EC Number |
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1.1.3.13 | - |
1.1.3.13 | AUG, MOD1, and AUG, MOD2 of Pichia methanolica are genes encoding AO, the AUG2 gene encodes one of the subunits of the alcohol oxodase and the catalase CTA1 of the organism. Genotyping of isozymes in different strains, overview |
1.1.3.13 | expression in Escherichia coli |
1.1.3.13 | expression in Escherichia coli. The OcAOD productivity of the recombinant E. coli is 24fold higher than that of Ochrobactrum sp. AIU 033, and it is further enhanced by 1.2 times by the coexpression of additional tatABC from E. coli BL21 |
1.1.3.13 | expression in Pichia pastoris |
1.1.3.13 | expression of the enzyme in Saccharomyces cerevisiae results in an inactive, mislocated cytosolic enzyme, co-expression of Hansenula polymorpha pyruvate carboxylase HpPyc1p activates the enzyme but does not improve its import into the peroxisomes, sorting of the recombinant enzyme into Saccharomyces cerevisiae peroxisomes requires an alternative peroxisomal targeting signal that is only formed in case of co-expression of HpPyc1p, overview |
1.1.3.13 | functional recombinant expression of C-terminally His6-tagged enzyme in Pichia pastoris strain GS115 from the plasmid pPIC9K-AOX intracellularly and extracellularly, batch culture. Expression in Escherichia coli strain DH5alpha in batch culture. Method comparison and optimization, overview |
1.1.3.13 | gene AOX1, strong and tightly regulated promoter inducible by methanol |
1.1.3.13 | rerouting of peroxisomal alcohol oxidase to the secretory pathway of Hansenula polymorpha. Using the leader sequence of the Saccharomyces cerevisiae mating factor alpha as sorting signal, alcohol oxidase is correctly sorted to the endoplasmic reticulum. The MFalpha presequence, but not the prosequence, is cleaved from the protein. Alcohol oxidase protein is present in the endoplasmic reticulum as monomers that lack FAD, and hence is enzymatically inactive. The recombinant alcohol oxidase protein is subject to gradual degradation, possibly because the protein does not fold properly. When the Saccharomyces cerevisiae invertase signal sequence is used, secretion of AO protein is observed in conjunction with bulk of the protein being localized to the ER. The amount of secreted AO protein increases with increasing copy numbers of the alcohol oxidase expression cassette integrated into the genome. The secreted alcohol oxidase protein is correctly processed and displays enzyme activity |
1.1.3.13 | single gene AOD, DNA sequence determination and analysis |