e Viith International Biotechnology Symposium New Delhi, February 19-25, 1984 EXTENDED ABSTRACT FORM Bacterial kinetics and energetics ; Theoretical and experinental evaluation of bacterial growth in batch, fed-batch and continuous cultivation modes. A.A.ESENER , J.A. ROELS and N.W.F. KOSSEN, Bioprocesstechnology group, Department of Chemical Engineering, Delft University of Technology, Julianalaan 67, 2628 BC Delft,The Netherlands| Models in which the variation of biomass composition in response to environmental changes is totally ignored are defined as ‘unstructured nodels'. Although very simple in their nature these type of models are being used for describing microbial growth phenomena and for control and optimization purposes. In this study such a simple unstructured model based on Monod kinetics and the relation for ‘linear substrate consumption’ has been evaluated and tested critically with extensive simulation and experimental data obtained during batch, fed-batch and continuous cultivation of the bacterium; Klebsiella pneumoniae NCTC418 with no product formation. ! The four parameter model (¥U"s M, » Una.» k,) is based on the following pair of state equafions®; d/dt (C..) = (r+ 8) a) a/at (C..¥) = (4, +8.) V @) 8 578s . together with the relation of linear substrate removal; toon! We ta. C, @) and the relation for Monod kinetics: Una Cy Sg / (ky + C4) @ Here # is the net flow term per unit volume to the system and is determined by the mode and configuration of the process. The volume term, V,is included to be able to account fgr processes where significant volume changes are encountered. With the aid of the model similations and experimental data it has been] shown that during growth in batch mode, system behaviour is rigidly Hixed by the kinetic parameter; the maximum specific growth rate. The energetic parameters have minimal influence. In continuous cultivation the behaviour is fixed by the energetic parameters; the maximum yield coefficient and the coefficient of maintenance. Implications of these observations have been discussed with reference to the design of experiments for parameter estimation. Comparison of model predictions with experimental data has igdicated that, for batch growth the endogeneous metabolism model’ provides a more comprehensive description] of the real behaviour than eq.(3). The ‘linear substrate removal’, NAME AND ADDRESS OF AUTHOR PRESENTING POSTER Name:...Dr...Ay. As. ESENER | wish fo present this ADDRESS: Koninklfike Shell. Lab. Ansterdan poster paper at the Shell Research B.V., Dept. HCP VILIBS. preferably at Badhuisweg.3;-1034CM-Amsteréan sessions] P-5 The NetherlandsEXTENDED ABSTRACT FORM connnueo eq(3), has also been tested with continuous culture data. It is show that above specific growth rate of 0.1 hr-! this relation agreed very well with the experimental findings.At lover growth rates, hovever, a systematic deviation, which could not be accounted for by the loss of| viability, has been observed. The ‘declining feed fed-batch’ technique” has also been used in connection with the simple model to estimate model paraneters. The experimental results suggest that the sane values of unstructured nodel parameters may be used in all modes of cultivation ive., no significant differences exist between the parameter values obtained from experiments carried out in different modes. References: 1, A.A.Esener, Ph.D. Thesis, Delft University of Technology,Delft (1981) 2. S.J. Pirt, Proc. Roy. Soc. London B,163, 224(1965). 3, A.A. Esener, J.A, Roels and N.W.P. Kossen, Biotechnol.Bioeng. ,24, 1877 (1982) 4. D. Herbert, Symp. Int. Cong. Microbiol. ,6,381(1958) 5. AsA. Esener, N.W.F. Kossen and J.A. Roels, Second European Congress on Biotechnology, Eastbourne, England,Abs. no:88 (1981) Nomenclature: concentration Monod saturation constant rate of production/consumption substrate biomass volume yield coefficient specific growth rate net flow into the system Sew aneaRrOEXTENDED ABSTRACT FORM Bacterial kinetics and energetics ; Theoretical and experimental evaluation of bacterial growth in batch, fed-batch and continuous cultivation modes. A.A.ESENER , J.A, ROELS and N.W.F. KOSSEN, Bioprocesstechnology group, Department of Chemical Engineering, Delft University of Technology, Julianalaan 67, 2628 BC Delft,The Netherlands Models in which the variation of biomass composition in response to environmental changes is totally ignored are defined as ‘unstructured models". Although very simple in their nature these type of models are being used for describing microbial growth phenomena and for control and optimization purposes. In this study such a simple unstructured model based on Monod kinetics and the relation for ‘linear substrate consumption’ has been evaluated and tested critically with extensive simulation and experimental data obtained during batch, fed-batch and continuous cultivation of the bacterium; Klebsiella pneumoniae NCTC4I8 with no product formation.! The four paraneter model (Y™%, a, , uggs k,) is based on the following pair of state equations’; alae (CV) = (r+ 0) V a d/at (C.V) = Cre +9) V @ together with the relation of linear substrate removals” fee) ee ce @) and the relation for Monod kinetics: Pea a @) Here @ is the net flow term per unit volume to the system and is determined by the mode and configuration of the process. The volume term, V,is included to be able to account fgr processes where significant volume changes are encountered. With the aid of the model simations and experimental data it has been shown that during growth in batch mode, system behaviour is rigidly fixed by the kinetic parameter; the maximim specific growth rate. The energetic paraneters have minimal influence. In continuous cultivation the behaviour is fixed by the energetic parameters; the maximum yield coefficient and the coefficient of maintenance. Implications of these observations have been discussed with reference to the design of experiments for parameter estimation. Comparison of model predictions with experimental data has ipdicated that, for batch growth the endogeneous metabolism model” provides a more comprehensive description of the real behaviour than eq.(3). The ‘linear substrate removal’, NAME AND ADDRESS OF AUTHOR PRESENTING POSTER Name:...DE..As. As, BSENER | wish to present this ADDRESS: Koninklijke Shell. Lab...Amsterdam poster paper at the Shell Research 3.V., Dept. HCP MURS, preferably at Badhuisweg:3;--1031-€M-Ansterdam ceosee sesionB—] PHS The NetherlandsEXTENDED ABSTRACT FORM conmnnuso. eq(3), has also been tested with continuous culture data. It is shown that above specific growth rate of 6.1 hré! this relation agreed very well with the experimental findings.At lower growth rates, however, a systematic deviation, which could not be accounted for by the loss of| viability, has been observed. ‘The ‘declining feed fed-batch' technique? has also been used in connection with the simple model to estimate model paraneters. The experimental results suggest that the sane values of unstructured model paraneters may be used in all modes of cultivation i.e., no significant differences exist between the parameter values obtained from experiments carried out in different modes. References: 1. A.A.Esener, Ph.D. Thesis, Delft University of Technology,Delft (1981) 2. S.J. Pirt, Proc. Roy. Soc. London B,163, 224(1965). 3. ALA. Esener, J.A. Roels and N.W.F. Kossen, Biotechnol.Bioeng. ,24, 187 (1982) 4, D. Herbert, Symp. Int. Cong. Microbiol. ,6,381 (1958) 5. A.A. Esener, N.W.F. Kossen and J.A. Roels, Second European Congress ‘on Biotechnology, Eastbourne, England,Abs. no:88 (1981) Nomenclature: concentration Monod saturation constant rate of production/consumption substrate biomass volume yield coefficient specific growth rate net flow into the system een aKonra