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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N.-J. Kim and J. Kang contributed equally to this work.
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Chang, H.N., Kim, NJ., 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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DOI: https://doi.org/10.1007/s00449-010-0485-8