In Silico Analysis of Glucose Oxidase from <i>Aspergillus niger</i>: Potential Cysteine Mutation Sites for Enhancing Protein Stability

Glucose oxidase (GOx) holds considerable advantages for various applications. Nevertheless, the thermal instability of the enzyme remains a grand challenge, impeding the success in applications outside the well-controlled laboratories, particularly in practical bioelectronics. Many strategies to mod...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Sirawit Ittisoponpisan, Itthipon Jeerapan
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	glucose oxidase glucose stability protein engineering disulfide bond mutagenesis Technology T Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/60e493618c4146a3ad3ac51349eda776
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:60e493618c4146a3ad3ac51349eda776
record_format	dspace
spelling	oai:doaj.org-article:60e493618c4146a3ad3ac51349eda7762021-11-25T16:46:42ZIn Silico Analysis of Glucose Oxidase from <i>Aspergillus niger</i>: Potential Cysteine Mutation Sites for Enhancing Protein Stability10.3390/bioengineering81101882306-5354https://doaj.org/article/60e493618c4146a3ad3ac51349eda7762021-11-01T00:00:00Zhttps://www.mdpi.com/2306-5354/8/11/188https://doaj.org/toc/2306-5354Glucose oxidase (GOx) holds considerable advantages for various applications. Nevertheless, the thermal instability of the enzyme remains a grand challenge, impeding the success in applications outside the well-controlled laboratories, particularly in practical bioelectronics. Many strategies to modify GOx to achieve better thermal stability have been proposed. However, modification of this enzyme by adding extra disulfide bonds is yet to be explored. This work describes the in silico bioengineering of GOx from <i>Aspergillus niger</i> by judiciously analyzing characteristics of disulfide bonds found in the Top8000 protein database, then scanning for amino acid residue pairs that are suitable to be replaced with cysteines in order to establish disulfide bonds. Next, we predicted and assessed the mutant GOx models in terms of disulfide bond quality (bond length and α angles), functional impact by means of residue conservation, and structural impact as indicated by Gibbs free energy. We found eight putative residue pairs that can be engineered to form disulfide bonds. Five of these are located in less conserved regions and, therefore, are unlikely to have a deleterious impact on functionality. Finally, two mutations, Pro149Cys and His158Cys, showed potential for stabilizing the protein structure as confirmed by a structure-based stability analysis tool. The findings in this study highlight the opportunity of using disulfide bond modification as a new alternative technique to enhance the thermal stability of GOx.Sirawit IttisoponpisanItthipon JeerapanMDPI AGarticleglucose oxidaseglucosestabilityprotein engineeringdisulfide bondmutagenesisTechnologyTBiology (General)QH301-705.5ENBioengineering, Vol 8, Iss 188, p 188 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	glucose oxidase glucose stability protein engineering disulfide bond mutagenesis Technology T Biology (General) QH301-705.5
spellingShingle	glucose oxidase glucose stability protein engineering disulfide bond mutagenesis Technology T Biology (General) QH301-705.5 Sirawit Ittisoponpisan Itthipon Jeerapan In Silico Analysis of Glucose Oxidase from <i>Aspergillus niger</i>: Potential Cysteine Mutation Sites for Enhancing Protein Stability
description	Glucose oxidase (GOx) holds considerable advantages for various applications. Nevertheless, the thermal instability of the enzyme remains a grand challenge, impeding the success in applications outside the well-controlled laboratories, particularly in practical bioelectronics. Many strategies to modify GOx to achieve better thermal stability have been proposed. However, modification of this enzyme by adding extra disulfide bonds is yet to be explored. This work describes the in silico bioengineering of GOx from <i>Aspergillus niger</i> by judiciously analyzing characteristics of disulfide bonds found in the Top8000 protein database, then scanning for amino acid residue pairs that are suitable to be replaced with cysteines in order to establish disulfide bonds. Next, we predicted and assessed the mutant GOx models in terms of disulfide bond quality (bond length and α angles), functional impact by means of residue conservation, and structural impact as indicated by Gibbs free energy. We found eight putative residue pairs that can be engineered to form disulfide bonds. Five of these are located in less conserved regions and, therefore, are unlikely to have a deleterious impact on functionality. Finally, two mutations, Pro149Cys and His158Cys, showed potential for stabilizing the protein structure as confirmed by a structure-based stability analysis tool. The findings in this study highlight the opportunity of using disulfide bond modification as a new alternative technique to enhance the thermal stability of GOx.
format	article
author	Sirawit Ittisoponpisan Itthipon Jeerapan
author_facet	Sirawit Ittisoponpisan Itthipon Jeerapan
author_sort	Sirawit Ittisoponpisan
title	In Silico Analysis of Glucose Oxidase from <i>Aspergillus niger</i>: Potential Cysteine Mutation Sites for Enhancing Protein Stability
title_short	In Silico Analysis of Glucose Oxidase from <i>Aspergillus niger</i>: Potential Cysteine Mutation Sites for Enhancing Protein Stability
title_full	In Silico Analysis of Glucose Oxidase from <i>Aspergillus niger</i>: Potential Cysteine Mutation Sites for Enhancing Protein Stability
title_fullStr	In Silico Analysis of Glucose Oxidase from <i>Aspergillus niger</i>: Potential Cysteine Mutation Sites for Enhancing Protein Stability
title_full_unstemmed	In Silico Analysis of Glucose Oxidase from <i>Aspergillus niger</i>: Potential Cysteine Mutation Sites for Enhancing Protein Stability
title_sort	in silico analysis of glucose oxidase from <i>aspergillus niger</i>: potential cysteine mutation sites for enhancing protein stability
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/60e493618c4146a3ad3ac51349eda776
work_keys_str_mv	AT sirawitittisoponpisan insilicoanalysisofglucoseoxidasefromiaspergillusnigeripotentialcysteinemutationsitesforenhancingproteinstability AT itthiponjeerapan insilicoanalysisofglucoseoxidasefromiaspergillusnigeripotentialcysteinemutationsitesforenhancingproteinstability
_version_	1718412957852893184

In Silico Analysis of Glucose Oxidase from <i>Aspergillus niger</i>: Potential Cysteine Mutation Sites for Enhancing Protein Stability

Ejemplares similares