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Lipase diffusion on solid surfaces
KTH, School of Engineering Sciences (SCI), Applied Physics, Cell Physics.
2005 (English)Licentiate thesis, comprehensive summary (Other scientific)
Place, publisher, year, edition, pages
Stockholm: KTH , 2005. , vii, 38 p.
Series
Trita-FYS, ISSN 0280-316X ; 2005:28
National Category
Physical Chemistry
Identifiers
URN: urn:nbn:se:kth:diva-381ISBN: 91-7178-073-4 (print)OAI: oai:DiVA.org:kth-381DiVA: diva2:9664
Presentation
2005-05-30, Sal FA31, AlbaNova, Roslagstullsbacken 21, Stockhlm, 10:00
Supervisors
Note
QC 20101221Available from: 2005-08-09 Created: 2005-08-09 Last updated: 2010-12-21Bibliographically approved
List of papers
1. A FRAP-based method to study protein surface diffusion
Open this publication in new window or tab >>A FRAP-based method to study protein surface diffusion
(English)In: Journal of the American Chemical Society, ISSN 0002-7863, E-ISSN 1520-5126Article in journal (Other academic) Submitted
National Category
Physical Chemistry
Identifiers
urn:nbn:se:kth:diva-27728 (URN)
Note
QC 20101221Available from: 2010-12-21 Created: 2010-12-21 Last updated: 2017-12-11Bibliographically approved
2. Adsorption and mobility of a lipase at a hydrophobic surface in the presence of surfactants
Open this publication in new window or tab >>Adsorption and mobility of a lipase at a hydrophobic surface in the presence of surfactants
2006 (English)In: Langmuir, ISSN 0743-7463, E-ISSN 1520-5827, Vol. 22, no 13, 5810-1817 p.Article in journal (Refereed) Published
Abstract [en]

With the aim of being able to manipulate the processes involved in interfacial catalysis, we have studied the effects of a mixture of nonionic/anionic surfactants, C12E6/LAS (1: 2 mol %), on the adsorption and surface mobility of a lipase obtained from Thermomyces lanuginosus (TLL). Surface plasmon resonance (SPR) and ellipsometry were used to analyze the competitive adsorption process between surfactants and TLL onto hydrophobic model surfaces intended to mimic an oily substrate for the lipase. We obtained the surface diffusion coefficient of a fluorescently labeled TLL variant on silica silanized with octadecyltrichlorosilane (OTS) by fluorescence recovery after photobleaching (FRAP) on a confocal laser scanning microscope. By means of ellipsometry we calibrated the fluorescence intensity with the surface density of the lipase. The TLL diffusion was measured at different surface densities of the enzyme and at two time intervals after coadsorption with different concentrations of C12E6/LAS. The surfactant concentrations were chosen to represent concentrations below the critical micelle concentration (CMC), in the CMC region, and above the CMC. The apparent TLL surface diffusion was extrapolated to infinite surface dilution, D-0. We found that the presence of surfactants strongly modulated the surface mobility of TLL: with D-0 = 0.8 x 10(-11) cm(2)/s without surfactants and D-0 = 13.1 x 10(-11) cm(2)/s with surfactants above the CMC. The increase in lipase mobility on passing the CMC was also accompanied by a 2- fold increase in the mobile fraction of TLL. SPR analysis revealed that surface bound TLL was displaced by C12E6/LAS in a concentration-dependent manner, suggesting that the observed increase in surface mobility imparts bulk-mediated diffusion and so-called rebinding of TLL to the surface. Our combined results on lipase/surfactant competitive adsorption and lipase surface mobility show how surfactants may play an important role in regulating interfacial catalysis from physiological digestion to technical applications such as detergency.

Keyword
Adsorption; Hydrophobicity; Mathematical models; Physiology; Surface active agents; Surface tension; Interfacial catalysis; Oily substrate; Surface plasmon resonance (SPR; Thermomyces lanuginosus; Enzymes; immobilized enzyme; surfactant; triacylglycerol lipase; adsorption; article; Ascomycetes; chemical structure; chemistry; enzymology; fluorescence recovery after photobleaching; hydrophobicity; surface plasmon resonance; surface property; surface tension; Adsorption; Ascomycota; Enzymes, Immobilized; Fluorescence Recovery After Photobleaching; Hydrophobicity; Lipase; Models, Molecular; Surface Plasmon Resonance; Surface Properties; Surface Tension; Surface-Active Agents
National Category
Physical Sciences
Identifiers
urn:nbn:se:kth:diva-6930 (URN)10.1021/la0531244 (DOI)000238217000048 ()2-s2.0-33745760732 (Scopus ID)
Note
QC 20100818Available from: 2007-03-27 Created: 2007-03-27 Last updated: 2017-12-14Bibliographically approved

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