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Team:Tianjin/project12
From 2010.igem.org
Project I -- Lignin Terminator
We are aimed at making some Saccharomyces cerevisiae which could synthesize lignin degrading enzymes and put them on the out surface of the yeast through the yeast surface display technology. There are four kinds of ligninolytic enzymes, lignin peroxidase(LiP), Meng peroxidase(MnP), glyoxal oxidase(GLOX), laccase(Lac). In recent days, our work is to display these four kinds of ligninolytic enzymes on four cells.
But our final goal is to display four enzymes on a cell. Those cells, which propagate themselves rapidly, could affix to the wood cellulose material and begin to degrade the lignin quickly with the enzymes of their surface. This yeast will make the lignin degradation pretreatment more secure to our environment and lower the cost. The products of this process are celluase and hemicellulase, which could be used to produce alcohol fuels and papermaking in the next steps.
Laccase is a polyphenol oxidase, which belongs to the family of blue multicopper oxidases. These enzymes catalyze the one-electron oxidation of four reducing-substrate molecules concomitant with the four-electron reduction of molecular oxygen to water. Laccases oxidize a broad range of substrates, preferably phenolic compounds. Geneart optimized the codon (without terminator condon) and synthetized the DNA sequence of laccase.
MGLQRFSFFVTLALVARSLAAIGPVASLVVANAPVSPDGFLRDAIVVNGVVPSPLITGKKGDRFQLNVVDTLTNHSMLKSTSIHWHGFFQAGTNWADGPAFVNQCPIASGHSFLYDFHVPDQAGTFWYHSHLSTQYCDGLRGPFVVYDPKDPHASRYDVDNESTVITLTDWYHTAARLGPRFPLGADATLINGLGRSASTPTAALAVINVQHGKRYRFRLVSISCDPNYTFSIDGHNLTVIEVDGINSQPLLVDSIQIFAAQRYSFVLNANQTVGNYWVRANPNFGTVGFAGGINSAILRYQGAPVAEPTTTQTTSVIPLIETNLHPLARMPVPGSPTPGGVDKALNLAFNFNGTNFFINNATFTPPTVPVLLQILSGAQTAQDLLPAGSVYPLPAHSTIEITLPATALAPGAPHPFHLHGHAFAVVRSAGSTTYNYNDPIFRDVVSTGTPAAGDNVTIRFQTDNPGPWFLHCHIDFHLDAGFAIVFAEDVADVKAANPVPKAWSDLCPIYDGLSEANQ
Three-dimensional Configuration of Lacase.
Confocal microscopic images of yeast displaying Lacc.
