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Enzymatic Activity of Lipase-Nanoparticle Conjugates and the Digestion of Lipid Liquid Crystalline Assemblies

Brennan, Jennifer L.; Kanaras, Antonios G.; Nativo, Paola; Tshikhudo, T. Robert; Rees, Claire; Fernandez, Laura Cabo; Dirvianskyte, Nijole; Razumas, Valdemaras; Skjot, Michael and Svendsen, Allan, et al. (2010) In Langmuir 26(16). p.13590-13599
Abstract
Variants of lipase were attached to gold nanoparticles (NPs) and their enzymatic activity was studied. The two bioengineered lipase variants have been prepared with biotin groups attached to different residues on the protein outer surface. The biotinylation was evidenced by denaturing polyacrylamide gel electrophoresis and quantified by the ([2-(4'-hydroxyazobenzene)]benzoic acid spectrophotometric test. NPs of 14 +/- 1 nm diameter coated with thiolated-polyethylene glycol ligands containing controlled proportions of biotin moieties have been prepared and characterized by transmission electron microscopy, UV-vis spectroscopy, small angle neutron scattering, and elemental analysis. These biotin-functionalized NPs were conjugated to lipase... (More)
Variants of lipase were attached to gold nanoparticles (NPs) and their enzymatic activity was studied. The two bioengineered lipase variants have been prepared with biotin groups attached to different residues on the protein outer surface. The biotinylation was evidenced by denaturing polyacrylamide gel electrophoresis and quantified by the ([2-(4'-hydroxyazobenzene)]benzoic acid spectrophotometric test. NPs of 14 +/- 1 nm diameter coated with thiolated-polyethylene glycol ligands containing controlled proportions of biotin moieties have been prepared and characterized by transmission electron microscopy, UV-vis spectroscopy, small angle neutron scattering, and elemental analysis. These biotin-functionalized NPs were conjugated to lipase using streptavidin as a linker molecule. Enzyme activity assays on the lipase-nanoparticle conjugates show that the lipase loading and activity of the NPs can be controlled by varying the percentage of biotin groups in the particle protecting coat. The lipase-NP conjugates prepared using one variant display higher activity than those prepared using the other variant, demonstrating orientation-dependent enzyme activity. Cryogenic transmission electron microscopy was used to visualize the enzymatic activity of lipase-NP on well-defined lipid substrates. It was found that lipase-coated NPs are able to digest the substrates in a different manner in comparison to the free lipase. (Less)
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Contribution to journal
publication status
published
subject
in
Langmuir
volume
26
issue
16
pages
13590 - 13599
publisher
The American Chemical Society
external identifiers
  • wos:000280667900076
  • scopus:77955534688
ISSN
0743-7463
DOI
10.1021/la1018604
language
English
LU publication?
yes
id
22bff80f-1d11-4681-9b19-86066d715b16 (old id 1675386)
date added to LUP
2010-09-22 08:59:02
date last changed
2018-05-29 10:17:47
@article{22bff80f-1d11-4681-9b19-86066d715b16,
  abstract     = {Variants of lipase were attached to gold nanoparticles (NPs) and their enzymatic activity was studied. The two bioengineered lipase variants have been prepared with biotin groups attached to different residues on the protein outer surface. The biotinylation was evidenced by denaturing polyacrylamide gel electrophoresis and quantified by the ([2-(4'-hydroxyazobenzene)]benzoic acid spectrophotometric test. NPs of 14 +/- 1 nm diameter coated with thiolated-polyethylene glycol ligands containing controlled proportions of biotin moieties have been prepared and characterized by transmission electron microscopy, UV-vis spectroscopy, small angle neutron scattering, and elemental analysis. These biotin-functionalized NPs were conjugated to lipase using streptavidin as a linker molecule. Enzyme activity assays on the lipase-nanoparticle conjugates show that the lipase loading and activity of the NPs can be controlled by varying the percentage of biotin groups in the particle protecting coat. The lipase-NP conjugates prepared using one variant display higher activity than those prepared using the other variant, demonstrating orientation-dependent enzyme activity. Cryogenic transmission electron microscopy was used to visualize the enzymatic activity of lipase-NP on well-defined lipid substrates. It was found that lipase-coated NPs are able to digest the substrates in a different manner in comparison to the free lipase.},
  author       = {Brennan, Jennifer L. and Kanaras, Antonios G. and Nativo, Paola and Tshikhudo, T. Robert and Rees, Claire and Fernandez, Laura Cabo and Dirvianskyte, Nijole and Razumas, Valdemaras and Skjot, Michael and Svendsen, Allan and Jorgensen, Christian I. and Schweins, Ralf and Zackrisson Oskolkova, Malin and Nylander, Tommy and Brust, Mathias and Barauskas, Justas},
  issn         = {0743-7463},
  language     = {eng},
  number       = {16},
  pages        = {13590--13599},
  publisher    = {The American Chemical Society},
  series       = {Langmuir},
  title        = {Enzymatic Activity of Lipase-Nanoparticle Conjugates and the Digestion of Lipid Liquid Crystalline Assemblies},
  url          = {http://dx.doi.org/10.1021/la1018604},
  volume       = {26},
  year         = {2010},
}