Industrial

microbiology 1
 Industrial microbiology and biotechnology involve the
use of microorganisms to achieve specific goals

 It is the commercial use of microorganisms to

produce valuable economic, environmental and
socially important products, or to carry out
important chemical transformations

 Biotechnology has developed rapidly due to the

genetic modification of microorganisms, particularly
by recombinant DNA technology
Fermentation Products

Food, beverage, food additives and supplements

 Dairy products (yogurt, cheese)

 Alcoholic beverages (beer, wine)
 Amino acids, vitamins
Food, beverages, food additives and supplements
► A wide range of fermented foods and beverages
products used worldwide and are of vast economic
importance

► Fermented dairy products resulting from the

activities of lactic acid bacteria in milk, which
modify flavour and texture,and increase long-term
product stability.

► Yeasts are exploited in the production of alcoholic

beverages e.g. beer and wine, due to their ability
to ferment sugars, derived from various plant
sources, to ethanol
► The first human use was for acetic acid, as vinegar,
produced as a result of the oxidation of alcoholic
beverages by acetic acid bacteria

► A further aerobic fermentation involves citric acid

production by the filamentous fungus, A. niger, which
has become a major industrial fermentation product, as
it has numerous food and non-food applications.
► amino acids and vitamins used as supplements in
human food and animal feed are produced most
economically by microorganisms, particularly if high-
yielding overproducing strains are developed.

► Some microorganisms contain high levels of protein

with good nutritional characteristics suitable for
both human and animal consumption.

► This so-called ‘single-cell protein’ (SCP) can be

produced from a wide range of microorganisms
cultivated on low-cost carbon sources.
 Health Care Products
► Antibiotics – over 4000 isolated, only 50 used regularly

► β-lactams, penicillins, and cephalosporins Aminoglycosides

(streptomysin), Tetracyclins

► Important to develop new antibiotics due to abuse/misuse of

current antibiotics
 Alkaloids, steroids, vaccines

 Therapeutic recombinant human proteins (insulin, interferons,

blood-clotting factors, human growth hormone)

 More recombinant therapeutic products to be developed

Production of microbial enzymes

► particularly extracellular hydrolytic enzymes, have numerous

roles----in the production of a wide range of specific food and
nonfood products.

► Proteases----are extensively used as additives to washing powders,

in the removal of protein hazes from beer and as microbial rennets
for the production of cheese.
► Immobilization of enzymes or whole cells
by their attachment to inert polymeric supports,
allows easier recovery and reuse of the
biocatalyst, and some enzymes are much more
stable in this form.

► the product does not become contaminated with

the enzyme.

► Applications of immobilized biological catalysts

include the production of amino acids, organic
acids and sugar syrups.

 Proteases, carbohydrases, Taq polymerase

Industrial chemicals and fuels
 Industrial feedstock chemicals supplied through fermentation
include various alcohols, solvents such as acetone, organic acids,
polysaccharides, lipids and raw materials for the production of
plastics

 Some of these fermentation products also have applications in

food manufacture.
After exhaustion of fossil fuels…Biological fuel generation may
make an increasing contribution, particularly in the conversion of
renewable plant biomass to liquid and gaseous fuels

► This plant biomass can be in the form of cultivated energy crops,

natural vegetation, and agricultural, industrial and domestic
organic wastes.
► methane and ethanol---main products
► other potential fuels can be generated using microorganisms,
including hydrogen, ethane, propane and butanol.
Overview of a Fermentation Process
Fermentation process – Upstream Processing

1. Fermentation Organism
► Key factors relating to this aspect are:

► the strategy for initially obtaining a suitable industrial

microorganism,
► strain improvement to enhance productivity and yield
► maintenance of strain purity
► preparation of a reliable inoculum
► continuing development of selected strains to improve the
economic efficiency of the process

 need suitable cells to produce desired products (bacteria,

fungi, yeast, animal cells)

 improve strain to enhance productivity and yield

 maintain purity of cultures

► Some microbial products are primary metabolites,
produced during active growth (the trophophase),
which include amino acids, organic acids, vitamins
and industrial solvents such as alcohols and acetone

► However, many of the most important industrial

products are secondary metabolites, which are
not essential for growth, e.g. alkaloids and
antibiotics

► These compounds are produced in the stationary

phase of a batch culture, after microbial biomass
production has peaked (the idiophase).
Characteristics of cells

 produce usable products or effects

 be available in pure culture
 be genetically stable, or genetically mutated
 produce spores or other reproductive structures to
allow easy inoculation

 grow rapidly and produce product quickly in large

scale culture
 be easily separated from products
 not be harmful to humans, plants, animals, etc
2. Fermentation Medium

 need cost-effective carbon and energy sources,

essential nutrients

 media often wastes from other processes, such as sugar

processing wastes, lignocellulosic wastes, cheese whey
and corn steep liquor

► The selection of suitable cost-effective carbon and

energy sources

► essential nutrients, along with overall media

optimization are vital aspects of process development
to ensure maximization of yield and profit.
► In many instances, the basis of industrial media
are waste products from other industrial
processes, notably sugar processing wastes,
lignocellulosic wastes, cheese whey and corn
steep liquor.
 Fermentation

 industrial microorganisms cultivated under

controlled conditions to optimize growth of
organism and production of microbial products

 Consequently, in order to obtain high yields Must

avoid environmental conditions that trigger regulatory
mechanisms (repression, feedback inhibition)

► The synthesis of microbial metabolites is usually tightly

regulated by the microbial cell.
► Fermentations are performed in large fermenters often
with capacities of several thousand litres

► These range from simple tanks, which may be stirred or

unstirred, to complex integrated systems involving
varying levels of computer control

► The fermenter and associated pipework, etc., must be

constructed of materials, usually stainless steel, that
can be repeatedly sterilized and that will not react
adversely with the microorganisms or with the target
products
 Fermentation process – Downstream Processing

 includes all processes after fermentation

 involve cell harvesting, cell disruption, product

purification from cell extracts or the growth medium

 must be rapid and efficient to purify product and to

maintain stability of product

 safe and inexpensive to dispose of wastes

► The mode of fermenter operation (batch, fed-batch or
continuous systems), the method of its aeration and
agitation, where necessary, and the approach taken to
process scale-up have major influences on
fermentation performance

► Conventional DSP includes all unit processes that follow

fermentation

► They involve cell harvesting, cell disruption, product

purification from cell extracts or the growth medium, and
finishing steps

► attempts are now being made to integrate

fermentation with DSP operations, which often
increases process productivity
► DSP must employ rapid and efficient methods for the
purification of the product, while maintaining it in a
stable form

► This is especially important where products are unstable

in the impure form or subject to undesirable
modifications if not purified rapidly

► For some products, especially enzymes, retention of

their biological activity is vital

► Finally, there must be safe and inexpensive disposal

of all waste products generated during the process
 Fermentation Products
Primary Metabolites:
 produced during active growth (trophophase)
 Amino acids, organic acids, alcohol fermentation
products, vitamins , Acetone

Secondary Metabolites
 produced during stationary phase after microbial
biomass production has peaked (idiophase)
 generally not essential for growth or reproduction

 antibiotics, citric acid , Alkaloids

Fermentation process
Genetic manipulation of
microorganisms
1) Mutation—once a promising culture is found,
it can be improved by mutagenesis with
chemical agents and UV light

2) Protoplast fusion
 Widely used with yeasts and molds,
especially if the microorganism is asexual or
of a single mating type; involves removal of
cell walls, mixing two different solutions of
protoplasts, and growth in selective media

 Can be done using species that are not

closely related
3) Insertion of short DNA sequences

 site-directed mutagenesis is used to insert short

lengths of DNA into specific sites in genome of a
microorganism

 leads to small changes in amino acid sequence that

can result in unexpected changes in protein
characteristics;

 site-directed mutagenesis is important to the field of

protein engineering
4) Transfer of genetic information between different
organisms

 Combinatorial biology—transfer of genes from one

organism to another….phage

 Can improve production efficiency and minimize

purification of the product

 Numerous vectors are available for transfer of genes

5) Modification of gene expression

 Can involve modifying gene regulation to

overproduce a product

 Pathway architecture and metabolic pathway

engineering—intentional alteration of pathways by
inactivating or deregulating specific genes

 Metabolic control engineering—intentional

alteration of controls for synthesis of a product
6) Natural genetic engineering

 employs forced evolution and adaptive mutations;

 specific environmental stresses are used to force

microorganism to mutate and adapts

 creates microorganism with new biological

capabilities
 Environmental roles of microorganisms

► Microorganisms are particularly important in wastewater

treatment, which utilizes the metabolic activities of diverse
mixed microbial populations capable of degrading any compound
that may be presented to them
Objectives:
► The two main objectives are to destroy all pathogenic microbes
present in the sewage, particularly the causal organisms of the
water-borne diseases cholera, dysentery and typhoid

► The second objective is to break down the organic matter in waste-

water to mostly methane and carbon dioxide, thereby producing a
final effluent (outflow) that can be safely discharged into the
environment.
► Microbial-based ‘clean technology’ is also being
increasingly used in the desulphurization of fuels and
the leaching of metals (e.g. copper, iron, uranium and
zinc) from low-grade mineral ores and wastes using
species of Thiobacillus and Sulfolobus

► Environmental biological control is a further area

where microorganisms are employed in an effort to
reduce our reliance on synthetic chemical pesticides.

► Bacteria, fungi, protozoa and viruses are cultivated to

produce biomass or cell products for the control of
fungal, insect and nematode pests of agricultural
crops, along with some vectors of human and animal
diseases.
► Microbial activities can also be employed
in the degradation of man-made
xenobiotic compounds within waste
streams and in the bioremediation of
environments contaminated by these
materials