Bioinformatics study of recombinant glucose oxidase expressed in Yarrowia lipolytica and optimization of its production by Taguchi experiment design method

  Glucose oxidase has a wide range of application in medical, pharmaceutical, food, textile, and environmental industries. Heterologous expression of the enzyme has been investigated in yeast hosts due to several industrial-scale limitations in enzyme production by Aspergillus niger. The present study aimed to investigate bioinformatics characteristics of recombinant glucose oxidase in Yarrowia lipolytica Po1g-GOX and to optimize the production conditions of the enzyme by Taguchi experimental design method.

Protein sequence, primary, secondary and tertiary structures and glycosylation rate of recombinant glucose oxidase enzyme were studied via bioinformatics tools. Optimization of recombinant glucose oxidase for glucose, peptone, yeast extract and pH at four levels in YPD media with thiamine were defined to Qualitek-4 software. The level of enzyme production was measured and analyzed by the software. Recombinant glucose oxidase had 605 amino acids, with 29% α-helix, 16% β sheet and 54% coil in its secondary structure showing similarity to that of native glucose oxidase of A. niger. Eight glycosylation sites have been located in this recombinant protein, and the tertiary structure had one domain showing high similarity to native glucose oxidase. The highest level of recombinant glucose oxidase production was obtained in medium containing 30 g/L glucose, 5 g/L yeast extract, 15 g/L peptone, and pH 7. Recombinant glucose oxidase produced in Yarrowia has a suitable potential to be used in pharmaceutical and food industries due to high similarity to native glucose oxidase.