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Enzyme production in a cell recycle fermentation system evaluated by computer simulations

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Abstract

Enzyme production in a cell recycle fermentation system was studied by computer simulations, using a mathematical model of α-amylase production by Bacillus amyloliquefaciens. The model was modified so as to enable simulation of enzyme production by hypothetical organisms having different production kinetics at different fermentation conditions important for growth and production. The simulations were designed as a two-level factorial assay, the factor studied being fermentation with or without cell recycling, repression of product synthesis by glucose, kinetic production constants, product degradation by a protease, mode of fermentation, and starch versus glucose as the substrate carbon source.

The main factor of importance for ensuring high enzyme production was cell recycling. Product formation kinetics related to the stationary growth phase combined with continuous fermentation with cell recycling also had a positive impact. The effect was greatest when two or more of these three factors were present in combinations, none of them alone guaranteeing a good result. Product degradation by a protease decreased the amount of product obtained; however, when combined with cell recycling, the protease effect was overshadowed by the increased production. Simulation of this type should prove a useful tool for analyzing troublesome fermentations and for identifying production organisms for further study in integrated fermentation systems.

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Abbreviations

a :

proportionality constant relating the specific growth rate to the logarithm of G (h)

a 1 :

reaction order with respect to starch concentration

a 2 :

reaction order with respect to glucose concentration

c :

starch concentration (g/l)

c 0 :

starch concentration in the feed (g/l)

D :

dilution rate (h−1)

e :

intrinsic intracellular amylase concentration (g product/g cell mass)

E :

extracellular amylase concentration (g/l)

F :

volumetric flow rate (l/h)

G :

average number of genome equivalents of DNA/cell

K 1 :

intracellular repression constant

K 2 :

intracellular repression constant

K s :

Monod saturation constant (g/l)

k 3 :

product excretion rate constant (h−1)

k I :

translation constant (g product/g mRNA/h)

k d :

first order decay constant (h−1)

k dw :

first order decay constant (h−1)

k gl :

rate constant for glucose production (g/l/h)

k m, dgr :

saturation constant for product degradation (g/l)

k st :

rate constant for starch hydrolysis (g/l/h)

k t1 :

proportionality constant for amylase production (g mRNA/g substrate)

k t2 :

proportionality constant for amylase production (g mRNA *h/g substrate)

k w :

protease excretion rate constant (h−1)

k wt1 :

proportionality constant for protease production (g mRNA/g substrate)

k wt2 :

proportionality constant for protease production (g mRNA *h/g substrate)

k wI :

translation constant (g protease/g mRNA/h)

m :

maintenance coefficient (g substrate/g cell mass/h)

n :

number of binding sites for the co-repressor on the cytoplasmic repressor

Q :

repression function, K1/K2 less than or equal to 1.0

Q w :

repression function, K1/K2 less than or equal to 1.0

r :

intrinsic amylase mRNA concentration (g mRNA/g cell mass)

r m :

intrinsic protease mRNA concentration (g mRNA/g cell mass)

R ex :

retention by the filter of the compounds x=: C starch, E amylase, or S glucose

R t :

amylase transport rate (g product/g cell mass/h)

R wt :

protease transport rate (g protease/g cell mass/h)

R s :

rate of glucose production (g/l/h)

R c :

rate of starch hydrolysis (g/l/h)

S 0 :

feed concentration of free reducing sugar (g/l)

s :

extracellular concentration of reducing sugar (g/l)

t :

time (h)

V :

volume (1)

w :

intracellular protease concentration (g/l)

W :

extracellular protease concentration (g/l)

X :

cell mass concentration (dry weight) (g/l)

Y :

yield coefficient (g cell mass/g substrate)

σ :

substrate uptake (g substrate/g cell mass/h)

μ :

specific growth rate of cell mass (h−1)

μ d :

specific death rate of cells (h−1)

μ m :

maximum specific growth rate of cell mass (h−1)

μ m,dgr :

maximum specific rate of amylase degradation (h−1)

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Authors and Affiliations

  1. Department of Biotechnology, Building 223, The Technical University of Denmark, DK-2800, Lyngby, Denmark

    S. Grøn, C. Emborg & K. Biedermann

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  1. S. Grøn
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  2. C. Emborg
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  3. K. Biedermann
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Additional information

This study was supported by the Nordic Industrial Foundation Bioprocess Engineering Programme and the Center for Process Biotechnology, The Technical University of Denmark.

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Grøn, S., Emborg, C. & Biedermann, K. Enzyme production in a cell recycle fermentation system evaluated by computer simulations. Bioprocess Engineering 13, 59–68 (1995). https://doi.org/10.1007/BF00420431

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