Part:BBa_K4382013
Ligninase Gene Construct
This gene construct contains 2 dye-decolorizing peroxidase genes(BBa_K1336003,BBa_K4382002),a strong RBS(BBa_K4382009),and a signal peptide(BBa_K1074014).
Usage and Biology
The most eminent feature of this part is that it acts like a polycistronic gene sequence as it consists of a Bacillus subtilis RBS.The Ribosome Binding site consists of a consensus sequence, which ensures that both the proteins are encoded from a single gene sequence.The presence of a SamyQ signaling peptide on the N-terminus of the protein ensures that the protein is secreted out of the cell. The proteins which are secreted outside are the enzymes BsDyP(BBa_K1336003) and DyP1B(BBa_K4382002). Both these enzymes are dye-decolorizing peroxidases which, in the context of our project, work in tandem - DyP1B breaks down Lignin into Benzylic ketones, which in turn is converted to vanillin by BsDyP.
The chemical production of vanillin is highly resource intensive and hence highly expensive in markets. Through our solution, we aim to produce vanillin from wheat straw by degrading the lignin which is present in the plant cell wall. Since we are using raw material which is easily available, which could potentially bring down the cost of vanillin production and hence its market price. This would be important, as vanillin is an important industrial and commercial chemical compound.
In addition to this, both enzymes can degrade a wide variety of compounds, which is described in detail in their individual part pages.
References
1)Rahmanpour, R., & Bugg, T. D. H. (2015). Characterisation of DYP-type peroxidases from pseudomonas fluorescens PF-5: Oxidation of mn(ii) and polymeric lignin by DYP1B. Archives of Biochemistry and Biophysics, 574, 93–98. https://doi.org/10.1016/j.abb.2014.12.022
2)Sugano, Y., & Yoshida, T. (2021). DYP-type peroxidases: Recent advances and perspectives. International Journal of Molecular Sciences, 22(11), 5556. https://doi.org/10.3390/ijms22115556
3)Colpa, D. I., Fraaije, M. W., & van Bloois, E. (2014). DYP-type peroxidases: A promising and versatile class of enzymes. Journal of Industrial Microbiology and Biotechnology, 41(1), 1–7. https://doi.org/10.1007/s10295-013-1371-6
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 1795
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 217
Illegal BsaI site found at 226
Illegal BsaI site found at 1192
Illegal SapI site found at 622
Illegal SapI site found at 980
Illegal SapI.rc site found at 1136
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