phleomycin-d1 and alcohol-oxidase

The heterologous protein expression in Pichia pastoris under the control of alcohol oxidase (AOX1)promoter comprises two steps, the growth and induction phases, which are time-consuming and technically demanding. Here, we describe an alternate method where expression is carried out directly in the methanol-containing medium. Using this method, we were successful in screening high-activity laccase clones from a library of laccase mutants generated by random mutagenesis. This simplified method not only saves time but also is highly efficient and can be used for screening a large number of clones.

Topics: Alcohol Oxidoreductases; Benzothiazoles; Bleomycin; Clone Cells; Colorimetry; Coloring Agents; Enzyme Induction; Fungal Proteins; Gene Library; Kinetics; Laccase; Methanol; Mitochondrial Proteins; Mutagenesis; Mutant Proteins; Oxidoreductases; Pichia; Plant Proteins; Polymerase Chain Reaction; Promoter Regions, Genetic; Protein Engineering; Recombinant Proteins; Sulfonic Acids