This document provides an overview of industrial microbiology. It discusses the history of industrial microbiology beginning with Pasteur's work demonstrating the role of microbes in fermentation. It also describes characteristics of industrial microbes, fermentation processes, sterilization, genetic manipulation techniques, major products including primary and secondary metabolites, and waste disposal methods. The key applications of microbes include production of enzymes, organic acids, antibiotics, and other pharmaceuticals on an industrial scale.
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1. KWARA STATE UNIVERSITY, MALETE, ILORIN, NIGERIA
COLLEGE OF PURE AND APPLIED SCIENCE
DEPARTMENT OF MICROBIOOGY
INDUSTRIAL MICROBIOLOGY
A TERM PAPER
BY
ABIOYE MAYOWA JOHNSON
14D/57MB/446
IN FULFILMENT OF THE REQUIREMENT OF THE COURSE MCB 201
(GENERAL MICROBIOLOGY I)
SEPTEMBER 2014.
2. Contents
Abstracts and Introduction to Industrial Microbiology Page 1
History Page 2-3
Characteristics of Industrial Microbes and Feature of Microbes Page 3
Fermentation Medium Page 4
Fermentor & Fermentation Process Page 4
Sterilization of culture media and fermenter Page 5
Microorganisms used in industrial microbiology Page 5
Genetic Manipulation of Microorganisms Page 5
Protoplast Fusion Page 6
Preservation of Microorganisms Page 6
Natural Genetic Engineering Page 6
Growth of microorganisms in an industrial setting Page 7
Major products of industrial microbiology Page 7
Primary metabolites Page 7
Secondary metabolites Page 8
Microbial biomass Page 8
Recombinant products Page 9
Important microbial products Page 9
3. Waste disposal and treatment of waste in industry Page 9
Systems for the treatment of wastes Page 10
Waste water disposal in the pharmaceutical industry Page 10
Deduction Page 11
Conclusion Page 11
References Page 12
ABSTRACT
The genetic diversity that abounds in the microbial world represents a vast, untapped of
medically and industrially important molecules. Industrial microbiology harnesses the capacity
of microbes to synthesis compound that have important application in medicine, food
preparation, and other industrial process. These compounds are commonly called natural
products. Industrial Microbiology deals with the uses of microorganism to assist in the
manufacture of product used in treating or preventing disease. Today many industrial and
pharmaceutical processes make us of genetic engineering.
INTRODUCTION TO INDUSTRIAL MICROBIOLOGY
Industrial microbiology is basically associated with the commercial exploitation of microbes for
the benefit of mankind. These microbial products may have direct or indirect impact on the
economics, environment and social parameters of the society. The use of microbes for the
production of industrially important metabolites is not a recent event. Mankind has been
4. producing alcoholic beverages and dairy products since the beginning of the civilization but they
were oblivious of the role of microbes in the production of these products. The exact role of
microbes in the fermentation process was convincingly shown for the first time by Louis Pasteur
in 1850s. With the refinement of technology to obtain pure cultures, the association of specific
microbes with specific products gained momentum. There are many definitions of
biotechnology. One of the broadest is the one given at the United Nations Conference on
Biological Diversity (also called the Earth Summit) at the meeting held in Rio de Janeiro, Brazil
in 1992. That conference defined biotechnology as “any technological application that uses
biological systems, living organisms, or derivatives thereof, to make or modify products or
processes for specific use.” Many examples readily come to mind of living things being used to
make or modify processes for specfic use. Some of these include the use of microorganisms to
make the antibiotic, penicillin or the dairy product, yoghurt; the use of microorganisms to
produce amino acids or enzymes are also examples of biotechnology. Developments in
molecular biology in the last two decades or so, have vastly increased our understanding of the
nucleic acids in the genetic processes. This has led to applications of biological manipulation at
the molecular level in such technologies as genetic engineering.
HISTORY
Traditional fermentation processes, such as those involved in the production of fermented dairy
products and alcoholic beverages, have been performed for thousands of years. However, it is
less than 150 years ago that the scientific basis of these processes was first examined. The birth
of industrial microbiology largely began with the studies of Pasteur. In 1857 he finally
demonstrated beyond doubt that alcoholic fermentation in beer and wine production was the
5. result of microbial activity, rather than being a chemical process. Prior to this, Cagniard-
Latour, Schwann and several other notable scientists had connected yeast activities with
fermentation processes, but they had largely been ignored. Pasteur also noted that certain
organisms could spoil beer and wine, and that some fermentations were aerobic, whereas others
were anaerobic. He went on to devise the process of pasteurization, a major contribution to food
and beverage preservation, which was originally developed to preserve wine. In fact, many of the
early advances of both pure and applied microbiology were through studies on beer brewing and
wine making. Pasteur’s publications, Études sur le Vin (1866), Études sur la Bière (1876) and
others, were important catalysts for the progress of industrial fermentation processes. Of the
further advances that followed, none were more important than the development of pure culture
techniques by Hansen at the Carlsberg rewery in Denmark. Pure strain brewing was carried out
here for the first time in 1883, using a yeast isolated by Hansen, referred to as Carlsberg Yeast
No. 1 (Saccharomyces carlsbergensis, now classified as a strain of Saccharomyces cerevisiae).
During the early part of the 20th century, further progress in this field was relatively slow.
Around the turn of the century there had been major advancements in the large-scale treatment of
sewage, enabling significant improvement of public health in urban communities. However, the
first novel industrial-scale fermentation process to be introduced was the acetone–butanol
fermentation, developed by Weizmann (1913–15) using the bacterium Clostridium
acetobutylicum. In the early 1920s an industrial fermentation process was also introduced for the
manufacture of citric acid, employing a filamentous fungus (mould), Aspergillus niger. Further
innovations in fermentation technology were greatly accelerated in the 1940s through efforts to
produce the antibiotic penicillin, stimulated by the vital need for this drug during World War II.
Not only did production rapidly move from small-scale surface culture to large-scale submerged
6. fermentations, but it led to great advances in both media and microbial strain development. The
knowledge acquired had a great impact on the successful development of many other
fermentation industries. More recent progress includes the ability to produce monoclonal
antibodies for analytical, diagnostic, therapeutic and purification purposes, pioneered by
Milstein and Kohler in the early 1970s. However, many of the greatest advances have followed
the massive developments in genetic engineering (recombinant DNA technology) over the last
20 years.
CHARACTERISTICS OF INDUSTRIAL MICROBIOLOGY
The discipline of microbiology is often divided into sub-disciplines such as medical
microbiology, environmental microbiology, food microbiology and industrial microbiology. The
boundaries between these sub-divisions are often blurred and are made only for convenience.
Bearing this qualification in mind, the characteristics of industrial microbiology can be
highlighted by comparing its features with those of another sub-division of microbiology,
medical microbiology
FEATURES OF MICROBES
These are tiny organisms which require the use of microscopes for their visualization. Microbes
are highly versatile organisms. They have many positive features which are responsible for their
uses in the field of industrial microbiology.
• Microbes grow and multiply very rapidly. Billions of cells can be produced in a single
day e.g. E.coli doubles itself in 15-20 minutes under optimal conditions of growth.
• Microbes require cheap nutrients for growth which are available throughout year.
7. • Microbes have great biodiversity. They can grow under extreme conditions of
temperature, pH, salts, pressure etc.
• The genotype of industrially important microbes is fully known. This aspect helps us to
understand the biosynthetic and regulation of a desired gene product. In addition,
scientists can either shut off undesirable biosynthetic pathways or boost the yield of
desired gene product.
FERMENTATION MEDIUM
The growth medium (liquid or solid) in which microbes grow and multiply is called fermentation
medium. The selected microbe should be able to utilize and grow on cheap sources of carbon and
nitrogen. Usually these sources are waste products of industrial processes e.g. molasses, whey,
corn steep liquor etc. Care is taken to avoid the use of such microbes which require expensive
nutrients like vitamins for their growth.
Fermentor & Fermentation Process
A fermenter, also called a bioreactor, is a vessel in which a particular microbe is grown under
controlled conditions to produce a desired byproduct or biomass. The aim of the fermenters is to
provide a stabilized condition for growth of cells and better production of a desired byproduct.
Fermentation processes
There are two groups of fermentation processes: liquid fermentation and solid fermentation. In
former case, cells are suspended in aqueous medium where as in latter case, the volume of free
liquid is minimal and the cells are adsorbed to a solid and nutrient rich material.
Liquid fermentation
8. Solid State Fermentation (SSF)
Sterilization of culture media and fermenter
Sterility is of utmost importance to avoid growth of unwanted contaminants and in achieving
maximal yield of a bioproduct by the use of fermentation process using known cultures as
inoculum. The most common means of achieving sterilization is moist heat. All forms of
vegetative microbes are killed by applying heat (60oC) for 10-15 min. The spores are heat
resistant and majority of them are destroyed only if exposed to 100oC for 10 min. However, in a
few cases, like spores of Bacillus stearothermophilus, temperature over 120oC is needed for
their destruction. Complete sterility of culture medium is achieved by subjecting the medium to
121oC for 15 min. or equivalent time temperature combination for small volumes of sample.
MICROORGANISMS USED IN INDUSTRIAL MICROBIOLOGY
Microbes used in Industrial Microbiology were cultured from natural materials like soil samples,
waters, and spoilt bread and fruit. Samples from all areas of the world continue to be examined
to identify new strain with diserable characteristics. However, because most of these microbes
will resist growth under standard laboratory conditions, techniques for enrichment of natural
samples and further purification are constantly evolving and here are the approaches used to
optimize microbes for industrial purpose.
Genetic Manipulation of Microorganisms
Genetic manipulations are used to produce microorganisms with new and desirable
characteristics. The classical methods of microbial genetics play a vital role in the development
9. of cultures for industrial microbiology. Mutation Once a promising culture is found, a variety of
techniques can be example, the first cultures of Penicillium notatum, which could be grown
only under static conditions, yielded low concentrations of penicillin. In 1943 a strain of
Penicillium chrysogenum was isolated
Protoplast Fusion
Protoplast fusion is now widely used with yeasts and molds. Most of these microorganisms are
asexual or of a single mating type, which decreases the chance of random mutations that could
lead to strain degeneration. To carry out genetic studies with these microorganisms, protoplasts
are prepared by growing the cells in an isotonic solution while treating them with enzymes,
including cellulase and beta-galacturonidase. Transfer of Genetic Information between Different
Organisms. New alternatives have arisen through the transfer of nucleic acids between different
organisms, which is part of the rapidly developing field of combinatorial biology. This involves
the transfer of genes for the synthesis of a specific product from one organism into another,
giving the recipient varied capabilities such as an increased capacity to carry out hydrocarbon
degradation. An important early example of this approach was the creation of the “superbug,”
patented by A. M. Chakarabarty in 1974, which had an increased capability of hydrocarbon
degradation. The genes for antibiotic production can be transferred to a microorganism that
produces another antibiotic, or even to a non-antibiotic-producing microorganism.
Natural Genetic Engineering
The newest approach for creating new metabolic capabilities in a given microorganism is the
area of natural genetic engineering, which employs forced evolution and adaptive mutations
10. Preservation of Microorganisms
Once a microorganism or virus has been selected or created to serve a specific purpose, it must
be preserved in its original form for further use and study. Periodic transfers of cultures have
been used in the past, although this can lead to mutations and phenotypic changes in
microorganisms. To avoid these problems, a variety of culture preservation techniques may be
used to maintain desired culture characteristics. Lyophilization, or freeze-drying, and storage in
liquid nitrogen are frequently employed with microorganisms
GROWTH OF MICROORGANISMS IN AN INDUSTRIAL SETTING
Once a medium is developed, the physical environment for microbial functioning in the mass
culture system must be defined. This often involves precise control of agitation, temperature, pH
changes, and oxygenation. Phosphate buffers can be used to control pH while also functioning as
a source of phosphorus. Oxygen limitations, especially, can be critical in aerobic growth
processes. The O2 concentration and flux rate must be sufficiently high to have O2 in excess
within the cells so that it is not limiting. This is especially true when a dense microbial culture is
growing. When filamentous fungi and actinomycetes are cultured, aeration can be even further
limited by filamentous growth. Such filamentous growth results in a viscous, plastic medium,
known as a non-Newtonian broth, which offers even more resistance to stirring and aeration.
To minimize this problem, cultures can be grown as pellets or flocs or bound to artificial
particles.
MAJOR PRODUCTS OF INDUSTRIAL MICROBIOLOGY
11. A vast range of industrial products which were earlier made by chemical processes are now
being made with the help of microbes. Contrary to the belief that microbes are harmful to
humans, majority of microbes are either harmless or provide beneficial products to humans.
Microbe based industrial products have made inroads in all walks of life i.e. health sector, food
sector, agriculture sector, industrial chemical sector and environmental sector. These products
can be categorized into four main groups
Microbial products of industrial importance
Category Product
Primary metabolites Enzymes, amino acids, nucleotides, organic acids, ethanol, butanediol
Secondary metabolites Antibiotics, gibberellins, hormones, pigments, alkaloids
Microbial biomass Baker’s yeast, single cell protein (SCP), probiotics, vaccines
Recombinant products Insulin, streptokinase, interferons, Interleukins, growth hormones,
vaccines
Primary metabolites
These are those metabolites which are produced when the cells are growing actively in the
presence of sufficient amount of nutrients. This phase of growth of cells is called trophophase.
Most of the primary metabolites (amino acids, nucleotides, proteins, nucleic acids, lipids,
carbohydrates) are essential or almost essential for the growth and survival of the organisms.
Some of the primary metabolites have found industrial applications e.g. ethanol, citric acid,
glutamic acid, lysine, vitamins etc.
12. Secondary metabolites
Secondary metabolites are produced by the cells after the active growth of cells has ceased. This
phase of growth of cells is called idiophase. Cells do not divide but are metabolically active.
Secondary metabolites are not essential for the survivability of the cells. Not all classes of
microbes exhibit secondary metabolism. It is commonly seen in fungi, yeast, actinomyces but is
absent in a few bacteria like E.coli, Salmonella, Shigella, Proteus, Klebsiella etc. The microbes
produce secondary metabolites when their growth rate either slows down or there is complete
cessation of growth. Both conditions result in the production of secondary metabolites by
microbes. In the cells there is correlation between primary and secondary metabolism. Secondary
metabolism succeeds primary metabolism. Alternatively, secondary metabolites are produced
from intermediates and end products of primary metabolites.
Microbial biomass
In a few instances the cells i.e. biomass of microbes, has industrial application as listed in Table
3. The prime example is the production of single cell proteins (SCP) which are in fact whole
cells of Spirullina (an algae), Saccharomyces (a yeast) and Lactobacillus (a bacterium). SCP is
essentially rich in amino acids which are either absent in vegetarian food or present in low
amounts e.g. lysine, threonine, methionine, leucine, isoleucine etc.
Recombinant products
With the advent of gene cloning techniques, many industrially important genes from plants,
animals and microbes have been cloned in a few selected microbes like E.coli and
Saccharomyces and Pichia. The basic idea behind all these clonings is to produce large amount
13. of proteins and scientists have been highly successful in achieving this goal. Table 4 lists some
of the recombinant products.
IMPORTANT MICROBIAL PRODUCTS
• Microbial enzymes: Enzymes are biocatalysts which are primarily protein in nature
though certain RNA molecules have also been shown to possess catalytic activity.
• Enzymatic processes are fast replacing chemical processes because such technology is
eco-friendly, stereo specific and generate less undesirable waste products.
WASTE DISPOSAL
TREATMENT OF WASTE IN INDUSTRY
The activities of industrial microorganisms usually occur in large volumes of water; the resulting
wastes are therefore transported in aqueous medium. This chapter will examine briefly the
treatment of waste water. The subject is of interest, not only from the intrinsic need to dispose of
wastes in industry, but especially because the basis for ultimate waste disposal is microbial.
Waste carried in water, whether from industry or from domestic activity is known as sewage.
Waste water disposal constitutes a peculiar branch of industrial microbiology. The methods to be
discussed were evolved originally to handle domestic sewage, but they have been extended for
use in those industries, such as the food and fermentation industries, which yield wastes
degradable by microorganisms. Sewage emanating from some chemical industries especially
those dealing with manmade chemicals are not only less degradable but are sometimes toxic to
microorganisms and man.
14. SYSTEMS FOR THE TREATMENT OF WASTES
The basic microbiological phenomenon in the treatment of wastes in aqueous environments is as
follows:
• The degradable organic compounds in the waste water (carbohydrates, proteins, fats,
etc.) are broken down by aerobic micro-organisms mainly bacteria and to some
extent, fungi. The result is an effluent with drastically reduced organic matter content.
• The materials difficult to digest form a sludge which must be removed from time to
time and which is also treated separately. And thereby fall under two headings:
aerobic breakdown of raw waste-water and anaerobic breakdown of sludge
WASTE WATER DISPOSAL IN THE PHARMACEUTICAL INDUSTRY
The treatment of wastes from a pharmaceutical industry is chosen to illustrate industrial waste
treatment because the wastes are representative of a broad range of materials and include easily
degradable organic materials, as well as sometimes some inorganic and even toxic compounds.
Which of the various methods of disposal is used by a particular firm will depend on a number of
factors foremost among which are:
(a) The cost of the disposal method;
(b) The location of the industry;
(c) The nature of the industry and hence of its waste materials,
(d) The governmental regulations operating in the locality.
The above factors are all inter-related. For example, in siting the industry in the first place, space
for, and the type of method of, waste disposal would have been considered. The cost of the
disposal will be influenced not only by the nature and quantity of the waste and consequently the
15. method adopted to handle it, but also what distance needs to be covered to have it disposed of.
EPA regulations may for example dictate that the BOD of the wastes be reduced to a certain
level before being discharged into a stream; any BOD reduction ultimately involves the
expenditure of funds.
Nature of Wastes: The wastes from pharmaceutical firms may include easily degradable
materials such as emulsion syrup, malt and tablet preparations.
.
Pre-treatment: Before treatment acid (or alkali) is neutralized, dissolved salts are removed
usually by precipitation as calcium salts through lime addition, which also neutralizes acidity.
Chloride and sulfate may be removed by ion exchange or rendered innocuous by dilution with
water.
Treatment: Before a routine is used within a treatment method, laboratory experiments would
have been carried out to determine how much of the wastes may be efficiently handled within a
given period. It may often be necessary to segregate the wastes, treating the more easily
biodegradable organic forms separately from those wastes rich in inorganic materials
DEDUCTION
A wide variety of compounds are produced in industrial microbiology that impart our lives in
many ways. These include antibiotics, amino acids, organic acids, biopolymers and
biosurfactant, microorganism can be used as biocatalyst to carry out specific chemical reaction,
16. and microorganism can be grown in controlled environments of various types using fermenters
and other culture systems.
CONCLUSION
Numerous impressive accomplishments have been made in the engineering and production of
microbial factories for synthesis of value-added products in the past few years. However,
continuous efforts towards exploring new production hosts, creating novel enzymes that catalyze
unnatural reactions, and developing more powerful tools for functional genomics and proteomics
will be necessary to expand the range of products that can be synthesized by microbial factories.
REFERENCES
. Jacquelyn G. Black page 736, (Industrial Microbiology History)
Michael J. Waites BSc, PhD, CBiol, MIBiol Neil L. Morgan BSc, PhD, MIFST John S. Rockey
BSc, MSc, PhD Gary Higton BSc, PhD Page 15 of 302, Industrial Microbiology an introduction
Industrial Microbiology: An Introduction. Editors: M. J. Waites, N.L. Morgan, J.S. Rockey,
G. Higton. Publishers: Blackwell Science, Oxford U.K
Manual of Industrial Microbiology and Biotechnology. Editors: A.L. Demain, J.E.Davies.
Publishers: ASM Press, Washington D.C., USA. 1999.
Biotechnology. Editor: U. Satyanarayanan. Publishers: Books and Allied (P) Ltd. Calcutta.
India. 2005.
Industrial Microbiology Prof. Rupinder Tewari Professor Department of Biotechnology Panjab
University
Prescott’s miciobiology 5th edition Lanson M. Prescott (Major Product of Industrial
Microbiology and Genetic Manipulation of Microorganisms)
Nduka Okafor Department of Biological Sciences Clemson University, Clemson South Carolina
USA (Characteristic of Industrial Microbiology)