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Recombinant Escherichia coli cultivation in a pressurized airlift bioreactor: assessment of the influence of temperature on oxygen transfer and uptake rates

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Abstract

Temperature influences the rates of oxygen transfer (OTR) and uptake (q O2) in aerobic bioprocesses. Hence, joint analysis of q O2 and OTR at variable temperature is essential for bioprocess optimization and control. However, no such analyses have yet been reported for cultures of engineered E. coli producing recombinant proteins. E. coli cultivations at different temperatures (27–37 °C) were performed using a 5-L stirred tank bioreactor (STB), and a 5-L airlift bioreactor (ALB) was used to measure k L a and validate models of q O2 and OTR. The equations were then employed to evaluate the cultivation process in the ALB at different pressures (0.1–0.4 MPa) and temperatures (27–37 °C). The results showed that the positive effect of temperature on k L a was more pronounced than the negative influence on oxygen solubility, increasing the OTR in the ALB. The specific growth rate and temperature influenced q O2. In contrast to previous reports, the results showed that q O2 was not explicitly affected by recombinant protein synthesis. In addition, model predictions revealed that biomass concentration and productivity were greatly improved by pressurization of the system and use of a lower temperature.

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Abbreviations

a :

Gas–liquid interfacial area per unit of liquid volume (m−1)

a k :

Parameter of Eq. (12) (h−1)

a q :

Parameter of Eq. (7) (°C−3 h−1)

a µ :

Parameter of Eq. (3) (°C−1 h−1)

ALB:

Airlift bioreactor

b k :

Parameter of Eq. (12) (min)

b q :

Parameter of Eq. (7) (h−1)

b µ :

Parameter of Eq. (3) (h−1)

C :

Dissolved oxygen concentration in the bulk liquid (g L−1)

c j :

Concentration of non-electrolyte j (g L−1)

c k :

Parameter of Eq. (12) (min−1)

C * :

Oxygen solubility in the culture medium (g L−1)

C *0 :

Oxygen solubility in water (g L−1)

C x :

Cell concentration (gDCW L−1)

C x0 :

Cell concentration at the start of exponential growth phase (gDCW L−1)

C xmax :

Maximum cell concentration (gDCW L−1)

D :

Oxygen diffusion coefficient (m2 h−1)

d b :

Bubble diameter (m)

DO:

Dissolved oxygen

DOT:

Dissolved oxygen tension (% of saturation)

H i :

Salting-out parameter of ion i in Eq. (15) (L mol−1)

h D :

Height of the aerated liquid (cm)

h L :

Height of the non-aerated liquid (cm)

I i :

Ionic strength of ion i (mol L−1)

IPTG:

Isopropyl-β-d-thiogalactopyranoside

K j :

Solubility parameter for non-electrolyte j in Eq. (15) (L g−1)

k e :

Electrode sensitivity (h−1)

k L :

Mass transfer coefficient (m h−1)

k L a :

Volumetric oxygen transfer coefficient (h−1)

q O2 :

Specific oxygen transfer rate (gO2 g −1DCW h−1)

q p :

Specific production rate of PspA (h−1)

OTR:

Oxygen transfer rate (g L−1 h−1)

OUR:

Oxygen uptake rate (g L−1 h−1)

PspA:

Pneumococcal surface protein A

P x :

Biomass productivity (gDCW L−1 h−1)

p :

Absolute pressure (MPa)

p O2 :

Absolute partial pressure of oxygen (MPa)

m O2 :

Oxygen consumption coefficient for maintenance (h−1)

M O2 :

Molar mass of molecular oxygen (g mol−1)

ṅ in :

Molar flow rate of the gas inlet stream (mol L−1 h−1)

ṅ out :

Molar flow rate of the exhaust gas (mol L−1 h−1)

STB:

Stirred tank bioreactor

t :

Time (h)

t e :

Exposure time (h)

T :

Temperature (°C)

y CO2 :

Mole fraction of carbon dioxide

y O2 :

Mole fraction of oxygen

Y x/O2 :

Biomass yield on oxygen (gDCW g −1O2 h−1)

V :

Working volume of the bioreactor (L)

Δt :

Time of cultivation (h)

ε :

Gas hold-up

ϕ :

Specific gas flow rate (min−1)

µ :

Specific growth rate (h−1)

µ max :

Maximum specific growth rate (h−1)

θ :

Parameter of Eq. (12) (dimensionless)

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Acknowledgements

The authors would like to thank CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico, Brazil) (Grant Process 142482/2014-5), FAPESP (Fundação de Amparo à Pesquisa do Estado de São Paulo, Brazil) (Grant Process 2015/10291-8), and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior, Brazil) for funding this work. We also acknowledge Amadeus Azevedo for HPLC analyses and Prof. Alberto C. Badino for the 5-L airlift bioreactor used.

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Authors and Affiliations

  1. Chemical Engineering Graduate Program, Federal University of São Carlos (PPEQ-UFSCar), Rodovia Washington Luís, km 235, São Carlos, SP, 13565-905, Brazil

    Gilson Campani, Gabriel Gonçalves da Silva, Teresa Cristina Zangirolami & Marcelo Perencin de Arruda Ribeiro

  2. Department of Chemical Engineering, Federal University of São Carlos, Rodovia Washington Luís, km 235, São Carlos, SP, 13565-905, Brazil

    Teresa Cristina Zangirolami & Marcelo Perencin de Arruda Ribeiro

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  1. Gilson Campani
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Correspondence to Marcelo Perencin de Arruda Ribeiro.

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Campani, G., Gonçalves da Silva, G., Zangirolami, T.C. et al. Recombinant Escherichia coli cultivation in a pressurized airlift bioreactor: assessment of the influence of temperature on oxygen transfer and uptake rates. Bioprocess Biosyst Eng 40, 1621–1633 (2017). https://doi.org/10.1007/s00449-017-1818-7

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