Biotechnology
Biotechnology is development of processes and their scale-up to produce drugs such as
penicillin, erythromycin (antibiotics), chemicals such as citric acid and ethanol used as foods
and fuels. These chemicals are made from micro-organisms—bacteria, viruses, fungi and
mammalian cells.
The aim again is to produce chemicals using techniques that are technically and economically
viable.
Similar to manufacture of chemicals on large scale as discussed until now, biotechnology
includes kinetics, transport phenomena, reactor design and unit operations for separations.
Our objectives are:
- Develop block diagrams for production of chemicals/microbes using biotechnology.
- Explain how biotech processes are different from core chemical processes.
- List and apply different strategies available for downstream processing of biotech.
Let‘s discuss penicillin production:
In general, for microbes, growth is most essential response to physiochemical environment. In
a suitable nutrient medium, organisms grow and also convert the medium to useful products.
𝑆𝑢𝑏𝑠𝑡𝑟𝑎𝑡𝑒 + 𝐶𝑒𝑙𝑙𝑠 → 𝑚𝑜𝑟𝑒 𝑐𝑒𝑙𝑙𝑠 + 𝑝𝑟𝑜𝑑𝑢𝑐𝑡𝑠
Typical growth Vs. time curve for bacteria population if sufficient nutrients are supplied
Lag phase: time for microbes to adjust to environment
Growth phase: microbes grow consuming the nutrients
Stationary phase: less nutrients, growth rate ~ death.
Death phase: nutrient deficient phase, death of cells
The growth of cells could be inhibited by substrate, product.
Product could be produced during growth/stationary phases
Cells produce products such as penicillin by consuming substrate. During the production, cells
multiply too. Should the substrate be used to make more of cells or penicillin?
Plot the curve for cells and product for non-growth associated systems.
Penicillin is produced from a strain penicillin chrysogenum. If this strain/microorganism
supplied with nutrients (food) and oxygen, it grows and produces penicillin (non-growth
associated). Penicillin production is an aerobic process (oxygen is needed); they are processes
that are anaerobic also (oxygen not needed).
Sufficient amount of the strain is grown in small fermentors and this strain is fed to a large
aerobic fermentor.
List the factors that are important in the design and operation of the fermentor.
Draw the following graph and explain the trend. How do you choose the operating point? Show
this point in the graph.
As a chemical engineer, how do you propose to operate at this point?
It is suggested that the fermentor is made deficient in one of the nutrients, say, nitrogen source,
to convert other nutrients to penicillin. The organism produces penicillin to kill other
organisms from growing—a strategy for survival of its population.
Discuss the consequences of surplus nutrients and starved cells on penicillin production.
Typical fermentor reactions are exothermic and temperature of its contents rises if generated
heat is not removed. How to do this?
Heat generation depends on rate of reactions, which is volume-based; heat removal is surface
area-based. Do you see any issues in scale-up?
If everything goes well, fermentor produces 50 g/L of penicillin (high or low?: compare it with
solubility of salt in water, ~360 g/L). Fermentor ‘broth’ also contains cell mass (solid) and
hotchpotch of nutrients (liquid).
What is the aim of steps downstream of fermentor?
The first aim is to separate the solid biomass and “beer” from the broth. This separation is
usually done by a rotary vacuum filter, shown below.
Explain the functioning of the filter.
After filtration, the filtrate contains ~50 g/L of penicillin. The liquid is essentially water.
How is the dissolved penicillin to be recovered?
- Distillation
- Crystallization
- Liquid-liquid extraction using a solvent such as butyl acetate
- Chromatography (this is an adsorption process)
The penicillin distribution co-efficient, K, defined as ratio of mass fraction of penicillin in butyl
acetate to mass fraction of penicillin in water depends strongly on the aqueous phase.
pH 2.1 4.4 5.8
K 25.0 1.38 0.10
Two options are available for penicillin extraction:
Option 1: use large quantities of butyl acetate and extract penicillin almost completely.
Option 2: explain this option before choosing between options 1 and 2
Butyl
Acid
acetate
Fermentation EXTRACTION Aqueous
MIXING TANK
broth UNIT I raffinate, pH 2.1
Organic
extract
Alkaline EXTRACTION Aqueous extract
solution UNIT 2 (product solution, pH 5.8)
Organic
raffinate
Penicillin is to be recovered from its concentrated aqueous phase. How?
Typical steps involved in the manufacture of chemicals by biochemical route are:
1. Fermentation 2. Downstream processing
Filtration Centrifugation Crystallization Liq-liq Chromatography Drying
extraction
Sterilization of process fluids
Fermenters require large amounts of liquids (media) and gas. These have to be sterilized so
the process is effective; contamination could kill the host organism; contaminants could grow
on media. Typically heat is used to decimate the contaminants below detectable levels.
Did you know the milk supplied to our houses, beverages such as Coke, Pepsi and fruit juices
are pasteurized, heat-treated to improve their shelf-life?
Ethanol is made by anaerobic fermentation of molasses by yeast cells.
𝐶𝑒𝑙𝑙𝑠 + 𝐶6 𝐻12 𝑂6 → 2𝐶2 𝐻5 𝑂𝐻 + 𝐶𝑂2 + 𝑚𝑜𝑟𝑒 𝑐𝑒𝑙𝑙𝑠
Ethanol is used as a fuel; it is blended with petrol and diesel in 5-10 % range. Such blended
fuel is used widely in Brazil. The idea is to convert molasses from sugar industry into ethanol.
Typical conditions: T = 303 K, pH = 4.5, residence time = 40 h, final ethanol conc = 8-12 %
Turns out ethanol inhibits yeast cell growth. If sugar concentration is high, yeast cells make
ethanol for ensuring their survival. They produce ethanol to kill other microbes.
Ethanol and water form a minimum boiling azeotrope of 351 K at 96 wt% ethanol. We need to
make very high purity ethanol (> 99.5 %) say for it is used as a fuel.
Draw a block diagram for ethanol production.
