Thermal Stabilization of an Endoglucanase by Cyclization
An intein-driven protein splicing approach allowed for the covalent linkage between the N- and C-termini of a polypeptide chain to create circular variants of the endo-ß-1,3-1,4-glucanase, LicA, from Bacillus licheniformis. Two circular variants, LicA-C1 and LicA-C2, which have connecting loops of 20 and 14 amino acids, respectively, showed catalytic activities that are approximately two and three times higher, respectively, compared to that of the linear LicA (LicA-L1). The thermal stability of the circular variants was significantly increased compared to the linear form. Whereas the linear glucanase lost half of its activity after 3 min at 65 °C, the two circular variants have 6-fold (LicA-C1) and 16-fold (LicA-C2) increased half-life time of inactivation. In agreement with this, fluorescence spectroscopy and differential scanning calorimetry studies revealed that circular enzymes undergo structural changes at higher temperatures compared to that of the linear form. The effect of calcium on the conformational stability and function of the circular LicAs was also investigated, and we observed that the presence of calcium ions results in increased thermal stability. The impact of the length of the designed loops on thermal stability of the circular proteins is discussed, and it is suggested that cyclization may be an efficient strategy for the increased stability of proteins
Main Authors: | , , , , , , |
---|---|
Format: | Article/Letter to editor biblioteca |
Language: | English |
Subjects: | bacillus-licheniformis, beta-glucosidase celb, circular proteins, crystal-structure, in-vivo, intein, pi-pfui, pyrococcus-furiosus, site-directed mutagenesis, stability, |
Online Access: | https://research.wur.nl/en/publications/thermal-stabilization-of-an-endoglucanase-by-cyclization |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
id |
dig-wur-nl-wurpubs-427293 |
---|---|
record_format |
koha |
spelling |
dig-wur-nl-wurpubs-4272932024-06-25 van Lieshout, J.F.T. Gutierrez, O. Vroom, W. Planas, A. de Vos, W.M. van der Oost, J. Koutsopoulos, S. Article/Letter to editor Applied Biochemistry and Biotechnology 167 (2012) 7 ISSN: 0273-2289 Thermal Stabilization of an Endoglucanase by Cyclization 2012 An intein-driven protein splicing approach allowed for the covalent linkage between the N- and C-termini of a polypeptide chain to create circular variants of the endo-ß-1,3-1,4-glucanase, LicA, from Bacillus licheniformis. Two circular variants, LicA-C1 and LicA-C2, which have connecting loops of 20 and 14 amino acids, respectively, showed catalytic activities that are approximately two and three times higher, respectively, compared to that of the linear LicA (LicA-L1). The thermal stability of the circular variants was significantly increased compared to the linear form. Whereas the linear glucanase lost half of its activity after 3 min at 65 °C, the two circular variants have 6-fold (LicA-C1) and 16-fold (LicA-C2) increased half-life time of inactivation. In agreement with this, fluorescence spectroscopy and differential scanning calorimetry studies revealed that circular enzymes undergo structural changes at higher temperatures compared to that of the linear form. The effect of calcium on the conformational stability and function of the circular LicAs was also investigated, and we observed that the presence of calcium ions results in increased thermal stability. The impact of the length of the designed loops on thermal stability of the circular proteins is discussed, and it is suggested that cyclization may be an efficient strategy for the increased stability of proteins en application/pdf https://research.wur.nl/en/publications/thermal-stabilization-of-an-endoglucanase-by-cyclization 10.1007/s12010-012-9674-z https://edepot.wur.nl/222446 bacillus-licheniformis beta-glucosidase celb circular proteins crystal-structure in-vivo intein pi-pfui pyrococcus-furiosus site-directed mutagenesis stability Wageningen University & Research |
institution |
WUR NL |
collection |
DSpace |
country |
Países bajos |
countrycode |
NL |
component |
Bibliográfico |
access |
En linea |
databasecode |
dig-wur-nl |
tag |
biblioteca |
region |
Europa del Oeste |
libraryname |
WUR Library Netherlands |
language |
English |
topic |
bacillus-licheniformis beta-glucosidase celb circular proteins crystal-structure in-vivo intein pi-pfui pyrococcus-furiosus site-directed mutagenesis stability bacillus-licheniformis beta-glucosidase celb circular proteins crystal-structure in-vivo intein pi-pfui pyrococcus-furiosus site-directed mutagenesis stability |
spellingShingle |
bacillus-licheniformis beta-glucosidase celb circular proteins crystal-structure in-vivo intein pi-pfui pyrococcus-furiosus site-directed mutagenesis stability bacillus-licheniformis beta-glucosidase celb circular proteins crystal-structure in-vivo intein pi-pfui pyrococcus-furiosus site-directed mutagenesis stability van Lieshout, J.F.T. Gutierrez, O. Vroom, W. Planas, A. de Vos, W.M. van der Oost, J. Koutsopoulos, S. Thermal Stabilization of an Endoglucanase by Cyclization |
description |
An intein-driven protein splicing approach allowed for the covalent linkage between the N- and C-termini of a polypeptide chain to create circular variants of the endo-ß-1,3-1,4-glucanase, LicA, from Bacillus licheniformis. Two circular variants, LicA-C1 and LicA-C2, which have connecting loops of 20 and 14 amino acids, respectively, showed catalytic activities that are approximately two and three times higher, respectively, compared to that of the linear LicA (LicA-L1). The thermal stability of the circular variants was significantly increased compared to the linear form. Whereas the linear glucanase lost half of its activity after 3 min at 65 °C, the two circular variants have 6-fold (LicA-C1) and 16-fold (LicA-C2) increased half-life time of inactivation. In agreement with this, fluorescence spectroscopy and differential scanning calorimetry studies revealed that circular enzymes undergo structural changes at higher temperatures compared to that of the linear form. The effect of calcium on the conformational stability and function of the circular LicAs was also investigated, and we observed that the presence of calcium ions results in increased thermal stability. The impact of the length of the designed loops on thermal stability of the circular proteins is discussed, and it is suggested that cyclization may be an efficient strategy for the increased stability of proteins |
format |
Article/Letter to editor |
topic_facet |
bacillus-licheniformis beta-glucosidase celb circular proteins crystal-structure in-vivo intein pi-pfui pyrococcus-furiosus site-directed mutagenesis stability |
author |
van Lieshout, J.F.T. Gutierrez, O. Vroom, W. Planas, A. de Vos, W.M. van der Oost, J. Koutsopoulos, S. |
author_facet |
van Lieshout, J.F.T. Gutierrez, O. Vroom, W. Planas, A. de Vos, W.M. van der Oost, J. Koutsopoulos, S. |
author_sort |
van Lieshout, J.F.T. |
title |
Thermal Stabilization of an Endoglucanase by Cyclization |
title_short |
Thermal Stabilization of an Endoglucanase by Cyclization |
title_full |
Thermal Stabilization of an Endoglucanase by Cyclization |
title_fullStr |
Thermal Stabilization of an Endoglucanase by Cyclization |
title_full_unstemmed |
Thermal Stabilization of an Endoglucanase by Cyclization |
title_sort |
thermal stabilization of an endoglucanase by cyclization |
url |
https://research.wur.nl/en/publications/thermal-stabilization-of-an-endoglucanase-by-cyclization |
work_keys_str_mv |
AT vanlieshoutjft thermalstabilizationofanendoglucanasebycyclization AT gutierrezo thermalstabilizationofanendoglucanasebycyclization AT vroomw thermalstabilizationofanendoglucanasebycyclization AT planasa thermalstabilizationofanendoglucanasebycyclization AT devoswm thermalstabilizationofanendoglucanasebycyclization AT vanderoostj thermalstabilizationofanendoglucanasebycyclization AT koutsopouloss thermalstabilizationofanendoglucanasebycyclization |
_version_ |
1813202819333750784 |