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dc.contributor.authorBorrero-de Acuña, José Manuel
dc.contributor.authorRohde, Manfred
dc.contributor.authorSaldias, Cesar
dc.contributor.authorPoblete-Castro, Ignacio
dc.date.accessioned2021-05-12T11:18:23Z
dc.date.available2021-05-12T11:18:23Z
dc.date.issued2021-03-16
dc.identifier.citationFront Bioeng Biotechnol. 2021 Mar 16;9:642023. doi: 10.3389/fbioe.2021.642023.en_US
dc.identifier.issn2296-4185
dc.identifier.pmid33796510
dc.identifier.doi10.3389/fbioe.2021.642023
dc.identifier.urihttp://hdl.handle.net/10033/622869
dc.description.abstractCrude glycerol has emerged as a suitable feedstock for the biotechnological production of various industrial chemicals given its high surplus catalyzed by the biodiesel industry. Pseudomonas bacteria metabolize the polyol into several biopolymers, including alginate and medium-chain-length poly(3-hydroxyalkanoates) (mcl-PHAs). Although P. putida is a suited platform to derive these polyoxoesters from crude glycerol, the attained concentrations in batch and fed-batch cultures are still low. In this study, we employed P. putida KT2440 and the hyper-PHA producer ΔphaZ mutant in two different fed-batch modes to synthesize mcl-PHAs from raw glycerol. Initially, the cells grew in a batch phase (μ max 0.21 h-1) for 22 h followed by a carbon-limiting exponential feeding, where the specific growth rate was set at 0.1 (h-1), resulting in a cell dry weight (CDW) of nearly 50 (g L-1) at 40 h cultivation. During the PHA production stage, we supplied the substrate at a constant rate of 50 (g h-1), where the KT2440 and the ΔphaZ produced 9.7 and 12.7 gPHA L-1, respectively, after 60 h cultivation. We next evaluated the PHA production ability of the P. putida strains using a DO-stat approach under nitrogen depletion. Citric acid was the main by-product secreted by the cells, accumulating in the culture broth up to 48 (g L-1) under nitrogen limitation. The mutant ΔphaZ amassed 38.9% of the CDW as mcl-PHA and exhibited a specific PHA volumetric productivity of 0.34 (g L-1 h-1), 48% higher than the parental KT2440 under the same growth conditions. The biosynthesized mcl-PHAs had average molecular weights ranging from 460 to 505 KDa and a polydispersity index (PDI) of 2.4-2.6. Here, we demonstrated that the DO-stat feeding approach in high cell density cultures enables the high yield production of mcl-PHA in P. putida strains using the industrial crude glycerol, where the fed-batch process selection is essential to exploit the superior biopolymer production hallmarks of engineered bacterial strains.en_US
dc.language.isoenen_US
dc.publisherFrontiersen_US
dc.rightsAttribution 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectPHA depolymeraseen_US
dc.subjectcrude glycerolen_US
dc.subjectfed-batchen_US
dc.subjectmcl-poly(3-hydroxyalkanoates)en_US
dc.subjectpseudomonas putidaen_US
dc.titleFed-Batch - Polyhydroxyalkanoates Production in Pseudomonas putida KT2440 and Δ phaZ KT2440 and Δ Mutant on Biodiesel-Derived Crude Glycerol.en_US
dc.typeArticleen_US
dc.contributor.departmentHZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany.en_US
dc.identifier.journalFrontiers in bioengineering and biotechnologyen_US
dc.source.volume9
dc.source.beginpage642023
dc.source.endpage
refterms.dateFOA2021-05-12T11:18:24Z
dc.source.journaltitleFrontiers in bioengineering and biotechnology
dc.source.countrySwitzerland


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Attribution 4.0 International
Except where otherwise noted, this item's license is described as Attribution 4.0 International