Cloning Glucose Oxidase Gene And Its High Production

The gene for Penicillium notatum glucose oxidase(EC1.1.3.4)has been cloned from its genome. The gene, isolated by PCR consisted of 1815 bp encoding 604 amino acid residues, and is preceded by a 18-amino acid presequence. The enzyme contains 4 potential sites for N-linked glycosylation.The DNA fragment encoding Penicillium notatum glucose oxidase, was cloned into vector of pPIC9 for expression in Pichia pastoris. When transformed into methylotrophic yeast Pichiapastoris GS115, The constructed plasmid pPIC9-sgod directed the synthesis and secretion of functionally active GOD. After induction in 3L fermentor for 132h, the GOD activity in the medium reached to 111.3U/mL.