Dmytruk K. Identification of structural and regulatory genes for riboflavin biosynthesis in the flavinogenic yeast Candida famata

The objeсt: riboflavin synthesis and regulation in the flavinogenic yeast C. famata. The aim: to identify of structural and regulatory genes for riboflavin synthesis in the flavinogenic yeast C. famata. Methods of research are molecular genetics, gene-engineering, microbiology, biochemistry. Genes RIB1, RIB2, RIB5, RIB6 and RIB7 of C. famata (coding for GTP cyclohydrolase II, 5-amino-6-(5-phosphoribosylamino)uracil reductase, 6,7-dimethyl-8-ribityllumazine synthase, 3,4-dihydroxy-2-butanone-4-phosphate synthase and riboflavin synthetase, respectively) were cloned using functional complementation method. The gene RIB3 of D. hansenii (coding for 2,5-diamino-6-ribitylamino-4(3H)-pyrimidinone -5'-phosphate deaminase) was PCR-isolated and identified. There was demonstrated that the alpha' subunit of casein kinase 2 is involved in the regulation of riboflavin synthesis. Using the developed method of insertional mutagenesis, the two positive regulatory genes of riboflavin synthesis were identified. Such genesare CfMET2 (homoserine-O-acetyltransferase) and SEF1 (putative transcription factor). Field of application: biotechnology.