EC Number   |
|---|
    4.4.1.15 | cloning into the pET30a (+) vector at the EcoRV/HindIII sites, all single and double mutants are constructed using a Phusion Site Directed Mutagenesis Kit. |
| 4.4.1.15 | epression of His6-tagged enzyme in Escherichia coli |
| 4.4.1.15 | expression in Escherichia coli |
| 4.4.1.15 | Expression of all recombinant proteins and mutants is carried out in Escherichia coli BL21. Altering of only two amino acid residues within the predicted active site served to change the enzyme from D-cysteine desulfhydrase to ACC deaminase. |
| 4.4.1.15 | overexpressed in host strain, enables Escherichia coli to utilize D-cysteine as sole sulfur source |
| 4.4.1.15 | semi-quantitative reverse transcription-PCR analysis |
