为了解重组酵母菌N6076不同发酵时间的差异表达基因信息及甲羟戊酸(mevalonate,MVA)代谢状况,以重组酵母菌N6076及其原始菌株 Kh08为研究对象,基于转录组测序和MVA测定,分析和比较了两者在3个发酵时间点(0、48和96 h)的差异表达基因的功能。GO功能分析结果表明,重组菌株N6076新增了14项生物学过程、2项细胞组分和2项分子功能,涉及1140个差异表达基因。KEGG代谢分析结果表明,重组菌株N6076新增了13条代谢通路,涉及77个差异表达基因,其中新增的萜类骨架生物合成途径涉及5个差异表达基因。甲羟戊酸激酶(mevalonate kinase,MVK)基因表达与MVA的LC-MS/MS测定结果表明,在整个发酵过程中MVK基因表达量与MVA产量的变化趋势一致。该研究为进一步认识重组酵母菌N6076的基因表达与MVA代谢调控提供了依据。
In order to understand the information of differentially expressed genes (DEGs) and the metabolism of mevalonate (MVA) in recombinant Saccharomyces cerevisiae strain N6076 at different fermentation phases, based on transcriptomic sequencing and MVA assay, its functions of DEGs at three fermentation time points (0 h, 48 h, 96 h) were analyzed and compared. The results of gene ontology (GO) terms showed that 14 biological processes (BP), 2 cell components (CC) and 2 molecular functions (MF) were added, involving 1 140 DEGs in recombinant strain N6076. The results of Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that there were 13 new metabolic pathways involving 77 DEGs, of which 5 DEGs were related to the new increased terpenoid skeleton biosynthesis pathway in recombinant strain N6076. The results of mevalonate kinase (MVK) gene expression and quantitative analysis of MVA by liquid chromatography-tandem mass spectrometry (LC-MS/MS) showed that the change trend of MVK gene expression was consistent with that of MVA production during the whole fermentation process. The results can provide a basis for further understanding the gene expression and MVA metabolism regulation in recombinant yeast strain N6076.
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