Yatsyshyn Valentyna Y, Ishchuk Olena P, Voronovsky Andriy Y, Fedorovych Daria V, Sibirny Andriy A
Institute of Cell Biology, National Academy of Sciences of Ukraine, Drahomanov Street, 14/16, Lviv 79005, Ukraine.
Metab Eng. 2009 May;11(3):163-7. doi: 10.1016/j.ymben.2009.01.004. Epub 2009 Feb 4.
Recombinant strains of the flavinogenic yeast Candida famata able to overproduce flavin mononucleotide (FMN) that contain FMN1 gene encoding riboflavin (RF) kinase driven by the strong constitutive promoter TEF1 (translation elongation factor 1alpha) were constructed. Transformation of these strains with the additional plasmid containing the FMN1 gene under the TEF1 promoter resulted in the 200-fold increase in the riboflavin kinase activity and 100-fold increase in FMN production as compared to the wild-type strain (last feature was found only in iron-deficient medium). Overexpression of the FMN1 gene in the mutant that has deregulated riboflavin biosynthesis pathway and high level of riboflavin production in iron-sufficient medium led to the 30-fold increase in the riboflavin kinase activity and 400-fold increase in FMN production of the resulted transformants. The obtained C. famata recombinant strains can be used for the further construction of improved FMN overproducers.
构建了产黄素酵母法夫酵母（Candida famata）的重组菌株，这些菌株能够过量生产黄素单核苷酸（FMN），它们含有由强组成型启动子TEF1（翻译延伸因子1α）驱动的编码核黄素（RF）激酶的FMN1基因。用含有在TEF1启动子控制下的FMN1基因的附加质粒转化这些菌株，与野生型菌株相比，核黄素激酶活性提高了200倍，FMN产量提高了100倍（后一特性仅在缺铁培养基中发现）。在铁充足培养基中具有失调的核黄素生物合成途径和高水平核黄素产量的突变体中过量表达FMN1基因，导致所得转化体的核黄素激酶活性提高了30倍，FMN产量提高了400倍。所获得的法夫酵母重组菌株可用于进一步构建改良的FMN过量生产菌株。
