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Engineering a predatory bacterium as a proficient killer agent for intracellular bio-products recovery: The case of the polyhydroxyalkanoates.

Martínez V, Herencias C, Jurkevitch E, Prieto MA - Sci Rep (2016)

Bottom Line: This work examines the potential of the predatory bacterium Bdellovibrio bacteriovorus HD100, an obligate predator of other Gram-negative bacteria, as an external cell-lytic agent for recovering valuable intracellular bio-products produced by prey cultures.The bio-product targets to be recovered were polyhydroxyalkanoates (PHAs) produced naturally by Pseudomonas putida and Cupriavidus necator, or by recombinant Escherichia coli strains.B. bacteriovorus with a mutated PHA depolymerase gene to prevent the unwanted breakdown of the bio-product allowed the recovery of up to 80% of that accumulated by the prey bacteria, even at high biomass concentrations.

View Article: PubMed Central - PubMed

Affiliation: Environmental Biology Department, Centro de Investigaciones Biológicas, CSIC, C/Ramiro de Maeztu 9, 28040 Madrid, Spain.

ABSTRACT
This work examines the potential of the predatory bacterium Bdellovibrio bacteriovorus HD100, an obligate predator of other Gram-negative bacteria, as an external cell-lytic agent for recovering valuable intracellular bio-products produced by prey cultures. The bio-product targets to be recovered were polyhydroxyalkanoates (PHAs) produced naturally by Pseudomonas putida and Cupriavidus necator, or by recombinant Escherichia coli strains. B. bacteriovorus with a mutated PHA depolymerase gene to prevent the unwanted breakdown of the bio-product allowed the recovery of up to 80% of that accumulated by the prey bacteria, even at high biomass concentrations. This innovative downstream process highlights how B. bacteriovorus can be used as a novel, biological lytic agent for the inexpensive, industrial scale recovery of intracellular products from different Gram-negative prey cultures.

No MeSH data available.


Related in: MedlinePlus

B. bacteriovorus Bd2637 preying on C. necator H16 accumulating PHB.(a) Phase-contrast microscopy of the co-culture at the onset of predation (time zero) and (b) after 24 h of incubation with B. bacteriovorus. (c) Number of viable cells of C. necator H16 accumulating PHB at time zero and after 24 h of predation without and with B. bacteriovorus Bd2637. (d) Number of viable cells of B. bacteriovorus Bd2637 at time zero and after 24 h of predation upon C. necator H16 accumulating PHB. (e) Total PHB content in the co-cultures. In all graphs, error bars indicate the standard deviation of the mean (n = 3). Asterisk indicate significant differences (*P ≤ 0.05) between control prey culture (i.e., without the predator) and B. bacteriovorus Bd2637 co-cultures, as determined by ANOVA-test.
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f6: B. bacteriovorus Bd2637 preying on C. necator H16 accumulating PHB.(a) Phase-contrast microscopy of the co-culture at the onset of predation (time zero) and (b) after 24 h of incubation with B. bacteriovorus. (c) Number of viable cells of C. necator H16 accumulating PHB at time zero and after 24 h of predation without and with B. bacteriovorus Bd2637. (d) Number of viable cells of B. bacteriovorus Bd2637 at time zero and after 24 h of predation upon C. necator H16 accumulating PHB. (e) Total PHB content in the co-cultures. In all graphs, error bars indicate the standard deviation of the mean (n = 3). Asterisk indicate significant differences (*P ≤ 0.05) between control prey culture (i.e., without the predator) and B. bacteriovorus Bd2637 co-cultures, as determined by ANOVA-test.

Mentions: Finally, other model PHB producer like C. necator H16 was grown under PHB production conditions and infected with B. bacteriovorus Bd2637. C. necator H16 (Fig. 6) felt prey to the predator demonstrating the versatility of this lytic system. Specifically, predation upon C. necator H16 let to an increase of 1-log in the viable cell number of B. bacteriovorus (Fig. 6d) and the recovery of 80% of the prey’s PHB after 24 h of predation (Fig. 6e). These results demonstrated the predator’s capability to prey upon other natural PHB producing bacteria, although scaling up would request to optimize the conditions of PHB production and recovery for each particular process.


Engineering a predatory bacterium as a proficient killer agent for intracellular bio-products recovery: The case of the polyhydroxyalkanoates.

Martínez V, Herencias C, Jurkevitch E, Prieto MA - Sci Rep (2016)

B. bacteriovorus Bd2637 preying on C. necator H16 accumulating PHB.(a) Phase-contrast microscopy of the co-culture at the onset of predation (time zero) and (b) after 24 h of incubation with B. bacteriovorus. (c) Number of viable cells of C. necator H16 accumulating PHB at time zero and after 24 h of predation without and with B. bacteriovorus Bd2637. (d) Number of viable cells of B. bacteriovorus Bd2637 at time zero and after 24 h of predation upon C. necator H16 accumulating PHB. (e) Total PHB content in the co-cultures. In all graphs, error bars indicate the standard deviation of the mean (n = 3). Asterisk indicate significant differences (*P ≤ 0.05) between control prey culture (i.e., without the predator) and B. bacteriovorus Bd2637 co-cultures, as determined by ANOVA-test.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834554&req=5

f6: B. bacteriovorus Bd2637 preying on C. necator H16 accumulating PHB.(a) Phase-contrast microscopy of the co-culture at the onset of predation (time zero) and (b) after 24 h of incubation with B. bacteriovorus. (c) Number of viable cells of C. necator H16 accumulating PHB at time zero and after 24 h of predation without and with B. bacteriovorus Bd2637. (d) Number of viable cells of B. bacteriovorus Bd2637 at time zero and after 24 h of predation upon C. necator H16 accumulating PHB. (e) Total PHB content in the co-cultures. In all graphs, error bars indicate the standard deviation of the mean (n = 3). Asterisk indicate significant differences (*P ≤ 0.05) between control prey culture (i.e., without the predator) and B. bacteriovorus Bd2637 co-cultures, as determined by ANOVA-test.
Mentions: Finally, other model PHB producer like C. necator H16 was grown under PHB production conditions and infected with B. bacteriovorus Bd2637. C. necator H16 (Fig. 6) felt prey to the predator demonstrating the versatility of this lytic system. Specifically, predation upon C. necator H16 let to an increase of 1-log in the viable cell number of B. bacteriovorus (Fig. 6d) and the recovery of 80% of the prey’s PHB after 24 h of predation (Fig. 6e). These results demonstrated the predator’s capability to prey upon other natural PHB producing bacteria, although scaling up would request to optimize the conditions of PHB production and recovery for each particular process.

Bottom Line: This work examines the potential of the predatory bacterium Bdellovibrio bacteriovorus HD100, an obligate predator of other Gram-negative bacteria, as an external cell-lytic agent for recovering valuable intracellular bio-products produced by prey cultures.The bio-product targets to be recovered were polyhydroxyalkanoates (PHAs) produced naturally by Pseudomonas putida and Cupriavidus necator, or by recombinant Escherichia coli strains.B. bacteriovorus with a mutated PHA depolymerase gene to prevent the unwanted breakdown of the bio-product allowed the recovery of up to 80% of that accumulated by the prey bacteria, even at high biomass concentrations.

View Article: PubMed Central - PubMed

Affiliation: Environmental Biology Department, Centro de Investigaciones Biológicas, CSIC, C/Ramiro de Maeztu 9, 28040 Madrid, Spain.

ABSTRACT
This work examines the potential of the predatory bacterium Bdellovibrio bacteriovorus HD100, an obligate predator of other Gram-negative bacteria, as an external cell-lytic agent for recovering valuable intracellular bio-products produced by prey cultures. The bio-product targets to be recovered were polyhydroxyalkanoates (PHAs) produced naturally by Pseudomonas putida and Cupriavidus necator, or by recombinant Escherichia coli strains. B. bacteriovorus with a mutated PHA depolymerase gene to prevent the unwanted breakdown of the bio-product allowed the recovery of up to 80% of that accumulated by the prey bacteria, even at high biomass concentrations. This innovative downstream process highlights how B. bacteriovorus can be used as a novel, biological lytic agent for the inexpensive, industrial scale recovery of intracellular products from different Gram-negative prey cultures.

No MeSH data available.


Related in: MedlinePlus