SEF1 and VMA1 Genes Regulate Riboflavin Biosynthesis in the Flavinogenic Yeast Candida Famata
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and VMA1 Genes Regulate Riboflavin Biosynthesis in the Flavinogenic Yeast Candida famata Y. Andreievaa, O. Lyzakb, Wen Liua, c, Yingqian Kangc, K. Dmytruka, and A. Sibirnya, d, * a
Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv, 79005 Ukraine bMax Planck Institute of Biochemistry, Martinsried, 82152 Germany c Guizhou Medical University, Guiyang, Guizhou, 550025 China dUniversity of Rzeszow, Rzeszow, 35-601 Poland *e-mail: [email protected] Received March 10, 2020; revised March 25, 2020; accepted September 18, 2020
Abstract—Riboflavin (vitamin B2) is an important component of the diet of living organisms since it is a precursor of flavin coenzymes FMN (flavin mononucleotide) and FAD (flavin adenine dinucleotide) involved in numerous enzymatic reactions. It is known that flavinogenic yeast C. famata is able to perform riboflavin overproduction under conditions of iron deficiency, but the regulation of this process remains unknown. It was shown that the deletion of the SEF1 gene (encoding transcription activator) blocked the ability for riboflavin overproduction under conditions of iron deficiency. It was determined that SEF1 promoters of other flavinogenic yeasts (Candida albicans and Candida tropicalis) fused with SEF1 ORF of C. famata can restore the overproduction of riboflavin in the sef1Δ mutant. The disruption of the VMA1 gene (encoding the vacuolar ATPase subunit A) led to overproduction of riboflavin in C. famata in iron complete medium. Keywords: riboflavin, Candida famata, VMA1, SEF1, yeast DOI: 10.3103/S0095452720050023
INTRODUCTION Vitamin B2 (riboflavin, or lactoflavin) plays an important role in humans and animals. Riboflavin is a precursor to FMN and FAD and is involved in numerous enzymatic reactions. Animals, unlike plants, fungi, and most prokaryotes, are not able to synthesize this compound [1, 2]. Therefore, the study and improvement of riboflavin production processes on an industrial scale is an urgent task. Riboflavin can be obtained by both chemical and microbial synthesis. The latter method has certain advantages, in particular, the reduction of waste, energy consumption, and the use of renewable resources, such as sugars or vegetable oil [2–4]. Riboflavin is produced in significant quantities. In 2012, the amount of synthesized riboflavin reached 9000 t. The main producer of riboflavin is the Chinese company Hubei Guangji Pharmaceutical. Vitamin B2 is also produced in other Asian and European countries by BASF, Aventis, Daicel, DSM, Kyowa, Mitsui, Roche, and Takeda. Approximately 70% of riboflavin is used in animal husbandry as a supplement to farm animal feed (pigs and poultry). The remaining 30% of riboflavin is used in the food industry as a dye as well as in medicine as part of multivitamin mixtures and drugs for the treatment of migraines, malaria, eye and skin diseases, and nervous disorders. The world market for riboflavin is 200–230 million dollars per
year, depending on the price of various preparations of this vitamin (the cost of feed riboflavin
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