Composite

Part:BBa_K3739109

Designed by: Shichen Geng   Group: iGEM21_XMU-China   (2021-09-30)
Revision as of 00:43, 22 October 2021 by Master123 (Talk | contribs)


K525998(T7-RBS)-LMT-his-CBM-hutH-B0010

LMT is a signal peptide from the lytic murein transglycosylase of V.natriegens, his-tag enable us to purify the linking protein, CBM can bind to cellulose and hutH is a histidine transaminase which converts histidine into urocanic acid.

Biology

LMT

Lytic murein transglycosylase (LMT) is an enzyme which is able to degrade murein, a component of cell wall of bacteria. There are two kind of LMTs existing in E.coli: the membrane-binding one and the soluble one. The gene coding LMT homolog is also incorporated in the genome of Vibrio natriegens. The LMT signal peptide (named LMT in our parts) is from the LMT homolog, which can lead the fused protein secreted out of Vibrio natriegens.

CBM

Cellulose enzymes have two domains, and the one that helps bind to cellulose is called cellulose binding module (CBM), and therefore it helps our fusion protein bind to cellulose-rich cell wall. Here we choose the CBM of CenA from Cellulomonas fimi, which has been successfully expressed in Escherichia coli.

hutH

The HutH comes from Pseudomonas putida. Under natural conditions, many microorganisms can use the histidine ammonia-lyase (HutH) to change L-histidine into urocanic acid. HutH catalyzes the first step in the degradation of histidine, and the product urocanic acid is further metabolized to glutamate. This enzyme could be found in the liver of vertebrates and in bacteria such as Escherichia coli, Salmonella and Pseudomonas. It is specific for L-histidine and can be inhibited by D-histidine or imidazole. The active center of the enzyme is thought to be dehydroalanine.

Usage

Used to construct the composite part BBa_K3739039 and BBa_K3739109.

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 635
    Illegal NgoMIV site found at 1071
    Illegal NgoMIV site found at 1806
    Illegal NgoMIV site found at 2042
    Illegal AgeI site found at 381
    Illegal AgeI site found at 471
    Illegal AgeI site found at 708
    Illegal AgeI site found at 1535
    Illegal AgeI site found at 2031
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//awards/composite_part/nominee
Parameters
None