Methylotrophic yeast Pichia pastoris is an object of modern biotechnology. Decisive understanding of gene regulation mechanisms is essential for successful protein production. In this study, we investigated the effect...Methylotrophic yeast Pichia pastoris is an object of modern biotechnology. Decisive understanding of gene regulation mechanisms is essential for successful protein production. In this study, we investigated the effect of deletions in P. pastoris genes encoding proteins, homologous to S. serevisiae Rtg1p, Rtg 2p, Msn2p and Msn4p. It was shown, that deletion in PpRTG1 gene results in inability of P. pastoris to grow on medium with methanol as a carbon source and ammonium sulfate as a source of nitrogen. We also demonstrate that deletions in PpRTG1 and PpRTG2 decrease activity of AOX1 promoter.展开更多
Methanol is a promising substrate for sustainable biomanufacturing,and Pichia pastoris has become a commonly used yeast for methanol utilization due to its powerful methanol metabolic pathways and methanol inducible p...Methanol is a promising substrate for sustainable biomanufacturing,and Pichia pastoris has become a commonly used yeast for methanol utilization due to its powerful methanol metabolic pathways and methanol inducible promoter.Previous reconstruction of gene circuits highly improved transcriptional activity,but excessive expression of chimeric transactivator damaged cell growth on methanol.Here we employed transcriptome analysis to investigate the effects of chimeric transactivator overexpression on cellular metabolism and regula-tory networks.The results showed that strong expression of chimeric transactivator unexpectedly downregulated methanol metabolism,especially the alcohol oxidase 1(AOX1),but without remarkable changes in expression of transcriptional factors.Meanwhile,the synthesis of peroxisomes also varied with chimeric transactivator expression.In addition,the enrichment analysis of differentially expressed genes revealed their impact on cellular metabolism.The gene expression patterns caused by different expression levels of chimeric trans-activators have also been clarified.This work provides useful information to understand the transcriptional regulation of the AOX1 promoter and methanol signaling.It revealed the importance of balancing transcription factor expression for the host improvement.展开更多
文摘Methylotrophic yeast Pichia pastoris is an object of modern biotechnology. Decisive understanding of gene regulation mechanisms is essential for successful protein production. In this study, we investigated the effect of deletions in P. pastoris genes encoding proteins, homologous to S. serevisiae Rtg1p, Rtg 2p, Msn2p and Msn4p. It was shown, that deletion in PpRTG1 gene results in inability of P. pastoris to grow on medium with methanol as a carbon source and ammonium sulfate as a source of nitrogen. We also demonstrate that deletions in PpRTG1 and PpRTG2 decrease activity of AOX1 promoter.
基金supported by National Key Research and Development Program of China(2022YFC2805102)Young Scientist Fund of National Natural Science Foundation of China(32201206)China Postdoctoral Science Foundation(2022M711146)。
文摘Methanol is a promising substrate for sustainable biomanufacturing,and Pichia pastoris has become a commonly used yeast for methanol utilization due to its powerful methanol metabolic pathways and methanol inducible promoter.Previous reconstruction of gene circuits highly improved transcriptional activity,but excessive expression of chimeric transactivator damaged cell growth on methanol.Here we employed transcriptome analysis to investigate the effects of chimeric transactivator overexpression on cellular metabolism and regula-tory networks.The results showed that strong expression of chimeric transactivator unexpectedly downregulated methanol metabolism,especially the alcohol oxidase 1(AOX1),but without remarkable changes in expression of transcriptional factors.Meanwhile,the synthesis of peroxisomes also varied with chimeric transactivator expression.In addition,the enrichment analysis of differentially expressed genes revealed their impact on cellular metabolism.The gene expression patterns caused by different expression levels of chimeric trans-activators have also been clarified.This work provides useful information to understand the transcriptional regulation of the AOX1 promoter and methanol signaling.It revealed the importance of balancing transcription factor expression for the host improvement.
