Substrate cycles in <it<Penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis

<p<Abstract</p< <p<Background</p< <p<<it<Penicillium chrysogenum</it<, the main production strain for penicillin-G, has a high content of intracellular carbohydrates, especially reduced sugars such as mannitol, arabitol, erythritol, as well as trehalose and...
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Autor*in:	Zhao Zheng [verfasserIn] ten Pierick Angela [verfasserIn] de Jonge Lodewijk [verfasserIn] Heijnen Joseph J [verfasserIn] Wahl S Aljoscha [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2012

Übergeordnetes Werk:	In: Microbial Cell Factories - BMC, 2003, 11(2012), 1, p 140
Übergeordnetes Werk:	volume:11 ; year:2012 ; number:1, p 140

Links:	Link aufrufen Link aufrufen Link aufrufen Journal toc

DOI / URN:	10.1186/1475-2859-11-140

Katalog-ID:	DOAJ072620706

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allfields	10.1186/1475-2859-11-140 doi (DE-627)DOAJ072620706 (DE-599)DOAJ8440abeef41f4b149d025291f5e5678a DE-627 ger DE-627 rakwb eng QR1-502 Zhao Zheng verfasserin aut Substrate cycles in <it<Penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis 2012 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier <p<Abstract</p< <p<Background</p< <p<<it<Penicillium chrysogenum</it<, the main production strain for penicillin-G, has a high content of intracellular carbohydrates, especially reduced sugars such as mannitol, arabitol, erythritol, as well as trehalose and glycogen. In previous steady state <sup<13</sup<C wash-in experiments a delay of labeling enrichments in glycolytic intermediates was observed, which suggests turnover of storage carbohydrates. The turnover of storage pools consumes ATP which is expected to reduce the product yield for energy demanding production pathways like penicillin-G.</p< <p<Results</p< <p<In this study, a <sup<13</sup<C labeling wash-in experiment of 1 hour was performed to systematically quantify the intracellular flux distribution including eight substrate cycles. The experiments were performed using a mixed carbon source of 85% Cmol<sub<Glc</sub</Cmol<sub<Glc+EtOH</sub< labeled glucose (mixture of 90% [1-<sup<13</sup<C<sub<1</sub<] and 10% [U-<sup<13</sup<C<sub<6</sub<]) and 15% ethanol [U-<sup<13</sup<C<sub<2</sub<]. It was found, that (1) also several extracellular pools are enriched with <sup<13</sup<C labeling rapidly (trehalose, mannitol, and others), (2) the intra- to extracellular metabolite concentration ratios were comparable for a large set of metabolites while for some carbohydrates (mannitol, trehalose, and glucose) the measured ratios were much higher.</p< <p<Conclusions</p< <p<The fast enrichment of several extracellular carbohydrates and a concentration ratio higher than the ratio expected from cell lysis (2%) indicate active (e.g. ATP consuming) transport cycles over the cellular membrane. The flux estimation indicates, that substrate cycles account for about 52% of the gap in the ATP balance based on metabolic flux analysis.</p< Microbiology ten Pierick Angela verfasserin aut de Jonge Lodewijk verfasserin aut Heijnen Joseph J verfasserin aut Wahl S Aljoscha verfasserin aut In Microbial Cell Factories BMC, 2003 11(2012), 1, p 140 (DE-627)355987651 (DE-600)2091377-1 14752859 nnns volume:11 year:2012 number:1, p 140 https://doi.org/10.1186/1475-2859-11-140 kostenfrei https://doaj.org/article/8440abeef41f4b149d025291f5e5678a kostenfrei http://www.microbialcellfactories.com/content/11/1/140 kostenfrei https://doaj.org/toc/1475-2859 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2012 1, p 140
spelling	10.1186/1475-2859-11-140 doi (DE-627)DOAJ072620706 (DE-599)DOAJ8440abeef41f4b149d025291f5e5678a DE-627 ger DE-627 rakwb eng QR1-502 Zhao Zheng verfasserin aut Substrate cycles in <it<Penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis 2012 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier <p<Abstract</p< <p<Background</p< <p<<it<Penicillium chrysogenum</it<, the main production strain for penicillin-G, has a high content of intracellular carbohydrates, especially reduced sugars such as mannitol, arabitol, erythritol, as well as trehalose and glycogen. In previous steady state <sup<13</sup<C wash-in experiments a delay of labeling enrichments in glycolytic intermediates was observed, which suggests turnover of storage carbohydrates. The turnover of storage pools consumes ATP which is expected to reduce the product yield for energy demanding production pathways like penicillin-G.</p< <p<Results</p< <p<In this study, a <sup<13</sup<C labeling wash-in experiment of 1 hour was performed to systematically quantify the intracellular flux distribution including eight substrate cycles. The experiments were performed using a mixed carbon source of 85% Cmol<sub<Glc</sub</Cmol<sub<Glc+EtOH</sub< labeled glucose (mixture of 90% [1-<sup<13</sup<C<sub<1</sub<] and 10% [U-<sup<13</sup<C<sub<6</sub<]) and 15% ethanol [U-<sup<13</sup<C<sub<2</sub<]. It was found, that (1) also several extracellular pools are enriched with <sup<13</sup<C labeling rapidly (trehalose, mannitol, and others), (2) the intra- to extracellular metabolite concentration ratios were comparable for a large set of metabolites while for some carbohydrates (mannitol, trehalose, and glucose) the measured ratios were much higher.</p< <p<Conclusions</p< <p<The fast enrichment of several extracellular carbohydrates and a concentration ratio higher than the ratio expected from cell lysis (2%) indicate active (e.g. ATP consuming) transport cycles over the cellular membrane. The flux estimation indicates, that substrate cycles account for about 52% of the gap in the ATP balance based on metabolic flux analysis.</p< Microbiology ten Pierick Angela verfasserin aut de Jonge Lodewijk verfasserin aut Heijnen Joseph J verfasserin aut Wahl S Aljoscha verfasserin aut In Microbial Cell Factories BMC, 2003 11(2012), 1, p 140 (DE-627)355987651 (DE-600)2091377-1 14752859 nnns volume:11 year:2012 number:1, p 140 https://doi.org/10.1186/1475-2859-11-140 kostenfrei https://doaj.org/article/8440abeef41f4b149d025291f5e5678a kostenfrei http://www.microbialcellfactories.com/content/11/1/140 kostenfrei https://doaj.org/toc/1475-2859 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2012 1, p 140
allfields_unstemmed	10.1186/1475-2859-11-140 doi (DE-627)DOAJ072620706 (DE-599)DOAJ8440abeef41f4b149d025291f5e5678a DE-627 ger DE-627 rakwb eng QR1-502 Zhao Zheng verfasserin aut Substrate cycles in <it<Penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis 2012 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier <p<Abstract</p< <p<Background</p< <p<<it<Penicillium chrysogenum</it<, the main production strain for penicillin-G, has a high content of intracellular carbohydrates, especially reduced sugars such as mannitol, arabitol, erythritol, as well as trehalose and glycogen. In previous steady state <sup<13</sup<C wash-in experiments a delay of labeling enrichments in glycolytic intermediates was observed, which suggests turnover of storage carbohydrates. The turnover of storage pools consumes ATP which is expected to reduce the product yield for energy demanding production pathways like penicillin-G.</p< <p<Results</p< <p<In this study, a <sup<13</sup<C labeling wash-in experiment of 1 hour was performed to systematically quantify the intracellular flux distribution including eight substrate cycles. The experiments were performed using a mixed carbon source of 85% Cmol<sub<Glc</sub</Cmol<sub<Glc+EtOH</sub< labeled glucose (mixture of 90% [1-<sup<13</sup<C<sub<1</sub<] and 10% [U-<sup<13</sup<C<sub<6</sub<]) and 15% ethanol [U-<sup<13</sup<C<sub<2</sub<]. It was found, that (1) also several extracellular pools are enriched with <sup<13</sup<C labeling rapidly (trehalose, mannitol, and others), (2) the intra- to extracellular metabolite concentration ratios were comparable for a large set of metabolites while for some carbohydrates (mannitol, trehalose, and glucose) the measured ratios were much higher.</p< <p<Conclusions</p< <p<The fast enrichment of several extracellular carbohydrates and a concentration ratio higher than the ratio expected from cell lysis (2%) indicate active (e.g. ATP consuming) transport cycles over the cellular membrane. The flux estimation indicates, that substrate cycles account for about 52% of the gap in the ATP balance based on metabolic flux analysis.</p< Microbiology ten Pierick Angela verfasserin aut de Jonge Lodewijk verfasserin aut Heijnen Joseph J verfasserin aut Wahl S Aljoscha verfasserin aut In Microbial Cell Factories BMC, 2003 11(2012), 1, p 140 (DE-627)355987651 (DE-600)2091377-1 14752859 nnns volume:11 year:2012 number:1, p 140 https://doi.org/10.1186/1475-2859-11-140 kostenfrei https://doaj.org/article/8440abeef41f4b149d025291f5e5678a kostenfrei http://www.microbialcellfactories.com/content/11/1/140 kostenfrei https://doaj.org/toc/1475-2859 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2012 1, p 140
allfieldsGer	10.1186/1475-2859-11-140 doi (DE-627)DOAJ072620706 (DE-599)DOAJ8440abeef41f4b149d025291f5e5678a DE-627 ger DE-627 rakwb eng QR1-502 Zhao Zheng verfasserin aut Substrate cycles in <it<Penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis 2012 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier <p<Abstract</p< <p<Background</p< <p<<it<Penicillium chrysogenum</it<, the main production strain for penicillin-G, has a high content of intracellular carbohydrates, especially reduced sugars such as mannitol, arabitol, erythritol, as well as trehalose and glycogen. In previous steady state <sup<13</sup<C wash-in experiments a delay of labeling enrichments in glycolytic intermediates was observed, which suggests turnover of storage carbohydrates. The turnover of storage pools consumes ATP which is expected to reduce the product yield for energy demanding production pathways like penicillin-G.</p< <p<Results</p< <p<In this study, a <sup<13</sup<C labeling wash-in experiment of 1 hour was performed to systematically quantify the intracellular flux distribution including eight substrate cycles. The experiments were performed using a mixed carbon source of 85% Cmol<sub<Glc</sub</Cmol<sub<Glc+EtOH</sub< labeled glucose (mixture of 90% [1-<sup<13</sup<C<sub<1</sub<] and 10% [U-<sup<13</sup<C<sub<6</sub<]) and 15% ethanol [U-<sup<13</sup<C<sub<2</sub<]. It was found, that (1) also several extracellular pools are enriched with <sup<13</sup<C labeling rapidly (trehalose, mannitol, and others), (2) the intra- to extracellular metabolite concentration ratios were comparable for a large set of metabolites while for some carbohydrates (mannitol, trehalose, and glucose) the measured ratios were much higher.</p< <p<Conclusions</p< <p<The fast enrichment of several extracellular carbohydrates and a concentration ratio higher than the ratio expected from cell lysis (2%) indicate active (e.g. ATP consuming) transport cycles over the cellular membrane. The flux estimation indicates, that substrate cycles account for about 52% of the gap in the ATP balance based on metabolic flux analysis.</p< Microbiology ten Pierick Angela verfasserin aut de Jonge Lodewijk verfasserin aut Heijnen Joseph J verfasserin aut Wahl S Aljoscha verfasserin aut In Microbial Cell Factories BMC, 2003 11(2012), 1, p 140 (DE-627)355987651 (DE-600)2091377-1 14752859 nnns volume:11 year:2012 number:1, p 140 https://doi.org/10.1186/1475-2859-11-140 kostenfrei https://doaj.org/article/8440abeef41f4b149d025291f5e5678a kostenfrei http://www.microbialcellfactories.com/content/11/1/140 kostenfrei https://doaj.org/toc/1475-2859 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2012 1, p 140
allfieldsSound	10.1186/1475-2859-11-140 doi (DE-627)DOAJ072620706 (DE-599)DOAJ8440abeef41f4b149d025291f5e5678a DE-627 ger DE-627 rakwb eng QR1-502 Zhao Zheng verfasserin aut Substrate cycles in <it<Penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis 2012 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier <p<Abstract</p< <p<Background</p< <p<<it<Penicillium chrysogenum</it<, the main production strain for penicillin-G, has a high content of intracellular carbohydrates, especially reduced sugars such as mannitol, arabitol, erythritol, as well as trehalose and glycogen. In previous steady state <sup<13</sup<C wash-in experiments a delay of labeling enrichments in glycolytic intermediates was observed, which suggests turnover of storage carbohydrates. The turnover of storage pools consumes ATP which is expected to reduce the product yield for energy demanding production pathways like penicillin-G.</p< <p<Results</p< <p<In this study, a <sup<13</sup<C labeling wash-in experiment of 1 hour was performed to systematically quantify the intracellular flux distribution including eight substrate cycles. The experiments were performed using a mixed carbon source of 85% Cmol<sub<Glc</sub</Cmol<sub<Glc+EtOH</sub< labeled glucose (mixture of 90% [1-<sup<13</sup<C<sub<1</sub<] and 10% [U-<sup<13</sup<C<sub<6</sub<]) and 15% ethanol [U-<sup<13</sup<C<sub<2</sub<]. It was found, that (1) also several extracellular pools are enriched with <sup<13</sup<C labeling rapidly (trehalose, mannitol, and others), (2) the intra- to extracellular metabolite concentration ratios were comparable for a large set of metabolites while for some carbohydrates (mannitol, trehalose, and glucose) the measured ratios were much higher.</p< <p<Conclusions</p< <p<The fast enrichment of several extracellular carbohydrates and a concentration ratio higher than the ratio expected from cell lysis (2%) indicate active (e.g. ATP consuming) transport cycles over the cellular membrane. The flux estimation indicates, that substrate cycles account for about 52% of the gap in the ATP balance based on metabolic flux analysis.</p< Microbiology ten Pierick Angela verfasserin aut de Jonge Lodewijk verfasserin aut Heijnen Joseph J verfasserin aut Wahl S Aljoscha verfasserin aut In Microbial Cell Factories BMC, 2003 11(2012), 1, p 140 (DE-627)355987651 (DE-600)2091377-1 14752859 nnns volume:11 year:2012 number:1, p 140 https://doi.org/10.1186/1475-2859-11-140 kostenfrei https://doaj.org/article/8440abeef41f4b149d025291f5e5678a kostenfrei http://www.microbialcellfactories.com/content/11/1/140 kostenfrei https://doaj.org/toc/1475-2859 Journal toc kostenfrei GBV_USEFLAG_A SYSFLAG_A GBV_DOAJ GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_31 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_224 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2005 GBV_ILN_2009 GBV_ILN_2011 GBV_ILN_2014 GBV_ILN_2055 GBV_ILN_2111 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 11 2012 1, p 140
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title	Substrate cycles in <it<Penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis
ctrlnum	(DE-627)DOAJ072620706 (DE-599)DOAJ8440abeef41f4b149d025291f5e5678a
title_full	Substrate cycles in <it<Penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis
author_sort	Zhao Zheng
journal	Microbial Cell Factories
journalStr	Microbial Cell Factories
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author_browse	Zhao Zheng ten Pierick Angela de Jonge Lodewijk Heijnen Joseph J Wahl S Aljoscha
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doi_str_mv	10.1186/1475-2859-11-140
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title_sort	substrate cycles in <it<penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis
callnumber	QR1-502
title_auth	Substrate cycles in <it<Penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis
abstract	<p<Abstract</p< <p<Background</p< <p<<it<Penicillium chrysogenum</it<, the main production strain for penicillin-G, has a high content of intracellular carbohydrates, especially reduced sugars such as mannitol, arabitol, erythritol, as well as trehalose and glycogen. In previous steady state <sup<13</sup<C wash-in experiments a delay of labeling enrichments in glycolytic intermediates was observed, which suggests turnover of storage carbohydrates. The turnover of storage pools consumes ATP which is expected to reduce the product yield for energy demanding production pathways like penicillin-G.</p< <p<Results</p< <p<In this study, a <sup<13</sup<C labeling wash-in experiment of 1 hour was performed to systematically quantify the intracellular flux distribution including eight substrate cycles. The experiments were performed using a mixed carbon source of 85% Cmol<sub<Glc</sub</Cmol<sub<Glc+EtOH</sub< labeled glucose (mixture of 90% [1-<sup<13</sup<C<sub<1</sub<] and 10% [U-<sup<13</sup<C<sub<6</sub<]) and 15% ethanol [U-<sup<13</sup<C<sub<2</sub<]. It was found, that (1) also several extracellular pools are enriched with <sup<13</sup<C labeling rapidly (trehalose, mannitol, and others), (2) the intra- to extracellular metabolite concentration ratios were comparable for a large set of metabolites while for some carbohydrates (mannitol, trehalose, and glucose) the measured ratios were much higher.</p< <p<Conclusions</p< <p<The fast enrichment of several extracellular carbohydrates and a concentration ratio higher than the ratio expected from cell lysis (2%) indicate active (e.g. ATP consuming) transport cycles over the cellular membrane. The flux estimation indicates, that substrate cycles account for about 52% of the gap in the ATP balance based on metabolic flux analysis.</p<
abstractGer	<p<Abstract</p< <p<Background</p< <p<<it<Penicillium chrysogenum</it<, the main production strain for penicillin-G, has a high content of intracellular carbohydrates, especially reduced sugars such as mannitol, arabitol, erythritol, as well as trehalose and glycogen. In previous steady state <sup<13</sup<C wash-in experiments a delay of labeling enrichments in glycolytic intermediates was observed, which suggests turnover of storage carbohydrates. The turnover of storage pools consumes ATP which is expected to reduce the product yield for energy demanding production pathways like penicillin-G.</p< <p<Results</p< <p<In this study, a <sup<13</sup<C labeling wash-in experiment of 1 hour was performed to systematically quantify the intracellular flux distribution including eight substrate cycles. The experiments were performed using a mixed carbon source of 85% Cmol<sub<Glc</sub</Cmol<sub<Glc+EtOH</sub< labeled glucose (mixture of 90% [1-<sup<13</sup<C<sub<1</sub<] and 10% [U-<sup<13</sup<C<sub<6</sub<]) and 15% ethanol [U-<sup<13</sup<C<sub<2</sub<]. It was found, that (1) also several extracellular pools are enriched with <sup<13</sup<C labeling rapidly (trehalose, mannitol, and others), (2) the intra- to extracellular metabolite concentration ratios were comparable for a large set of metabolites while for some carbohydrates (mannitol, trehalose, and glucose) the measured ratios were much higher.</p< <p<Conclusions</p< <p<The fast enrichment of several extracellular carbohydrates and a concentration ratio higher than the ratio expected from cell lysis (2%) indicate active (e.g. ATP consuming) transport cycles over the cellular membrane. The flux estimation indicates, that substrate cycles account for about 52% of the gap in the ATP balance based on metabolic flux analysis.</p<
abstract_unstemmed	<p<Abstract</p< <p<Background</p< <p<<it<Penicillium chrysogenum</it<, the main production strain for penicillin-G, has a high content of intracellular carbohydrates, especially reduced sugars such as mannitol, arabitol, erythritol, as well as trehalose and glycogen. In previous steady state <sup<13</sup<C wash-in experiments a delay of labeling enrichments in glycolytic intermediates was observed, which suggests turnover of storage carbohydrates. The turnover of storage pools consumes ATP which is expected to reduce the product yield for energy demanding production pathways like penicillin-G.</p< <p<Results</p< <p<In this study, a <sup<13</sup<C labeling wash-in experiment of 1 hour was performed to systematically quantify the intracellular flux distribution including eight substrate cycles. The experiments were performed using a mixed carbon source of 85% Cmol<sub<Glc</sub</Cmol<sub<Glc+EtOH</sub< labeled glucose (mixture of 90% [1-<sup<13</sup<C<sub<1</sub<] and 10% [U-<sup<13</sup<C<sub<6</sub<]) and 15% ethanol [U-<sup<13</sup<C<sub<2</sub<]. It was found, that (1) also several extracellular pools are enriched with <sup<13</sup<C labeling rapidly (trehalose, mannitol, and others), (2) the intra- to extracellular metabolite concentration ratios were comparable for a large set of metabolites while for some carbohydrates (mannitol, trehalose, and glucose) the measured ratios were much higher.</p< <p<Conclusions</p< <p<The fast enrichment of several extracellular carbohydrates and a concentration ratio higher than the ratio expected from cell lysis (2%) indicate active (e.g. ATP consuming) transport cycles over the cellular membrane. The flux estimation indicates, that substrate cycles account for about 52% of the gap in the ATP balance based on metabolic flux analysis.</p<
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title_short	Substrate cycles in <it<Penicillium chrysogenum</it< quantified by isotopic non-stationary flux analysis
url	https://doi.org/10.1186/1475-2859-11-140 https://doaj.org/article/8440abeef41f4b149d025291f5e5678a http://www.microbialcellfactories.com/content/11/1/140 https://doaj.org/toc/1475-2859
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