EC Number   |
|---|
   1.10.3.2 | - |
| 1.10.3.2 | attempts of heterologous expression of the wild-type laccase using a Pichia pastoris secretory expression system are unsuccessful most likely because the enzyme is too unstable and degrades immediately after production. Therefore the stability of the laccase is improved by using a phylogeny-based design method. A mutant laccase is created in which sixteen original residues are replaced with those found in the phylogenetically inferred ancestral sequence. The resulting mutant protein is successfully produced using the Pichia pastoris secretory expression system and then purified |
| 1.10.3.2 | by signal peptide prediction, the enzyme is assumed to be a secretory protein starting from Gln23. The DNA encoding the mature protein is then cloned and expressed in Escherichia coli BL21 (DE3). The recombinant enzyme, expressed as an apoprotein, is dialyzed against copper-containing buffer to yield a holoprotein |
| 1.10.3.2 | cloned and expressed in Pichia pastoris |
| 1.10.3.2 | CotA-type laccase, DNA and amino acid sequence determination and analysis, recombinant expression in Escherichia coli strain JM109 |
| 1.10.3.2 | DNA and amino acid sequence determination and analysis |
| 1.10.3.2 | DNA and amino acid sequence determination and analysis, expression in Escherichia coli |
| 1.10.3.2 | DNA and amino acid sequence determination and analysis, primary structure of the gene, sequence comparisons |
| 1.10.3.2 | DNA and amino acid sequence determination and analysis, sequence comparison |
| 1.10.3.2 | DNA and amino acid sequence determination and analysis, subcloning in Escherichia coli strain Top10, expression of wild-type and mutant enzymes in Saccharomyces cerevisiae strain W303-1A |
