Biochemical Engineering

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Referenc es

1. Bioc hemical Engineering, 2nd edition (1973) by Shuichi Aiba, Arthur

Humhrey and Na ncy Mills, Ac ademic press

2. Principles of Fermentation Tec hnology (1984) by Stranbury P.F. and

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Perga mon Press

3. Bioc hemical engineering, unit processes in Fermentation (1958) by

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Steel R. (editor) Heywood and C o. Ltd

4. Bioc hemical Engineering (1964) by Webb F.C., Van Nostrand

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1.0 Introduction
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1.1 Historical Background

$ About 150 years ago, Louis Pasteur pointed out the important role of
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living micro orga nisms in bioc hemical proc esses. In 1928, a major
breakthrough was achieved by Alexander Fleming in discovering
%
penicillin. The urgent need of penicillin throughout the 2nd world war led
to microbiologists, biochemists and chemical engineers in a “crash”
programme of developing and designing processes in areas which
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hitherto unfamiliar to them. Three American companies led the
way-Merck, Pfizer, and Squibb. Sinc e then micro organisms have been
known to be used in the manufacture of a host of complex chemicals,
antibiotics, enzymes and vitamins.

In the presidential address to the institutions of C hemical Engineers in

1952, Sir Harold Hartley ma de a first public referenc e to the term
Biochemical Engineering. He recognized Biochemical Engineering as
an emerging branch of chemical engineering. He further underscored
the imminent contribution of biological substances and processes in
industrialisation. This inevitably led to among the first series of
Biochemical lectures being offered at the University of Manchester,
where George Davis gave his first lectures in Chemical engineering.
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However, most of the Biochemical fundamental principles had been in

app lica tion since a ntiquity.
And so, what is Biochemical Engineering? Biochemical engineering is
the interaction of two disciplines-Biological sciences and chemical
engineering. Biochemical engineering is concerned with conducting
biological processes on an industrial scale, providing the link between
Biological and Chemical engineering. Biological sciences will include
microbiology, biochemistry and genetics. The heart of Biochemical
engineering lies in the scale up and management of the cellular
processes. Thus, therefore there is a precise need for the chemical
engineer to understand the relative dynamic nature of the biological
catalysts.

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In order to develop biological units on a n industrial scale, three distinct
disciplines of chemica l engineering, biochemistry and microbiology are
required combined with specific trade technology. The experiences
gained from chemical engineering are handy at the development of
this rather fast growing branch of engineering. Some interesting
comparisons can be drawn between chemical engineering and
Biochemica l engineering:
!
Basic concepts of material and energy transfer and fluid flow are
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common in both but biologica l proc esses are notoriously
associated with a narrow range of temperature and non-
Newtonian materials
!
Ma ny unit of operation a re c ommon o nly for the d etails to differ

The scope of biochemical engineering

1. Industrial fields in fermentation
2. Operations of food processing; pasteurization, sterilization
and preservation
3. Industrial solvents, organic acids
4. Ma nufac ture of sera a nd vac cines
5. C ommercial enzymes
6. Extraction processes for insulin and other hormones
7. Proc essing of forests and crop products
8. Effluent disposal
Among the above activities are some that may attract the prophet
and visionary.

However, it is important to note that a broad base of technologies

must be included in Biochemical engineering, which might require
additional information and knowledge. For example, vaccine
manufacture requires in depth medical information while aspects of

hygiene
engineer.and sterilization
The awareness areofrather unfamiliar
Biological to a traditional
background chemical
is essential in

Biochemical engineering. This course assumes that the students have

ba sic knowledge in biologica l sciences.

The organization of this course is in such a way that it prepares students

to work in the existing industries as well as applying their basic
knowledge to the unborn industries of the future.
 Role of a biochemical engineer

!
Design and operation o f absolutely pure and mixed cultures
!
Prevention of contamination – providing sterile conditions as well
as “contaminant proof” environment
!
Design of auxiliary systems of air compression, delivery systems,
instrumentation and control
!
Demonstration of pilot plant equipment and its subsequent
scale-up to the production stage
!
Separation and isolation of the product using well known
techniques such as filtration, extraction, adsorption and
concentrations

Primarily, the lab oratory scientists, microbiologists, biochemists,

genecists must continue in the discovery and advancement of
desirable interac tions betwe en micro orga nisms and their environment.
The bioc hemical engineer must control and translate the laboratory

) * + ,
results to the produc tion scale operation in an economic manner.

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1.2 Exploitation of Biolog ic al procDocuments
esses

!
Potential source of proteins e.g. the use of bacteria to
breakdown n-alkane
!
C o-oxidation of substrates that do not supp ort their growth
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resulting into more useful ones e.g. p-xylene can be oxidized to
dimethyl-cis and cis-muconic acid
!
Production of enzymes-some of the enzymes are used in
(
detergent manufacture, hydrolytic reactions and as analytic

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tools
!
Production of polymers- Microbial cells excrete little slime outside
the cell wall. For example glucan which is used as plasma

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!
substitute
Pollution control-treatment of wastes with high BOD(Biological
oxygen demand)
Types of Biologica l materials

It is not easy to define a living process and it may become

increasing difficult as further biochemical advances are made. This
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course is designed primarily for the micro-orga nisms and enzymes

but it is important to gain a broader sense of other forms of life-the
similarities and differences between the unicellular organisms and
larger animals and plants.

Industrial applications of micro-organisms can be broadly classified

into:
Those in which great care is taken to use only one selected strain of
a particular organism, typified by manufacture of antibiotics and
some vac cines
Those that effort is made to maintain a fairly constant mixture of
strains of the same species e.g. in brewing and wine making
Processes which are chiefly depend on adventitious flora of mixed
species e.g. curing bacon in the manufacture of margarine, tea,
coffee etc. these activities are by large dependent on tradition a nd
experience.

1.3 MIC RO- ORG A NISMS

Types of micro-organisms

Micro organisms exist as single cells or at most in relatively

unspecialized multi-cellular colonies, with no capacity to control
cellular temperatures.

Micro organisms may be classified into four main groups

!
Bacteria
!
Viruses
!
Fungi including yeast and actinomycetes
!
Protozoa including algae

Bacteria

These a re single c ells, in the form of coc ci, rods and spirals capa ble
of independent growth.

Bacteria are ubiquitous in nature, in aerobic and anaerobic

environments containing water. In the genera, synthetic abilities
range from those of autotrophic species, which require only
inorganic compounds for growth, to those of heterotrophic species,
which may have synthetic ability and must be grown in tissue
culture.
 Due to the diverse abilities, bacteria may be exploited industrially to
ac cumulate both intermediate and end products of metabolism.

Viruses

Viruses are the smallest microbes, obligate intracellular parasites of

animals, plants, insects, fungi, algae or bacteria. They contain no
water and have little or no synthetic or metabolic activity
themselves. Growth and multiplic ations take plac e intracellular. This
frequently results in the damage and subsequent death of the host
cells.

The genetic material of viruses may either be ribonucleic (RNA) acid

or the deoxyribonucleic acid (DNA).

Myxoviruses show helical symmetry, with nucleic protein helix

enc losed by a lipoprotein sheath. Bac teriopha ges are viruses that
are parasitic on bacteria. These phages are possible contaminants
of biologic al systems.

Fungi

Fungi are widely spread in environments of lower relative humidity

than which they

anearobic, favourform
bacteria.
long The metabolism
filamentous, of fungi cells(Hyphae)
nucleated is essentially
between 4 u to 20µ wide.

Generally fungi are free living saprophytes but a few are pa rasitic
on animals and many are serious pathogens of plants.
Ac tinomycetes are intermediate between fungi and bacteria.

Industrially, this group is extremely important a s a source of powerful

antibiotics.
Protozoa

These are widely distributed in fresh and salty water, in soil and in
animals. They may be unicellular or multi cellular and exhibit a wide
range of morphologica l forms. Protozoa are either photosynthetic or
non-photosythetic while algae are capable of photosynthesis.

Protozoa are handy in removing bacteria from waste water in

trickling filters and activated sludge plants.

1.4 Requirements for G rowth and Formulation of Me dia

Requirements for growth

Detailed investigation is prerequisite to establish the most suitable

medium for individual biological processes, but certain requirements
must be met by any such medium. All micro organisms require
water, sources of energy, carbon, nitrogen, mineral elements and
possibly vitamins plus oxygen if aerobic. It is important to appreciate
that the c ultural conditions that achieve maximum cell mass may
not be necessarily those that give maximum yield of some products
of metabo lism. On a small scale, it is relatively simple to devise
medium, although supporting satisfactory growth may not be
suitable for use in large sc ale.

On large scale, one must normally use resources of cheap nutrients

to create medium which will meet as many as possible of the
following criteria:
!
It will prod uce maximum yield of produc ts and biomass per
gram of substrate used
!
It will prod uce ma ximum conc entration of the products
!
It will permit maximum rate of product formation
!
There will be minimum yield of undesirable products
!
It will be cheap and of consistent quality and readily
available throughout the yea r
!
It will cause minimum problems in other aspec ts of production
process particularly aeration and agitation, extraction,
 process particularly aeration and agitation, extraction,
purification a nd waste mana gement

Some of the c hea p sources of nutrients include cane molasses, beet

molasses, c ereal grains, glucose, sucrose and lactose(C arbon sources)
while ammonium salts, urea, nitrates, corn steep liquor, Soya bean
meal, slaughter house waste and fermentation residues(nitrogen
sources).The medium selected will affect the design of equipment and
processes.

The best temperature for cultivation varies with species but orga nisms
occurring in the soil naturally grow best at temperatures between 25°c
and 30°c while those isolated from a nimals grow best at 37°c . Some
organisms are actually thermophilic e.g. those used in bio-digesters
(Lactobacillus ), which grow best at 40-45°c .

The products of microbial metabolism often cause major shifts in PH. It is

important to maintain desirable PH level. Sometimes the optimum PH
for product formation may not be the optimum PH for growth. When
acid by-products accumulate in the medium, causing unwanted fall in
the PH, ammonia is slowly fed to the culture, so supplying nitrogen for
growth while maintaining PH. Calcium ca rbonate may also be used for
the PH control in conditions where the required product is water
soluble. When cell mass or some insoluble metabolite is required, acidic
products are conveniently neutralised by a dd ing sodium hydroxide.

Micro organisms vary in their need for oxygen. On one hand fungi,
algae and a few bacteria are obligate anaerobes. On the other hand,
a few bacteria are strict anaerobes and many bacteria and many
bacteria can grow in both situations (facultative anaerobes).

The physical c onditions of temperature and PH will have a profound

effect on microbial growth. The useful range of temperature over
which the metabolic processes of a micro-organism proceed at

significant rate
temperature is quite innarrow,
is important usually
controlling not more
the flavour thanof many
balance -20°c.
fermented beverages and food. There is usually an optimum PH range,
which is normally limited with complete inactivation or dea th on either
extreme.

1.5 FORMULA TIO N O F MEDIA

This is an essential pa rt in the design of succ essful laboratory

experiments, pilot-scale development and the manufacturing
proc esses. The constituent of media must satisfy the elemental
requirements for the biomass and metabolite production and there
must be adequate supply of energy for the biosynthesis and cell
maintenance. The first step to consider is an equation based on the
stoichiometry for growth and product formation.

Thus,

Carbon + energy + nitrogen + other requirements ⎯environmen

⎯⎯⎯ ⎯ tal
→ cell biomass + products

+ CO2 + H 2 O + Heat

A quantitative treatment of the above equation is desirable in the

economical design of the media if compound wastage is to be
minimized. This calls for the elemental composition (K, N, O, Mg etc),
whose
excess data is not
quantity ofreadily available. Sometimes,
some elements. For example,it the
is important to have
concentration of
phosphorous is deliberately raised in some c ulture to offer a buffering
effect in addition.

The carbon substrate has dual role in the biosynthesis and energy
generation. The carbon requirement under aerobic conditions may be
estimated from the cellular yield coefficient(Y) which is defined as

γ = quantity of cell dry matter produced

quantity of carbon substrates utilised
Analyses made to determine how the observed
carbon sources to the product compares
maximum yield.

1.6 CONSTITUENTOF CULTURE MEDIA

WATER

Water is a major component. Clea n wa ter of

therefore required in large quantities from a
the factors needed to c onsider include PH, dis
contamination. For example, the mineral content
in the brewing industry; with hard wa ters
suitable for pilsner type lagers. It is advisable
make it more adaptable for the biological processes

Energy sources

Energy for growth comes from either the

components or from light. Most industrial micro-org
organotrophs, therefore the commonest sources
carbon sources such as carbohydrates, lipids a

Carbon sources
!
Loc al laws especially in some countries where
The following
the useare some ingredients.
of some of the examples of carbon
For instance,
may only be called by a certain name
!
Starch from
within maizegeographical
a limited grains, potatoes, areacassor
!
Sucrose vin
include fromde suga r ca ne or
bordeaux suga r beet
, which is cultivate
!
Lactose
East from milk whey powder
of France
!
! Commercial
Method vegetable
of media oils-both
preparation as ita is
e.g.
foaming agent
! sugars
C
and separately
ornamino
steep liquor-
a because
cids toa they
by-product
form blac may
of react
starch
k nitrogen
!
Simple organic acids and alkanes
The influence of carbon sources on product for
Factors influencing the choice of carbon sources
Of great significance is the rate at which
!
The main
metabolized produc
. This ts of the
will influenc fermentation-
e the formation
proc esses
of primary metabolites.
of primary metabolites.
!
Impurities of the carbohydrate sources
!
Government legislation e.g. in the EU,
Nitrogen sourcesare encouraged compared
molasses
molasses used micro organisms can utilise
Most industrially
sources of nitrogen. In organic sources may
ammonium salts and nitrates. Most orga nic
mainly supplied as amino acids, protein or urea.
relatively expensive.

Fac tors influenc ing the sources of nitrogen sour

The nitrogen sources have been shown to infl

pattern. Antibiotic production may be inhibited
nitrogen source. For example, in the prod ucti
soybean meal is considered a good nitrogen
ba lanc e of nutrients.

Minerals
In many media ma gnesium, phosphorous
chlorine are considered as essential. These
component. Others such as cobalt, copper,
molybdenum and zinc are also essential
impurities in major ingredients.

Vitamins sources
While many of the natural carbon and nitrogen
some of the required vitamin, any vitamin
eliminated b y a c areful blend of materials.

Nutrient recycle
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