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Batch and fed-batch growth of Pichia pastoris under increased air pressure

Abstract

Pichia pastoris CBS 2612 behavior under air pressures of 1, 3 and 5 bar in culture media of glycerol (pure and crude) and methanol was studied. Generally, the increase in oxygen transfer rate due to the increase of total pressure improved cellular growth for all carbon sources and for batch and fed-batch processes with different feeding rate strategies. In batch cultures, 1.4-, 1.2-, and 1.5-fold improvement in biomass production was obtained with the increase of air pressure up to 5 bar, using methanol, pure glycerol, and crude glycerol, respectively. The increase of air pressure to 5 bar using exponential feeding rate led to 1.4-fold improvement in biomass yield per glycerol mass consumed, for crude and pure glycerol. The current low cost of crude glycerol from the biodiesel production together with the present results shows the possibility of improving cell mass production of P. pastoris using increased air pressure.

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Abbreviations

C :

Dissolved oxygen concentration in the liquid (mg O2/L)

C* :

Solubility of oxygen in the liquid (mg O2/L)

CDW:

Cell dry weight

D :

Dilution rate (h−1)

DO:

Dissolved oxygen tension (%)

F :

Feed rate (mL/min)

\( H_{{{\text{O}}_{2} }} \) :

Henry constant for oxygen

k L a :

Volumetric oxygen mass transfer coefficient (h−1 or s−1)

NAD:

Nicotinamide adenine dinucleotide

OTR:

Oxygen transfer rate (mg O2/L h)

qO2 :

Specific oxygen uptake rate (mg O2/g h)

q s :

Maximum specific substrate consumption rate (g/g h)

\( p_{{{\text{O}}_{ 2} }} \) :

Oxygen partial pressure (bar)

P :

Absolute pressure (bar)

\( P_{\text{T}} \) :

Total air pressure (bar)

t :

Time (h)

V 0 :

Initial culture volume (mL)

\( y_{{{\text{O}}_{ 2} }} \) :

Oxygen molar fraction in the gas

Y x/O :

Cell mass yield per oxygen mass consumed (g/g)

Y x/s :

Cell mass yield per carbon source mass consumed (g/g)

μ :

Specific growth rate (h−1)

υ :

Superficial gas velocity (m/s)

O:

Oxygen

S:

Substrate

T:

Total

X:

Biomass

0:

Initial value

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Acknowledgments

The authors acknowledge the financial support provided by “Fundação para a Ciência e Tecnologia” (Grant SFRH/BD/47371/2008).

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Correspondence to Isabel Belo.

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Lopes, M., Belo, I. & Mota, M. Batch and fed-batch growth of Pichia pastoris under increased air pressure. Bioprocess Biosyst Eng 36, 1267–1275 (2013). https://doi.org/10.1007/s00449-012-0871-5

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Keywords

  • Pichia pastoris
  • Increased air pressure
  • Oxygen transfer rate (OTR)
  • Crude glycerol