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ACTIVITY BRIEF
Enzyme technology
The science at work
Enzymes
Enzymes have become big business. They are used in many industrial processes to catalyse
biological reactions. Enzymes are exploited in a variety of manufacturing processes such as
food processing and for the synthesis of medicines such as antibiotics like artificial penicillin.
They are also used to clean up factory effluents and pollution in water and soil. Many
processes can be made faster and cheaper by using the right enzyme and conditions.
Optimum conditions are maintained during factory production by use of bioreactors. These
are vessels which are designed to provide the ideal environment for reactions involving
enzymes or living organisms.
Your brief
Most people in the UK produce an enzyme called lactase in their small intestine to break
down lactose, the sugar found in milk. However, many people do not produce enough
lactase and therefore have problems digesting the milk. They may get diarrhoea, wind and
stomach cramps if they consume milk or milk products. This is caused by the action of
bacteria in the gut, fermenting the lactose. This problem has been addressed by a number of
companies who produce lactose free milk. Such companies have to address the problem of
establishing the optimum conditions for their milk treatment process.
STUDY SHEET
Investigating a bioreactor
Introduction
Simple bioreactors can be constructed in the laboratory, using basic apparatus. These can be
used to study the feasibility of processes and to establish the most favourable reaction
conditions. The most promising processes can then be trialled using more sophisticated
equipment. Eventually pilot studies yield the information that scientists and engineers need
to be able to take the expensive step of designing and building chemical manufacturing plant
on an industrial scale.
In this task, you design, carry out and write up an experimental test of a bioreactor. In the
next task, you will be asked to write a report to discuss your results in relation to industrial
practice.
The experiment
Lactose reduced milk can be made by using a bioreactor to treat skimmed milk with
immobilised lactase enzyme. Using the information provided and other sources, design and
carry out a practical investigation of the effect of temperature on the activity of
the enzyme lactase, when immobilised and used in a bioreactor.
Summary
You will need to:
1 Write a clear plan of action of your own to include:
the method/procedure to be followed, with measurements of appropriate precision,
repeated when appropriate
a list of equipment and reagents, with quantities and concentrations
timing and laboratory/other requirements
detailed risk assessments consistent with COSHH guidelines, using secondary sources
an explanation of the use of a range of techniques and equipment.
2 Obtain data:
working with appropriate accuracy
using appropriate precision
making a detailed set of relevant observations
collecting sufficient data to complete simple statistics.
3 Display and analyse the data accurately:
using a range of methods
illustrating trends clearly.
4 Draw simple conclusions:
basing them solely on your data and observations
relating them to your tables, charts and/or calculations.
Doing it
1 Agree a time scale with your teacher. You must work to a deadline, but establish how
much class time you will have.
2 Carry out research and decide on a suitable method. Use Fact sheet: Bioreactor
techniques and other suitable resources to help you.
3 Write a provisional plan in outline and have it approved by your teacher. Use the
Summary to check what you need to include. This first draft need not be in detail, just
establish that you have chosen a workable safe method.
4 After you have consulted your teacher, modify or develop your plan. It would be a good
idea to plan a short pilot study to test your chosen methods and establish the quantities
and concentrations of the materials that you need. Include:
requests for materials and resources
risk assessments
planned times for supervised laboratory work.
Health and safety: Do not undertake any practical work until you have had
your plans agreed by your teacher.
Before carrying out any practical work a risk assessment must be carried out.
Wear eye protection. Some people are allergic to lactase solution; avoid
getting it on your skin or rubbing eyes after handling. Wash your hands before
and after your practical work.
5 Carry out your investigation. You may need to repeat some things or modify your
approach as you go along. Continue to submit revised plans as necessary and do not
undertake any work without prior approval. Keep a record of all the work that you
do as evidence of your planning, including any rough notes and references to sources
that you use (paper, electronic or human).
6 It is good practice to plot results in a graph or calculate averages as you go along, so you
can see if results are reliable. If you have inconsistent or anomalous results, you should
repeat measurements and parts of procedures to check for possible causes of errors and
to get more accurate results. Make notes of any relevant observations.
7 Complete the analysis of your data and write a report of your investigation to meet the
criteria shown in the Summary. Include:
introduction
method
results
conclusions.
Do not include a detailed discussion of your findings. You will do this in your next task.
FACT SHEET
Bioreactor techniques
Lactose reduced milk can be made by using a bioreactor to treat skimmed milk with
immobilised lactase enzyme. The following techniques may be modified for investigating this
process.
3 For batch processing, close the tap and add enough distilled water to the bioreactor to
cover the alginate beads. Run this off into a measuring cylinder and record the volume.
This indicates the volume of substrate that may be processed in one batch in the
bioreactor.
Add enough substrate solution to the column/body of the bioreactor to cover the alginate
beads. Leave for a measured period of time, then open the screw clip or tap fully and
collect the product.
STUDY SHEET
Using bioreactors
Introduction
You should carry out the work in Study sheet: Investigating a bioreactor before starting this
activity. You need to have carried out your experiment using a bioreactor to investigate the
effect of temperature on the activity of lactase. You should have written up the first half of
your report and formed conclusions based on your data. You now need to carry out some
research on the use of bioreactors in industry and write a detailed discussion.
Discusssion
To conclude your report you will need to write a detailed discussion. Start by making notes
on the following, keeping a record of any sources that you use:
1 List and make short notes on the uses of enzyme technology in medicine and industry,
including named examples.
2 What reaction is used to reduce the lactose content of milk?
3 What affect did temperature have on the rate of this reaction? Explain why temperature
had this effect.
4 What other factors might affect the rate of this reaction and how were they kept
constant during this experiment?
5 For the industrial process of removing lactose from milk
(i) Why would a company want to immobilise lactase and not just add it directly to milk?
(ii) What are any disadvantages of immobilising an enzyme?
6 How does your bioreactor relate to:
(i) batch-system
(ii) continuous-system?
Which of these is better for producing large amounts of lactose-free milk? Explain your
answer.
7 How does your bioreactor compare to those used in industry?
Now write the final part of your report under the sub-heading Discussion. Use information
from your findings and your notes to meet the marking criteria which include:
interpreting the results in detail using secondary sources to support your findings
drawing conclusions relating your results to the use of bioreactors and enzyme
immobilisation, specifying named examples in either medicine or industry
discussing the significance of your findings in terms of how enzymes work and the
advantage of enzyme technology to industry
Resources
Resources on the production of lactose-free milk
http://www.lactolite.co.uk/lactofree/
http://www.nutraingredients.com/news/ng.asp?id=36761-valio-extends-lactose
Resources on enzymes
http://www.bbc.co.uk/schools/gcsebitesize/chemistry/chemicalreactions/5enzymesrev5.shtml
http://biotech.about.com/od/casestudies/tp/dairyenzymes.htm
http://urc.cc/bio_r.htm
Companies that produce enzymes for use in industry
Advanced Enzymes: http://www.enzymeindia.com/
Baxel: http://www.microtack.com/index.html
Teacher notes
This activity gives students an opportunity to meet OCR assessment objective. Students
design an investigation and make a simple bioreactor containing an immobilised enzyme
(lactase) and investigate the effect of increased temperature on the enzyme.
It does not cover introductory aspects of Unit 15.2.4: Enzyme Technology. There is already a
wealth of information available. For example, there are many good practicals that allow
students to find out how enzymes work by investigating rates of reaction and limiting factors
(see other resources).
Information about the lock and key and induced fit hypothesises can be found in most A-
level biology text books. You may wish to cover some of these aspects before using the
investigation for assessment.
Organisation
Students will need to begin their investigation by reading through the activity and planning
the experiment. In particular they will need to carry out risk assessments and plan which
temperatures they wish to investigate.
Students should preferably carry out the investigation by themselves. However, quite a lot of
Diastix may be required per experiment. The exact number of Diastix required will depend
upon the number of temperatures tested and the number of repeats. Diastix cost around £4
to £5 for 50.
The same column should be used for collecting each complete set of results. This will
eliminate discrepancies in results due to variations in the concentration of enzyme in
different columns (dependent on accurate measurement and the number/size of beads put
into each column). As long as students carry out sufficient individual work, class results could
be pooled for the final analysis, providing results using separate columns are not combined
for calculations.
Students who have difficulty designing their investigation may be given extra help (taken
into account for marking). If necessary, ideas from the following protocol could be provided,
up to giving the whole method to follow. Students will still need to carry out risk
assessments and decide on the temperatures to investigate.
Resources
A number of enzyme experiments are available from the NCBE:
http://www.ncbe.reading.ac.uk/NCBE/PROTOCOLS/pracbiotech.html
NCBE also supply lactase (as lactozym) at about £14 per 100 cm3. Details and background
information on the commercial use of lactose reduction in dairy products can be found at:
http://www.ncbe.reading.ac.uk/NCBE/MATERIALS/ENZYMES/lactozym.html
Investigating the effect of competitive and non-competitive inhibitors on the enzyme
ß-galactosidase from SAPs may be useful to investigate how enzymes work:
http://www-saps.plantsci.cam.ac.uk/worksheets/scotland/inhibit.htm
Many practicals on enzymes may be found in: King, T., Reiss, M. & Roberts M. (2001).
Practical Advanced Biology Nelson Thornes Ltd. ISBN 0-17-448308-2.
Technical notes
Make sure distilled or deionised water is used when making up solutions, since calcium salts
in tap water will cause the sodium alginate to set.
Sodium alginate doesn’t dissolve easily. The best way is to mix the sodium alginate and
distilled water in a beaker, then place the beaker on a magnetic stirrer/heater on a low heat
(just to warm slightly) and leave for about 1-2 hours. The solution can go mouldy after
about 1 week, so a fairly fresh solution is required.
Lactase needs to be kept refrigerated.