Dynamic in vivo metabolome response of Saccharomyces cerevisiae to a stepwise perturbation of the ATP requirement for benzoate export
Although much information is available on in vitro role of ATP in regulation, the in vivo kinetics of reactions in which ATP plays a role are only partly known. In order to study such reactions, it is therefore necessary to study the role of ATP in vivo. This study presents an in vivo, targeted pert...
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description | Although much information is available on in vitro role of ATP in regulation, the in vivo kinetics of reactions in which ATP plays a role are only partly known. In order to study such reactions, it is therefore necessary to study the role of ATP in vivo. This study presents an in vivo, targeted perturbation of the ATP flux in aerobic glucose-limited chemostat cultures of Saccharomyces cerevisiae, which was accomplished by transiently (20 min) changing the extracellular undissociated benzoic acid concentration via the pH of the culture. The performed pH shifts resulted in, within about 20 s, a 40% decrease (pH upshift) or a 23% increase (pH downshift) of the calculated ATP consumption rate while the specific glucose uptake rate did not change because of the glucose-limited condition. The pH upshift resulted in a strong decrease in the glycolytic and TCA cycle fluxes; carbon and energy balances indicated an increased flux toward storage carbohydrates. As expected, the pH downshift leads to the opposite effects. Overall, consistent responses were observed in the metabolic fluxes, the off gas concentrations of O₂ and CO₂ and intracellular metabolite concentrations, except for the concentrations of adenosine nucleotides which unexpectedly only showed minor dynamics. This demonstrates that our knowledge of the regulation of the ATP level, the storage metabolism, and central carbon metabolism of yeast is still incomplete. The new dynamic metabolite datasets obtained in this study will prove of great value in developing kinetic models. Biotechnol. Bioeng. 2008;99: 421-441. © 2007 Wiley Periodicals, Inc. |
doi_str_mv | 10.1002/bit.21557 |
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In order to study such reactions, it is therefore necessary to study the role of ATP in vivo. This study presents an in vivo, targeted perturbation of the ATP flux in aerobic glucose-limited chemostat cultures of Saccharomyces cerevisiae, which was accomplished by transiently (20 min) changing the extracellular undissociated benzoic acid concentration via the pH of the culture. The performed pH shifts resulted in, within about 20 s, a 40% decrease (pH upshift) or a 23% increase (pH downshift) of the calculated ATP consumption rate while the specific glucose uptake rate did not change because of the glucose-limited condition. The pH upshift resulted in a strong decrease in the glycolytic and TCA cycle fluxes; carbon and energy balances indicated an increased flux toward storage carbohydrates. As expected, the pH downshift leads to the opposite effects. Overall, consistent responses were observed in the metabolic fluxes, the off gas concentrations of O₂ and CO₂ and intracellular metabolite concentrations, except for the concentrations of adenosine nucleotides which unexpectedly only showed minor dynamics. This demonstrates that our knowledge of the regulation of the ATP level, the storage metabolism, and central carbon metabolism of yeast is still incomplete. The new dynamic metabolite datasets obtained in this study will prove of great value in developing kinetic models. Biotechnol. 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Bioeng</addtitle><description>Although much information is available on in vitro role of ATP in regulation, the in vivo kinetics of reactions in which ATP plays a role are only partly known. In order to study such reactions, it is therefore necessary to study the role of ATP in vivo. This study presents an in vivo, targeted perturbation of the ATP flux in aerobic glucose-limited chemostat cultures of Saccharomyces cerevisiae, which was accomplished by transiently (20 min) changing the extracellular undissociated benzoic acid concentration via the pH of the culture. The performed pH shifts resulted in, within about 20 s, a 40% decrease (pH upshift) or a 23% increase (pH downshift) of the calculated ATP consumption rate while the specific glucose uptake rate did not change because of the glucose-limited condition. The pH upshift resulted in a strong decrease in the glycolytic and TCA cycle fluxes; carbon and energy balances indicated an increased flux toward storage carbohydrates. As expected, the pH downshift leads to the opposite effects. Overall, consistent responses were observed in the metabolic fluxes, the off gas concentrations of O₂ and CO₂ and intracellular metabolite concentrations, except for the concentrations of adenosine nucleotides which unexpectedly only showed minor dynamics. This demonstrates that our knowledge of the regulation of the ATP level, the storage metabolism, and central carbon metabolism of yeast is still incomplete. The new dynamic metabolite datasets obtained in this study will prove of great value in developing kinetic models. Biotechnol. Bioeng. 2008;99: 421-441. © 2007 Wiley Periodicals, Inc.</description><subject>Adenosine triphosphatase</subject><subject>Adenosine Triphosphate - metabolism</subject><subject>Aerobiosis</subject><subject>ATP</subject><subject>Benzoates - metabolism</subject><subject>benzoic acid</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Carbon - metabolism</subject><subject>Cell culture</subject><subject>Fermentation</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Glucose</subject><subject>Kinetics</subject><subject>Metabolism</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - metabolism</subject><subject>Studies</subject><subject>Systems Biology</subject><subject>targetted perturbation</subject><subject>Yeast</subject><issn>0006-3592</issn><issn>1097-0290</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0U1v1DAQBuAIgehSOPAHwEICicO2tpPY8bEsUArlQ-oWuFkTZ0xdkji1s9sud_43XnahEhLqyRrpmRmP3ix7yOgeo5Tv127c46ws5a1swqiSU8oVvZ1NKKVimpeK72T3YjxPpayEuJvtMClYkeflJPv5ctVD5wxxPVm6pScdjlD71ndIAsbB9xGJt-QEjDmD4LuVwUgMBly66ADJ6AmQOOJw6ZIcMIyLUMPofL9uG8-QHMw_pVEXCxeww34k1gdSY__Dw4gErwYfxvvZHQttxAfbdzc7ff1qPnszPf54eDQ7OJ6akhZyarkSRlCQiolC1ZWlhS2YsKqGHCpJgZuGNVYaKgzYpilASKbAqrJBRpHlu9mzzdwh-IsFxlF3LhpsW-jRL6KWlJVVKfMbYc6YUBWvboRMCcp5WSb45B947hehT9dqzvKUh-Trtc83yAQfY0Crh-A6CCvNqF5HrVPU-nfUyT7aDlzUHTbXcpttAk-3AKKB1gbojYvXTimpZMGT29-4S9fi6v8b9Yuj-Z_V002HS8Ff_e2A8F0LmctSf_lwqL--nX1-_47N9No_3ngLXsO3kH5xesIpyymtCkbT_b8AcUbZaQ</recordid><startdate>20080201</startdate><enddate>20080201</enddate><creator>Kresnowati, M.T.A.P</creator><creator>van Winden, W.A</creator><creator>van Gulik, W.M</creator><creator>Heijnen, J.J</creator><general>Wiley Subscription Services, Inc., A Wiley Company</general><general>Wiley</general><general>Wiley Subscription Services, Inc</general><scope>FBQ</scope><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QF</scope><scope>7QO</scope><scope>7QQ</scope><scope>7SC</scope><scope>7SE</scope><scope>7SP</scope><scope>7SR</scope><scope>7T7</scope><scope>7TA</scope><scope>7TB</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>C1K</scope><scope>F28</scope><scope>FR3</scope><scope>H8D</scope><scope>H8G</scope><scope>JG9</scope><scope>JQ2</scope><scope>KR7</scope><scope>L7M</scope><scope>L~C</scope><scope>L~D</scope><scope>P64</scope><scope>M7N</scope><scope>7X8</scope></search><sort><creationdate>20080201</creationdate><title>Dynamic in vivo metabolome response of Saccharomyces cerevisiae to a stepwise perturbation of the ATP requirement for benzoate export</title><author>Kresnowati, M.T.A.P ; van Winden, W.A ; van Gulik, W.M ; Heijnen, J.J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5047-f296c60a791649b8f04f416f9ba3a870a2cd1df7c06cafdd4a6719af95de10e13</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Adenosine triphosphatase</topic><topic>Adenosine Triphosphate - metabolism</topic><topic>Aerobiosis</topic><topic>ATP</topic><topic>Benzoates - metabolism</topic><topic>benzoic acid</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Carbon - metabolism</topic><topic>Cell culture</topic><topic>Fermentation</topic><topic>Fundamental and applied biological sciences. 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Bioeng</addtitle><date>2008-02-01</date><risdate>2008</risdate><volume>99</volume><issue>2</issue><spage>421</spage><epage>441</epage><pages>421-441</pages><issn>0006-3592</issn><eissn>1097-0290</eissn><coden>BIBIAU</coden><abstract>Although much information is available on in vitro role of ATP in regulation, the in vivo kinetics of reactions in which ATP plays a role are only partly known. In order to study such reactions, it is therefore necessary to study the role of ATP in vivo. This study presents an in vivo, targeted perturbation of the ATP flux in aerobic glucose-limited chemostat cultures of Saccharomyces cerevisiae, which was accomplished by transiently (20 min) changing the extracellular undissociated benzoic acid concentration via the pH of the culture. The performed pH shifts resulted in, within about 20 s, a 40% decrease (pH upshift) or a 23% increase (pH downshift) of the calculated ATP consumption rate while the specific glucose uptake rate did not change because of the glucose-limited condition. The pH upshift resulted in a strong decrease in the glycolytic and TCA cycle fluxes; carbon and energy balances indicated an increased flux toward storage carbohydrates. As expected, the pH downshift leads to the opposite effects. Overall, consistent responses were observed in the metabolic fluxes, the off gas concentrations of O₂ and CO₂ and intracellular metabolite concentrations, except for the concentrations of adenosine nucleotides which unexpectedly only showed minor dynamics. This demonstrates that our knowledge of the regulation of the ATP level, the storage metabolism, and central carbon metabolism of yeast is still incomplete. The new dynamic metabolite datasets obtained in this study will prove of great value in developing kinetic models. Biotechnol. Bioeng. 2008;99: 421-441. © 2007 Wiley Periodicals, Inc.</abstract><cop>Hoboken</cop><pub>Wiley Subscription Services, Inc., A Wiley Company</pub><pmid>17614335</pmid><doi>10.1002/bit.21557</doi><tpages>21</tpages></addata></record> |
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subjects | Adenosine triphosphatase Adenosine Triphosphate - metabolism Aerobiosis ATP Benzoates - metabolism benzoic acid Biological and medical sciences Biotechnology Carbon - metabolism Cell culture Fermentation Fundamental and applied biological sciences. Psychology Glucose Kinetics Metabolism Saccharomyces cerevisiae Saccharomyces cerevisiae - metabolism Studies Systems Biology targetted perturbation Yeast |
title | Dynamic in vivo metabolome response of Saccharomyces cerevisiae to a stepwise perturbation of the ATP requirement for benzoate export |
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