EC Number   |
Protein Variants |
Reference |
|---|
    1.3.99.4 | more |
development of a synthetic 3-ketosteroid DELTA1-dehydrogenase (DELTA1-KstDR) for the generation of a catabolic pathway enabling cholesterol degradation in human cells. Coexpression with cholesterol-3-hydroxy-dehydrogenase (CholD) and 3-oxosteroid-9alpha-hydroxylase (Kst-9alphaH). CholD activity catalyzes the generation of a C-3 ketone, which is needed for DELTA1-KstD activity, which introduces a double bond between the C-1 and C-2 atoms of 3-oxosteroids. Cholesterol catabolism pathway in Actinomycetes, overview. The downstream enzymes need to have equal or greater activity when compared to the enzymes acting upstream to allow the pathway to flow and to prevent the accumulation of toxic intermediates, kinetic study |
763735 |
| 1.3.99.4 | more |
enzyme from strain ST-095 only differs in residue V366 from strain JC-12 (S366). Mutation results in almost 3fold higher catalytic efficiency in strain JC-12 |
-, 743031 |
| 1.3.99.4 | more |
enzyme mutants are constructed based on in silico protein docking modeling using site-directed mutagenesis and exogenous expression |
-, 763772 |
| 1.3.99.4 | more |
targeted disruption of the MSMEG5898 (ksdD-1) gene, but not the MSMEG4855 (ksdD-2) gene, results in partial inactivation of the cholesterol degradation pathway and accumulation of the intermediate 4-androstene-3,17-dione, reversible by the introduction of the wild-type ksdD-1 gene into deltaksdD-1 or overexpression of ksdD-2 |
-, 675829 |
| 1.3.99.4 | R256A |
site-directed mutagenesis, almost inactive mutant |
-, 763180 |
| 1.3.99.4 | R256H |
site-directed mutagenesis, the mutant shows over 70% reduced activity compare to wild-type enzyme |
763180 |
| 1.3.99.4 | R273A |
site-directed mutagenesis, inactive mutant |
763180 |
| 1.3.99.4 | S138A |
site-directed mutagenesis, no significant change in enzyme expression is caused by the mutation, the mutant shows reduced activity with 4-androstene-3,17-dione compared to wild-type enzyme |
-, 763772 |
| 1.3.99.4 | S325F |
mutant enzyme has no detectable KSTD enzyme activity |
-, 656722 |
| 1.3.99.4 | T503I |
mutant enzyme has no detectable KSTD enzyme activity |
-, 656722 |
