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Enzymatic activity of Lipase-Nanoparticle conjugates and the digestion of lipid liquid crystalline assemblies

Enzymatic activity of Lipase-Nanoparticle conjugates and the digestion of lipid liquid crystalline assemblies
Enzymatic activity of Lipase-Nanoparticle conjugates and the digestion of lipid liquid crystalline assemblies
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.
0743-7463
13590-13599
Brennan, Jennifer L.
caafcaba-0fae-4102-8b85-672f6d031d1e
Kanaras, Antonios G.
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Nativo, Paola
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Tshikhudo, T. Robert
ab2e8ec8-8f13-4267-b285-f89024197880
Rees, Claire
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Fernandez, Laura Cabo
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Dirvianskyte, Nijole
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Razumas, Valdemaras
dee3e9b3-a9b3-4531-b497-7dba90382e71
Skjøt, Michael
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Svendsen, Allan
ab8f9ae4-d9d0-4488-aa51-98be07f03775
Jørgensen, Christian I.
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Schweins, Ralf
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Zackrisson, Malin
f57da4e9-198b-4504-a0f8-2e78a110f42f
Nylander, Tommy
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Brust, Mathias
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Barauskas, Justas
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Brennan, Jennifer L.
caafcaba-0fae-4102-8b85-672f6d031d1e
Kanaras, Antonios G.
667ecfdc-7647-4bd8-be03-a47bf32504c7
Nativo, Paola
bce6035f-4ac6-4b95-a8d9-da22c21835c3
Tshikhudo, T. Robert
ab2e8ec8-8f13-4267-b285-f89024197880
Rees, Claire
8a2f3aea-e19b-4646-b920-cc7c02b34bee
Fernandez, Laura Cabo
700cbbb0-1258-4b70-8c0f-bc62a9119f18
Dirvianskyte, Nijole
10324375-7247-442f-817e-bfbc0fb94f42
Razumas, Valdemaras
dee3e9b3-a9b3-4531-b497-7dba90382e71
Skjøt, Michael
764c3bd2-4a20-455f-9422-91020272c525
Svendsen, Allan
ab8f9ae4-d9d0-4488-aa51-98be07f03775
Jørgensen, Christian I.
c8bb04eb-4448-4d92-80a6-9f1c53f77aee
Schweins, Ralf
9f5a7be2-6240-4511-8a6c-86e9eba92124
Zackrisson, Malin
f57da4e9-198b-4504-a0f8-2e78a110f42f
Nylander, Tommy
18d94ea0-6456-4eaf-b552-844261a37283
Brust, Mathias
b1f6f579-7c99-4529-824d-8c182c8c525d
Barauskas, Justas
97991d13-4310-42a1-919b-0399b56edc0d

Brennan, Jennifer L., Kanaras, Antonios G., Nativo, Paola, Tshikhudo, T. Robert, Rees, Claire, Fernandez, Laura Cabo, Dirvianskyte, Nijole, Razumas, Valdemaras, Skjøt, Michael, Svendsen, Allan, Jørgensen, Christian I., Schweins, Ralf, Zackrisson, Malin, Nylander, Tommy, Brust, Mathias and Barauskas, Justas (2010) Enzymatic activity of Lipase-Nanoparticle conjugates and the digestion of lipid liquid crystalline assemblies. Langmuir, 26 (16), 13590-13599. (doi:10.1021/la1018604).

Record type: Article

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.

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Published date: 26 July 2010

Identifiers

Local EPrints ID: 162521
URI: https://eprints.soton.ac.uk/id/eprint/162521
ISSN: 0743-7463
PURE UUID: 9d71f9a8-2343-4eaa-8833-a9e660f9a960
ORCID for Antonios G. Kanaras: ORCID iD orcid.org/0000-0002-9847-6706

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Date deposited: 23 Aug 2010 15:44
Last modified: 14 Dec 2018 01:33

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Contributors

Author: Jennifer L. Brennan
Author: Paola Nativo
Author: T. Robert Tshikhudo
Author: Claire Rees
Author: Laura Cabo Fernandez
Author: Nijole Dirvianskyte
Author: Valdemaras Razumas
Author: Michael Skjøt
Author: Allan Svendsen
Author: Christian I. Jørgensen
Author: Ralf Schweins
Author: Malin Zackrisson
Author: Tommy Nylander
Author: Mathias Brust
Author: Justas Barauskas

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