Müller, Franziska Maria; Marke, Gina; Drees, Steffen Lorenz; Wöhlbrand, Lars; Rabus, Ralf; Philipp, Bodo: Substrate Inhibition of 5β-Δ4-3-Ketosteroid Dehydrogenase in Sphingobium sp. Strain Chol11 Acts as Circuit Breaker During Growth With Toxic Bile Salts
Inhalt
- Materials and Methods
- Cultivation of Bacteria
- Growth Experiments
- Cell Suspension Experiments
- Cloning Techniques and Construction of Unmarked Gene Deletions
- Preparation of Cell Extracts
- Purification of 5β-Δ 4 -KSTD1
- Enzyme Assays
- Preparation of Steroid Compounds
- Analytical Methods
- Bioinformatical Methods
- Results
- Proteome Analysis Reveals a Candidate for 5β-Δ 4 -KSTD
- Nov2c019 Displays KSTD Activity When Expressed Heterologously
- Purified 5β-Δ 4 -KSTD1 Efficiently Oxidizes 5β-3-Ketosteroids to Respective Δ 4 -3-Ketosteroids
- 5β-Δ 4 -KSTD1 Displays Substrate Inhibition
- Phenotype of the Unmarked Deletion Mutant of 5β-Δ 4 -kstd1 Shows Reduced Oxidation of 3-Ketocholate to Δ 4 -3-Ketocholate
- 5β-Δ 4 -KSTD1 Homologs Are Present in Many Bile-Salt Degrading Bacteria
- Discussion
- Data Availability Statement
- Author Contributions
- References