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Uses of enzymes and immobilized enzymes
BIOCHEMICAL EDUCATION
April 1979 Vol. 7 No. 2
Cellular Degradative Processes
developed a n u m b e r of review journals and book series such as Advances in Biochemical Engineering, Topics in Enzyme and Fermentation Biotechnology and Enzyme and Microbial Technology. With almost 100 papers it is not possible to convey the direction of all the studies described. The main areas of activity are presented in separate sections. The first is concerned with the production and extraction of microbial enzymes. Two papers seem especially significant. The first describes how batch experiments may be used to optimize continuous culture for the production of an intracellular enzyme. Now that continuous culture for enzyme production is done commercially such an approach has considerable importance. The second paper describes the use of two-phase polymer solutions for the large-scale fractionation of enzymes and their separation from cell debris. The approach, based on Albertsson's pioneering work, is probably the most significant innovation in large-scale enzyme purification since affinity chromatography. The invited papers on engineering aspects of the application of enzymes do not seem to contain any novel ideas but in the related poster papers is an intriguing account of enzymic regeneration of ATP using a composite triple m e m b r a n e , one to bind enzyme, plus a cation and anion exchanger for selective retention. This is a system interesting to the biologist and the engineer. The sections on modified and immobilized enzymes, industrial applications for enzymes and affinity chromatography each contain important papers for those who must keep up-to-date with the field, but the areas in which striking research developments are apparent therapeutic applications and cell immobilization. That an entire section can be devoted to new medical applications itself reflects the greatly increased activity in this field recently. The papers show the diversity of methods by which the enzymes are presented in order to trick the i m m u n e system: by combination with polymers, in liposomes and microcapsules, in fibres and within membranes. At the first meeting of the series immobilized cells were discussed in only one isolated but prophetic paper. Now they challenge immobilized enzymes strongly and attract probably greater research interest. They offer the prospect of a cheaper biological cat.alyst than immobilized enzymes and one well suited to more complex conversions. Extraordinary operational stabilities reported have been explained recently on the basis of cell replication of the immobilized cells. This is a necessary book for anyone who wants to be active in the field of enzyme and immobilized cell technology or comprehend its importance to medicine, industry or even cell physiology. It is not. like volume 1, an easy introduction to the subject and it is rather light on American work. However, it does give the best account of world-wide activity up to 1977. P. Dunnill Department of Chemical and Biochemical Engineering University College London Torrington Place, London WC1E 7JE
By R. T . D e a n . P p 80. A v o l u m e in t h e Outline Studies in Biology Series. C h a p m a n a n d H a l l , L o n d o n , 1978. P a p e r back £1.75. This slim volume is designed to provide 'a concise conceptual outline of cellular degradative processes' for undergraduates, and for graduates preparing for studies in this field. It succeeds. Roger Dean is clearly familiar with all the aspects of cellular degradation that he describes in the book, and the introduction in Chapter 1, which sets out the roles of degradative processes in cells, is particularly well written. The second chapter, dealing with the methods of turnover studies and the problems these entail, is subtitled 'A very brief survey'. I feel that the outline has been made a little too concise here. This valuable section would have been more comprehensible to undergraduates if it had been expanded at least to 'A brief survey'. Chapter 3 describes the structures of biological macromolecules and the enzymes available for their catabolism. An introduction to the lysosome, the specialized degradative organelle of eukaryotes, follows in Chapter 4. This is probably the best of all the sections, containing a particularly lucid description of the dynamics of the lysosome system. The patterns of degradation and their control are described in Chapters 5 and 6. Although the degradation of proteins has been fairly well explored, much of the discussion of other topics, such as the turnover of organelles, is speculative. Overall, the book gives a clear and concise guide to the mechanics of cellular degradation and is particularly useful in stressing the importance of degradative processes in cellular control and, surprisingly, biosynthesis. There are a few irritations in the presentation - - numerous misprints (including, on the front cover, the author's initials), a severely mutilated structural diagram of pepstatin, a lack of acknowledgement of the source of many of the borrowed figures. However, while research workers will regret the necessary limitation to the bibliography, both they and undergraduates will find this informative account of cellular degradative processes very good value. M. F. Heath Strangeways Research Laboratory Wort's Causeway Cambridge CB1 4RN, U.K.
Enzyme Engineering. Volume 4 E d i t e d by G e o r g e s B. B r o u n , G e o r g M a n e c k e a n d L e m u e l B. W i n g a r d , Jr. Pp 502. P l e n u m P r e s s : New Y o r k a n d L o n d o n , 1978. £ 2 4 . 8 8 . The latest volume in this series reports the fourth of a sequence of meetings held biennially since 1971 on areas of enzyme and more recently immobilized cell technology. This was the first time that the meeting bad been held outside the United States. It therefore reflected the activities of European groups overshadowed at previous conferences by presentations from the massive American enzyme engineering programmes funded in the early '70s by the National Science Foundation. This was also the first meeting in the series where poster papers were accepted and where the programme committee had asked regular contributors to the plenary sessions of earlier conferences to give a lead in presenting just poster papers. Their inclusion as brief presentations in the proceedings gives a particularly wide coverage of current studies and the editors of this volume are to be congratulated on having this account of current research activity published within 12 months of the meeting. The style of the present volume has changed greatly from volume 1. where a few lengthy reviews and personal research presentations were collected. Now the plenary lectures are not reviews and have been edited down to brief articles so that the distinction between them and the written versions of poster papers is small. In some ways this is appropriate because it is in the poster papers that many of the most original ideas and techniques are to be found. One does miss authoritative reviews but since the first meeting there have
47
Uses of Enzymes and Immobilized Enzymes By F r a n c i s X. H a s s e l b e r g e r . Pp P u b l i s h e r s , Inc. C h i c a g o , 1978. $ 1 4 . 9 5 .
220.
Nelson-Hall
This is a relatively brief account of enzymes and related subjects in chemistry, biochemistry, medicine - - and quite a few other subjects! The author digresses often into personal reminiscence and allusion to well-known figures in the field now and in the past. This, however, makes it fascinating reading for the relative layman, and beginning students of biochemistry and related disciplines. The mainly uncritical acceptance of some findings in the medical applications of enzymes, detracts however from the scientific merit of the book. Nevertheless. the author deals in an accurate and readily understandable way with enzymes and a diversity of enzyme applications, including those in the food and dairying industries, and in analysis. The advantages of immobilized enzymes are well presented, including types of enzyme reactors. Other subjects covered include fermentation, production of enzymes, enzyme-
48
BIOCHEMICAL EDUCATION
deficiency diseases and the assay of enzymes during clinical di~agnosis. This book is well worth a quick read through by lecturers and others familiar with the areas covered. Most biochemistry students would benefit from reading the book, especially as enzyme biotechnology is not usually covered in courses in, for example, medical biochemistry. The book is highly recommended to students of home economics, food science, food technology, microbiology and related subjects, where there is a biochemical component to the course. Alan W i s e m a n Department of Biochemistry, University of Surrey Guildford Surrey, U.K.
Topics in Enzyme and Fermentation Biotechnology. Volume 2 E d i t e d by A l a n W i s e m a n . P p . 308. P u b l i s h e d by Ellis H o r w o o d L t d . , C h i c h e s t e r a n d d i s t r i b u t e d by J o h n W i l e y a n d S o n s , C h i c h e s t e r a n d New Y o r k . 1978. £ 1 5 . 0 0 . Five of the six chapters of this new volume concern the importance of enzymes in technology. They are therefore very properly a subject of interest to all those engaged in biochemical education. Each subhect provides a challenge to the biochemist in terms of fundamental enzymology. The chapter on the stabilization of enzymes by the editor of the series illustrates this point well. The technical significance of achieving additional stabilization of enzymes is considerable. It ranges from improved industrial enzyme-reactor operation to the better shelf life of laboratory enzyme reagents. However the fundamental reasons for enzyme stability are of just as much interest to the research biochemist and to the theoretician. At present the subject remains largely a collection of empirical observations. Nevertheless this review is most welcome. A contribution on enzyme electrodes and enzyme based sensors again demonstrates the inseparable relationship between fundamental biochemistry and biochemical technology. To sense say amino acids an immobilized L-amino acid oxidase enzyme is coupled to an a m m o n i u m specific electrode. Its degree of selectivity can be determined by the choice of enzyme. The enzyme-based glucose electrode is becoming an invaluable hospital tool coping with the ever increasing load and cost of blood glucose determinations. The review illustrates the considerable biochemical and chemical ingenuity now being applied to new assays by these approaches. An article about enzymes immobilized on inorganic supports is more specialist in nature. The attachment of enzymes to solid supports has permitted the development of major new industrial processes where the immobilized enzyme acts very m u c h in the m a n n e r previously associated with reactors in the chemical industry. To date the supports used for immobilization in major industrial processes have been organic in nature despite great research investment by the Corning Company on glass and ceramic supports. The authors, themselves engaged in developing inorganic supports at Harwell, present their advantages and it is useful to have a view other than that of the Corning Group who have so totally dominated the literature. The last article on enzymes shows them in quite a different light, as a potential nuisance. It surveys the important antibioticinactivating enzymes. From the early days of penicillin production these microbial enzymes have occupied scientists and with the recognition of their importance in drug resistance this interest must remain strong. Dr. Melling of the Microbiological Research Establishment deals with /3-1actamases and the chloramphenicoland aminoglycoside-inactivating enzymes. He absolves the enzymes of being totally 'hostile' by noting their uses in, for example, screening new antibiotics and treating penicillin allergy.
April 1979 Voi. 7 No. 2
The one .remaining chapter in this volume seems out of place. It concerns the biological treatment of wastes and is a lengthy article of a kind suitable as a teaching text. No one is more enthusiastic than the reviewer for showing the links between different sections of biochemical technology but these are not evident in this instance. Over many volumes of a book series perhaps it does not matter to library purchasers whether single volumes represent a coherent theme. However publishers annoyance with photocopying can receive little sympathy if they continue to ignore the individual purchaser of scientific books. Today he is too often forced to pay for articles in which his interest is very slight in order to obtain several of real interest. To be fair the present volume has a great deal to interest the biochemist and perhaps his dipping into the microbiology and technology of waste treatment will bring forth unexpected advantages. At a time when biochemistry students are perhaps particularly under pressure to show awareness of the direct uses of their discipline to the community this is certainly a book that can be recommended for biochemistry libraries. P. Dunnill Department of Chemical & Biochemical Engineering University College London Torrington Place, London W C I E 7JE
Ion Chromatographic Analysis of Environmental Pollutants E d i t e d by E u g e n e Sawicki, J. D. M u l i k a n d E. W i t t g e n stein. Pp 210. P u b l i s h e d by A n n A r b o r Science P u b l i s h e r s I n c . , A n n A r b o r , M i c h i g a n , U S A (1978) a n d d i s t r i b u t e d by J o h n W i l e y a n d S o n s , C h i c h e s t e r a n d New Y o r k . £ 1 7 . 4 5 The use of chromatographic analysis for the quantitative analysis of mixtures is limited firstly by the ability to separate the components, and secondly by the methods available to detect and to quantify them. This book introduces a development of ion exchange chromatography which permits the use of a non-specific detection system, electrical conductance, for the analysis of a wide range of anions and cations within the same sample, thus greatly extending the usefulness of ion exchange chromatography. In the preface the authors suggest that the book should be seen only as the beginning of the application of 'Ion Chromatography' to environmental problems. Certainly it is a beguiling technique with its potential sensitivity and freedom from interference. The ability to analyse chloride, sulphate, sulphite and phosphate in a single sample is most attractive. Chapter one is speculative - - suggesting a whole range of possible applications. The second chapter, written by one of the originators of this elegant procedure gives a very lucid account of the principles involved. In the limited space available the author establishes the potential of the technique whilst at the same time drawing attention to its limitations. The chapters that follow present a series of applications concerned mainly with the measurement of atmospheric pollution. For the dedicated reader there is useful information to be gleaned here: for most of us stronger editing would have been useful. Some of the chapters attempt a reasoned critique of the method, particularly that which describes the analysis of airborne particulate matter. Here it is shown that at present the variability of 'Ion Chromatography' may be greater than for more conventional methods of analysis. In other examples there is no evidence that the technique, per se. has real advantages over existing techniques. Perhaps a book for the committed. In the absence of a satisfactory chapter giving a critical discussion of the results obtained in the described procedures, the reader will need to make a considerable effort to determine on the basis of this book whether 'Ion Chromatography' represents a justifiable investment. Gordon Jones Water Research Centre Stevenage, Herts, U.K.
April 1979 Vol. 7 No. 2
Cellular Degradative Processes
developed a n u m b e r of review journals and book series such as Advances in Biochemical Engineering, Topics in Enzyme and Fermentation Biotechnology and Enzyme and Microbial Technology. With almost 100 papers it is not possible to convey the direction of all the studies described. The main areas of activity are presented in separate sections. The first is concerned with the production and extraction of microbial enzymes. Two papers seem especially significant. The first describes how batch experiments may be used to optimize continuous culture for the production of an intracellular enzyme. Now that continuous culture for enzyme production is done commercially such an approach has considerable importance. The second paper describes the use of two-phase polymer solutions for the large-scale fractionation of enzymes and their separation from cell debris. The approach, based on Albertsson's pioneering work, is probably the most significant innovation in large-scale enzyme purification since affinity chromatography. The invited papers on engineering aspects of the application of enzymes do not seem to contain any novel ideas but in the related poster papers is an intriguing account of enzymic regeneration of ATP using a composite triple m e m b r a n e , one to bind enzyme, plus a cation and anion exchanger for selective retention. This is a system interesting to the biologist and the engineer. The sections on modified and immobilized enzymes, industrial applications for enzymes and affinity chromatography each contain important papers for those who must keep up-to-date with the field, but the areas in which striking research developments are apparent therapeutic applications and cell immobilization. That an entire section can be devoted to new medical applications itself reflects the greatly increased activity in this field recently. The papers show the diversity of methods by which the enzymes are presented in order to trick the i m m u n e system: by combination with polymers, in liposomes and microcapsules, in fibres and within membranes. At the first meeting of the series immobilized cells were discussed in only one isolated but prophetic paper. Now they challenge immobilized enzymes strongly and attract probably greater research interest. They offer the prospect of a cheaper biological cat.alyst than immobilized enzymes and one well suited to more complex conversions. Extraordinary operational stabilities reported have been explained recently on the basis of cell replication of the immobilized cells. This is a necessary book for anyone who wants to be active in the field of enzyme and immobilized cell technology or comprehend its importance to medicine, industry or even cell physiology. It is not. like volume 1, an easy introduction to the subject and it is rather light on American work. However, it does give the best account of world-wide activity up to 1977. P. Dunnill Department of Chemical and Biochemical Engineering University College London Torrington Place, London WC1E 7JE
By R. T . D e a n . P p 80. A v o l u m e in t h e Outline Studies in Biology Series. C h a p m a n a n d H a l l , L o n d o n , 1978. P a p e r back £1.75. This slim volume is designed to provide 'a concise conceptual outline of cellular degradative processes' for undergraduates, and for graduates preparing for studies in this field. It succeeds. Roger Dean is clearly familiar with all the aspects of cellular degradation that he describes in the book, and the introduction in Chapter 1, which sets out the roles of degradative processes in cells, is particularly well written. The second chapter, dealing with the methods of turnover studies and the problems these entail, is subtitled 'A very brief survey'. I feel that the outline has been made a little too concise here. This valuable section would have been more comprehensible to undergraduates if it had been expanded at least to 'A brief survey'. Chapter 3 describes the structures of biological macromolecules and the enzymes available for their catabolism. An introduction to the lysosome, the specialized degradative organelle of eukaryotes, follows in Chapter 4. This is probably the best of all the sections, containing a particularly lucid description of the dynamics of the lysosome system. The patterns of degradation and their control are described in Chapters 5 and 6. Although the degradation of proteins has been fairly well explored, much of the discussion of other topics, such as the turnover of organelles, is speculative. Overall, the book gives a clear and concise guide to the mechanics of cellular degradation and is particularly useful in stressing the importance of degradative processes in cellular control and, surprisingly, biosynthesis. There are a few irritations in the presentation - - numerous misprints (including, on the front cover, the author's initials), a severely mutilated structural diagram of pepstatin, a lack of acknowledgement of the source of many of the borrowed figures. However, while research workers will regret the necessary limitation to the bibliography, both they and undergraduates will find this informative account of cellular degradative processes very good value. M. F. Heath Strangeways Research Laboratory Wort's Causeway Cambridge CB1 4RN, U.K.
Enzyme Engineering. Volume 4 E d i t e d by G e o r g e s B. B r o u n , G e o r g M a n e c k e a n d L e m u e l B. W i n g a r d , Jr. Pp 502. P l e n u m P r e s s : New Y o r k a n d L o n d o n , 1978. £ 2 4 . 8 8 . The latest volume in this series reports the fourth of a sequence of meetings held biennially since 1971 on areas of enzyme and more recently immobilized cell technology. This was the first time that the meeting bad been held outside the United States. It therefore reflected the activities of European groups overshadowed at previous conferences by presentations from the massive American enzyme engineering programmes funded in the early '70s by the National Science Foundation. This was also the first meeting in the series where poster papers were accepted and where the programme committee had asked regular contributors to the plenary sessions of earlier conferences to give a lead in presenting just poster papers. Their inclusion as brief presentations in the proceedings gives a particularly wide coverage of current studies and the editors of this volume are to be congratulated on having this account of current research activity published within 12 months of the meeting. The style of the present volume has changed greatly from volume 1. where a few lengthy reviews and personal research presentations were collected. Now the plenary lectures are not reviews and have been edited down to brief articles so that the distinction between them and the written versions of poster papers is small. In some ways this is appropriate because it is in the poster papers that many of the most original ideas and techniques are to be found. One does miss authoritative reviews but since the first meeting there have
47
Uses of Enzymes and Immobilized Enzymes By F r a n c i s X. H a s s e l b e r g e r . Pp P u b l i s h e r s , Inc. C h i c a g o , 1978. $ 1 4 . 9 5 .
220.
Nelson-Hall
This is a relatively brief account of enzymes and related subjects in chemistry, biochemistry, medicine - - and quite a few other subjects! The author digresses often into personal reminiscence and allusion to well-known figures in the field now and in the past. This, however, makes it fascinating reading for the relative layman, and beginning students of biochemistry and related disciplines. The mainly uncritical acceptance of some findings in the medical applications of enzymes, detracts however from the scientific merit of the book. Nevertheless. the author deals in an accurate and readily understandable way with enzymes and a diversity of enzyme applications, including those in the food and dairying industries, and in analysis. The advantages of immobilized enzymes are well presented, including types of enzyme reactors. Other subjects covered include fermentation, production of enzymes, enzyme-
48
BIOCHEMICAL EDUCATION
deficiency diseases and the assay of enzymes during clinical di~agnosis. This book is well worth a quick read through by lecturers and others familiar with the areas covered. Most biochemistry students would benefit from reading the book, especially as enzyme biotechnology is not usually covered in courses in, for example, medical biochemistry. The book is highly recommended to students of home economics, food science, food technology, microbiology and related subjects, where there is a biochemical component to the course. Alan W i s e m a n Department of Biochemistry, University of Surrey Guildford Surrey, U.K.
Topics in Enzyme and Fermentation Biotechnology. Volume 2 E d i t e d by A l a n W i s e m a n . P p . 308. P u b l i s h e d by Ellis H o r w o o d L t d . , C h i c h e s t e r a n d d i s t r i b u t e d by J o h n W i l e y a n d S o n s , C h i c h e s t e r a n d New Y o r k . 1978. £ 1 5 . 0 0 . Five of the six chapters of this new volume concern the importance of enzymes in technology. They are therefore very properly a subject of interest to all those engaged in biochemical education. Each subhect provides a challenge to the biochemist in terms of fundamental enzymology. The chapter on the stabilization of enzymes by the editor of the series illustrates this point well. The technical significance of achieving additional stabilization of enzymes is considerable. It ranges from improved industrial enzyme-reactor operation to the better shelf life of laboratory enzyme reagents. However the fundamental reasons for enzyme stability are of just as much interest to the research biochemist and to the theoretician. At present the subject remains largely a collection of empirical observations. Nevertheless this review is most welcome. A contribution on enzyme electrodes and enzyme based sensors again demonstrates the inseparable relationship between fundamental biochemistry and biochemical technology. To sense say amino acids an immobilized L-amino acid oxidase enzyme is coupled to an a m m o n i u m specific electrode. Its degree of selectivity can be determined by the choice of enzyme. The enzyme-based glucose electrode is becoming an invaluable hospital tool coping with the ever increasing load and cost of blood glucose determinations. The review illustrates the considerable biochemical and chemical ingenuity now being applied to new assays by these approaches. An article about enzymes immobilized on inorganic supports is more specialist in nature. The attachment of enzymes to solid supports has permitted the development of major new industrial processes where the immobilized enzyme acts very m u c h in the m a n n e r previously associated with reactors in the chemical industry. To date the supports used for immobilization in major industrial processes have been organic in nature despite great research investment by the Corning Company on glass and ceramic supports. The authors, themselves engaged in developing inorganic supports at Harwell, present their advantages and it is useful to have a view other than that of the Corning Group who have so totally dominated the literature. The last article on enzymes shows them in quite a different light, as a potential nuisance. It surveys the important antibioticinactivating enzymes. From the early days of penicillin production these microbial enzymes have occupied scientists and with the recognition of their importance in drug resistance this interest must remain strong. Dr. Melling of the Microbiological Research Establishment deals with /3-1actamases and the chloramphenicoland aminoglycoside-inactivating enzymes. He absolves the enzymes of being totally 'hostile' by noting their uses in, for example, screening new antibiotics and treating penicillin allergy.
April 1979 Voi. 7 No. 2
The one .remaining chapter in this volume seems out of place. It concerns the biological treatment of wastes and is a lengthy article of a kind suitable as a teaching text. No one is more enthusiastic than the reviewer for showing the links between different sections of biochemical technology but these are not evident in this instance. Over many volumes of a book series perhaps it does not matter to library purchasers whether single volumes represent a coherent theme. However publishers annoyance with photocopying can receive little sympathy if they continue to ignore the individual purchaser of scientific books. Today he is too often forced to pay for articles in which his interest is very slight in order to obtain several of real interest. To be fair the present volume has a great deal to interest the biochemist and perhaps his dipping into the microbiology and technology of waste treatment will bring forth unexpected advantages. At a time when biochemistry students are perhaps particularly under pressure to show awareness of the direct uses of their discipline to the community this is certainly a book that can be recommended for biochemistry libraries. P. Dunnill Department of Chemical & Biochemical Engineering University College London Torrington Place, London W C I E 7JE
Ion Chromatographic Analysis of Environmental Pollutants E d i t e d by E u g e n e Sawicki, J. D. M u l i k a n d E. W i t t g e n stein. Pp 210. P u b l i s h e d by A n n A r b o r Science P u b l i s h e r s I n c . , A n n A r b o r , M i c h i g a n , U S A (1978) a n d d i s t r i b u t e d by J o h n W i l e y a n d S o n s , C h i c h e s t e r a n d New Y o r k . £ 1 7 . 4 5 The use of chromatographic analysis for the quantitative analysis of mixtures is limited firstly by the ability to separate the components, and secondly by the methods available to detect and to quantify them. This book introduces a development of ion exchange chromatography which permits the use of a non-specific detection system, electrical conductance, for the analysis of a wide range of anions and cations within the same sample, thus greatly extending the usefulness of ion exchange chromatography. In the preface the authors suggest that the book should be seen only as the beginning of the application of 'Ion Chromatography' to environmental problems. Certainly it is a beguiling technique with its potential sensitivity and freedom from interference. The ability to analyse chloride, sulphate, sulphite and phosphate in a single sample is most attractive. Chapter one is speculative - - suggesting a whole range of possible applications. The second chapter, written by one of the originators of this elegant procedure gives a very lucid account of the principles involved. In the limited space available the author establishes the potential of the technique whilst at the same time drawing attention to its limitations. The chapters that follow present a series of applications concerned mainly with the measurement of atmospheric pollution. For the dedicated reader there is useful information to be gleaned here: for most of us stronger editing would have been useful. Some of the chapters attempt a reasoned critique of the method, particularly that which describes the analysis of airborne particulate matter. Here it is shown that at present the variability of 'Ion Chromatography' may be greater than for more conventional methods of analysis. In other examples there is no evidence that the technique, per se. has real advantages over existing techniques. Perhaps a book for the committed. In the absence of a satisfactory chapter giving a critical discussion of the results obtained in the described procedures, the reader will need to make a considerable effort to determine on the basis of this book whether 'Ion Chromatography' represents a justifiable investment. Gordon Jones Water Research Centre Stevenage, Herts, U.K.