Part:BBa_K1092005
T7-RBS-Laccase
This is a Biobrick of laccase containing a T7 promoter and ribosomal binding site (RBS).
The laccase (cotA) found in Bacillus sp. HR03 is a bacterial laccase that have the potential ability to degrade a variety of aromatic compounds (Mohammadian et al., 2010). Previous study showed that it can be successfully expressed in Escherichia coli strain BL-21(DE3), and was correctly folded (Mollania et al., 2013). Moreover, the complete gene sequence of this laccase was readily recorded in GenBank, with the reference code of FJ663050.1. It is thus used as one of the two major enzymes for PAH degradation.
Properties of the laccase:
Optimal temperature: ~70°C;
Optimal pH: ~7 (Mohammadian et al., 2010).
Laccases (benzenediol oxygen oxidoreductase; EC 1.10.3.2) are a type of oxidative enzymes that show the ability of oxidizing a variety of compounds ranging from phenolic compounds to polycyclic aromatic hydrocarbons (PAHs) (Hadibarata et al., 2012, Mohammadian et al., 2010).
They can be found both in bacteria and fungi, especially white-rot fungi that have the ability to degrade lignin, an important structural component of plants (Hadibarata et al., 2012). The experiment of Hadibarata et al. suggested that laccase could play a very significant role in benzo[a]pyrene degradation, a reaction that hardly occurs (Hadibarata et al., 2012). And the previous research done by Hadibarata et al. and Zeng et al. suggested that laccase degraded PAHs by adding oxygen atoms into the ring, forming quinone-like intermediates. After such transformation, hydrophilic quinones can be further converted into other metabolites by other enzymes.
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 1386
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1547
| n/a | T7-RBS-Laccase |