Abstract
The deletion of PHO13 (pho13δ) in Saccharomyces cerevisiae, encoding a phosphatase enzyme of unknown specificity, results in the transcriptional activation of genes related to the pentose phosphate pathway (PPP) such as TAL1 encoding transaldolase. It has been also reported that the pho13Δ mutant of S. cerevisiae expressing a heterologous xylose pathway can metabolize xylose efficiently compared to its parental strain. However, the interaction between the pho13Δ-induced transcriptional changes and the phenotypes of xylose fermentation was not understood. Thus we investigated the global metabolic changes in response to pho13Δ when cells were exponentially growing on xylose. Among the 134 intracellular metabolites that we identified, the 98% reduction of sedoheptulose was found to be the most significant change in the pho13Δ mutant as compared to its parental strain. Because sedoheptulose-7-phosphate (S7P), a substrate of transaldolase, reduced significantly in the pho13Δ mutant as well, we hypothesized that limited transaldolase activity in the parental strain might cause dephosphorylation of S7P, leading to carbon loss and inefficient xylose metabolism. Mutants overexpressing TAL1 at different degrees were constructed, and their TAL1 expression levels and xylose consumption rates were positively correlated. Moreover, as TAL1 expression levels increased, intracellular sedoheptulose concentration dropped significantly. Therefore, we concluded that TAL1 upregulation, preventing the accumulation of sedoheptulose, is the most critical mechanism for the improved xylose metabolism by the pho13Δ mutant of engineered S. cerevisiae.
| Original language | English |
|---|---|
| Pages (from-to) | 88-96 |
| Number of pages | 9 |
| Journal | Metabolic Engineering |
| Volume | 34 |
| DOIs | |
| Publication status | Published - 2016 Mar 1 |
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Keywords
- Cas9-guided genome editing technique
- GC-TOF/MS
- Metabolomics
- NMR
- RNA-seq
ASJC Scopus subject areas
- Bioengineering
- Biotechnology
- Applied Microbiology and Biotechnology
Cite this
PHO13 deletion-induced transcriptional activation prevents sedoheptulose accumulation during xylose metabolism in engineered Saccharomyces cerevisiae. / Xu, Haiqing; Kim, Sooah; Sorek, Hagit; Lee, Yongsuk; Jeong, Dukyeol; Kim, Jungyeon; Oh, Eun Joong; Yun, Eun Ju; Wemmer, David E.; Kim, Kyoung Heon; Kim, Soo Rin; Jin, Yong Su.
In: Metabolic Engineering, Vol. 34, 01.03.2016, p. 88-96.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - PHO13 deletion-induced transcriptional activation prevents sedoheptulose accumulation during xylose metabolism in engineered Saccharomyces cerevisiae
AU - Xu, Haiqing
AU - Kim, Sooah
AU - Sorek, Hagit
AU - Lee, Yongsuk
AU - Jeong, Dukyeol
AU - Kim, Jungyeon
AU - Oh, Eun Joong
AU - Yun, Eun Ju
AU - Wemmer, David E.
AU - Kim, Kyoung Heon
AU - Kim, Soo Rin
AU - Jin, Yong Su
PY - 2016/3/1
Y1 - 2016/3/1
N2 - The deletion of PHO13 (pho13δ) in Saccharomyces cerevisiae, encoding a phosphatase enzyme of unknown specificity, results in the transcriptional activation of genes related to the pentose phosphate pathway (PPP) such as TAL1 encoding transaldolase. It has been also reported that the pho13Δ mutant of S. cerevisiae expressing a heterologous xylose pathway can metabolize xylose efficiently compared to its parental strain. However, the interaction between the pho13Δ-induced transcriptional changes and the phenotypes of xylose fermentation was not understood. Thus we investigated the global metabolic changes in response to pho13Δ when cells were exponentially growing on xylose. Among the 134 intracellular metabolites that we identified, the 98% reduction of sedoheptulose was found to be the most significant change in the pho13Δ mutant as compared to its parental strain. Because sedoheptulose-7-phosphate (S7P), a substrate of transaldolase, reduced significantly in the pho13Δ mutant as well, we hypothesized that limited transaldolase activity in the parental strain might cause dephosphorylation of S7P, leading to carbon loss and inefficient xylose metabolism. Mutants overexpressing TAL1 at different degrees were constructed, and their TAL1 expression levels and xylose consumption rates were positively correlated. Moreover, as TAL1 expression levels increased, intracellular sedoheptulose concentration dropped significantly. Therefore, we concluded that TAL1 upregulation, preventing the accumulation of sedoheptulose, is the most critical mechanism for the improved xylose metabolism by the pho13Δ mutant of engineered S. cerevisiae.
AB - The deletion of PHO13 (pho13δ) in Saccharomyces cerevisiae, encoding a phosphatase enzyme of unknown specificity, results in the transcriptional activation of genes related to the pentose phosphate pathway (PPP) such as TAL1 encoding transaldolase. It has been also reported that the pho13Δ mutant of S. cerevisiae expressing a heterologous xylose pathway can metabolize xylose efficiently compared to its parental strain. However, the interaction between the pho13Δ-induced transcriptional changes and the phenotypes of xylose fermentation was not understood. Thus we investigated the global metabolic changes in response to pho13Δ when cells were exponentially growing on xylose. Among the 134 intracellular metabolites that we identified, the 98% reduction of sedoheptulose was found to be the most significant change in the pho13Δ mutant as compared to its parental strain. Because sedoheptulose-7-phosphate (S7P), a substrate of transaldolase, reduced significantly in the pho13Δ mutant as well, we hypothesized that limited transaldolase activity in the parental strain might cause dephosphorylation of S7P, leading to carbon loss and inefficient xylose metabolism. Mutants overexpressing TAL1 at different degrees were constructed, and their TAL1 expression levels and xylose consumption rates were positively correlated. Moreover, as TAL1 expression levels increased, intracellular sedoheptulose concentration dropped significantly. Therefore, we concluded that TAL1 upregulation, preventing the accumulation of sedoheptulose, is the most critical mechanism for the improved xylose metabolism by the pho13Δ mutant of engineered S. cerevisiae.
KW - Cas9-guided genome editing technique
KW - GC-TOF/MS
KW - Metabolomics
KW - NMR
KW - RNA-seq
UR - http://www.scopus.com/inward/record.url?scp=84953774938&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84953774938&partnerID=8YFLogxK
U2 - 10.1016/j.ymben.2015.12.007
DO - 10.1016/j.ymben.2015.12.007
M3 - Article
C2 - 26724864
AN - SCOPUS:84953774938
VL - 34
SP - 88
EP - 96
JO - Metabolic Engineering
JF - Metabolic Engineering
SN - 1096-7176
ER -