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The single berberine bridge enzyme homolog of Physcomitrella patens is a cellobiose oxidase
Author(s) -
Toplak Marina,
Wiedemann Gertrud,
Ulićević Jelena,
Daniel Bastian,
Hoernstein Sebastian N. W.,
Kothe Jennifer,
Niederhauser Johannes,
Reski Ralf,
Winkler Andreas,
Macheroux Peter
Publication year - 2018
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/febs.14458
Subject(s) - biochemistry , enzyme , biosynthesis , cellobiose , biology , chemistry , stereochemistry , cellulase
The berberine bridge enzyme from the California poppy Eschscholzia californica ( Ec BBE ) catalyzes the oxidative cyclization of ( S )‐reticuline to ( S )‐scoulerine, that is, the formation of the berberine bridge in the biosynthesis of benzylisoquinoline alkaloids. Interestingly, a large number of BBE ‐ like genes have been identified in plants that lack alkaloid biosynthesis. This finding raised the question of the primordial role of BBE in the plant kingdom, which prompted us to investigate the closest relative of Ec BBE in Physcomitrella patens ( Pp BBE 1), the most basal plant harboring a BBE ‐ like gene. Here, we report the biochemical, structural, and in vivo characterization of Pp BBE 1. Our studies revealed that Pp BBE 1 is structurally and biochemically very similar to Ec BBE . In contrast to Ec BBE , we found that Pp BBE 1 catalyzes the oxidation of the disaccharide cellobiose to the corresponding lactone, that is, Pp BBE 1 is a cellobiose oxidase. The enzymatic reaction mechanism was characterized by a structure‐guided mutagenesis approach that enabled us to assign a catalytic role to amino acid residues in the active site of Pp BBE 1. In vivo experiments revealed the highest level of Pp BBE 1 expression in chloronema, the earliest stage of the plant's life cycle, where carbon metabolism is strongly upregulated. It was also shown that the enzyme is secreted to the extracellular space, where it may be involved in later steps of cellulose degradation, thereby allowing the moss to make use of cellulose for energy production. Overall, our results suggest that the primordial role of BBE ‐like enzymes in plants revolved around primary metabolic reactions in carbohydrate utilization. Database Structural data are available in the PDB under the accession numbers 6EO4 and 6EO5 .