Increased thermal stability of a glucose oxidase biosensor under high hydrostatic pressure

Yang, Daoyuan; Olstad, Hanna E.; Reyes‐De‐Corcuera, José I.

doi:10.1016/j.enzmictec.2019.109486

Cited by 9 publications

(3 citation statements)

References 60 publications

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“…Experimental efforts to tailor the external protective microenvironment of the immobilized enzyme and the efforts on random mutagenesis to engineer the enzymatic structure itself have been devoted to enhancing the thermal stability when using GOx [ 7 , 8 , 9 ]. To effectively find an alternative way, the modification of enzymes by protein-structure engineering can judiciously tailor protein behaviors and secure the protein structure.…”

Section: Introductionmentioning

confidence: 99%

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

Ittisoponpisan

Jeerapan

2021

Bioengineering

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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 Aspergillus niger 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.

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Section: Introductionmentioning

confidence: 99%

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

Ittisoponpisan

Jeerapan

2021

Bioengineering

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“…Most structural studies of the effects of HHP in enzymes in solution have been done on small proteins (< ~30 kDa) in part because they are amenable to a variety of structural analyses including nuclear magnetic resonance (NMR) (Pastore & Temussi, 2023). We have studied the effects of HHP on the thermal stability of glucose oxidase (GOx) a 160 kDa dimer (Yang et al, 2020), alcohol oxidase (AOx) a 675 kDa octamer (Buchholz-Afari et al, 2019), and xanthine oxidase (XOx), a 283 kDa dimer (Halalipour et al, 2017a(Halalipour et al, , 2017b) by studying the decrease in the rate of inactivation. The optimal pressure for stability and the extent of stabilization were different for each oxidase.…”

Section: Introductionmentioning

confidence: 99%

Stabilization of galactose oxidase by high hydrostatic pressure: Insights on the role of cavities size

Kang,

Reyes‐De‐Corcuera

2024

Biotech & Bioengineering

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High hydrostatic pressure stabilized galactose oxidase (GaOx) at 70.0–80.0°C against thermal inactivation. The pseudo‐first‐order rate constant of inactivation kinact decreased by a factor of 8 at 80°C and by a factor of 44 at 72.5°C. The most pronounced effect of pressure was at the lowest studied temperature of 70.0°C with an activation volume of inactivation ΔV‡ of 78.8 cm3 mol−1. The optimal pressure against thermal inactivation was between 200 and 300 MPa. Unlike other enzymes, as temperature increased the ΔV‡ of inactivation decreased, and as pressure increased the activation energy of inactivation Eai increased. Combining the results for GaOx with earlier research on the pressure‐induced stabilization of other enzymes suggests that ΔV‡ of inactivation correlates with the total molar volume of cavities larger than ~100 Å3 in enzyme monomers for enzymes near the optimal pH and whose thermal unfolding is not accompanied by oligomer dissociation.

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“…Nesta técnica, a modificação da superfície da matriz com agentes bifuncionais são requeridos para aumentar a eficiência da imobilização, uma vez que sem os agentes, as ligações entre matriz e enzima são fracas. O biocatalisador é formado com alta concentração de enzima e devido aos agentes bifuncionais, este se torna estável (Resende et al, 2017;Capecchi et al, 2018;Olstad;Reyes-de-Corcuera, 2020).…”

Section: Imobilizaçãounclassified

Produção de biocatalisador através da técnica de adsorção física a partir da lipase de Burkholderia cepacia e o estudo da influência de aditivos no processo de imobilização

Moreira Correa

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Increased thermal stability of a glucose oxidase biosensor under high hydrostatic pressure

Cited by 9 publications

References 60 publications

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

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

Stabilization of galactose oxidase by high hydrostatic pressure: Insights on the role of cavities size

Produção de biocatalisador através da técnica de adsorção física a partir da lipase de Burkholderia cepacia e o estudo da influência de aditivos no processo de imobilização

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