Fed-Batch <i>mcl</i>- Polyhydroxyalkanoates Production in <i>Pseudomonas putida</i> KT2440 and Δ<i>phaZ</i> Mutant on Biodiesel-Derived Crude Glycerol

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dc.contributor.authorBorrero-de Acuna, Jose Manuel
dc.contributor.authorRohde, Manfred
dc.contributor.authorSaldias, Cesar
dc.contributor.authorPoblete-Castro, Ignacio
dc.date.accessioned2025-01-20T23:51:37Z
dc.date.available2025-01-20T23:51:37Z
dc.date.issued2021
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 Delta phaZ mutant in two different fed-batch modes to synthesize mcl-PHAs from raw glycerol. Initially, the cells grew in a batch phase (mu(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 Delta phaZ 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 Delta phaZ 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.
dc.fuente.origenWOS
dc.identifier.doi10.3389/fbioe.2021.642023
dc.identifier.issn2296-4185
dc.identifier.urihttps://doi.org/10.3389/fbioe.2021.642023
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/94827
dc.identifier.wosidWOS:000634719300001
dc.language.isoen
dc.revistaFrontiers in bioengineering and biotechnology
dc.rightsacceso restringido
dc.subjectmcl-poly(3-hydroxyalkanoates)
dc.subjectfed-batch
dc.subjectcrude glycerol
dc.subjectpseudomonas putida
dc.subjectPHA depolymerase
dc.subject.ods12 Responsible Consumption and Production
dc.subject.odspa12 Producción y consumo responsable
dc.titleFed-Batch <i>mcl</i>- Polyhydroxyalkanoates Production in <i>Pseudomonas putida</i> KT2440 and Δ<i>phaZ</i> Mutant on Biodiesel-Derived Crude Glycerol
dc.typeartículo
dc.volumen9
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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