CHAPTER 1

Introduction

1.1 Biotechnology: A Definition and Overview

In a general sense, biotechnology is a multi- and interdisciplinary field of
activities dealing with biological and biochemical processes carried out on
a broad range of scale (10 I to 106 m 3 ) as a technique for production.
Biotechnology has been defined differently in different countries and con-
tinents. Based on a definition by the European Federation of Biotechnology
(EFB), a broader sense is discussed in Europe recently:
"Biotechnology is the integrated use of natural sciences (biology inc!. molecular biology,
biochemistry, but also chemistry and physics) and engineering sciences (chemical
reaction engineering, electronics) in order to achieve the application of (technological,
industrial) organisms, cells (microbes, plant and animal cells), parts thereof and molec-
ular analogues in order to provide biosociety desirable goods and services"
Thus, the main characteristics of biotechnology are integration, orientation
towards applications but also a considerably impact on human society in the
future ("Biosociety", FAST, 1980; Moser, 1988).
Significant fields of applications are
Industrial Biotechnology
Food- and Agricultural (Phyto-, Zoo-) Biotechnology
Healthcare (Pharma-) Biotechnology and
Environmental Biotechnology.
This definition excludes medical engineering, which is concerned with the
construction of instruments for biological or health care example,
heart-lung machines. Biomedical engineering is also commonly referred to as
biotechnology and/or bioengineering, and no clear-cut distinction is apparent
from the name alone. Further, technological activities in the field of agricul-
ture, for example, traditional breeding (phytotechnology and zootechnology),
belong to biotechnology only in the broadest sense.
One systematic treatment of biotechnology results from a logical considera-
tion of the catalysts responsible for the conversions in biochemical reactions
2 1. Introduction

and other biologically active agents. In this sense, biocatalysts are not only
naturally or synthetically produced enzymes and biomolecules respectively
analogues but also cells or subcellular fractions from microorganisms and
from plant and animal tissues. The type of catalyst used to carry out a bio-
logical process delineates the individual technology: enzyme or (simple and
complex) fermentation technology or tissue culture with suspended or im-
mobilized catalysts. In addition to the production techniques, there is also a
group of sterilization technique used to inactivate or kill biological materials.
For historical reasons, precise separation of biotechnology from other fields
is often difficult. There is overlap among various areas, such as conventional
food technology and biotechnology (Fig. 1.1). The production of yogurt or

TECHNICAL
B lOP ROC E SSE S

--------
(BIOTECHNOLOGY and FOOD TECHNOLOGY)

Production .
Destructlon
of of
biological material biological material

J
'\ "'"
STERILIZATION-
TECHNOLOGY
TECHNOLOGY
FOOD BEVERAGE WASTE WATER-,WASTE- TISSUES-
PROCESSING INDUSTRY TECHNOLOGY CULTURE
e.g.yoghurt e.g. animal,plant
beer,wine cell s
t
t
, I •
I
t
I I
I

,I t + +
MICROBIAL REACTORS ENZYME
t REACTORS STERILIZERS

VAT
BARREL "jEHTER I

f
B lOR E C TOR S

H I 0 LOG I CAL REA C TOR S

FIGURE 1.1. Biological reactors and their areas of application for "taming" many
industrial-scale bioprocesses. (After Moser, 1978, 1981.)
 1.1 Biotechnology: A Definition and Overview 3

cheese, for example, would be considered as bioprocesses of food technology.

The high degree of complexity of bioprocessing in food technology and
the difficulty of obtaining quantitative measurements (a difficulty shared by
many other divisions of fermentation technology) usually retard or inhibit
a systematic approach. In addition, special expectations exist for the quality
of the foodstuff resulting from bioprocessing and the techniques of large-scale
production are only slowly being introduced. Despite the fundamentally
different orientation of food technology, however, progress is being made in
introducing scientific methods. Because of commercial considerations, sooner
or later the working principles of biotechnology will be applied in complex
processes.
Precisely the same fate can be seen in neighboring area, the biological
treatment of waste water and solid wastes (Fig. 1.1). The pressure of environ-
mental problems has grown so great that here too the methods of thinking
that have been successfully used in conventional biotechnical processes are
being applied. One can now speak of the biotechnology of waste water
treatment.
Figure 1.1 shows the correspondence between the various technologies of
bioprocessing and the containers in which the reactions are carried out on
an engineering scale. These "bioreactors" range from sterilizers to proper
bioreactors (microbiological reactors such as fermenters, waste water treat-
ment units, vessels for cultivation of plant & animal cells, enzyme reactors) to
miscellaneous containers such as bottles, barrels, and so on used primarily for
carrying out "arts and crafts"-type complex biological processes.
What are the future prospects of biotechnology? The range of products
is being extended from antibiotics, steroids, vitamins, diagnostics, vaccines,
enzymes, and polysaccharides to such fine chemicals as organic acids, alcohols,
and single-cell protein (Rehm and Reed, 1982ff). Figure 1.2 shows the course
of development of prices in relation to the scale of production facilities for
the period 1970 to 2000 (Hines, 1980). From this figure one can conclude that,
at present, biotechnical methods are more economical than chemical ones
only in the case of pharmaceuticals that are complex molecules. The prediction
is that soon single-cell proteins can be economically produced by biotechnical
methods, and that by 1995 even inexpensive chemicals may be produced
in the same way. Economic feasibility in processes for producing simple
materials is first reached only in large-scale production units. At present,
small-scale, high-price processes are being carried out, mainly on a dis-
continuous basis. For the future, one looks toward large-scale, low-price
processes carried out on a continuous basis. Additional facts concerning
economics are discussed by Hamer (1985).
Some general conclusions can now be drawn from experience in fermentation,
food, and waste water treatment engineering:
1. The development of processes from the laboratory stage to the stage of tech-
nical maturity where they can be of service to mankind must be accelerated
and made more reliable. Three stages are involved in "taming" a bioprocess.
4 1. Introduction

PInJI{SS
IN

SElllrfJ
PRICE
t
BlnTEDffiIlXiY

t PHARMACEUT ICALS
1970

........,...

-... - ...... --.:..-

:"'-:;:.:-;, ;.--"".-=r
1980
SIMPLE CHEMICALS t 1990
b --,---

- PROIXJCTION SCALE

FIGURE 1.2_ Predicted development of biotechnical processes (---) compared with

development of chemical processes (--) for three classes of products (pharmaceuticals,
single-cell protein, simple chemicals) as examples of products with different molecular
complexity. The date (t) shows when it is predicted that the bioprocess will be more
economical than the chemical process. Region a: small-scale, high-price processes.
Region b: large-scale, low-price processes. (After Hines, 1980. With permission of
Butterworth scientific Ltd.)

The first is that of nature itself. In the earth, -in organisms, and in water,
biological processes operate on their own. They are often first discovered when
human intervention disturbs them. This stage was, and is, the school in which
human beings learn through trial and error to reproduce the processes and
use them for their own ends. The protocols for doing this are strictly observed
and are often kept secret. This was, and is, the stage of the artist and crafts-
person in, for example, the making of beer, wine, and sauerkraut. In part, it
is still the foundation upon which modern technology builds to create complex
products. Modern technology goes beyond this foundation to ensure repro-
ducibility of products through quantitative methods. In the field of chemical
engineering, such reproducibility was accomplished several decades ago.
2. Developing completely new methods of production demands a new organi-
zation of scientific and technical work. In recent years the problems of raw
material supplies, energy, and the environment have posed new questions.
With processes based on biotechnology, previously unused or underused
natural sources of energy and raw materials can be used or used more
intensively, and disturbed ecological cycles can be stabilized (for example,
CO2 , sunlight, cellulose, waste materials, and the cycles of the self-cleaning
rivers).
Industrial use of biotechnology requires new technical knowledge, new
 1.2 Bioprocess Technology 5

working methods, and valid principles of operation. The new processes extend
well beyond the framework of traditional technology in both complexity and
scale. Neither simple fermentation technology nor waste water treatment
technology, nor the technologies of food processing, pharmaceuticals, or water
purification, provide a model adequate to serve as the basis for organization
of the new types of production. Traditional technologies developed largely in
isolation; they were product oriented. The current existence within each field
of independent terms describing technical measures and quantities common
to them all is a sign of this product orientation.
To foster the rapid and reliable development of processes based on bio-
technology, and especially to foster new production methods applicable on a
usable scale, a more unified view is needed. The many similar biotechnical
methods used by all of the scientific disciplines represented and by all of the
industries involved must be placed on a more universal basis, both from neces-
sity and from consideration of the advantages of universality. The product-
oriented way of thinking should be woven into a "process-oriented" viewpoint
that unites processes having similar working principles (Dechema, 1974; A.
Moser, 1977; Ringpfeil, 1977). In the chemical industry, such process orienta-
tion led to the development of chemical engineering. Figure 1.1 shows the
process-oriented way of thinking with regard to the food, beverage, and waste
water treatment industries. The different bioreactors must be reduced to a few
basic types. Finally, general working principles are needed. It is necessary and
advantageous to universalize concepts, definitions, and nomenclature.

1.2 Bioprocess Technology

The circumstances just described are the starting point of this book and the
initiative for the book's objectives. The term "bioprocess technology," created
by the author in 1972 includes the reaction engineering of biochemical pro-
cesses (Reuss, 1977). The term is now increasingly accepted for the integrated
disciplines of engineering sciences in biotechnology (i.e., kinetics, reactors,
measurement and control, upstream and downstream processing). Thus, it
includes the general working principles for introducing, executing, and opti-
mizing biological processes on a technical scale. The term "biotechnology"
fulfills its comprehensive role only when it is understood to mean the scientific
discipline of general applicability associated with engineering-scale biopro-
cesses made reproducible through the utilization of systematic, quantitative,
and system-analytical methods. The inter and trans-disciplinary character of
biotechnology that is reflected in the hybrid name is also found in other areas
of engineering. No doubt, the purely technical component of reactors and their
layout is fundamentally the same as in chemical engineering. The special
feature of biotechnology lies in the nature of the biological processes, in the
interaction of biochemistry and biology with technical capabilities. This book
reflects this special feature in that it discusses less the problems of reactors
6 1. Introduction

and more the problems involved in integrated quantification of bioprocesses.

The emphasis here, then, will be on bioprocess kinetics, because kinetics
represents, so to speak, the heart of bioprocess technology including inter-
actions with physics.
A further limitation on the contents of this book is the interdisciplinary
structure of biotechnology. Complete coverage of all of the component areas
of biotechnology exceeds the capabilities of one author. Such a book would
also duplicate information already presented in readily available textbooks.
Among these are
- Aiba, S., Humphrey, A., and Millis, N.F. (1973,1976). Biochemical Engineer-
ing. New York, Academic Press.
- Atkinson, B. (1974). Biochemical Reactors. London: Pion, Ltd.
- Atkinson, B., and Mavituna, F. (1983). Biochemical Engineering and Bio-
technology Handbook. New York: Nature Press; London: MacMillan.
- Bailey, 1.E., and Ollis, D.F. (1977). Biochemical Engineering Fundamentals.
New York: McGraw-Hill
- Bergter, F. (1983). Wachstum, von und
Modelle. lena: G. Fischer.
- Biryukov, V.V. (1985). Optimization of Periodic Processes for Microbial
Synthesis. Moscow. (in russ.)
- Kafarow, W.W., Winarow, A.l., and Gordiejew, L.S. (1979). Lesnaja
Promyshlenost (Modeling of Biochemical Reactors). Moscow. (in russ.)
- Malek, I., and Fencl, Z. (1966). Theoretical and Methodological Basis of
Continuous Culture of Microorganisms. New York: Academic Press.
- Moo-Young, M. (ed.-in-chief) (1985). Comprehensive Biotechnology, Vol.
1-4. Oxford: Pergamon Press.
- Moser, A. (1981) BioprozeBtechnik, Springer-Verlag, Vienna, New York.
- Pirt, S.1. (1975). Principles of Microbe and Cell Cultivation. Oxford: Blackwell
Scientific.
- Prave, P., Faust, U., Sittig, W., and Sukatsch, D.A. (1982). Handbuch der
Biotechnologie. Wiesbaden: Akademische Verlagsgesellschaft.
- Rehm, H.l., and Reed, G. (eds.) (1982ff). Comprehensive
Treatise, Vol. 1-8, Deerfield Beach, Fl. and Basel: Verlag Chemie Weinheim.
- Schiigerl, K. (1985). Bioreaktionstechnik. Vol. 1. Aarau Salle u Sauerlander.
Figure 1.3 schematically illustrates the procedures and different fields that
contribute to process development from concept to technological maturity.
The important component parts (in circles in the figure) are
1. The biochemical-biological basis for the preparation ofthe catalyst (isolation,
culture of pure cell lines, maintenance of strains, enzyme preparation), all
the problems of raw materials (the composition of the culture media), and all
the problems of analysis (chemical analysis; biochemical, microbiological,
and physical methods).
2. The stage of "upstream" processing dealing with the sterilization of raw
materials, with cell culture, and with equipment and instrumentation for
 1.2 Bioprocess Technology 7

/
/

DESIGN

FIGURE 1.3. Areas of biotechnology potentially contributing to the development of

a process. (From Moser, 1977.)

process control including techniques for carrying out the process on an

experimental scale (measurement and control).
3. Bioprocess technology and its working principles used for planning, eval-
uating, and calculating (modeling) bioprocesses (kinetics and reactors).
4. "Downstream" processing, which includes methods for the separation of
cells, cell rupture, product isolation, and purification.
5. Costs and other economic calculations.
6. Genetics, used for the development of biocatalysts with new characteristics
through mutation, selection, and adaptation.
Understanding of the extent of the problems involved in biotechnology
can best be reached through examining the professionalliter'ature, found in
various journals and review series. The most important of these are
Journals
Acta Biotechnologica
Applied Microbiology and Biotechnology
Biochemical Engineering Journal
Bioengineering News
Biofuture
Bioprocess Engineering
Biosensors
Biotech News
Biotechnologie in Nederland
8 1. Introduction

Bio/Technology
Biotechnology Abstracts (Derwent)
Biotechnology and Applied Biochemistry
Biotechnology and Bioengineering
Biotechnology Laboratory
Biotechnology Letters
Biotechnology News
Biotechnology Newswatch
Biotechnology Progress
Chemical and Biochemical Engineering Quarterly
Chemical Engineering Journal
Chemie-Ingenieur-Technik
Current Biotechnology Abstracts
Enzyme and Microbial Technology
European Journal of Applied Microbiology and Biotechnology
Forum Biotechnologie
Journal of Applied Chemistry and Biotechnology
Journal of Biotechnology
Journal of Chemical Technology and Biotechnology
Journal of Fermentation Technology
Journal of Industrial Microbiology
Pascal Biotechnology-Bulletin Signaletique
Practical Biotechnology
Process Biochemistry
Swiss Biotech
Trends in Biotechnology
Review Series
Advances in Biochemical Engineering
Advances in Biotechnological Processes
Annual Reports on Fermentation Processes
Biotechnology Advances
Biotechnology and Bioengineering Symposia
Developments in Industrial Microbiology
Methods in Microbiology
Progress in Industrial Microbiology
The process orientation ofthis book is illustrated by a comparison between
chemical processes and bioprocesses such as simple fermentation or waste
water treatment as a case of complex fermentation process from a process
engineering viewpoint (Table 1.1). The advantages and disadvantages of the
two processes are based on the characteristics of the catalysts. In comparison
with the usual chemical catalysts, enzyme catalysts are both highly active and
highly selective. In the last decades, even more active chemical catalysts have
been developed; however, this has been costly. Since isolation and purification
of enzymes also have costs, enzymes in pure form are also not inexpensive.
 1.2 Bioprocess Technology 9

TABLE 1.1. Comparison ofbioprocessing and chemical processing.

Criterion of
comparison Chemical process
Catalyst More or less active and selective
Expensive if selective
Expensive regeneration
Reaction conditions Mostly high temperature and
high pressure
Raw materials Pure if poorly selective catalyst
Process Often multistage processing with
recovery of intermediates,
resulting in low yields
Environmental crisis
Fast reaction rates at high
concentration level and high
yields
Bioprocess
Enzymes: highly active and
selective
Regeneration easy due to
microbial growth
"Construction" of new catalysts
with genetic engineering
Mainly - 25°C and - 1 bar
pressure
Biological demands of nutrients,
biological optimum (cf. Fig. 5.1),
maintenance, mutation
Impure, inactive, and diluted
("unconventional" raw
materials: cellulose, starch, oil)
One-stage possible without
intermediary product recovery
(e.g., steroid transformation)
Potentially better for environment
Mainly slow reaction rates
Low concentrations
High demand in sophisticated
apparatus/equipment,
supplementary education
"Demon of nature" (biological
material, infections, mutations)

Today, use of enzymes in the whole cell as the catalysts is preferred; in general,
fermentation technology is still the least costly option. In addition, catalyst
regeneration is easily attained through cell growth and division.
All of the facts noted in Table 1.1 result from the nature of the catalyst.
A quantitative treatment of the kinetics of biochemical-biological systems,
given hereinafter, will mirror these process engineering characteristics. Thus,
biological process kinetics will have fundamental formulas for processes of
growth and product accumulation which operate with a typically biological
optimum in a relatively narrow range of concentrations, pH and temperature.
The formulas must, of course, also take into account the metabolism, the
effects of the interaction between organism and environment, and the nutri-
tional requirements of the cell.
The process-engineering comparison between simple fermentation and a
complex bioprocess such as that used for waste water treatment, shown in
detail in Table 1.2, brings out the problems involved in quantifying practices
used in complex cases: multiple substrate kinetics operating in either sequential
or parallel form; mixed populations; dependence on pH and 'temperature; the
influence of homogeneous or heterogeneous reactor operation in discontin-
10 1. Introduction

TABLE 1.2. Process engineering comparison between simple fermentation and

complex fermentation bioprocess (e.g., biological waste water treatment).
Complex
Criterion of Simple fermentation fermentation bioprocessing
comparison (lab scale) (technical scale)
Catalyst (population) Mainly pure cultures Mixed population "biocoenosis"
Raw materials Synthetic media with S Complex media with unknown
limitations known and even toxic components
(optimized media)
Analytical methods Expensive, modern (e.g., Cheap and simple, often global
enzymatic analyses, (e.g., chemical, biological oxy-
research oriented) gen demand); need for stability
(long time) and reproducibility
Temperature, pH Constant Gradients
Reactors, mode of Mainly homogeneous type Heterogeneous types dominate
operation (e.g., stirred tanks) in batch or (gradients in homogeneous
continuous mode of operation reactors, transport limitation
Pseudo homogeneous approach in three-phase systems, biofilm
preferred reactors)
Batch, semicontinuous operation
Inflow Constant Changes
Kinetics Idealized conditions: I-S Multi-S limitation
limitation Microbial interactions (multi-X)
Transient operation (dynamics)
Interactions between biology and
physics
Structured approach needed
(for biological cell as well as
for reactors)

From A. Moser, 1981.

uous, semidiscontinuous, or fully continuous operations; unstable behavior

with respect to changes in the quantity or quality of input materials, and
special effects such as multiphase operation (heterogeneities) and the problems
of applying global methods of measurement that reflect only overall kinetics.
Identification of the separate tasks in the area of bioprocess technology
clearly shows that in every case the reaction is closely coupled to the reactor.
The quantification of processes involves the kinetics, the transport phenomena,
and the interactions between them. This fact influences the whole way of
thinking and the whole mode of working. Figure 1.4 illustrates a general
strategy for bioprocess kinetic analysis on the basis of this mode of thinking-
an "integration strategy" (A. Moser, 1982)-which will be outlined in more
detail in Sect. 2.3.1. At the same time, the scheme shown in Fig. 1.4 can serve
as a guide to this book: The sequential steps represent the concepts that will
be developed in the following chapters.
The strategy followed in the book is in agreement with the theory of
deduction (Popper, 1972, 1976) which recently was stressed again for economic-
 1.2 Bioprocess Technology 11

FIGURE 1.4. Flow chart sheet of the strategy of bioprocess kinetic analysis for different
process situations: stationary/instationary, homogeneous/heterogeneous, differential!
integral, and true dynamic/balanced (frozen) reactor operation. rds, rate-determining
step; qss, quasi-steady-state, (From Moser, A. 1983.)
12 1. Introduction

ecological evaluation (F. Moser, 1981, 1983). This scientific strategy is most
helpful to analyze and synthetize the complex network of integrated phenom-
ena of biology and physics. As integration means more than summing-up, the
success of engineering design work depends on the elucidation of interactions.

BIBLIOGRAPHY
Dechema (1974). Biotechnologie, Studie uber Forschung und Entwicklung. Bonn:
Bundesministerium fiir Forschung und Technologie.
FAST (1980) Sub-programme Bio-society, Research activities Commission of the
European Communities EUR 7105. Brussels (M. Cantley)
Hamer, G. (1985). Trends Biotechnol., 3, 73.
Hines, D.A. (1980). Enzyme Microb. Technol., 2, 327-329.
Moser, A. (1977). Habilitationsschrift, Technical University, Graz, Austria.
Moser, A. (1978). Erniihrung/Nutrit., 2, 505.
Moser, A. (1981). Bioprozesstechnik. Vienna and New York: Springer-Verlag.
Moser, A. (1982). Biotechnol. Lett., 4, 73.
Moser, F. (1981). Paper presented at 2nd World Conference of Chemical Engineering,
4-9 Oct., Montreal.
Moser, A. (1988). Trends in Biotechnology, Vol. 6, No.8, August.
Moser, F. (1983). Chern. Eng. (Lond), August/September, 13.
Popper, K.R. (1972). The Logic of Scientific Discovery. London: Hutchinson.
Popper, K.R. (1976). Die Logik der Forschung. 6th ed. Mohr Studienausgabe.
Rehm, H.J., and Reed, G. (eds.) (1982ff). Biotechnology-A Comprehensive Treatise.
Deerfield Beach, Fla. and Basel Vol. 1-8 Verlag Chemie Weinheim.
Reuss, M. (1977). Fort. Verfahrenstechnik, 15F, 549-566.
Ringpfeil, M. (1977). Chern. Tech., 29, 424-428.
Moo-Young, M. (ed.-in-chief) (1985). Comprehensive Biotechnology, Vol. 1-4, Oxford:
Pergamon Press.