Industrial Microbiology

302 Pages·2010·2.08 MB·English

Industrial Microbiology:
An Introduction

Michael J. Waites

BSc, PhD, CBiol, MIBiol
Neil L. Morgan

BSc, PhD, MIFST
John S. Rockey

BSc, MSc, PhD
Gary Higton

BSc, PhD

All:
School of Applied Science, South Bank University,
London, UK

Industrial Microbiology: An Introduction

Industrial Microbiology:
An Introduction

Michael J. Waites

BSc, PhD, CBiol, MIBiol
Neil L. Morgan

BSc, PhD, MIFST
John S. Rockey

BSc, MSc, PhD
Gary Higton

BSc, PhD

All:
School of Applied Science, South Bank University,
London, UK

© 2001 by
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A catalogue record for this title
is available from the British Library

ISBN 0-632-05307-0

Library of Congress
Cataloging-in-Publication Data

Introduction to Industrial Microbiology/

Michael J. Waites . . . [et al.].

p.
cm.

ISBN 0-632-05307-0
(pbk.)

1. Industrial Microbiology: an Introduction.
I. Waites, Michael J.

QR53 .I522 2001
660.6¢2—dc21
2001025874

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Dedication

This book is dedicated to the memory of our great friend and co-author Gary Higton
who, at the age of only 40 years, died unexpectedly during the final stages of its prepara-
tion. Gary was a knowledgeable microbiologist, a fine teacher, supportive colleague and
a loyal friend. He is greatly missed by us all.

Contents

Preface, ix
Acknowledgements, xi
Introduction to industrial microbiology, 1

Part 1
Microbial physiology

1
Microbial cell structure and function, 7

2
Microbial growth and nutrition, 21

3
Microbial metabolism, 46

Part 2
Bioprocessing

4
Industrial microorganisms, 75

5
Fermentation media, 86

6
Fermentation systems, 94

7
Downstream processing, 109

8
Product development, regulation and safety, 124

Part 3
Industrial processes and products

9
Microbial enzymes, 133

10
Fuels and industrial chemicals, 144

11
Health care products, 165

12
Food and beverage fermentations, 179

13
Food additives and supplements, 210

14
Microbial biomass production, 218

15
Environmental biotechnology, 229

16
Microbial biodeterioration of materials and its control, 247

17
Animal and plant cell culture, 258

Index, 265

vii

Preface

isms, their ability to grow on a wide range of substrates
and to produce an extensive array of products, many of
which are commercially available.

Part 2 is devoted to bioprocessing. It examines the

commercial fermentation operations and requirements
for large-scale cultivation of microorganisms. This
involves the acquisition and development of suitable
production strains that must then be provided with
nutrients, especially appropriate carbon and energy
sources. The object of any industrial fermentation is
then to optimize either growth of the organism or
the production of a target microbial product. This is
normally achieved by performing fermentations under
rigorously controlled conditions in large fermenters
with culture capacities often in excess of several
thousand litres. The design and operation of these
fermentation processes is discussed, along with the
downstream processing operations necessary for the re-
covery and purification of the target products. Aspects
of safety and good manufacturing practices are also
examined.

Over the last twenty years, many traditional and es-

tablished industrial fermentation processes have been
advanced through the contribution of genetic engineer-
ing (in vitro recombinant DNA technology). This tech-
nology has also facilitated the development of many
novel processes and products. It not only accelerates
strain development, leading to improvements in the
production microorganisms, but can aid downstream
processing and other elements of the process. Initially, it
involved the manipulation of bacteria, but has moved
to cloning in yeasts, filamentous fungi, and plant and
animal cells. Developments in this field continue to
grow rapidly.

There are several good texts that cover the fundamen-

tal aspects of genetic manipulation of microorganisms.
Consequently, we have not attempted to give a detailed
account of such techniques here, they are merely intro-
duced and further reading is suggested. Nevertheless,
many examples of the application and potential of

Industrial microbiology is primarily associated with
the commercial exploitation of microorganisms, and
involves processes and products that are of major
economic, environmental and social importance
throughout the world. There are two key aspects of in-
dustrial microbiology, the first relating to production of
valuable microbial products via fermentation processes.
These include traditional fermented foods and bever-
ages, such as bread, beer, cheese and wine, which have
been produced for thousands of years. In addition, over
the last hundred years or so, microorganisms have been
further employed in the production of numerous chemi-
cal feedstocks, energy sources, enzymes, food ingredi-
ents and pharmaceuticals. The second aspect is the role
of microorganisms in providing services, particularly
for waste treatment and pollution control, which uti-
lizes their abilities to degrade virtually all natural and
man-made products. However, such activities must be
controlled while these materials are in use, otherwise
consequent biodeterioration leads to major economic
losses.

This textbook is intended to be an introduction to in-

dustrial microbiology. In writing it, the authors have
drawn on their experience teaching industrial micro-
biology and other aspects of applied microbiology
to undergraduates and masters students on a range of
applied biology, microbiology, biotechnology, food sci-
ence and chemical engineering courses. It is assumed
that the reader will have an elementary knowledge of
microbiology and biochemistry. Nevertheless, even
those students with only a basic knowledge of chemistry
and cell biology, and those who are not specialist micro-
biologists, should find the material accessible.

The book is divided into three sections. Part 1 is

designed to underpin the main content. Its aim is to
provide a sufficient, albeit brief, overview of microbial
structure, physiology and biochemistry to enable the
student to fully appreciate the diversity of microorgan-
isms and their metabolic capabilities. The reader should
soon come to recognize the versatility of microorgan-

ix

genetically engineered microorganisms within industri-
al processes are discussed.

Part 3 explores the wide range of industrial microbial

processes and products, including human food and
animal feed production, the provision of chemical
feedstocks, alternative energy sources, enzymes and
products for application in human and animal health.
We consider that the economic and scientific impor-
tance of traditional and long-established fermentation
processes can be somewhat overlooked, as attention is
often dominated by more recent developments. There-
fore we have aimed to give a balanced and comprehen-
sive coverage of current industrial processes and their

products. The production of valuable commodities
from animal and plant cell culture is also included,
primarily because the culture and handling of such
cells involves techniques similar to those used for the
propagation of microorganisms.

An additional aspect of industrial microbiology, ex-

amined here, is the application of extensive degradative
abilities of microorganisms, particularly the harnessing
of these properties in waste treatment and pollution
control. The necessity to limit these activities in inap-
propriate situations, i.e. the prevention of biodeteriora-
tion of materials while still in use, is also discussed.

x
Preface

Acknowledgements

We would like to thank the following authors and the publishers for allowing us to use figures
and tables from their publications:
Figs 1.1 and 3.12 are from Dawes, I. W. & Sutherland, I. W. (1992) Microbial Physiology,
2nd Edition. Blackwell Scientific Publications.
Fig. 1.2 is from Poxton, I. R. (1993) Journal of Applied Bacteriology Supplement 74, 1S–11S.
Figs 4.3, 7.8 and 7.9 are from Brown, C. M., Campbell, I. & Priest, F. G. (1987) Introduction
to Biotechnology. Blackwell Scientific Publications.
Fig. 12.7 is from Lewis, M. & Young, T. W. (1995) Brewing. Chapman & Hall.
Tables 12.3 and 12.4 are based on tables in Bamforth, C. W. (1985) The use of enzymes in
brewing. Brewers’ Guardian 114, 21–26.
In addition, we wish to thank our colleague Dr Tom Coultate for his advice and enthusiastic
support.

xi

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 fermenta-
tion in beer and wine production was the 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 contribu-
tion to food and beverage preservation, which was orig-
inally 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 Brewery 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 Saccha-
romyces 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 advance-
ments 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 Weiz-

mann (1913–15) using the bacterium Clostridium ace-
tobutylicum. In the early 1920s an industrial fermenta-
tion 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
fermentations, but it led to great advances in both media
and microbial strain development. The knowledge ac-
quired 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, thera-
peutic and purification purposes, pioneered by Milstein
and Kohler in the early 1970s. However, many of the
greatest advances have followed the massive develop-
ments in genetic engineering (recombinant DNA tech-
nology) over the last 20 years. This technology has had,
and will continue to have, a tremendous influence on
traditional, established and novel fermentation process-
es and products. It allows genes to be transferred from
one organism to another and allows new approaches to
strain improvement. The basis of gene transfer is the in-
sertion of a specific gene sequence from a donor organ-
ism, via an expression vector, into a suitable host. Hosts
for expression vectors can be prokaryotes such as
the bacterium Escherichia coli; alternatively, where
post-translational processing is required, as with some
human proteins, a eukaryotic host is usually required,
e.g. a yeast.

A vast range of important products, many of which

were formerly manufactured by chemical processes, are
now most economically produced by microbial fermen-
tation and biotransformation processes. Microorgan-
isms also provide valuable services. They have proved to
be particularly useful because of the ease of their mass
cultivation, speed of growth, use of cheap substrates

Introduction to industrial microbiology

1