Abstract
DyP-type peroxidases are heme-containing enzymes that have received increasing attention over recent years with regards to their potential as biocatalysts. A novel DyP-type peroxidase (CboDyP) was discovered from the alkaliphilic cellulomonad, Cellulomonas bogoriensis, which could be overexpressed in Escherichia coli. The biochemical characterization of the recombinant enzyme showed that it is a heme-containing enzyme capable to act as a peroxidase on several dyes. With the tested substrates, the enzyme is most active at acidic pH values and is quite tolerant towards solvents. The crystal structure of CboDyP was solved which revealed atomic details of the dimeric heme-containing enzyme. A peculiar feature of CboDyP is the presence of a glutamate in the active site which in most other DyPs is an aspartate, being part of the DyP-typifying sequence motif GXXDG. The E201D CboDyP mutant was prepared and analyzed which revealed that the mutant enzyme shows a significantly higher activity on several dyes when compared with the wild-type enzyme.
Highlights
DyP-type peroxidases (DyPs) are heme-containing enzymes known for their ability to degrade dyes through their peroxidase activity
Except for typical class A sequence features such as a Tat-signal sequence and the presence of a conserved His to interact with the iron in the heme cofactor, the CboDyP sequence has an aberrant sequence in the region where normally a GXXDG motif is found
It has been reported to play a role as a proton acceptor during the heterolytic cleavage of hydrogen peroxide. It forms a hydrogen bond with the distal solvent species and was shown to be essential for catalysis. [1,3,4] In this paper, we present a full characterization of CboDyP concerning its peroxidase activity on several dyes and its thermostability using different solvents
Summary
DyP-type peroxidases (DyPs) are heme-containing enzymes known for their ability to degrade dyes through their peroxidase activity. Recent studies have shown that DyPs are involved in lignin degradation and can be used for oxidation of various compounds such as carotenoids, phenols, and aromatic sulfides. An example of a type A DyP is the peroxidase from Thermobifida fusca known as TfuDyP [1]. While this was the first reported bacterial DyP, several other DyPs have been reported in the last decade. Except for typical class A sequence features such as a Tat-signal sequence and the presence of a conserved His to interact with the iron in the heme cofactor, the CboDyP sequence has an aberrant sequence in the region where normally a GXXDG motif is found. We elucidated its crystal structure, and probed the role of the aberrant glutamate in its active site
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