Photocontrolled reversible self-assembly of dodecamer nitrilase
Abstract Background Naturally photoswitchable proteins act as a powerful tool for the spatial and temporal control of biological processes by inducing the formation of a photodimerizer. In this study, a method for the precise and reversible inducible self-assembly of dodecamer nitrilase in vivo (in...
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doaj-5a0b44c6bb7f403a8878690a61190f5a2020-11-25T02:19:06ZengSpringerOpenBioresources and Bioprocessing2197-43652017-08-01411810.1186/s40643-017-0167-3Photocontrolled reversible self-assembly of dodecamer nitrilaseQiao Yu0Yong Wang1Shengyun Zhao2Yuhong Ren3East China University of Science and TechnologyKey Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of SciencesFujian Key Laboratory of Eco-Industrial Green Technology, Wuyi UniversityState Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and TechnologyAbstract Background Naturally photoswitchable proteins act as a powerful tool for the spatial and temporal control of biological processes by inducing the formation of a photodimerizer. In this study, a method for the precise and reversible inducible self-assembly of dodecamer nitrilase in vivo (in Escherichia coli) and in vitro (in a cell-free solution) was developed by means of the photoswitch-improved light-inducible dimer (iLID) system which could induce protein–protein dimerization. Results Nitrilase was fused with the photoswitch protein AsLOV2-SsrA to achieve the photocontrolled self-assembly of dodecamer nitrilase. The fusion protein self-assembled into a supramolecular assembly when illuminated at 470 nm. Scanning electron microscopy showed that the assembly formed a circular sheet structure. Self-assembly was also induced by light in E. coli. Dynamic light scattering and turbidity assay experiments showed that the assemblies formed within a few seconds under 470-nm light and completely disassembled within 5 min in the dark. Assembly and disassembly could be maintained for at least five cycles. Both in vitro and in vivo, the assemblies retained 90% of the initial activity of nitrilase and could be reused at least four times in vitro with 90% activity. Conclusions An efficient method was developed for the photocontrolled assembly and disassembly of dodecamer nitrilase and for scaffold-free reversible self-assembly of multiple oligomeric enzymes in vivo and in vitro, providing new ideas and methods for immobilization of enzyme without carrier.http://link.springer.com/article/10.1186/s40643-017-0167-3DisassemblyiLIDNitrilasePhotoswitchSelf-assembly |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Qiao Yu Yong Wang Shengyun Zhao Yuhong Ren |
spellingShingle |
Qiao Yu Yong Wang Shengyun Zhao Yuhong Ren Photocontrolled reversible self-assembly of dodecamer nitrilase Bioresources and Bioprocessing Disassembly iLID Nitrilase Photoswitch Self-assembly |
author_facet |
Qiao Yu Yong Wang Shengyun Zhao Yuhong Ren |
author_sort |
Qiao Yu |
title |
Photocontrolled reversible self-assembly of dodecamer nitrilase |
title_short |
Photocontrolled reversible self-assembly of dodecamer nitrilase |
title_full |
Photocontrolled reversible self-assembly of dodecamer nitrilase |
title_fullStr |
Photocontrolled reversible self-assembly of dodecamer nitrilase |
title_full_unstemmed |
Photocontrolled reversible self-assembly of dodecamer nitrilase |
title_sort |
photocontrolled reversible self-assembly of dodecamer nitrilase |
publisher |
SpringerOpen |
series |
Bioresources and Bioprocessing |
issn |
2197-4365 |
publishDate |
2017-08-01 |
description |
Abstract Background Naturally photoswitchable proteins act as a powerful tool for the spatial and temporal control of biological processes by inducing the formation of a photodimerizer. In this study, a method for the precise and reversible inducible self-assembly of dodecamer nitrilase in vivo (in Escherichia coli) and in vitro (in a cell-free solution) was developed by means of the photoswitch-improved light-inducible dimer (iLID) system which could induce protein–protein dimerization. Results Nitrilase was fused with the photoswitch protein AsLOV2-SsrA to achieve the photocontrolled self-assembly of dodecamer nitrilase. The fusion protein self-assembled into a supramolecular assembly when illuminated at 470 nm. Scanning electron microscopy showed that the assembly formed a circular sheet structure. Self-assembly was also induced by light in E. coli. Dynamic light scattering and turbidity assay experiments showed that the assemblies formed within a few seconds under 470-nm light and completely disassembled within 5 min in the dark. Assembly and disassembly could be maintained for at least five cycles. Both in vitro and in vivo, the assemblies retained 90% of the initial activity of nitrilase and could be reused at least four times in vitro with 90% activity. Conclusions An efficient method was developed for the photocontrolled assembly and disassembly of dodecamer nitrilase and for scaffold-free reversible self-assembly of multiple oligomeric enzymes in vivo and in vitro, providing new ideas and methods for immobilization of enzyme without carrier. |
topic |
Disassembly iLID Nitrilase Photoswitch Self-assembly |
url |
http://link.springer.com/article/10.1186/s40643-017-0167-3 |
work_keys_str_mv |
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