PLKNPLC
We have contributed six new parts to the wiki this year.
First and foremost, panS (BBa_K4936005) is our plasmid backbone for our genes of interest. Our team has based this plasmid on the work of the HKSSC 2022 team, which has an E. coli origin of replication (pBR322 oriV–oriT) from the pANS-based shuttle vectors designed by Chen et al. flanked by a set of loxP sites. Prior to transformation into S. elongatus, the unnecessary bacterial ORI can be removed through the Cre-lox system to reduce the metabolic burden of replicating the shuttle vector.
Furthermore, we have selected 5 ligninolytic enzymes from three different fungal species, which are among the most-studied for ligninolytic ability, to compare their efficiency in decolourising MB. Their coding sequences are registered as BBa_K4936000 to BBa_K4936004. Our team has codon optimised all sequences for S. elongatus using reformats of the codon table generated by HKSSC 2021.
Lignin peroxidase H8 is found in P. chrysoporium strain BKMF-1767. Our team selected this enzyme due to reported high efficiency of oxidation with veratryl alcohol as the substrate [1]. Peroxidases in general possess attractive qualities for dye removal due to high oxidative potential and low substrate specificity.
This is a lignin peroxidase isozyme recently characterized in T. versicolor strain PRL572. Our team selected this enzyme as T. versicolor is well studied and agreed to be one of the most potent white-rot fungi in degradation of lignin.
Laccase 2 is a multicopper oxidase from T. versicolor. The broad substrate range of Lac2 has long made it an attractive choice for dye decolourization. Laccase in general is also an oxidoreductase with high levels of functionality as it does not require H2O2 for activity unlike other peroxidases.
This is manganese peroxidase 1 from P. chrysoporium.
This enzyme is versatile peroxidase 2 from P. eryngii. Our team selected this enzyme as P. eryngii had been found to have one of the highest percentages of final MB degradation in solution [2].