Rheological Behavior of High Cell Density Pseudomonas putida LS46 Cultures during Production of Medium Chain Length Polyhydroxyalkanoate (PHA) Polymers

Blunt, Warren; Gaugler, Marc; Collet, Christophe; Sparling, Richard; Gapes, Daniel J.; Levin, David B.; Çiçek, Nazim

doi:10.3390/bioengineering6040093

Cited by 8 publications

(6 citation statements)

References 85 publications

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“…This trend is in agreement with that observed for P. aeruginosa and Pseudomonas putida . A possible reason for such behavior can be ascribed to shear-induced alignment or breaking of cell aggregates, or eventually to cell deformation as reported in refs and .…”

Section: Resultssupporting

confidence: 92%

Forced Wetting Properties of Bacteria-Laden Droplets Experiencing Initial Evaporation

et al. 2023

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Microbial adhesion and spreading on surfaces are crucial aspects in environmental and industrial settings being also the early stage of complex surface-attached microbial communities known as biofilms. In this work, Pseudomonas fluorescens-laden droplets on hydrophilic substrates (glass coupons) are allowed to partially evaporate before running wetting measurements, to study the effect of evaporation on their interfacial behavior during spillover or splashing. Forced wetting is investigated by imposing controlled centrifugal forces, using a novel rotatory device (Kerberos). At a defined evaporation time, results for the critical tangential force required for the inception of sliding are presented. Microbe-laden droplets exhibit different wetting/spreading properties as a function of the imposed evaporation times. It is found that evaporation is slowed down in bacterial droplets with respect to nutrient medium ones. After sufficient drying times, bacteria accumulate at droplet edges, affecting the droplet shape and thus depinning during forced wetting tests. Droplet rear part does not pin during the rotation test, while only the front part advances and spreads along the force direction. Quantitative results obtained from the well-known Furmidge′s equation reveal that force for sliding inception increases as evaporation time increases. This study can be of support for control of biofilm contamination and removal and possible design of antimicrobial/antibiofouling surfaces.

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Section: Resultssupporting

confidence: 92%

Forced Wetting Properties of Bacteria-Laden Droplets Experiencing Initial Evaporation

et al. 2023

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“…Application of a fed-batch strategy to increase PHA productivity was proved by many other related studies [ 38 , 39 , 40 ]. Therefore, P. resinovorans was cultured with an optimized C/N balance on a flask scale, and cell growth and PHA production monitored ( Figure 6 A).…”

Section: Resultsmentioning

confidence: 97%

Two-Stage Bio-Hydrogen and Polyhydroxyalkanoate Production: Upcycling of Spent Coffee Grounds

et al. 2023

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Coffee waste is an abundant biomass that can be converted into high value chemical products, and is used in various renewable biological processes. In this study, oil was extracted from spent coffee grounds (SCGs) and used for polyhydroxyalkanoate (PHA) production through Pseudomonas resinovorans. The oil–extracted SCGs (OESCGs) were hydrolyzed and used for biohydrogen production through Clostridium butyricum DSM10702. The oil extraction yield through n–hexane was 14.4%, which accounted for 97% of the oil present in the SCGs. OESCG hydrolysate (OESCGH) had a sugar concentration of 32.26 g/L, which was 15.4% higher than that of the SCG hydrolysate (SCGH) (27.96 g/L). Hydrogen production using these substrates was 181.19 mL and 136.58 mL in OESCGH and SCGH media, respectively. The consumed sugar concentration was 6.77 g/L in OESCGH and 5.09 g/L in SCGH media. VFA production with OESCGH (3.58 g/L) increased by 40.9% compared with SCGH (2.54 g/L). In addition, in a fed–batch culture using the extracted oil, cell dry weight was 5.4 g/L, PHA was 1.6 g/L, and PHA contents were 29.5% at 24 h.

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“…The scale-up of agitation speed in IFSBs was validated using the principle of equality of the Froude number. This principle played a pivotal role in propelling the movement of the IFSB, significantly impacting flow characteristics, mixing effects, oxygen transfer, and shear force within the bioreactor (Blunt et al, 2019). Scaling up the total airflow rate in IFSBs was straightforward, as it was based on the working volume.…”

Section: Discussionmentioning

confidence: 99%

Improving product quality and productivity of an antibody-based biotherapeutic using inverted frustoconical shaking bioreactors

Wang,

Xu,

Guo

et al. 2024

Front. Bioeng. Biotechnol.

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The Chinese hamster ovarian (CHO) cells serve as a common choice in biopharmaceutical production, traditionally cultivated in stirred tank bioreactors (STRs). Nevertheless, the pursuit of improved protein quality and production output for commercial purposes demand exploration into new bioreactor types. In this context, inverted frustoconical shaking bioreactors (IFSB) present unique physical properties distinct from STRs. This study aims to compare the production processes of an antibody-based biotherapeutic in both bioreactor types, to enhance production flexibility. The findings indicate that, when compared to STRs, IFSB demonstrates the capability to produce an antibody-based biotherapeutic with either comparable or enhanced bioprocess performance and product quality. IFSB reduces shear damage to cells, enhances viable cell density (VCD), and improves cell state at a 5-L scale. Consequently, this leads to increased protein expression (3.70 g/L vs 2.56 g/L) and improved protein quality, as evidenced by a reduction in acidic variants from 27.0% to 21.5%. Scaling up the culture utilizing the Froude constant and superficial gas velocity ensures stable operation, effective mixing, and gas transfer. The IFSB maintains a high VCD and cell viability at both 50-L and 500-L scales. Product expression levels range from 3.0 to 3.6 g/L, accompanied by an improved acidic variants attribute of 20.6%–22.7%. The IFSB exhibits superior productivity and product quality, underscoring its potential for incorporation into the manufacturing process for antibody-based biotherapeutics. These results establish the foundation for IFSB to become a viable option in producing antibody-based biotherapeutics for clinical and manufacturing applications.

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Rheological Behavior of High Cell Density Pseudomonas putida LS46 Cultures during Production of Medium Chain Length Polyhydroxyalkanoate (PHA) Polymers

Cited by 8 publications

References 85 publications

Forced Wetting Properties of Bacteria-Laden Droplets Experiencing Initial Evaporation

Forced Wetting Properties of Bacteria-Laden Droplets Experiencing Initial Evaporation

Two-Stage Bio-Hydrogen and Polyhydroxyalkanoate Production: Upcycling of Spent Coffee Grounds

Improving product quality and productivity of an antibody-based biotherapeutic using inverted frustoconical shaking bioreactors

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