E3

2 Use of microorganisms

2.1 Using microorganisms for commercial purposes

1 Microorganisms are widely used to produce commercial products because:
- the use of microorganisms helps lower the production costs as they reproduce and grow
rapidly and have simple nutritional requirements. They are also efficient at converting
raw materials into products and do not need large areas of land to grow.
- there are no welfare issues to consider.
- microbial products can often be easily extracted and purified for use.
- there are a wide range of microorganisms that can carry out different reactions.
- microorganisms can be genetically modified relatively easily.
2 In industrial fermentation (工業發酵), microorganisms are cultured in fermenters (發酵器)
(also called bioreactors (生物反應器)) under growth conditions that are carefully monitored
and controlled to optimize microbial growth and production of desired products.

nutrients acid or alkali

- for metabolism and growth - adjusts the pH to the optimum value for
of microorganisms microbial growth
air filter

exhaust gas out

- avoids pressure buildup
- the gas may be a
valuable by-product,
cooling water jacket e.g. carbon dioxide for
water out
- carries cold water to remove use in soft drinks
heat produced by microbial
activities, preventing the stirrers
denaturation of their - mix nutrients and microorganisms well
enzymes - ensure an even temperature throughout
the culture
probes
- monitor temperature, pH stainless steel container
and oxygen concentration - can be sterilized by steam
- can resist acidic waste products of
water in microorganisms
air filter
- removes unwanted microorganisms
in the air to avoid contamination

air supply
- provides oxygen for
useful products are microorganisms that
collected and purified respire aerobically
and purified

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3 Any contamination of the microbial culture in the fermenter is undesirable because unwanted
microorganisms may
- compete with microorganisms in the culture for resources.
- produce harmful substances.
In either case, microbial growth in the fermenter and product yield will be reduced.
4 To avoid contamination of the microbial culture and product in the fermenter, it is necessary to
carry out aseptic techniques.

2.2 Food processing

Alcoholic fermentation
1 Yeast can carry out alcoholic fermentation (酒精發酵) when oxygen supply is insufficient.

glucose ethanol + carbon dioxide

2 Beer-brewing:

Barley (大麥) grains are The germinated barley, The malt is mixed with hot
soaked in water for called malt (麥芽), is crushed water of 65 °C. Starch is
germination. Water activates to increase the surface area broken down into simple
the enzymes in the barley for enzymatic actions. sugars by the enzymes.
grains.

Air is pumped into the The hops are removed. The crushed malt is
mixture for several hours to A small amount of yeast is removed. The sugary
allow the yeast to reproduce. added to the cooled wort in a solution, called wort
Then air supply is stopped fermenter. (麥芽汁), is boiled with
for alcoholic fermentation to hops (啤酒花藤).
take place.

As the ethanol content

reaches 4–6%, the
fermented liquid is
pasteurized (heated to 72 °C)
to kill the yeast. The beer is
bottled or canned.

3 Wine-brewing:

Grapes are Sugars in the grape Fermented grape After filtration, the
crushed and juice are fermented juice is kept in wine is put into
grape juice is into ethanol and barrels to develop bottles and sealed.
extracted. carbon dioxide by a good flavour.
yeast.

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4 Bread-making:

Flour, yeast, water, sugar The dough is pressed and The dough is covered and left
and salt are mixed to form stretched into the shape of in a warm place for alcoholic
a dough. a loaf. fermentation to take place.

Bread with a spongy The dough is baked in an The carbon dioxide produced
texture is ready to serve. oven. The bubbles forms bubbles inside the
expand and the ethanol dough, making the dough
evaporates in the heat. rise.

Lactic acid fermentation

5 Under anaerobic conditions, certain lactic acid bacteria (乳酸菌) can carry out lactic acid
fermentation (乳酸發酵).

lactose + water lactic acid

6 Making of yoghurt:

Milk is heated to 90 °C After the milk is cooled to 40 °C, The yoghurt is cooled to
for 20 minutes to kill all lactic acid bacteria are added to 5 °C to prevent further
microorganisms and carry out lactic acid fermentation and
drive away all dissolved fermentation. The lactic acid packaged.
oxygen. produced brings the pH of milk
down to 4. The milk proteins
therefore coagulate and the milk
thickens to become yoghurt.

7 Making of cheese:

Lactose in The lactic acid produced After the whey is The curd is left in
milk is used brings the pH of milk down, drained off, the a room for weeks
by lactic acid causing the milk to separate curd is salted or months to
bacteria for into a solid called the curd and pressed into mature.
lactic acid (凝乳) and a liquid called the moulds.
fermentation. whey (乳清). Sometimes,
an enzyme called rennin
(凝乳酶) is added to speed
up the coagulation of milk.

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2.3 Pharmaceutical industry

Antibiotic production
1 Some fungi and bacteria can be cultured in fermenters for the mass production of antibiotics.
2 Mass production of penicillin (青霉素):

Penicillium (青霉) is As the nutrients in the The culture is Purified penicillin

first cultured in a culture medium become then filtered to can be used in its
fermenter under limiting, the mould remove the original form or
optimal conditions to enters the stationary mould and modified by
allow rapid growth of phase and secretes penicillin is chemical
the mould. During this a large quantity of extracted from procedure to
time, very little penicillin into the the filtrate. increase its
penicillin is produced. culture medium. effectiveness.

Vaccine production
3 Attenuated or killed pathogenic microorganisms or viruses, their subunits or products
(e.g. inactivated toxins) can be used to produce vaccines.
4 Genetically modified (GM) microorganisms can also produce antigens for making into
recombinant vaccines.
- The production of recombinant vaccines poses less risk to workers because whole pathogens
are not grown.
- Recombinant vaccines are also safer for use because subunits of pathogens cannot reproduce
in the body or cause diseases.

2.4 Industrial enzyme production

1 Proteases, lipases and amylases in biological washing powder can help remove stains on
clothes. Most of them are produced by bacteria that can grow in alkaline environments.
2 Pectinase (果膠酶) is produced by certain moulds. The enzyme can break down pectin (果膠),
a polysaccharide which holds cellulose fibres together in plant cell walls.
3 Benefits of using pectinases in fruit juice extraction:
- Cell walls can be broken down at a lower temperature. This lowers the production costs
and avoids the colour, flavour and nutrients of the fruit juice from being affected.
- More juice can be released from fruit cells. This increases the yield of fruit juice.
- The fruit juice is more nutritious because the breakdown of the cell walls helps the release
of some nutrients.
- Pectinases can help break down some polysaccharides, clarifying the fruit juice to increase
its market value.
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2.5 Sewage treatment
1 Sewage treatment (污水處理) is carried out to reduce the harmful contents of sewage (污水)
before it is discharged to the water bodies.
2 Processes of sewage treatment:

 screening  primary  aerobic decomposition  final  disinfection

sedimentation sedimentation
grit channels primary aeration tank final ultraviolet light /
untreated (沉砂槽) sedimentation tank (曝氣池) sedimentation tank chlorine gas
sewage (初級沉降池) (最後沉降池)

liquid
effluent
bar (廢水)
screens
(隔篩) sludge air air air
activated
(污泥) sludge
activated sludge (活性污泥)

digester
(消化池)

methane (甲烷)
(used as fuel)
sludge
dried sludge
(to be disposed
of or as fertilizer)
 anaerobic decomposition
(sludge digestion)

3 Roles of microorganisms in sewage treatment:

Aerobic decomposition Under aerobic conditions,

in the aeration tank - bacteria, protozoans and fungi in the sewage break down organic
matter into inorganic compounds.
- nitrifying bacteria convert ammonium compounds into nitrates.

Anaerobic decomposition Under anaerobic conditions,

(sludge digestion) in the - anaerobic bacteria decompose organic matter in the sludge to
digester produce hydrogen, carbon dioxide and organic acids.
- methanogens (產甲烷菌) use hydrogen, carbon dioxide and organic
acids as substrates to produce methane.

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2.6 Biogas production
1 Biogas (沼氣) is produced during the decomposition of organic waste by anaerobic bacteria
under anaerobic conditions. It contains methane (55–70%) and can be used as fuel.
2 In some farms, digesters are built for biogas production. They are often built underground to:
- maintain an optimum temperature for biogas production.
- withstand the pressure built up by biogas.
- reduce their visual impact on the environment.

organic ground seal digested sludge

biogas
waste in surface out used as fertilizer

biogas

decomposition of
digester organic waste by
anaerobic bacteria

3 In Hong Kong, biogas is collected from landfills and produced by decomposing food waste.
Food waste is turned into biogas and compost (堆肥) as follows:

Food The suspension is Residues from the digesters are placed in

waste is carried to digesters. composting tunnels (堆肥隧道). At about 55 °C
crushed At about 35 °C and and under aerobic conditions, microorganisms
and mixed under anaerobic decompose the remaining organic matter at a
with water. conditions, fast rate, releasing carbon dioxide and heat.
microorganisms After about 20 days, the residues become
decompose the organic compost. Compost is rich in minerals and can
matter to produce be used as fertilizer.
biogas.

4 Advantages of using biogas:

- Using biogas can reduce the use of other sources of energy.
- Burning biogas is cleaner than burning fossil fuels.
- Using waste material to produce biogas reduces waste disposal.
- The production costs of biogas are low.
- Digested sludge and compost obtained from biogas production can be used as fertilizer.

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2.7 Genetically modified microorganisms
1 Significance of using genetically modified (GM) microorganisms (基因改造生物):
- GM microorganisms can act as ‘biological factories’ for producing a wide range of
pharmaceutical products, food additives and industrial enzymes.
2 Potential hazards of using GM microorganisms:

Examples

Hazards to - The long-term effects of GM microorganisms and their products on human

human health health are still unknown.
- GM microorganisms may transfer antibiotic resistance genes to
pathogens, producing ‘superbugs’ that are resistant to multiple antibiotics.
- The insertion of new genes may increase the ability of microorganisms
to cause diseases in humans.
- GM microorganisms may produce unknown substances that contaminate
their products and cause allergic reactions in humans.
- GM microorganisms may be wrongly used as biological weapons.

Hazards to - GM microorganisms may transfer their modified genes to the wild types,
the environment causing genetic pollution. These genes may have unexpected and
adverse effects on the wild types.
- GM microorganisms may out-compete the wild types and upset the
ecological balance, resulting in a decrease in biodiversity.

3 Measures to minimize the potential hazards of using GM microorganisms:

- Establishing strict guidelines for the use of aseptic techniques in laboratory procedures
to prevent GM microorganisms from infecting handlers or escaping into the environment.
- Testing the proteins produced by GM microorganisms for their likelihood of causing
allergic reactions.
- Modifying GM microorganisms so that they cannot exchange their genes with
microorganisms in the wild.

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