Improved production of medium-chain-length Polyhydroxyalkanotes in glucose-based fed-batch cultivations of metabolically engineered Pseudomonas putida strains.

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Hdl Handle:
http://hdl.handle.net/10033/306374
Title:
Improved production of medium-chain-length Polyhydroxyalkanotes in glucose-based fed-batch cultivations of metabolically engineered Pseudomonas putida strains.
Authors:
Poblete-Castro, Ignacio; Rodriguez, Andre Luis; Lam, Carolyn Ming Chi; Kessler, Wolfgang
Abstract:
One of the major challenges in metabolic engineering for enhanced synthesis of value-added chemicals is to design and develop new strains which can be translated into well-controlled fermentation processes using bioreactors. The aim of this study was to assess the influence of various fed-batch strategies in the performance of metabolically-engineered Pseudomonas putida strains, ∆gcd and ∆gcd-pgl, for improving production of medium-chain-length poly-hydroxyalkanoates (mcl-PHAs) using glucose as the only carbon source. First we developed a fed-batch process which comprised an initial phase of biomass accumulation based on an exponential feeding carbon-limited strategy. For the mcl-PHA accumulation stage, three induction techniques were tested under nitrogen limitation. The substrate-pulse feeding was more efficient than the constant-feeding approach to promote the accumulation of the desirable product. Nonetheless, the most efficient approach for maximum PHA synthesis was the application of a dissolved-oxygen-stat feeding strategy (DO-stat), where P. putida ∆gcd mutant strain showed a final PHA content and specific PHA productivity of 67%, and 0.83 [g•L(-1)•h(-1)], respectively. To our knowledge this mcl-PHA titer is the highest value that has been ever reported using glucose as the solely carbon and energy source. Our results also highlighted the effect of different fed-batch strategies upon the extent of realization of the intended metabolic modification of the mutant strains.
Affiliation:
Microbial Drugs Group, Helmholtz Centre for Infection Research, Inhoffenstraße 7, 38124 Braunschweig, Germany.
Citation:
Improved production of medium-chain-length Polyhydroxyalkanotes in glucose-based fed-batch cultivations of metabolically engineered Pseudomonas putida strains. 2013: J. Microbiol. Biotechnol.
Journal:
Journal of microbiology and biotechnology
Issue Date:
22-Oct-2013
URI:
http://hdl.handle.net/10033/306374
PubMed ID:
24150495
Type:
Article
ISSN:
1738-8872
Appears in Collections:
publications of the department of microbial drugs (MWIS)

Full metadata record

DC FieldValue Language
dc.contributor.authorPoblete-Castro, Ignacioen
dc.contributor.authorRodriguez, Andre Luisen
dc.contributor.authorLam, Carolyn Ming Chien
dc.contributor.authorKessler, Wolfgangen
dc.date.accessioned2013-12-05T13:13:01Z-
dc.date.available2013-12-05T13:13:01Z-
dc.date.issued2013-10-22-
dc.identifier.citationImproved production of medium-chain-length Polyhydroxyalkanotes in glucose-based fed-batch cultivations of metabolically engineered Pseudomonas putida strains. 2013: J. Microbiol. Biotechnol.en
dc.identifier.issn1738-8872-
dc.identifier.pmid24150495-
dc.identifier.urihttp://hdl.handle.net/10033/306374-
dc.description.abstractOne of the major challenges in metabolic engineering for enhanced synthesis of value-added chemicals is to design and develop new strains which can be translated into well-controlled fermentation processes using bioreactors. The aim of this study was to assess the influence of various fed-batch strategies in the performance of metabolically-engineered Pseudomonas putida strains, ∆gcd and ∆gcd-pgl, for improving production of medium-chain-length poly-hydroxyalkanoates (mcl-PHAs) using glucose as the only carbon source. First we developed a fed-batch process which comprised an initial phase of biomass accumulation based on an exponential feeding carbon-limited strategy. For the mcl-PHA accumulation stage, three induction techniques were tested under nitrogen limitation. The substrate-pulse feeding was more efficient than the constant-feeding approach to promote the accumulation of the desirable product. Nonetheless, the most efficient approach for maximum PHA synthesis was the application of a dissolved-oxygen-stat feeding strategy (DO-stat), where P. putida ∆gcd mutant strain showed a final PHA content and specific PHA productivity of 67%, and 0.83 [g•L(-1)•h(-1)], respectively. To our knowledge this mcl-PHA titer is the highest value that has been ever reported using glucose as the solely carbon and energy source. Our results also highlighted the effect of different fed-batch strategies upon the extent of realization of the intended metabolic modification of the mutant strains.en
dc.languageENG-
dc.rightsArchived with thanks to Journal of microbiology and biotechnologyen
dc.titleImproved production of medium-chain-length Polyhydroxyalkanotes in glucose-based fed-batch cultivations of metabolically engineered Pseudomonas putida strains.-
dc.typeArticleen
dc.contributor.departmentMicrobial Drugs Group, Helmholtz Centre for Infection Research, Inhoffenstraße 7, 38124 Braunschweig, Germany.en
dc.identifier.journalJournal of microbiology and biotechnologyen

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