ̶ 16.0 g/L), NaCl concentration (0.0 ̶ 25.0 g/L) and aqueous pH (6.0 ̶ 9.0) were investigated via statistical software. Solubilization of erythromycin into mixed reverse micelle could be easily evaluated by the measurement of erythromycin concentration in the organic phase. The results showed that AOT and zwitterion concentration governed the erythromycin solubilization by a factor of 4.562 based on the Pareto chart.

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“…4 The extraction process could be effectively used as a primary separation/purification step. 5 In most cases it is considered that the driving force for the mass transfer of biomolecules is electrostatic interaction between the protein and the charged surfactant head groups. However, other interactions such as hydrophobic interaction between protein and surfactant, have been demonstrated for the solubilization mechanism.…”

Section: Introductionmentioning

confidence: 99%

Reverse micellar extraction of bromelain from Ananas comosus L. Merryl

Hemavathi

Hebbar

Raghavarao

2007

J of Chemical Tech & Biotech

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BACKGROUND: A reverse micellar system (RMS) of ionic surfactants is used for the first time for the extraction and primary purification of fruit bromelain (EC 3.4.22.33) from the aqueous extract of pineapple (Ananas comosus L. Merryl). The effect of various process parameters on both forward and back extraction of bromelain is studied to improve the extraction efficiency of RMS. Most of the reverse micellar extraction (RME) studies reported so far are on model systems and its application to enzyme extraction from a natural source is rarely reported.

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Reverse Micellar Extraction for Downstream Processing of Proteins/Enzymes

Cited by 77 publications

References 282 publications

Ester Cross-Link Profiling of the Cutin Polymer of Wild-Type and Cutin Synthase Tomato Mutants Highlights Different Mechanisms of Polymerization

Ester Cross-Link Profiling of the Cutin Polymer of Wild-Type and Cutin Synthase Tomato Mutants Highlights Different Mechanisms of Polymerization

The solubilization and extraction parameters of erythromycin from aqueous phase into mixed AOT/SB3-18 reverse micelle phase

Reverse micellar extraction of bromelain from Ananas comosus L. Merryl

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