Heterologous Expression and Biochemical Characterization of a New Chloroperoxidase Isolated from the Deep-Sea Hydrothermal Vent Black Yeast Hortaea werneckii UBOCC-A-208029
Bastien Cochereau
(1, 2)
,
Yoran Le Strat
(1, 2)
,
Qiaolin Ji
(1, 2)
,
Audrey Pawtowski
(1)
,
Ludovic Delage
(3)
,
Amélie Weill
(1)
,
Lisa Mazéas
(3)
,
Cécile Hervé
(3)
,
Gaëtan Burgaud
(1)
,
Nina Gunde-Cimerman
(4)
,
Yves François Pouchus
(2)
,
Nathalie Demont-Caulet
(5)
,
Catherine Roullier
(2)
,
Laurence Meslet-Cladiere
(1)
1
LUBEM -
Laboratoire Universitaire de Biodiversité et Ecologie Microbienne
2 ISOMER - Institut Des Substances et Organismes de la Mer - UR 2160
3 LBI2M - Laboratoire de Biologie Intégrative des Modèles Marins
4 Molecular Genetics and Biology of Microorganisms
5 ECOSYS - Ecologie fonctionnelle et écotoxicologie des agroécosystèmes
2 ISOMER - Institut Des Substances et Organismes de la Mer - UR 2160
3 LBI2M - Laboratoire de Biologie Intégrative des Modèles Marins
4 Molecular Genetics and Biology of Microorganisms
5 ECOSYS - Ecologie fonctionnelle et écotoxicologie des agroécosystèmes
Bastien Cochereau
- Fonction : Auteur
- PersonId : 805863
- ORCID : 0000-0003-1073-8140
- IdRef : 269865004
Yoran Le Strat
- Fonction : Auteur
- PersonId : 1313285
- IdRef : 273350978
Cécile Hervé
- Fonction : Auteur
- PersonId : 2698
- IdHAL : cecile-herve
- ORCID : 0000-0001-6649-8137
- IdRef : 101613873
Yves François Pouchus
- Fonction : Auteur
- PersonId : 1358970
- ORCID : 0000-0001-8062-227X
- IdRef : 060117095
Catherine Roullier
- Fonction : Auteur
- PersonId : 739737
- IdHAL : catherine-roullier
- ORCID : 0000-0001-6486-6821
- IdRef : 113167393
Résumé
The initiation of this study relies on a targeted genome-mining approach to highlight the presence of a putative vanadium-dependent haloperoxidase-encoding gene in the deep-sea hydrothermal vent fungus Hortaea werneckii UBOCC-A-208029. To date, only three fungal vanadium-dependent haloperoxidases have been described, one from the terrestrial species Curvularia inaequalis, one from the fungal plant pathogen Botrytis cinerea , and one from a marine derived isolate identified as Alternaria didymospora . In this study, we describe a new vanadium chloroperoxidase from the black yeast H. werneckii , successfully cloned and overexpressed in a bacterial host, which possesses higher affinity for bromide (K m = 26 µM) than chloride (K m = 237 mM). The enzyme was biochemically characterized, and we have evaluated its potential for biocatalysis by determining its stability and tolerance in organic solvents. We also describe its potential three-dimensional structure by building a model using the AlphaFold 2 artificial intelligence tool. This model shows some conservation of the 3D structure of the active site compared to the vanadium chloroperoxidase from C. inaequalis but it also highlights some differences in the active site entrance and the volume of the active site pocket, underlining its originality.
Format du dépôt | Fichier |
---|---|
Type de dépôt | Article dans une revue |
Titre |
en
Heterologous Expression and Biochemical Characterization of a New Chloroperoxidase Isolated from the Deep-Sea Hydrothermal Vent Black Yeast Hortaea werneckii UBOCC-A-208029
|
Résumé |
en
The initiation of this study relies on a targeted genome-mining approach to highlight the presence of a putative vanadium-dependent haloperoxidase-encoding gene in the deep-sea hydrothermal vent fungus Hortaea werneckii UBOCC-A-208029. To date, only three fungal vanadium-dependent haloperoxidases have been described, one from the terrestrial species Curvularia inaequalis, one from the fungal plant pathogen Botrytis cinerea , and one from a marine derived isolate identified as Alternaria didymospora . In this study, we describe a new vanadium chloroperoxidase from the black yeast H. werneckii , successfully cloned and overexpressed in a bacterial host, which possesses higher affinity for bromide (K m = 26 µM) than chloride (K m = 237 mM). The enzyme was biochemically characterized, and we have evaluated its potential for biocatalysis by determining its stability and tolerance in organic solvents. We also describe its potential three-dimensional structure by building a model using the AlphaFold 2 artificial intelligence tool. This model shows some conservation of the 3D structure of the active site compared to the vanadium chloroperoxidase from C. inaequalis but it also highlights some differences in the active site entrance and the volume of the active site pocket, underlining its originality.
|
Auteur(s) |
Bastien Cochereau
1, 2
, Yoran Le Strat
1, 2
, Qiaolin Ji
1, 2
, Audrey Pawtowski
1
, Ludovic Delage
3
, Amélie Weill
1
, Lisa Mazéas
3
, Cécile Hervé
3
, Gaëtan Burgaud
1
, Nina Gunde-Cimerman
4
, Yves François Pouchus
2
, Nathalie Demont-Caulet
5
, Catherine Roullier
2
, Laurence Meslet-Cladiere
1
1
LUBEM -
Laboratoire Universitaire de Biodiversité et Ecologie Microbienne
( 120369 )
- ESIAB - Parvis Blaise Pascal - Technopôle Brest-Iroise - 29280 Plouzané
- France
2
ISOMER -
Institut Des Substances et Organismes de la Mer - UR 2160
( 1089893 )
- 2, rue de la Houssinière BP 81227 44322 NANTES CEDEX 3 France
- France
3
LBI2M -
Laboratoire de Biologie Intégrative des Modèles Marins
( 542078 )
- Station Biologique de Roscoff Place Georges Teissier 29680 Roscoff
- France
4
Molecular Genetics and Biology of Microorganisms
( 1175858 )
- Slovénie
5
ECOSYS -
Ecologie fonctionnelle et écotoxicologie des agroécosystèmes
( 1002424 )
- 22 place de l'Agronomie CS 20040 91123 Palaiseau cedex
- France
|
Langue du document |
Anglais
|
Nom de la revue |
|
Volume |
25
|
Numéro |
4
|
Page/Identifiant |
519-536
|
Vulgarisation |
Non
|
Comité de lecture |
Oui
|
Audience |
Internationale
|
Date de publication |
2023
|
Public visé |
Scientifique
|
Domaine(s) |
|
Mots-clés |
en
Marine fungi, Black yeast, Halogens, Enzymes, Vanadium chloroperoxidase
|
DOI | 10.1007/s10126-023-10222-7 |
Pubmed Id | 37354383 |
UT key WOS | 001016142600002 |
Origine :
Publication financée par une institution
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