The single berberine bridge enzyme homolog of Physcomitrella patens is a cellobiose oxidase

Marina Toplak, Gertrud Wiedemann, Jelena Ulićević, Bastian Daniel, Sebastian N W Hoernstein, Jennifer Kothe, Johannes Niederhauser, Ralf Reski, Andreas Winkler, Peter Macheroux

Research output: Contribution to journalArticleResearchpeer-review

Abstract

The berberine bridge enzyme from the California poppy Eschscholzia californica (EcBBE) 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 EcBBE in Physcomitrella patens (PpBBE1), the most basal plant harboring a BBE-like gene. Here, we report the biochemical, structural, and in vivo characterization of PpBBE1. Our studies revealed that PpBBE1 is structurally and biochemically very similar to EcBBE. In contrast to EcBBE, we found that PpBBE1 catalyzes the oxidation of the disaccharide cellobiose to the corresponding lactone, that is, PpBBE1 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 PpBBE1. In vivo experiments revealed the highest level of PpBBE1 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.

Original languageEnglish
Pages (from-to)1923-1943
Number of pages21
JournalThe FEBS journal
Volume285
Issue number10
DOIs
Publication statusPublished - May 2018

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Biosynthesis
Alkaloids
Cellulose
Eschscholzia
Genes
Benzylisoquinolines
Berberine
Cellobiose
Mutagenesis
Disaccharides
Cyclization
Lactones
Enzymes
Metabolism
Life cycle
Carbon
Bryopsida
Carbohydrates
Bryophyta
Amino Acids

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The single berberine bridge enzyme homolog of Physcomitrella patens is a cellobiose oxidase. / Toplak, Marina; Wiedemann, Gertrud; Ulićević, Jelena; Daniel, Bastian; Hoernstein, Sebastian N W; Kothe, Jennifer; Niederhauser, Johannes; Reski, Ralf; Winkler, Andreas; Macheroux, Peter.

In: The FEBS journal, Vol. 285, No. 10, 05.2018, p. 1923-1943.

Research output: Contribution to journalArticleResearchpeer-review

Toplak, M, Wiedemann, G, Ulićević, J, Daniel, B, Hoernstein, SNW, Kothe, J, Niederhauser, J, Reski, R, Winkler, A & Macheroux, P 2018, 'The single berberine bridge enzyme homolog of Physcomitrella patens is a cellobiose oxidase' The FEBS journal, vol. 285, no. 10, pp. 1923-1943. https://doi.org/10.1111/febs.14458
Toplak M, Wiedemann G, Ulićević J, Daniel B, Hoernstein SNW, Kothe J et al. The single berberine bridge enzyme homolog of Physcomitrella patens is a cellobiose oxidase. The FEBS journal. 2018 May;285(10):1923-1943. https://doi.org/10.1111/febs.14458
Toplak, Marina ; Wiedemann, Gertrud ; Ulićević, Jelena ; Daniel, Bastian ; Hoernstein, Sebastian N W ; Kothe, Jennifer ; Niederhauser, Johannes ; Reski, Ralf ; Winkler, Andreas ; Macheroux, Peter. / The single berberine bridge enzyme homolog of Physcomitrella patens is a cellobiose oxidase. In: The FEBS journal. 2018 ; Vol. 285, No. 10. pp. 1923-1943.
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title = "The single berberine bridge enzyme homolog of Physcomitrella patens is a cellobiose oxidase",
abstract = "The berberine bridge enzyme from the California poppy Eschscholzia californica (EcBBE) 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 EcBBE in Physcomitrella patens (PpBBE1), the most basal plant harboring a BBE-like gene. Here, we report the biochemical, structural, and in vivo characterization of PpBBE1. Our studies revealed that PpBBE1 is structurally and biochemically very similar to EcBBE. In contrast to EcBBE, we found that PpBBE1 catalyzes the oxidation of the disaccharide cellobiose to the corresponding lactone, that is, PpBBE1 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 PpBBE1. In vivo experiments revealed the highest level of PpBBE1 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.",
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AU - Toplak, Marina

AU - Wiedemann, Gertrud

AU - Ulićević, Jelena

AU - Daniel, Bastian

AU - Hoernstein, Sebastian N W

AU - Kothe, Jennifer

AU - Niederhauser, Johannes

AU - Reski, Ralf

AU - Winkler, Andreas

AU - Macheroux, Peter

N1 - © 2018 The Authors. The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.

PY - 2018/5

Y1 - 2018/5

N2 - The berberine bridge enzyme from the California poppy Eschscholzia californica (EcBBE) 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 EcBBE in Physcomitrella patens (PpBBE1), the most basal plant harboring a BBE-like gene. Here, we report the biochemical, structural, and in vivo characterization of PpBBE1. Our studies revealed that PpBBE1 is structurally and biochemically very similar to EcBBE. In contrast to EcBBE, we found that PpBBE1 catalyzes the oxidation of the disaccharide cellobiose to the corresponding lactone, that is, PpBBE1 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 PpBBE1. In vivo experiments revealed the highest level of PpBBE1 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.

AB - The berberine bridge enzyme from the California poppy Eschscholzia californica (EcBBE) 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 EcBBE in Physcomitrella patens (PpBBE1), the most basal plant harboring a BBE-like gene. Here, we report the biochemical, structural, and in vivo characterization of PpBBE1. Our studies revealed that PpBBE1 is structurally and biochemically very similar to EcBBE. In contrast to EcBBE, we found that PpBBE1 catalyzes the oxidation of the disaccharide cellobiose to the corresponding lactone, that is, PpBBE1 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 PpBBE1. In vivo experiments revealed the highest level of PpBBE1 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.

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