Part:BBa_K5117010
BhrPET
This part contains the gene of the uncultured bacterium HR29 encoding a polyethylene terephthalate hydrolase (PETase, EC 3.1.1.101). It is a homologue of the leaf-branch compost cutinase (LCC) and codon optimized for Bacillus subtilis (Xi et al. 2021). In addition to the coding sequence, the signal peptide of the aprE gene for secretion in Bacillus subtilis is included.
BhrPET only served for design purposes of the TU Dresden iGEM 2024 Team and was required for the construction of composite parts (see Contribution page).
Target organism: Bacillus subtilis
Main purpose of use: Gene expression and protein production using the host Bacillus subtilis
Design
For compatibility with the BioBrick RFC[10] standard, the restriction sites EcoRI, XbaI, SpeI, PstI and NotI were removed from the coding sequence. To make the part compatible with the Type IIS standard, BsaI and SapI sites were removed as well. This was achieved by codon exchange using the codon usage table of Bacillus subtilis (Codon Usage Database Kazusa).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 579
Illegal AgeI site found at 624 - 1000COMPATIBLE WITH RFC[1000]
References
Xi X., Ni K., Hao H., Shang Y., Zhao B., Qian Z. (2021): Secretory expression in Bacillus subtilis and biochemical characterization of a highly thermostable polyethylene terephthalate hydrolase from bacterium HR29. Enzyme and microbial technology 143, 109715.
| None |