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Multi-stage high cell continuous fermentation for high productivity and titer

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Abstract

We carried out the first simulation on multi-stage continuous high cell density culture (MSC-HCDC) to show that the MSC-HCDC can achieve batch/fed-batch product titer with much higher productivity to the fed-batch productivity using published fermentation kinetics of lactic acid, penicillin and ethanol. The system under consideration consists of n-serially connected continuous stirred-tank reactors (CSTRs) with either hollow fiber cell recycling or cell immobilization for high cell-density culture. In each CSTR substrate supply and product removal are possible. Penicillin production is severely limited by glucose metabolite repression that requires multi-CSTR glucose feeding. An 8-stage C-HCDC lactic acid fermentation resulted in 212.9 g/L of titer and 10.6 g/L/h of productivity, corresponding to 101 and 429% of the comparable lactic acid fed-batch, respectively. The penicillin production model predicted 149% (0.085 g/L/h) of productivity in 8-stage C-HCDC with 40 g/L of cell density and 289% of productivity (0.165 g/L/h) in 7-stage C-HCDC with 60 g/L of cell density compared with referring batch cultivations. A 2-stage C-HCDC ethanol experimental run showed 107% titer and 257% productivity of the batch system having 88.8 g/L of titer and 3.7 g/L/h of productivity. MSC-HCDC can give much higher productivity than batch/fed-batch system, and yield a several percentage higher titer as well. The productivity ratio of MSC-HCDC over batch/fed-batch system is given as a multiplication of system dilution rate of MSC-HCDC and cycle time of batch/fed-batch system. We suggest MSC-HCDC as a new production platform for various fermentation products including monoclonal antibody.

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Abbreviations

B :

Bleed ratio (dimensionless)

D :

Dilution rate (/h), D t for MSC-HCDC system, D i for each reactor of MSC-HCDC, D f dilution rate increase by feed stream for each reactor

k d :

Specific death rate (/h)

m x :

Maintenance coefficient

P :

Product titer (g/L)

PD:

Productivity (g/L/h)

PR:

Productivity ratio (dimensionless)

Q :

Feed flow rate (main stream) (L/h)

q :

Feed flow rate (branched stream) (L/h)

qp/x,\( \overline{q} \)p/x:

Specific, average productivity of cells (g/g/h)

r P :

Product formation rate (g-product/L/h)

S :

Substrate concentration (g/L)

V :

Reactor volume (L)

w :

Volume ratio (dimensionless)

X,\( \overline{X} \):

Cell concentration, average cell concentration (g/L), X = 100 g/L (lactic acid, ethanol) and X = 40 and 60 g/L (penicillin) were used

Y :

Yield value

μ :

Specific growth rate (/h)

β :

Flow ratio of the top effluent and filtrate (dimensionless)

γ :

Filtration ratio in the cell filtering unit (dimensionless)

θ :

Cycle or residence time of fermentation system (h)

0:

Initial value

b, fb:

Batch, fed-batch system

c:

Continuous

f:

Feed stream

hcdc:

High cell-density culture

i :

Number of optional stages

j :

Number of optional stages

n :

Number of stages

t :

Total value for D, P, PD, V, Q or θ

HRT:

Hydraulic retention time

SRT:

Solid retention time

Mab:

Monoclonal antibody

C-HCDC:

Continuous high cell-density culture

SSC-HCDC:

Single-stage continuous high cell-density culture

MSC-HCDC:

Multistage continuous high cell-density culture

PHB:

Polyhydroxybutyric acid

B/FB:

Batch or fed-batch

FB:

Fed-batch

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Author information

Correspondence to Ho Nam Chang.

Additional information

N.-J. Kim and J. Kang contributed equally to this work.

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Chang, H.N., Kim, N., Kang, J. et al. Multi-stage high cell continuous fermentation for high productivity and titer. Bioprocess Biosyst Eng 34, 419–431 (2011). https://doi.org/10.1007/s00449-010-0485-8

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Keywords

  • Multi-stage continuous fermentation
  • High cell density
  • Ethanol
  • Lactic acid
  • Antibody