Sequence-based bioprospecting of myo-inositol oxygenase (Miox) reveals new homologues that increase glucaric acid production in Saccharomyces cerevisiae.

•A sequence similarity network of 1579 Miox enzymes shows conservation within main life groups and high diversity among fungal sequences.•Thirty-five Miox homologues, including twenty-five uncharacterized sequences were successfully expressed in Saccharomyces cerevisiae.•Miox variants that have a defective “dynamic loop” protecting the substrate binding site showed no activity.•Miox stability throughout the fermentation is a major bottleneck to be surmounted for cost-effective glucaric acid synthesis in yeast.•Miox from Talaromyces marneffei allowed for 44 % higher specific glucaric acid production than the often-used Arabidopsis thaliana Miox4 variant.