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Characterization of lipase in reversed micelles formulated by Cibacron Blue F-3GA modified Span 85

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Characterization of lipase in reversed micelles formulated by Cibacron Blue F-3GA modified Span 85. / Zhang, Dong Hao; Guo, Zheng; Sun, Yan.

I: Biotechnology Progress, Bind 23, Nr. 1, 2007, s. 108-115.

Publikation: Bidrag til tidsskrift/Konferencebidrag i tidsskrift /Bidrag til avisTidsskriftartikelForskningpeer review

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Zhang, Dong Hao ; Guo, Zheng ; Sun, Yan. / Characterization of lipase in reversed micelles formulated by Cibacron Blue F-3GA modified Span 85. I: Biotechnology Progress. 2007 ; Bind 23, Nr. 1. s. 108-115.

Bibtex

@article{ff3cd9f02ddb11df9806000ea68e967b,
title = "Characterization of lipase in reversed micelles formulated by Cibacron Blue F-3GA modified Span 85",
abstract = "Sorbitan trioleate (Span 85) modified by Cibacron Blue F-3GA (CB) was prepared and used as an affinity surfactant to formulate a reversed micellar system for Candida rugosa lipase (CRL) solubilization. The system was characterized and evaluated by employing CRL-catalyzed hydrolysis of olive oil as a model reaction. The micellar hydrodynamic radius results reflected, to some extent, the redistribution of surfactant and water after enzyme addition, and the correlation between surfactant formulation, water content (W0), micellar size, and enzyme activity. An adequate modification density of CB was found to be important for the reversed micelles to retain enough hydration capacity and achieve high enzyme activity. Compared with the results in AOT-based reversed micelles, CRL in this micellar system exhibited a different activity behavior versus W0. The optimal pH and temperature of the encapsulated lipase remained unchanged, but the apparent activity was significantly higher than that of the native enzyme in bulk solution. Kinetic studies indicated that the encapsulated lipase in the reversed micelles of CB-formulated Span 85 followed the Michaelis-Menten equation. The Michaelis constant was found to decrease with increasing surfactant concentration, suggesting an increase of the enzyme affinity for the substrate. Stability of the lipase in the reversed micelles was negatively correlated to W0. Introduction Reversed micelles are nanometer-scale transparent aggregates of water and surfactant dispersed in a bulk apolar solvent where the surfactant molecules are arranged with their polar head toward the water pool and hydrophobic tail in contact with bulk apolar solvent (1). This has been associated to the idea that reversed micelles mimic those naturally occurring proteins located in the membrane with both aqueous and nonaqueous domains (2). The enzyme confined to the water pool of the reversed micelles is therefore supposed to be capable of retaining its conformation and activity, which is of particular importance for those enzymes with higher optimum working water activity (aw) (3). The microencapsulated enzymes are also assumed to be sheltered and protected from the detrimental effects of solvent (2, 4). Compared with other interfacial media, reversed micelles give a larger interfacial area, facilitating efficient contact between enzyme and substrate (5). Previous work has also shown that reversed micellar technology could increase the apparent activity of enzymes and even yield superactivity for specific enzymes (6). Lipases are ubiquitous enzymes that could find many important applications (7, 8). The natural substrate of lipases is triglyceride, and the majority of lipases are interfacial activated enzymes (9). This makes lipase an interesting model enzyme for reversed micelle system studies, in which lipase-catalyzed",
author = "Zhang, {Dong Hao} and Zheng Guo and Yan Sun",
year = "2007",
doi = "10.1021/bp060188b",
language = "English",
volume = "23",
pages = "108--115",
journal = "Biotechnology Progress",
issn = "8756-7938",
publisher = "Wiley-Blackwell Publishing, Inc",
number = "1",

}

RIS

TY - JOUR

T1 - Characterization of lipase in reversed micelles formulated by Cibacron Blue F-3GA modified Span 85

AU - Zhang, Dong Hao

AU - Guo, Zheng

AU - Sun, Yan

PY - 2007

Y1 - 2007

N2 - Sorbitan trioleate (Span 85) modified by Cibacron Blue F-3GA (CB) was prepared and used as an affinity surfactant to formulate a reversed micellar system for Candida rugosa lipase (CRL) solubilization. The system was characterized and evaluated by employing CRL-catalyzed hydrolysis of olive oil as a model reaction. The micellar hydrodynamic radius results reflected, to some extent, the redistribution of surfactant and water after enzyme addition, and the correlation between surfactant formulation, water content (W0), micellar size, and enzyme activity. An adequate modification density of CB was found to be important for the reversed micelles to retain enough hydration capacity and achieve high enzyme activity. Compared with the results in AOT-based reversed micelles, CRL in this micellar system exhibited a different activity behavior versus W0. The optimal pH and temperature of the encapsulated lipase remained unchanged, but the apparent activity was significantly higher than that of the native enzyme in bulk solution. Kinetic studies indicated that the encapsulated lipase in the reversed micelles of CB-formulated Span 85 followed the Michaelis-Menten equation. The Michaelis constant was found to decrease with increasing surfactant concentration, suggesting an increase of the enzyme affinity for the substrate. Stability of the lipase in the reversed micelles was negatively correlated to W0. Introduction Reversed micelles are nanometer-scale transparent aggregates of water and surfactant dispersed in a bulk apolar solvent where the surfactant molecules are arranged with their polar head toward the water pool and hydrophobic tail in contact with bulk apolar solvent (1). This has been associated to the idea that reversed micelles mimic those naturally occurring proteins located in the membrane with both aqueous and nonaqueous domains (2). The enzyme confined to the water pool of the reversed micelles is therefore supposed to be capable of retaining its conformation and activity, which is of particular importance for those enzymes with higher optimum working water activity (aw) (3). The microencapsulated enzymes are also assumed to be sheltered and protected from the detrimental effects of solvent (2, 4). Compared with other interfacial media, reversed micelles give a larger interfacial area, facilitating efficient contact between enzyme and substrate (5). Previous work has also shown that reversed micellar technology could increase the apparent activity of enzymes and even yield superactivity for specific enzymes (6). Lipases are ubiquitous enzymes that could find many important applications (7, 8). The natural substrate of lipases is triglyceride, and the majority of lipases are interfacial activated enzymes (9). This makes lipase an interesting model enzyme for reversed micelle system studies, in which lipase-catalyzed

AB - Sorbitan trioleate (Span 85) modified by Cibacron Blue F-3GA (CB) was prepared and used as an affinity surfactant to formulate a reversed micellar system for Candida rugosa lipase (CRL) solubilization. The system was characterized and evaluated by employing CRL-catalyzed hydrolysis of olive oil as a model reaction. The micellar hydrodynamic radius results reflected, to some extent, the redistribution of surfactant and water after enzyme addition, and the correlation between surfactant formulation, water content (W0), micellar size, and enzyme activity. An adequate modification density of CB was found to be important for the reversed micelles to retain enough hydration capacity and achieve high enzyme activity. Compared with the results in AOT-based reversed micelles, CRL in this micellar system exhibited a different activity behavior versus W0. The optimal pH and temperature of the encapsulated lipase remained unchanged, but the apparent activity was significantly higher than that of the native enzyme in bulk solution. Kinetic studies indicated that the encapsulated lipase in the reversed micelles of CB-formulated Span 85 followed the Michaelis-Menten equation. The Michaelis constant was found to decrease with increasing surfactant concentration, suggesting an increase of the enzyme affinity for the substrate. Stability of the lipase in the reversed micelles was negatively correlated to W0. Introduction Reversed micelles are nanometer-scale transparent aggregates of water and surfactant dispersed in a bulk apolar solvent where the surfactant molecules are arranged with their polar head toward the water pool and hydrophobic tail in contact with bulk apolar solvent (1). This has been associated to the idea that reversed micelles mimic those naturally occurring proteins located in the membrane with both aqueous and nonaqueous domains (2). The enzyme confined to the water pool of the reversed micelles is therefore supposed to be capable of retaining its conformation and activity, which is of particular importance for those enzymes with higher optimum working water activity (aw) (3). The microencapsulated enzymes are also assumed to be sheltered and protected from the detrimental effects of solvent (2, 4). Compared with other interfacial media, reversed micelles give a larger interfacial area, facilitating efficient contact between enzyme and substrate (5). Previous work has also shown that reversed micellar technology could increase the apparent activity of enzymes and even yield superactivity for specific enzymes (6). Lipases are ubiquitous enzymes that could find many important applications (7, 8). The natural substrate of lipases is triglyceride, and the majority of lipases are interfacial activated enzymes (9). This makes lipase an interesting model enzyme for reversed micelle system studies, in which lipase-catalyzed

U2 - 10.1021/bp060188b

DO - 10.1021/bp060188b

M3 - Journal article

C2 - 17269677

VL - 23

SP - 108

EP - 115

JO - Biotechnology Progress

JF - Biotechnology Progress

SN - 8756-7938

IS - 1

ER -