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Morton Coutts (1904-2004), father of continuous fermentation Morton Coutts' plan for continuous fermentation at Dominion Breweries' Waitemata Brewery was patented as a world first in 1956.

 A system at which;
Fresh medium is continuously being supplied Volume of culture is kept constant Removal of culture at same flow rate as the feeding rate
Fi, Xi, Si, Pi

An extension concept of fed-batch

D.V = Fi = Fo 0, V = constant D = Dilution rate (T-1)

F = flow rate X = biomass S = substrate P = product V = volume

F, X, S, P Fo, Xo, So, Po

 Prolong exponential growth of batch culture Condition of Medium;

Growth is substrate limited Growth is not toxin limited Exponential growth will proceed until the additional substrate is exhausted

Steady A steady will be achieved state state =Formation of new biomass is eventually balanced with the loss of cells
from the vessel

Under steady state condition; Specific growth rate is controlled by

Continuous Culture Distinguished Characteristics

Microbial growth takes place under steady-state condition Growth occurs in constant rate and constant environment Physical and chemical factors are maintained constant Factors can also be controlled independently by operator

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Growth curve of batch fermentation

Growth curve of continuous fermentation

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INTRODUCTION
An apparatus for the continuous cultivation of microorganisms or plant cells Made of two main parts : -a nutrient reservior -a growth-chamber Nutrients are supplied continuously to the culture vessel Residual nutrients and cells are removed from the vessel (fermenter) at the same rate by an overflow Volume of culture in the fermenter remain constant

TK5301 - 2008 The volume of the chemostat can be controlled either by using:

Figure 1: A pump

Figure 2: An overflow system

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The nutrient medium contains an excess of all growth factors except one, the growth-limiting nutrient The concentration of the cells (biomass) is dependent on the concentration of the growthlimiting nutrient in the medium feed Increase or decrease in the concentration of the growth limiting factor is correspondingly expressed by increase or decrease in the growth rate of cells Thus the desired rate of cell growth can be maintained by adjusting the level of concentrations with respect to the growth limiting factor and other constituents

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To grow microorganisms on very toxic nutrients To select mutants with a higher affinity to the growth-limiting nutrient To select the species that are optimally adapted to the growth limitation and culture conditions in a mixed population To study the properties of organisms at selected growth rates To gather steady state data about an organism in order to generate a mathematical model relating to its metabolic processes

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Case study 1 :
Chemostat Production of Plantaricin C by Lactobacillus plantarum LL441 By: Barcena et. al. (1998)
Lactobacillus plantarum LL441-isolated from homemade cheese whey Produces bacteriocin - Plantaricin C Characteristics of Plantaricin C: -3.5 kDa -resistant to harsh environmental conditions -active at pH values from 2.0 to 7.0 -induces the formation of pores in the plasma membrane of sensitive cells -considered to be a food preservatives

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Objective :

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Plantaricin C was optimally produced in chemostat Kept at pH 5.0, 300C,150 rpm Dilution rate for different carbon source: -glucose : 0.05h-1 -sucrose : 0.10h-1 -fructose : 0.12 h-1

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Introduction
A continuous culture device in which a bacterial culture is maintained at a constant volume and cell density (turbidity) by adjusting the flow rate of fresh medium into the growth tube by means of a photocell and appropriate electrical connections.
If the turbidity tends to increase, the feed rate is increased to dilute the turbidity back to its set point. When the turbidity tends to fall, the feed rate is lowered so that growth can restore the turbidity to its setpoint

Most turbidostats use a spectrophotometer or turbidometer to measure the optical density for control purposes.

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Sketch of Turbidostat

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Turbidometer
also known as a nephelometer Process;
light shines throughout sample of water light strikes a colloid
the light is scattered bounce off the colloid reflects upwards

light does not strike a colloid

shines through the water sample.

The meter measures how much light is reflected off colloids in the liquid medium.

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Case Study 2:
Algae and rotifer turbidostats: studies on stability of live feed cultures By: Walz et. al. (1997)
Highly improved turbidity sensors was used. Algal density was regulated by turbidity measurements and the growth rates were monitored. The regulation system allowed an effective online process control. The turbidostat is a very good tool for such experiments because it functions optimally at maximum growth rates The algal growth in the turbidostat is not limited by nutrients. This provides a better quality of live feed for rotifers in turbidostats than in chemostats Its possibility to monitoring the growth process of algae and rotifers

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Advantages of continuous fermentation

High productivity Ability to relieve repression under specific nutrient limitation The distillery can be run at steady state conditions and at set-points in control. Without fluctuations during operations, utility requirements are constant, giving a greater economy in usage. Reductions in manpower.

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Disadvantages
Prone to contamination. Cost of pumps, reservoirs, sterilizers, and controls is relatively high. Economic benefits of improved kinetics is small compared to cost of feed and preprocessing

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Continuous industrial microbial processes are much less common than batch processes, but most biological waste treatment steps are operated continuously I. Continuous cultivation of rumen microorganisms, a system with possible application to the anaerobic degradation of lignocellulosic waste materials (Huub et al., 1986) II.A bioprocessing mode for simultaneous fungal biomass protein production and wastewater treatment using an external air-lift bioreactor (Bo et al.,2001)

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Continuous fermentation has been successfully applied in

brewing industry ( Dennis et al., 2000) single cell protein production (King, 1982).

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References

Barcena, J.M.B., Sineriz F., De Llano, D.G., Rodriguez, A., and Suarez, J.E. (1998). Chemostat production of plantaricin C by Lactobacillus plantarum LL441. J. Appl. & Environ. Microbiol., 64:3512-3514 Walz, N., Hintze, T., and Rusche, R. (1997). Algae and rotifer turbidostats: studies on stability of live feed cultures. Hydrobiologia 358: 127132 Gijzen, H.J., Zwart, K.B., vanGelder, P.T. and Vogels, G.D.. (1986). Continuous cultivation of rumen microorganisms, a system with possible application to the anaerobic degradation of lignocellulosic waste materials. Applied Microbiology and Biotechnology. 22:155-162 Jin, B., Yu, Q., and van Leeuwen, J.H. (2001). A bioprocessing mode for simultaneous fungal biomass protein production and wastewater treatment using an external air-lift bioreactor. Journal of Chemical Technology & Biotechnology. 76 (10): 1041-1048 OBrien, D.J., Roth, L.H., and McAloon, A.J. (2000). Ethanol

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