METHODS in MICROBIOLOGY This Page Intentionally Left Blank METHODS in MICROBIOLOGY Edited by J. R. NORRIS Borden Microbiological Laboratory, Shell Research Limited, Sittingbourne, Kent, England D.W.RIBBONS Department of Biochemistry, University of Miami School of Medicine, and Howard Hughes Medical Institute, Miami, Florida, U.S.A. Volume 6B @ 1972 ACADEMIC PRESS London and New York ACADEMIC PRESS INC. (LONDON) L T D 24-28 Oval Road, London NW1 7DX U.S. Edition published by ACADEMIC PRESS INC. 111 Fifth Avenue New York, New York 10003 Copyright 0 1972 by ACADEMIC PRESS INC. (LONDON) LTD All Rights Reserved No part of this book may be reproduced in any form by photostat, microfilm, or any other means, without written permission from the publishers Library of Congress Catalog Card Number: 68-57745 ISBN: 0-12-521546-0 PRINTEDIN GREATBRITAINBY ADLARDAND SONLIMITED DORKING, SURREY LIST OF CONTRIBUTORS M. J . ALLEN,Shell Research Lid, Borden MicrobiologicalLaboratory, Sittingbourne, Kent, England R. B. BEECHEY, Shell Research Ltd, Woodstock Agricultural Research Centre, Sittingbourne, Kent, England G. W . CROSBIE, Department of Biochemistry, The University,Hull, England N . R. EATON,Department of Biology, Brooklyn College of the City University of New York, Brooklyn, New York, U.S.A. A. FERRARI, Division of Biomedical Sciences, Damon Corporation, Needham, Mass., U.S.A. W . W . FORREST, The Australian Wine Research Institute, Glen Osmond, South Australia P. B. GARLAND, Department of Biochemistry, University of Dundee, Dundee, Scotland J . H . HASH,Department of Microbiology, Vanderbilt University School of Mfdicine, Nashville, Tennessee, U.S.A J. MARTEN, Division of Biomedical Sciences, Damon Corporation, Needham, Mass., U.S.A. D. G. NICHOLLS, Department of Medicine, University of Nottingham, Nottingham, England J . L. PEEL,Agricultural Research Council Food Research Institute, Colney Lane, Norwich N O R 70F, England J . R. POSTGATE, S.R.C. Unit of Nitrogen Fixation, University of Sussex, Falmer, Sussex J. R. QUAYLE,Department of Microbiology, University of Shefield, Shefield, England D. W . RIBBONS,Department of Biochemistry, University of Miami School of Medicine, and Howard Hughes Medicul Institute, Miami, Florida, U.S.A. C. H. WANG,Radiation Center and Department of Chemistry, Oregon State University, Corvallis, Oregon, U.S.A. . V ACKNOWLEDGMENTS For permission to reproduce, in whole or in part, certain figures and diagrams we are grateful to the followingEkco Electronics Limited , Southend-on-Sea, England ; G.E.C. Limited, Kingsway, London; Master Instruments, Sidney, Australia; The Nuclear-Chicago Corporation, Des Plaines, Illinois, U.S.A. ; The Packard Instrument Company; Pergamon Press, London; 20th Century Electronics Limited, Croydon, Surrey, England. Detailed acknowledgments are given in the legends to figures. vi PREFACE The main theme in Volume 6 of “Methods in Microbiology” is the application of biochemical techniques to the study of micro-organisms. T h e topics covered include the use of biochemical and enzymic tests to characterize microbial types, the quantitative separation and analysis of fermentation products produced by micro-organisms, a critical appraisal of methods available to elucidate metabolic pathways and the control of enzyme synthesis, the assay of selected enzymes in crude extracts of cells as indicators of metabolic pathways, the use of antimetabolites to study biosynthesis and electron transport. The applications of radiotracer techniques are described in detail in five chapters: respiratory measurements with dyes and with electrodes for oxygen and carbon dioxide are included while manometric methods, which are so well described elsewhere (Umbreit, B