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By Peter F. The successful structure of the previous edition of Principles of Fermentation Technology has been retained in this third edition, which covers the key component parts of a fermentation process including growth kinetics, strain isolation and improvement, inocula development, fermentation media, fermenter design and operation, product recovery, and the environmental impact of processes.
This accurate and accessible third edition recognizes the increased importance of animal cell culture, the impact of the post-genomics era on applied science and the huge contribution that heterologous protein production now makes to the success of the pharmaceutical industry.
This title is ideally suited for both newcomers to the industry and established workers as it provides essential and fundamental information on fermentation in a methodical, logical fashion. Stanbury, Whitaker and Hall have integrated the biological and engineering aspects of fermentation to make the content accessible to members of both disciplines with a focus on the practical application of theory. This text collates all the fermentation fundamentals into one concise reference, making it a valuable resource for fermentation scientists, as well as those studying in the field.
No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. This book and the individual contributions contained in it are protected under copyright by the Publisher other than as may be noted herein. Knowledge and best practice in this field are constantly changing.
As new research and experience broaden our understanding, changes in research methods, professional practices, or medical treatment may become necessary. Practitioners and researchers must always rely on their own experience and knowledge in evaluating and using any information, methods, compounds, or experiments described herein.
In using such information or methods they should be mindful of their own safety and the safety of others, including parties for whom they have a professional responsibility. This book is dedicated to all the staff, past and present, of the Department of Biological and Environmental Sciences, University of Hertfordshire.
We wish to thank the authors, publishers, and manufacturing companies listed below for allowing us to reproduce either original or copyright material:. Abe Fig. Nienow Figs. Richards Figs. Shinskey Fig. Talcott Figs. American Society for Testing and Materials: Fig. Copyright ASTM, reprinted with permission. Chilton Book Company Ltd. Elsevier: Figs. Tables 2. Kluwer Academic Publications: Fig. John Wiley and Sons: Figs. Last but not least, we wish to express our thanks to Lesley Stanbury and Lorna Whitaker for their support, encouragement, and patience during the preparation of both this, and previous editions of " Principles of Fermentation Technology.
This chapter introduces the reader to the fermentation industry and lays the foundation for the rest of the book. A typical fermentation is then described in terms of its component parts, or unit operations, enabling the reader to place the content of the subsequent chapters in the context of the whole process.
Finally, the rationale adopted by the authors is explained and the reader is guided through the contents of the book. The term fermentation is derived from the Latin verb fervere, to boil, thus describing the appearance of the action of yeast on the extracts of fruit or malted grain. However, fermentation has come to have with different meanings to biochemists and to industrial microbiologists.
Its biochemical meaning relates to the generation of energy by the catabolism of organic compounds, whereas its meaning in industrial microbiology tends to be much broader. The catabolism of sugar is an oxidative process, which results in the production of reduced pyridine nucleotides, which must be reoxidized for the process to continue.
Under aerobic conditions, reoxidation of reduced pyridine nucleotide occurs by electron transfer, via the cytochrome system, with oxygen acting as the terminal electron acceptor. However, under anaerobic condition, reduced pyridine nucleotide oxidation is coupled with the reduction of an organic compound, which is often a subsequent product of the catabolic pathway.
In the case of the action of yeast on fruit or grain extracts, NADH is regenerated by the reduction of pyruvic acid to ethanol. Different microbial taxa are capable of reducing pyruvate to a wide range of end products, as illustrated in Fig. Thus, the term fermentation has been used in a strict biochemical sense to mean an energy-generation process in which organic compounds act as both electron donors and terminal electron acceptors.
Pyruvate formed by the catabolism of glucose is further metabolized by pathways which are characteristic of particular organisms and which serve as a biochemical aid to identification.
The production of ethanol by the action of yeast on malt or fruit extracts has been carried out on a large scale for many years and was the first industrial process for the production of a microbial metabolite. Thus, industrial microbiologists have extended the term fermentation to describe any process for the production of product by the mass culture of a microorganism. Brewing and the production of organic solvents may be described as fermentation in both senses of the word but the description of an aerobic process as a fermentation is obviously using the term in the broader, microbiological, context and it is in this sense that the term is used in this book.
The historical development of these processes will be considered in a later section of this chapter, but it is first necessary to include a brief description of the five groups. The commercial production of microbial biomass may be divided into two major processes: the production of yeast to be used in the baking industry and the production of microbial cells to be used as human food or animal feed single-cell protein.
However, it was not until the s that the production of microbial biomass as a source of food protein was explored to any great depth. However, the demise of the animal feed biomass fermentation was balanced by ICI plc and Rank Hovis McDougal establishing a process for the production of fungal biomass for human food.
Enzymes have been produced commercially from plant, animal, and microbial sources. However, microbial enzymes have the enormous advantage of being able to be produced in large quantities by established fermentation techniques.
Also, it is infinitely easier to improve the productivity of a microbial system compared with a plant or an animal one. Enzyme production is closely controlled in microorganisms and in order to improve productivity these controls may have to be exploited or modified. Also, the number of gene copies coding for the enzyme may be increased by recombinant DNA techniques.
After the inoculation of a culture into a nutrient medium there is a period during which growth does not appear to occur; this period is referred as the lag phase and may be considered as a time of adaptation. Following a period during which the growth rate of the cells gradually increases, the cells grow at a constant maximum rate and this period is known as the log, or exponential, phase. Eventually, growth ceases and the cells enter the so-called stationary phase. After a further period of time, the viable cell number declines as the culture enters the death phase.
As well as this kinetic description of growth, the behavior of a culture may also be described according to the products that it produces during the various stages of the growth curve. During the log phase of growth, the products produced are either anabolites products of biosynthesis essential to the growth of the organism and include amino acids, nucleotides, proteins, nucleic acids, lipids, carbohydrates, etc. The synthesis of anabolic primary metabolites by wild-type microorganisms is such that their production is sufficient to meet the requirements of the organism.
Thus, it is the task of the industrial microbiologist to modify the wild-type organism and to provide cultural conditions to improve the productivity of these compounds. This has been achieved very successfully, over many years, by the selection of induced mutants, the use of recombinant DNA technology, and the control of the process environment of the producing organism.
However, despite these spectacular achievements, microbial processes have only been able to compete with the chemical industry for the production of relatively complex and high value compounds.
In recent years, this situation has begun to change. The advances in metabolic engineering arising from genomics, proteomics, and metabolomics have provided new powerful techniques to further understand the physiology of over-production and to reengineer microorganisms to over-produce end products and intermediates of primary metabolism.
During the deceleration and stationary phases, some microbial cultures synthesize compounds which are not produced during the trophophase and which do not appear to have any obvious function in cell metabolism.
It is important to realize that secondary metabolism may occur in continuous cultures at low growth rates and is a property of slow-growing, as well as nongrowing cells. When it is appreciated that microorganisms grow at relatively low growth rates in their natural environments, it is tempting to suggest that it is the idiophase state that prevails in nature rather than the trophophase, which may be more of a property of microorganisms in culture.
The interrelationships between primary and secondary metabolism are illustrated in Fig. Although the primary biosynthetic routes illustrated in Fig. Thus, Fig.
Also, not all microorganisms undergo secondary metabolism—it is common amongst microorganisms that differentiate such as the filamentous bacteria and fungi and the sporing bacteria but it is not found, for example, in the Enterobacteriaceae.
Thus, the taxonomic distribution of secondary metabolism is quite different from that of primary metabolism. It is important to appreciate that the classification of microbial products into primary and secondary metabolites is a convenient, but in some cases, artificial system. To quote Bushell , the classification should not be allowed to act as a conceptual straitjacket, forcing the reader to consider all products as either primary or secondary metabolites.
It is sometimes difficult to categorize a product as primary or secondary and the kinetics of synthesis of certain compounds may change depending on the cultural conditions. Figure 1. The physiological role of secondary metabolism in the producer organism in its natural environment has been the subject of considerable debate and their functions include effecting differentiation, inhibiting competitors, and modulating host physiology.
However, the importance of these metabolites to the fermentation industry is the effects they have on organisms other than those that produce them. Thus, the products of secondary metabolism have formed the basis of a major section of the fermentation industry. The advent of recombinant DNA technology has extended the range of potential fermentation products.
Genes from higher organisms may be introduced into microbial cells such that the recipients are capable of synthesizing foreign proteins.
These proteins are described as heterologous meaning derived from a different organism. A wide range of microbial cells has been used as hosts for such systems including Escherichia coli, Saccharomyces cerevisiae, and filamentous fungi. Animal cells cultured in fermentation systems are also widely used for the production of heterologous proteins.
Although the animal cell processes were based on microbial fermentation technology, a number of novel problems had to be solved—animal cells were considered extremely fragile compared with microbial cells, the achievable cell density is very much less than in a microbial process and the media are very complex. Products produced by such genetically engineered organisms include interferon, insulin, human serum albumin, factors VIII and IX, epidermal growth factor, calf chymosin, and bovine somatostatin.
Important factors in the design of these processes include the secretion of the product, minimization of the degradation of the product, and control of the onset of synthesis during the fermentation, as well as maximizing the expression of the foreign gene. Microbial cells may be used to convert a compound into a structurally related, financially more valuable, compound.
Because microorganisms can behave as chiral catalysts with high positional specificity and stereospecificity, microbial processes are more specific than purely chemical ones and enable the addition, removal, or modification of functional groups at specific sites on a complex molecule without the use of chemical protection. The reactions, which may be catalyzed include dehydrogenation, oxidation, hydroxylation, dehydration and condensation, decarboxylation, animation, deamination, and isomerization.
Microbial processes have the additional advantage over chemical reagents of operating at relatively low temperatures and pressures without the requirement for potentially polluting heavy-metal catalysts. Although the production of vinegar is the oldest established microbial transformation process conversion of ethanol to acetic acid , the majority of these processes involve the production of high-value compounds including steroids, antibiotics, and prostaglandins.
A novel application of microbial transformation is the use of microorganisms to mimic mammalian metabolism. Humans and animals will metabolize drugs such that they may be removed from the body.
The resulting metabolites may be biologically active themselves—either eliciting a desirable effect or causing damage to the organism. Thus, in the development of a drug it is necessary to determine the activity of not only the administered drug but also its metabolites. These studies may require significant amount of the metabolites and while it may be possible to isolate them from tissues, blood, urine, or faeces of the experimental animal, their concentration is often very low resulting in such approaches being time-consuming, expensive, and far from pleasant.
Sime discussed the exploitation of the metabolic ability of microorganisms to perform these biotransformations. The anomaly of the transformation fermentation process is that a large biomass has to be produced to catalyze a single reaction.
Principles of Fermentation Technology / Edition 3
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Principles of Fermentation Technology
By Peter F. The successful structure of the previous edition of Principles of Fermentation Technology has been retained in this third edition, which covers the key component parts of a fermentation process including growth kinetics, strain isolation and improvement, inocula development, fermentation media, fermenter design and operation, product recovery, and the environmental impact of processes. This accurate and accessible third edition recognizes the increased importance of animal cell culture, the impact of the post-genomics era on applied science and the huge contribution that heterologous protein production now makes to the success of the pharmaceutical industry. This title is ideally suited for both newcomers to the industry and established workers as it provides essential and fundamental information on fermentation in a methodical, logical fashion. Stanbury, Whitaker and Hall have integrated the biological and engineering aspects of fermentation to make the content accessible to members of both disciplines with a focus on the practical application of theory. This text collates all the fermentation fundamentals into one concise reference, making it a valuable resource for fermentation scientists, as well as those studying in the field.