Coding

Part:BBa_K2718005

Designed by: Quentin PIPPO   Group: iGEM18_Aix-Marseille   (2018-08-27)


Methionine-γ-lyase-Histag

This DNA sequence codes a methionine-y-lyase-Histag (Brevibacteirum linens) which is an enzyme that converts L-methionine to methanethiol, ammonium and 2-oxobutanoate. When methanethiol is formed it gets further oxidezed to dimethyldisulfide(DMDS) and dimethyltrisufide(DMTS).

(picture of reaction)

Characterization

To characterize methionine-γ-lyase, activity tests were done :

  • DTNB (5,5′-Dithiobis 2-nitrobenzoic acid) test : DTNB reacts with thiol group so for methionine-y-lyase, DTNB reacts with thiol group of methanethiol
  • MTBH (3-Methyl-2-benzothiazolinone hydrazone hydrochloride hydrate) test : MBTH reacts with alpha-keto compounds so for methionine-y-lyase, MTBH reacts with 2 oxobutanoate.


More information can be found on our wiki.

Methionine-y-lyase is available without the histidine tag (BBa_1493300) and with a strong promoter (BBa_K2718006)


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 154
    Illegal NgoMIV site found at 667
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 460


[edit]
Categories
Parameters
None