A Three‐Enzyme Cascade Reaction through Positional Assembly of Enzymes in a Polymersome Nanoreactor
Abstract Porous polymersomes based on block copolymers of isocyanopeptides and styrene have been used to anchor enzymes at three different locations, namely, in their lumen (glucose oxidase, GOx), in their bilayer membrane ( Candida antarctica lipase B, CalB) and on their surface (horseradish peroxi...
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Online Access: | http://dx.doi.org/10.1002/chem.200802114 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fchem.200802114 https://onlinelibrary.wiley.com/doi/full/10.1002/chem.200802114 |
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crwiley:10.1002/chem.200802114 2024-10-13T14:03:13+00:00 A Three‐Enzyme Cascade Reaction through Positional Assembly of Enzymes in a Polymersome Nanoreactor van Dongen, Stijn F. M. Nallani, Madhavan Cornelissen, Jeroen J. L. M. Nolte, Roeland J. M. van Hest, Jan C. M. 2009 http://dx.doi.org/10.1002/chem.200802114 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fchem.200802114 https://onlinelibrary.wiley.com/doi/full/10.1002/chem.200802114 en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor Chemistry – A European Journal volume 15, issue 5, page 1107-1114 ISSN 0947-6539 1521-3765 journal-article 2009 crwiley https://doi.org/10.1002/chem.200802114 2024-09-23T04:34:07Z Abstract Porous polymersomes based on block copolymers of isocyanopeptides and styrene have been used to anchor enzymes at three different locations, namely, in their lumen (glucose oxidase, GOx), in their bilayer membrane ( Candida antarctica lipase B, CalB) and on their surface (horseradish peroxidase, HRP). The surface coupling was achieved by click chemistry between acetylene‐functionalised anchors on the surface of the polymersomes and azido functions of HRP, which were introduced by using a direct diazo transfer reaction to lysine residues of the enzyme. To determine the encapsulation and conjugation efficiency of the enzymes, they were decorated with metal‐ion labels and analysed by mass spectrometry. This revealed an almost quantitative immobilisation efficiency of HRP on the surface of the polymersomes and a more than statistical incorporation efficiency for CalB in the membrane and for GOx in the aqueous compartment. The enzyme‐decorated polymersomes were studied as nanoreactors in which glucose acetate was converted by CalB to glucose, which was oxidised by GOx to gluconolactone in a second step. The hydrogen peroxide produced was used by HRP to oxidise 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) to ABTS .+ . Kinetic analysis revealed that the reaction step catalysed by HRP is the fastest in the cascade reaction. Article in Journal/Newspaper Antarc* Antarctica Wiley Online Library Chemistry – A European Journal 15 5 1107 1114 |
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English |
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Abstract Porous polymersomes based on block copolymers of isocyanopeptides and styrene have been used to anchor enzymes at three different locations, namely, in their lumen (glucose oxidase, GOx), in their bilayer membrane ( Candida antarctica lipase B, CalB) and on their surface (horseradish peroxidase, HRP). The surface coupling was achieved by click chemistry between acetylene‐functionalised anchors on the surface of the polymersomes and azido functions of HRP, which were introduced by using a direct diazo transfer reaction to lysine residues of the enzyme. To determine the encapsulation and conjugation efficiency of the enzymes, they were decorated with metal‐ion labels and analysed by mass spectrometry. This revealed an almost quantitative immobilisation efficiency of HRP on the surface of the polymersomes and a more than statistical incorporation efficiency for CalB in the membrane and for GOx in the aqueous compartment. The enzyme‐decorated polymersomes were studied as nanoreactors in which glucose acetate was converted by CalB to glucose, which was oxidised by GOx to gluconolactone in a second step. The hydrogen peroxide produced was used by HRP to oxidise 2,2′‐azinobis(3‐ethylbenzothiazoline‐6‐sulfonic acid) (ABTS) to ABTS .+ . Kinetic analysis revealed that the reaction step catalysed by HRP is the fastest in the cascade reaction. |
format |
Article in Journal/Newspaper |
author |
van Dongen, Stijn F. M. Nallani, Madhavan Cornelissen, Jeroen J. L. M. Nolte, Roeland J. M. van Hest, Jan C. M. |
spellingShingle |
van Dongen, Stijn F. M. Nallani, Madhavan Cornelissen, Jeroen J. L. M. Nolte, Roeland J. M. van Hest, Jan C. M. A Three‐Enzyme Cascade Reaction through Positional Assembly of Enzymes in a Polymersome Nanoreactor |
author_facet |
van Dongen, Stijn F. M. Nallani, Madhavan Cornelissen, Jeroen J. L. M. Nolte, Roeland J. M. van Hest, Jan C. M. |
author_sort |
van Dongen, Stijn F. M. |
title |
A Three‐Enzyme Cascade Reaction through Positional Assembly of Enzymes in a Polymersome Nanoreactor |
title_short |
A Three‐Enzyme Cascade Reaction through Positional Assembly of Enzymes in a Polymersome Nanoreactor |
title_full |
A Three‐Enzyme Cascade Reaction through Positional Assembly of Enzymes in a Polymersome Nanoreactor |
title_fullStr |
A Three‐Enzyme Cascade Reaction through Positional Assembly of Enzymes in a Polymersome Nanoreactor |
title_full_unstemmed |
A Three‐Enzyme Cascade Reaction through Positional Assembly of Enzymes in a Polymersome Nanoreactor |
title_sort |
three‐enzyme cascade reaction through positional assembly of enzymes in a polymersome nanoreactor |
publisher |
Wiley |
publishDate |
2009 |
url |
http://dx.doi.org/10.1002/chem.200802114 https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fchem.200802114 https://onlinelibrary.wiley.com/doi/full/10.1002/chem.200802114 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Chemistry – A European Journal volume 15, issue 5, page 1107-1114 ISSN 0947-6539 1521-3765 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1002/chem.200802114 |
container_title |
Chemistry – A European Journal |
container_volume |
15 |
container_issue |
5 |
container_start_page |
1107 |
op_container_end_page |
1114 |
_version_ |
1812819646909251584 |