A2 Biology : Chapter 10 : Biotechnology

Biotechnology is the industrial use of living organisms (or parts of living organisms) to produce food, drugs or other products. Microorganisms are often used in biotechnological processes because: They have rapid life cycles so large populations can be built up quickly and many reproduce asexually so are genetically identical and carry out the same metabolic processes. They can be grown in fermenters with minimal attention as they have specific and simple requirements for growth. Many be grown using waste materials from industry which would otherwise have no use and may be costly to dispose of. Their use does not usually raise ethical concerns. As they have a single copy of each gene, if one is altered by gene technology there are no other copies of that gene to mask it. The expression of their genes is simple and therefore easier to genetically modify. Many have evolved to live in hot environments and the substances they produce (e.g. enzymes) are able to work at high temperatures.

The growth of a bacterial population follows a recognisable growth curve with four definable phases. Lag phase when genes are being switched on to enable enzymes to be synthesised to allow the absorption and metabolism of nutrients. In this phase no population growth is detected. Log or exponential growth phase bacterial cells are growing and dividing at their maximum rate for the conditions (nutrients and space) they are in. In this phase the population doubles in each generation. Stationary phase one or more of the nutrients are beginning to run out. The number of new cells produced is matched by the number of cells dying. Decline or death phase nutrient levels decline sufficiently to prevent reproduction occurring. Accumulation of waste products which may be toxic occurs and may also prevent further reproduction. At this stage the death rate is faster than the reproduction rate. Due to the high cost of enzymes used in industrial processes it is desirable to use them many times. One way of doing this is to immobilise them in one of four ways as shown in the diagram. Advantages of using immobilised enzymes include being able to re-use the same enzymes and being able to produce a product free of contamination by enzymes. Immobilised enzymes are also more tolerant of changes in temperature and pH than those in solution (i.e. less likely to denature) as their molecules are held firmly by the material in which they are embedded and because the embedded parts of the molecule are not fully exposed to the changes. An example of the use of immobilised enzymes is the production of lactosefree milk. Lactase enzyme is immobilised in alginate beads by dropping a solution of sodium alginate and lactase enzyme into a solution of calcium chloride. Milk is then poured over the beads held in a column and milk free of both lactose sugar and lactase enzyme is collected.

A2 Biology : Chapter 10 : Biotechnology

Microorganisms used to produce products used in medicines and foods are often grown in containers called fermenters. The growing conditions in the fermenter can be monitored and manipulated to maximise the yield of product required. This is summarised in the diagram. A primary metabolite is a substance which is produced by most of the cells in a culture most of the time. A secondary metabolite is a substance only produced by some cells or at a particular growth stage. Penicillin is an example of a secondary metabolite as its production only reaches high levels after the fungus (Penicillium) has been growing in the medium for a period of time. The fungus is grown in the fermenter until the maximum amount of penicillin has been produced. Fermentation is then stopped, the antibiotic harvested, the fermenter cleaned out and the fermentation process started again. Enzymes (e.g. proteases and lipases to be added to washing powders) are produced industrially. The microogranism which will produce the enzyme is first grown using a carbon source (often a waste product of the maize or sugar cane industry) and a nitrogen source (e.g. protein or urea). Bacteria living in hot springs are often selected as their enzymes function at a higher optimum temperature so they can be used in industrial processes employing higher temperatures. After fermentation the culture is heated to kill the cells which are then harvested by centrifugation and broken open by passing them through small opening at high pressure to release the enzyme. The enzyme is concentrated by removal of water and extracted from the cell debris, nucleic acids and larger proteins by centrifugation. Enzymes used in washing powders are often enclosed in capsules before adding to products to prevent irritation and allergies on skin contact with the user. Production of penicillin and industrially used enzymes are examples of batch culture. In a variant of this process, fed batch culture, a carbohydrate source is periodically added resulting in fermentation and product production continuing for a longer period of time. Mycoprotein is produced by continuous culture, where the fungus Fusarium which is made up of long, thin threads called hyphae is grown in a fermenter. Glucose is provided as a respiratory substrate and ammonium phosphate as a nitrogen source. No stirrer is used as this would break the fungal hyphae. Liquid culture is drained off from the fermenter, centrifuged to separate the hyphae and enzymes used to break down the RNA which otherwise results in an unpleasant taste. The resulting mycoprotein is high in protein and low in fat. Batch culture is easy to set up and requires minimal attention. Once cleaned out the fermenter can be used for another industrial process. If an error (e.g. contamination) occurs it is only the batch concerned which must be discarded. Continuous culture eliminates wasted production time whilst the fermenter is being emptied and cleaned and fermenter size may be reduced as product is continually removed. The inlet and outlet pipes of continuous fermenters can however sometimes become blocked by cells clumping together. Ensuring asepsis (i.e. without microorganisms) is important when growing industrial cultures as other microorganisms may metabolise the substrate in different ways to produce unwanted products or be pathogenic themselves. Asepsis can be achieved by ensuring fermenters, their attachments and substances which enter them are sterile. Maintaining a pressure difference in favour of airflow out of the room where a fermenter is housed reduces the probability of microorganisms entering on air currents from outside. Covering the clothing and skin of workers will also reduce the probability of contamination.