New design of a chimeric enzyme based on vanadium-dependent bromoperoxidase and glucose oxidase to highlight the microbicidal properties of the reaction products: HOBr and HOCl
Abstract
Vanadium-dependent haloperoxidases are enzymes found in bacteria, fungi and red or brown macroalgae. They are used as a defense system for the latter by producing bromoform. The intermediate product, HOBr or HOCl, is also a microbicidal compound that can be exploited in many domains as medical tools or disinfectant sprays for example. We show that the mutation identified in Ohshiro's work, which alters substrate specificity toward chloride, is transferable to the homologous enzyme from Chondrus crispus. We then applied genetic and enzymatic engineering to design a chimera with glucose oxidase, thereby enhancing antimicrobial properties by providing a local source of H 2 O 2 . We used the SpyTag/SpyCatcher technology to form the chimera and obtained homogenous objects, indicating that one oligomeric form is favored. We demonstrated the release of HOBr and HOCl thanks to NADH that reacts spontaneously with it outside the active site of the enzyme. We also measured the steady-state kinetic parameters of the wild-type or modified enzymes. The formation of the chimera increased the specificity of ccVHPO1 inside the chimera, towards KBr or H 2 O 2 , compared to the enzyme alone with the SpyCatcher. Finally, we showed a significant increase in the microbicidal effect between a coupled enzymatic system with the glucose oxidase (the two enzymes are free in solution) compared to the chimera system, which is completely bactericidal at concentrations around 20 nM.
Related
- Evaluation of the immunogenic properties of a chimeric protein against West Nile virus: in-silico analysis and assessment in mouse
- Highlight: Balancing selection, more frequent than anticipated?
- Tryptophan oxidation-induced dynamics highlight why stable γS-crystallin variants aggregate
- Chimeric archaeal BHB-human pre-tRNA scaffold HARBOR system enables versatile production of functional circular RNA modalities
- Seizure recruitment properties are dependent upon dynamotype: A modeling study
BibTeX
@article{amalric2026design,
title = {{New design of a chimeric enzyme based on vanadium-dependent bromoperoxidase and glucose oxidase to highlight the microbicidal properties of the reaction products: HOBr and HOCl}},
author = {Valentin Amalric and Sébastien Gounel and Aurélien Thureau and Gilles Pécastaings and Paul J Branham and Vinayak Agarwal and Claire Stines-Chaumeil},
journal = {Journal of Biotechnology},
year = {2026},
month = aug,
doi = {10.1016/j.jbiotec.2026.08.002},
url = {https://hal.science/hal-05732358v1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - New design of a chimeric enzyme based on vanadium-dependent bromoperoxidase and glucose oxidase to highlight the microbicidal properties of the reaction products: HOBr and HOCl
AU - Valentin Amalric
AU - Sébastien Gounel
AU - Aurélien Thureau
AU - Gilles Pécastaings
AU - Paul J Branham
AU - Vinayak Agarwal
AU - Claire Stines-Chaumeil
PY - 2026
DA - 2026/08/31
JO - Journal of Biotechnology
DO - 10.1016/j.jbiotec.2026.08.002
UR - https://hal.science/hal-05732358v1
AB - Vanadium-dependent haloperoxidases are enzymes found in bacteria, fungi and red or brown macroalgae. They are used as a defense system for the latter by producing bromoform. The intermediate product, HOBr or HOCl, is also a microbicidal compound that can be exploited in many domains as medical tools or disinfectant sprays for example. We show that the mutation identified in Ohshiro's work, which alters substrate specificity toward chloride, is transferable to the homologous enzyme from Chondrus crispus. We then applied genetic and enzymatic engineering to design a chimera with glucose oxidase, thereby enhancing antimicrobial properties by providing a local source of H 2 O 2 . We used the SpyTag/SpyCatcher technology to form the chimera and obtained homogenous objects, indicating that one oligomeric form is favored. We demonstrated the release of HOBr and HOCl thanks to NADH that reacts spontaneously with it outside the active site of the enzyme. We also measured the steady-state kinetic parameters of the wild-type or modified enzymes. The formation of the chimera increased the specificity of ccVHPO1 inside the chimera, towards KBr or H 2 O 2 , compared to the enzyme alone with the SpyCatcher. Finally, we showed a significant increase in the microbicidal effect between a coupled enzymatic system with the glucose oxidase (the two enzymes are free in solution) compared to the chimera system, which is completely bactericidal at concentrations around 20 nM.
SN - 0168-1656
ER -