FERMENTER DESIGN

INTRODUCTION Fermenter Guidelines for fermenter design Requirements of a fermenter Parameters for fermenter design Gas transfer Heat transfer Nutrient transfer Aeration & agitation References FERMENTER Fermenter provides a suitable environment in which an organism can efficiently produce a target product, that might be 1. Cell biomass 2. Metabolite 3. Bioconversion product

The sizes of bioreactor can vary over several orders of magnitudes 1. The microbial cell (few mm) 2. Shake flask (100-1000ml) 3. Laboratory fermenter (1-50L) 4. Pilot scale (0.3-10m) 5. Plant scale (2-500m)

PARTS OF FERMENTER 1. Vessel 2. Impellers 3. Baffle 4. Sparger 5. Drain point 6. Shaft 7. Aseptic inoculation pipe 8. Sampling point

GUIDELINES FOR FERMENTER DESIGN AND OPERATION Material : Stainless steel Height to diameter ratio of the vessel: 2 to 1 or 3 to 1 Impeller Two or three disk turbine impellers Diameter: 0.3 to 0.4 of tank diameter Agitation speed: 50 200 rpm Impeller - shaft enters either from the top or bottom. Baffle - Four equally spaced to prevent vortex formation Width : one tenth of the tank diameter Sparger Ring sparger (Single orifice for a small Fermenter) Heating or cooling coil For sterilization or to control the temperature - Agitation and aeration - Cell suspension -Enhance the aeration (oxygen limitation problem) -Mixing -Problem with shear sensitive cells -Heating and cooling -Sensors - pH Control

DESIGN OF A FERMENTER Factors to consider when designing a fermenter Aseptic and regulator capability, Long term reliability Adequate aeration and agitation Low power consumption Temperature and pH controls Sampling facilities Large volume & low value products High value & low volume products Productivity & yield Product purification Water management Energy requirements Waste treatment REQUIREMENTS OF AFERMENTORFERMENTOR The vessel must be strong enough to withstand pressure The vessel should not corrupt the fermentation product Prevention of growth of contaminating microorganism must be provided

Efficient O2 Supply if fermentation is aerobic Addition of anti-foaming agent as demanded by the foaming state of the medium Fermenter should posses temperature Control Fermenter should posses a mechanism for detecting pH values of culture media There must be drain in the bottom of the fermenter MIXING RELATED DESIGN ISSUES ISSUES Agitator selection Power draw and torque calculations Scale-up Mechanical design Blending performance (scale of agitation, turnovers- per-minute, blend time, homogeneity) Heat removal, temperature field, and possible heat damage Solid-liquid mixing (just-suspended speed, settled solids fraction, cloud height) Reaction performance (productivity, selectivity) Surface motion, solids and gas drawdown Shear rates and impact velocities, possible shear damage Optimum feed locations Substrate concentration field, nutrient starvation Oxygen starvation or poisoning (local or global) CO2 or other product poisoning (local or global) pH control Gas-liquid mixing (mass transfer, gas holdup, power factors)

PARAMETERS FOR THE FERMENTER DESIGN FERMENTER DESIGN Physical Parameters Chemical Parameters Biochemical Parameters Biological Parameters PHYSICAL PARAMETERS Agitation power & speed Broth volume Color Density Foaming Gas flow rate & humidity Heat generation rate & transfer rate Liquid flow rate Temperature Osmotic pressure

CHEMICAL PARAMETERS Amino acid CO2 Cation level Conductivity Ionic strength Malliard reaction products Nitrogen Nutrient composition O2 Phosphorous B IOCHEMICAL PARAMETERS Amino acids A T P / A D P/AMP Carbohydrates Cell mass composition Enzyme NAD/NADH Vitamins & nucleic acid B IOLOGICAL PARAMETERS Age distribution Aggregation & contamination Genetic instability Mutation Total cell count Degeneration Doubling time GAS TRANSFER The theory of gas transfer refers to a process where the gaseous form of a compound is eventually dissolved into or driven out of the water. The rate at which the gas is transferred from air to water or vice versa is proportional to the area of the gas-liquid interface and the difference between saturation concentration and the actual concentration in the water. These indictate both the direction and rate of gas transfer at the gas water interface The availability of the oxygen to the biological system depends upon: Solubility Mass transfer rate of oxygen in the fermentation broth3 Rate of utilization of DO by microbial biomass Solubility decreases as temperature or salinity increases For example, the saturation level of dissolved oxygen is lower in warm water than coldwater systems

Gas solubility increases as the total pressure (sum of atmospheric and hydrostatic pressure) or mole fraction increases. In other words, the saturation level increases as the total pressure of the system increases. The rate of dissolution of a gas into water is proportional to the difference between actual and saturation concentrations of the gas in solution, Cs C The area of the gas liquid interface, a(m) The thickness of the liquid film, d The diffusion coefficient, D dC/dt= KL.a . ( C s C ) Where dC/dt = rate of gas transfer KLa = overall mass transfer coefficient DETERMINATION OF OXYGEN TRANSFER COEFFICIENTS TRANSFER COEFFICIENTS KLa=no2/(C * C L )-------1.) Sodium sulfite oxidation method Dynamic method Direct method Oxygen yield coefficient method SODIUM SULFITE OXIDATION METHOD METHOD This method employs the oxidation of sodium sulfite by oxygen in the presence of copper or cobalt which act as catalyst. Na2SO3+O2=Na2SO4 To find KLa by this method, air is sparged through a1N Na2SO3 solution in the presence of copper ions at a conc. of 10 M & mechanical agitation. The conc. of dissolved oxygen in the liquid is nearly zero since the oxidation reaction is extremely fast. Therefore, KLa = no2/C* Where, no2 = rate of oxygen transfer C* = saturation dissolved oxygen conc. in solution DYNAMIC METHOD This method is the simplest one since it requires only the dynamic oxygen balance in a batch culture, which has the following form dC L /dt = KL . a . ( C * C L ) - Qo2 . X Where Qo2X = rate of oxygen consumption per unit mass of cell This method contains three versions: Addition of lethal agent The gassing out method Dynamic oxygen balance

Frequency response technique Therefore, KLa can be calculated as KLa = Q o 2 X/C * C DIRECT METHOD This method based upon oxygen balance in inlet & outlet gas streams around a fermenter. oxygen balance for the sparged air yields no2.T=1/VL(no2.ino2.o) Where no2.T= rate of oxygen transfer VL=volume of fermentation broth The dissolved & saturated oxygen conc., CL & C * , need to be measured experimentally in order to determine KLa C* can be calculated from the partial pressure of oxygen in the exit gas stream in small scale fermenter But in large scale fermenter, the assumption of a perfectly mixed gas stream may not be valid. So, the log mean of the driving force i.e., ( C * C L )log. mean, between the inlet & outlet of the fermenter should be used in the following equation determining KLa, KLa = no2.T/( C * C L ) log. mean OXYGEN YIELD COEFFICIENT METHOD METHOD The rate of oxygen transfer can be related to the growth rate of microorganism using the Oxygen yield coefficient according to the following eq.no2. T= uX/Yo2 Where U= specific growth rate of microorganism X=cell conc. Yo2=yield coefficient of oxygen no2.T=oxygen transfer rate Once no2.T is determined by this equation then equation 1. can be used to calculate the KLa provided that the dissolved oxygen conc. is measured. HEAT TRANSFER Efficient heat transfer is important in controlling the temperature during sterilization operations Heat generated in the fermentor needs to be dissipated by cooling Optimum temp. should be maintained in the fermentor This can be achieved by: External jackets Internal coils External surface heat exchanger TRANSPORT OF NUTRIENT Transport of reactants to & from phase to the biological component has a significant impact on the performance of the reactor.

It is affected if the rate of the transport of the limiting nutrients is slower than the rate of utilization by the cells. AERATION AND AGITATION The primary purpose of aeration is to provide microorganism in submerged culture with sufficient oxygen for metabolic requirements. Agitation should ensure that a uniform suspension of microbial cells is achieved in a homogenous nutrient medium. The structural components are: -- Impellers -- Baffles -- Spargers -- Stirrer gland IMPELLERS Impeller types can be radial, mixed flow, axial and peripheral and are selected on the basis of the pump design and the application. The number of vanes will affect the efficiency; in general more vanes are more efficient. The agitator is required to achieve a no. of mixing objectives ex. bulk fluid & gas phase mixing, oxygen transfer, heat transfer, air dispersion & maintaining a uniform environment throughout the vessel contents.

SPARGER

What is a Sparger? Sparger is a technical term for injecting gas into a liquid or for spraying a liquid onto a solid. Spargers are porous disc or tube assemblies that are also referred to as Bubblers, Carbonators, Aerators, Porous Stones and Diffusers. Porous Metal Spargers are widely used in Gas-Liquid contacting applications that affect everyday living. A common example of sparging can be seen in an aquarium where air is bubbled into the tank through a fine porous stone in order to maintain the level of the dissolved oxygen in the water . Types of spargers: Porous sparger Orifice Nozzle Combined sparger-agitator

BAFFLES Four baffles are normally incorporated into vessel of all sizes to prevent a vortex & to improve aeration efficiency. The agitation increased with wider baffles but drops sharply with narrow baffles. STIRRER GLAND The satisfactory sealing of the stirrer shaft assembly has been one of the most difficult problems to overcome in the construction of fermentation equipment which can be operated aseptically for long periods. Types of stirrer glands: Stuffing box Simple bush seal Mechanical seal Magnetic drive

REFERENCES Principles of fermentation technology, 2nd edition by P . F . S T A N B U R Y& A . W H I T A K E R nd Bioprocess engineering, basic concepts, 2 edition by M .L.SHULER&F.KARGI ebook.com www.google.com

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