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Untersuchungen zu rekombinanten eukaryotischen Baeyer-Villiger-Monooxygenasen / von Friedemann Leipold

Baeyer-Villiger-Oxidation, Monooxygenasen, Rhodococcus rhodochrous, Heterologe Genexpression, Enzymkatalyse, Substratspezifität, Butyrolactonderivate, Cycloalkanonmonooxygenase, Steroidmonooxygenase, Cylindrocarpon radicicola, CODEHOP, rationales Proteindesign, Baeyer-Villiger monooxygenase, cycloal... Full description

PPN (Catalogue-ID): 740825186
Personen: Leipold, Friedemann
Format: eBook eBook
Language: German
Published: 2012
Hochschule: Greifswald, Univ., Diss., 2012
Basisklassifikation: 35.74
Subjects:

Baeyer-Villiger-Oxidation / Monooxygenasen / Rhodococcus rhodochrous / Heterologe Genexpression / Enzymkatalyse / Substratspezifität / Butyrolactonderivate

Formangabe: Hochschulschrift
Physical Description: Online-Ressource (PDF-Datei: 174 S., 16,7 MB)
Technische Details: Systemvoraussetzungen: Acrobat reader.

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520 |a In this thesis, Baeyer-Villiger monooxygenase (BVMO) genes from Cylindrocarpon radicicola ATCC 11011 were identified and the corresponding enzymes characterized in comparison to prokaryotic BVMOs. Thus, one aim of this work was an evaluation of the enzymatic capability of this filamentous fungus with regard to the biocatalytic Baeyer-Villiger oxidation. Due to the fact that the genome of C. radicicola has not been sequenced yet, methods of protein purification as well as identification via molecular biological approaches were pursued for the discovery of new BVMO sequences. However, the application of a three-step purification protocol used for the cell-free extract of C. radicicola did not result in protein of sufficient purity which was mandatory for subsequent investigations. Therefore, the identification of BVMO sequences was carried out based on molecular biological methods. Using degenerate primers based on conserved sequence motifs of known BVMOs, and which were derived by means of the CODEHOP approach, three sequences homologous to BVMOs were successfully amplified and identified. Two of the identified sequences were homologous to putative steroid monooxygenases (STMOs) from fungal origins. However, these sequences were assigned as putative cryptic genes based on sequence comparison. For this reason, no further studies were carried out on these two sequences. However, one of the sequences newly identified from C. radicicola was functionally overexpressed ... 
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