Parts
Our collection includes six new basic parts and 2 composite parts. We wished to improve upon the design of the 2014 Braunschweig team which successfully expressed the soluble methane monooxygenase from Methylococcus capsulatus in E. coli. We adjusted the individual subunits of the sMMO by performing codon optimization for expression in E. coli. We have also created a composite part in which the promoter was replaced with the AraC-pBAD promoter as we believe this promoter allows more precise control of the expression. Secondly, our design also includes the co-transformation of the pGro7 plasmid from Takara, which encodes for the two chaperones GroES and GroEL. The 2014 Braunschweig team has identified these two chaperones as capable of holding sMMO properly. The expression of these two chaperones is also driven by L-arabinose. This will allow us to use one inducer to trigger the expression of all gene products.
Basic Parts
Part Number | Name | Description |
mmoX | Encodes for the alpha subunit of the soluble MMO. Codon optimized for E. coli expression. | |
mmoY | Encodes for the beta subunit of the hydroxylase of the soluble MMO. Codon optimized for E. coli expression. | |
mmoB | Encodes for the regulatory subunit B of the soluble MMO. Codon optimized for E. coli expression. | |
mmoZ | Encodes for the γ subunit of the hydroxylase of the soluble MMO. Codon optimized for E. coli expression. | |
mmoD | Encodes for a putative inhibitor but the exact function is unknown. Codon optimized for E. coli expression. | |
mmoC | Encodes for the reductase subunit of the soluble MMO. Codon optimized for E. coli expression. |
Composite Parts:
Part Number | Name | Description |
Soluble methane monooxygenase | Encodes for all 6 subunits of the soluble MMO. It is driven by the inducible lacI promoter. Each subunit is further driven by a weak ribosome binding site (B0032). | |
Soluble methane monooxygenase | Similar to the above construct except that the promoter has been replaced with the inducible AraBAD promoter. |