Identification of the genes affecting the regulation of riboflavin synthesis in the flavinogenic yeast Pichia guilliermondii using insertion mutagenesis.
Identification of the genes affecting the regulation of riboflavin synthesis in the flavinogenic yeast Pichia guilliermondii using insertion mutagenesis.
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DOI：
10.1111/j.1567-1364.2011.00720.x
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发表时间：
2011-05
影响因子：
3.2
通讯作者：
Sibirny AA
中科院分区：
文献类型：
--
作者：
Boretsky YR;Pynyaha YV;Boretsky VY;Fedorovych DV;Fayura LR;Protchenko O;Philpott CC;Sibirny AA
毕赤酵母（Pichia guilliermondii）是一类所谓的黄酮类酵母的代表，它们在铁限制下过量产生核黄素（维生素B2）。采用插入诱变技术，分离出过量产生核黄素的吉列蒙地假体。重组位点的核苷酸序列分析显示，插入盒整合到基因组中，破坏了P. guilliermondii与棉花Ashbya gossypii和酿酒酵母（Saccharomyces cerevisiae）的VMA1基因和酿酒酵母（S. cerevisiae）的FES1和FRA1基因相似的基因。与亲本菌株相比，构建的p.g illermondii Δvma1-17突变体核黄素产量提高了5 - 7倍，铁细胞含量降低了2倍。与亲本菌株相比，吉利蒙毕赤酵母Δfra1-45突变株在细胞中积累了1.8 - 2.2倍的铁，产生了5 - 7倍的核黄素。与野生型菌株和Δfra1-45突变体相比，Δvma1-17和Δfes1-77敲除菌株在37°C下都不能生长。在37℃下观察到野生型菌株核黄素产量增加。虽然Δfes1-77突变体没有过量产生核黄素，但当与先前分离的p.g illiermondii过量产生核黄素的突变体rib80-22杂交时，它表现出部分互补。互补分析显示，Δvma1-17和Δfra1-45突变体与先前报道的该酵母的核黄素产生突变体hit1-1、rib80-22和rib81-31不同。
Pichia guilliermondii is a representative of a group of so-called flavinogenic yeast species that overproduce riboflavin (vitamin B2) in response to iron limitation. Using insertion mutagenesis, we isolated P. guilliermondii mutants overproducing riboflavin. Analysis of nucleotide sequence of recombination sites revealed that insertion cassettes integrated into the genome disrupting P. guilliermondii genes similar to the VMA1 gene of Ashbya gossypii and Saccharomyces cerevisiae and FES1 and FRA1 genes of S. cerevisiae. The constructed P. guilliermondii Δvma1–17 mutant possessed five- to sevenfold elevated riboflavin production and twofold decreased iron cell content as compared with the parental strain. Pichia guilliermondii Δfra1–45 mutant accumulated 1.8–2.2-fold more iron in the cells and produced five- to sevenfold more riboflavin as compared with the parental strain. Both Δvma1–17 and Δfes1–77 knockout strains could not grow at 37 °C in contrast to the wild-type strain and the Δfra1–45 mutant. Increased riboflavin production by the wild-type strain was observed at 37 °C. Although the Δfes1–77 mutant did not overproduce riboflavin, it showed partial complementation when crossed with previously isolated P. guilliermondii riboflavin-overproducing mutant rib80–22. Complementation analysis revealed that Δvma1–17 and Δfra1–45 mutants are distinct from previously reported riboflavin-producing mutants hit1-1, rib80-22 and rib81-31 of this yeast.
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影响因子：
3.1
作者：
Boretsky, Yuriy R.;Protchenko, Olga V.;Sibirny, Andriy A.
通讯作者：
Sibirny, Andriy A.
影响因子：
56.9
作者：
TANNER, FW;VOJNOVICH, C;VANLANEN, JM
通讯作者：
VANLANEN, JM
影响因子：
4.8
作者：
Kumanovics, Attila;Chen, Opal S.;Kaplan, Jerry
通讯作者：
Kaplan, Jerry
影响因子：
4.4
作者：
Crossley, Rachel A.;Gaskin, Duncan J. H.;Walton, Nicholas J.
通讯作者：
Walton, Nicholas J.
影响因子：
4.8
作者：
Milgrom, Elena;Diab, Heba;Kane, Patricia M.
通讯作者：
Kane, Patricia M.