Identification of the genes affecting the regulation of riboflavin synthesis in the flavinogenic yeast Pichia guilliermondii using insertion mutagenesis

• DOI: 10.1111/j.1567-1364.2011.00720.x
• Publication Date: 2011-01-24T17:19:37Z
• Authors: Yuriy R. Boretsky; Yuriy V. Pynyaha; Volodymyr Y. Bore...; Dariya V. Fedorovych; Lyubov R. Fayura; Olha Protchenko; Caroline C. Philpott; Andriy A. Sibirny
• Keywords: Identification; Pichia

Pichia guilliermondii is a representative of group so-called flavinogenic yeast species that overproduce riboflavin (vitamin B(2)) in response to iron limitation. Using insertion mutagenesis, we isolated P. mutants overproducing riboflavin. Analysis nucleotide sequence recombination sites revealed cassettes integrated into the genome disrupting genes similar VMA1 gene Ashbya gossypii and Saccharomyces cerevisiae FES1 FRA1 S. cerevisiae. The constructed guilliermondiiΔvma1-17 mutant possessed five- sevenfold elevated production twofold decreased cell content as compared with parental strain. guilliermondiiΔfra1-45 accumulated 1.8-2.2-fold more cells produced Both Δvma1-17 Δfes1-77 knockout strains could not grow at 37 °C contrast wild-type strain Δfra1-45 mutant. Increased by was observed °C. Although did riboflavin, it showed partial complementation when crossed previously riboflavin-overproducing rib80-22. Complementation analysis are distinct from reported riboflavin-producing hit1-1, rib80-22 rib81-31 this yeast.