3.

ENZYME

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3.1 BIOLOGICAL CHARACTERISTICS OF
WASTEWATER

1. Microorganisms found in untreated wastewater include:

bacteria, archaea, fungi/yeast, algae, protozoa, rotifers,
algae and viruses.
2. Source of theses microorganism is from human and
animals waste.
3. The removal of OM, nutrients and specific trace of
organic constituents (in some cases) found in wastewater
is accomplished biologically using variety of
microorganisms, principally BACTERIA.
4. Microorganisms are used to transform/oxidize the OM,
nutrients and specific trace organic constituents (in some
cases) into acceptable end products and additional
BIOMASS.
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3.1.1 Bacteria

1. The interior of bacteria cell contains a colloidal

suspension of protein, carbohydrates and other
complex OM, called the cytoplasm.
2. Bacteria are classified into 5 groups according to their
basic shapes: spherical (cocci), rod (bacilli), spiral
(spirilla), comma (vibrios) or corkscrew (spirochaetes).
They can exist as single cells, in pairs, chains or
clusters.
3. Bacteria moves by means of long, whip-like structures
called flagella. Other bacteria secrete a slime layer and
ooze over surface like slugs. Others are fairly stationary.

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4. Bacteria can also be classified based on;
i. Metabolism:
Heterotrophs; use OM as carbon source for the
formation of new biomass.
Autotrophs; derived cell carbon from CO2.
ii. Energy sources:
 Phototrophs; use light as energy sources.
 Chemotrophs: Obtained energy from the oxidation of
reduced inorganic compounds, such as ammonia,
ferrous iron and sulfide

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3.1.2 Other microorganisms

1. Fungi or yeast: Useful in sludge composting.

2. Archaea: Important in anaerobic processes
3. Protozoa: Act as polishers of the effluent (SS and
turbidity) by removing a certain amount of non-
flocculated bacteria and fine suspended biomass
particles.
4. Rotifers: Their presence in effluent indicates a highly
efficient aerobic biological purification process.
5. Algae: Important for lagoon type treatment plant.

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Biomass/sludge formed by bacteria

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Example of protozoa in biomass/sludge

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3.2 UNDERSTANDING CELL GROWTH

1. The increase in the cell size and cell mass during the
development of an organism is termed as growth.
2. The growth of the organism is affected by both Physical
and Nutritional factors.
3. Physical factors; pH, temperature, Osmotic pressure,
Hydrostatic pressure, and Moisture content of the
medium in which the organism is growing.
4. Nutritional factors; the amount of Carbon, nitrogen,
Sulphur, phosphorous, and other trace elements
provided in the growth medium.

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5. Bacteria can also be classified based on; The dynamics
of the bacterial growth, and can be studied by plotting
the cell growth versus the incubation time or log of cell
number versus time.
6. The curve thus obtained is a sigmoid curve and is
known as a standard growth curve.

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Standard growth curve

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BIOMASS GROWTH PHASES FOR A BATCH REACTOR

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Credit Image: Metcalf and Eddy, 2014
3.2.1 Bacteria growth pattern

1. Lag Phase: The time required for the microorganism to

acclimate to new environment.
2. Exponential or Growth Phase; Bacteria cells are
multiplying at maximum rate due to the availability of
food or nutrients. Biomass growth curve increased
exponentially.
3. Declining Phase: The growth rate decrease and then
ceases.
4. Stationary Phases: The biomass concentration remains
relatively constant with time. Bacteria growth is offset by
the death of cells.

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5. Endogenous Phase: The microorganisms compete for
the limiting food or nutrient availability. Some
microorganism metabolize their own protoplasm. The
rate of metabolism decrease and starvation occurs.
6. Death Phase: No growth is occurring. The change in
biomass concentration is due to cell death or cell lysis.

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3.3 WHAT ARE ENZYMES

1. Enzymes are biological molecules (typically proteins)

that acts as a catalyst.
2. Enzyme is responsible for accelerating the rate of a
reaction in which various substrates are converted to
products through the formation of an enzymes-
substrate complex.
3. Each type of enzyme catalyzes only one type of
reaction and will operate on only one type of substrate.
4. As a consequence, enzymes are highly specific and are
able to discriminate between slightly different substrate
molecules.
5. Enzymes exhibit optimal catalytic activity over a narrow
range of temperature, ionic strength and pH. 14
3.3.1 Active site theory of enzyme reaction

1. Lock and key model:

– Enzyme has a rigid shape
– Substrate enters the active site and fits snugly
– Enzyme-substrate complex is formed
– Substrate is change into products
– Products exit the active site

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2. Induced Fit model:
– Substrate enters the active site
– Enzyme changes its shape slightly to accept substrate
– Enzyme-substrate complex is formed
– Substrate is change into products
– Products exit the active site upon which point the enzyme returns
to its original shape

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3.3.2 Bacteria and Enzyme

1. Bacteria have the capability of producing many different

types of enzymes. They are living organisms that
respond to their environment.
2. Bacteria are capable of producing enzymes that
degrade a wide variety of organic materials such as fats,
oils, cellulose, proteins and starches.
3. All of these materials are polymers that must be reacted
with more than one type of enzyme in order to be
efficiently degraded to their basic building blocks. Nature
provides a specific "team" of enzymes to attack each
type of polymer.

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4. For example, there are three different classes of
enzymes (endocellulases, exocellulases,
cellobiohydrolases) that are required to degrade a
cellulose polymer into basic glucose units.
5. All three types of enzymes are referred to as cellulases,
but each class attacks a specific structure or
substructure of the polymer. Acting individually, none of
the cellulases is capable of efficiently degrading the
polymer.
6. Bacteria can produce the complete "team" of enzymes
that are necessary to degrade and consume the organic
materials present in their environment at any given time.
Moreover, bacteria can produce multiple "teams" at the
same time.
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 Microbiology in wastewater treatment

https://www.youtube.com/watch?v=epAh6hHOq3c

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