Antibiotics

886 Pages·1975·45.55 MB·English

by  John H. Hash

Preface
In three decades or so of widespread use, antibiotics have wrought
a revolution in the medical, veterinary, and agricultural sciences; indeed,
in all of the biological sciences. Early research on antibiotics was neces-
sarily directed at the production, isolation, characterization, and phar-
macology of this important class of natural products. It was soon appar-
ent that microorganisms could develop resistance to antibiotics and that
this resistance was due, at least in part, to the possession of enzymes
that could, in some fashion, chemically modify the antibiotic. OT keep
ahead (or even abreast) of antibiotic resistance, it is necessary either
to constantly discover new antibiotics or to develop derivatives that are
insensitive to the enzymes that cause inactivation of the natural com-
pound. Both avenues have been tried. It is evident that the search for
new antibiotics must eventually reach the point of diminishing returns
and that the second approach offers the best hope of extendiag the life
of an antibiotic. Research on the biosyntheses of antibiotics as well as
on enzymatic means of degradation has received less emphasis than other
aspects of antibiotic chemistry, but, as it has become apparent that a
knowledge of biosynthetic pathways can assist in isolating intermediate
compounds capable of being modified chemically, this area has received
more attention. Similarly, degradative enzymes have been found that are
capable of providing antibiotic derivatives which can be chemically modi-
fied, allowing production of large numbers of semisynthetic antibiotics.
Other practical uses of antibiotic enzymology include the use of enzymes
as analytical reagents in determining the concentrations of antibiotics
in samples and in the effective removal of antibiotics from reaction mix-
tures by converting them to inactive compounds.
Aside from the utilitarian aspects of antibiotic enzymology a principal
driving force behind research on these enzymes is the intellectual curios-
ity as to the raison d'etre of antibiotic synthesis. The production of these
secondary metabolites, which serve no evident function in the producing
organism, requires large quantities of energy and considerable metabolic
machinery. In some instances very complicated pathways involving 20-30
enzymes are required to synthesize an antibiotic. As the pathways are
unraveled and the branch points with normal metabolic routes are estab-
lished, light may be shed on the mechanisms of antibiotic synthesis.
Even with the renewed interest in antibiotic enzymology, the extent
and scope of research in this area are uneven, and the published results
are scattered throughout the scientific literature. The aim of this volume
is to collate in one source as much information concerning antibiotic
iiix
xiv ECAFERP
enzymology as possible. The work is divided into three sections. The first
is concerned with methods used in the study of antibiotics, and covers
techniques from culturing the producing organism to various chromato-
graphic methods to sophisticated physical techniques. The second and
third sections are devoted to enzymes involved in antibiotic biosynthesis
and antibiotic degradatio n and modification, respectively. In some cases
the division between the second and third section is quite arbitrary be-
cause it is not always clear whether an enzyme belongs in a biosynthetic
or degradative pathway. The coverage of enzymes represents the state
of the art of antibiotic enzymology; the range extends from pure enzymes
that have been sequenced to enzymes that have been studied only in crude
extracts. Many other enzymes that act on antibiotics, antibiotic precur-
sors, or antibiotic derivatives have been detected in extracts or whole
cells. Most of these had to be omitted because of a paucity of information.
It is evident that only a small part of antibiotic enzymology has reached
the stage where it can be consolidated into a treatise of this kind, and
it is hoped that this volume will serve as a stimulus for further research
on the enzymes involved with this important class of compounds.
I am indebted to many people for many ideas and suggestions, but
I am especially indebted to investigators in pharmaceutical laboratories
throughout the world for their ideas and contributions.
JOHN H. HASH
Contributors to Volume XLIII
Article numbers are in parentheses following the names of contributors.
Affiliations listed are current.
DRANREB J. ABBOTT (55b), Fermentation Cambridge, Massachusetts
Products Research, Lilly Research JOHN E. GNIDWOD (48), Department o
Laboratories, Eli Lilly and Company, Biochemistry, The University o Wis-
Indianapolis, Indiana ,nisnoc Madison, Wisconsin
E. P. MAHARBA (29, 55a), Sir William ETTELOC DUEZ (53f), Institut de Bota-
Dunn School o Pathology, University nique, Service de Microbiologie, Uni-
o Ox/ord, Oxord, England versitg de Liege, Sart Tilman, Liege,
NAJRODA SOLAZSA (8), The Squibb Insti- Belgium
tute ro Medical Research, New E. F. RENTSLE (37), Department o Bio-
Brunswick, New Jersey, and Princeton logical Organic Chemistry, Albert
University, Princeton, New Jersey niet~,niE Medical Center, Philadelphia,
REDNIHOM S. ALAHTAB (17), College o/ Pennsylvania
Pharmacy, The Ohio State University, OE)IEMA A. FANTINI (2), Department o
Columbus, Ohio Microbiology, Lederle Laboratories,
RIMIDALV BETINA (7), Department o Pearl River, New York
Technical Microbiology and Biochem- AICIRTAP TTECWAF (29, 55a), riS William
istry, Faculty o Chemistry, Slovak Dunn School o Pathology, University
PoIytechnical University, Bratislava, o ,droxO ,droxO England
Czechoslovakia HEINZ G. FLOSS (34), Department o
DLANOD B. SRE)IROB (10), Department o Medicinal Chemistry and Pharmacog-
Fermentation and Isolation, Lederle nosy, School o Pharmacy and Phar-
Laboratories, Pearl River, New York macal Sciencies, Purdue University,
DRAHREBE REKUERB (41), Lohman and West Laayette, Indiana
Company AG, Neuelder Strasse, EIRAM-NAEJ FRERE (53f), Institut ed
912 Cuxhaven, Germany Botanique, Service de Microbiologie,
RICHA~ BaU~NEa (31), Institut iir Bio- ~tisrevinU ed Liege, Sart Tilman,
chemische Technologie und Mikro- Liege, Belgium
biologie, Technischen HochschuIe DAvm FROST (8), The Squibb Institute
Wien, Wien Getreidemarkt, Austria ro Medical Research, New Brunswick,
JOHN H. STAOC (58), Research Labora- New Jersey
tories, The Upjohn Company, Kala- DIVAD S. FUKUDA (55b), Lilly Research
mazoo, Michigan Laboratories, Eli Lilly and Company,
M. COLE (54a, 54b), Research Division, Indianapolis, Indiana
Beecham Pharmaceuticals Betch- STEN KCEBNETAG (30), Division o Pure
worth, Surrey, England and Applied Biochemistry, The Royal
JOHN W. NAROCROC (33), Department o Institute o Technology, Stockholm,
Biochemistry, Northwestern Univer- Swede~
sity School o Medicine, Chicago, G. M. REHCUAG (40), Department o
sionillI Chemistry, The University of ,yraglaC
NAMYL C. GIARC (16), The RockeeUer ,yraglaC Alberta, Canada
University, New York, New York EIRAM-NAEJ GHUYSEN (53f), Institnt ed
NAILUJ SEIVAD (3), Department o Bio- ,et~qinatoB Service ed Microbiologie,
chemistry, The University o/ Wiscon- gtisrevinU de Liege, Sart Tilman,
sin, Madison, Wisconsin Lihge, Belgium
DLONRA L. DEMAIN (52), Department o LEAHCIM J. HAAS (48), Department o
Nutrition and Food Science, Massa- Biochemistry, The University o Wis-
chusetts Institute of Technology, consin, Madison, Wisconsin
ix
X CONTRIBUTORS TO VOLUME XLIII
JOHN H. HASH (46, 47), Department o/ Inc., Nutley, New Jersey
Microbiology, Vanderbilt University, RETSEL A. MITSCHER (17), College o/
School o/ Medicine, Nashville, Pharmacy, The Ohio State University,
Tennessee Columbus, Ohio
DRAHREG HEINRICH (41), Case Western TREBRON NEUSS (20), Lilly Research
Reserve School ]o Medicine, Cleve- Laboratories, Eli Lilly and Company,
land, Ohio Indianapolis, Indiana
TREFLU NNAMENROH (34), Department AIHTNYC H. NAHGALLAC'O (5), Bacterial
]o Medicinal Chemistry and Pharma- Chemotherapy Unit, Glaxo Research
cognosy, School ]o Pharmacy and Limited, Green/ord, Middlesex,
Pharmacal Sciences, Purdue Univer- England
sity, West La]ayette, Indiana NAES C. RONNOC'O (14), Lilly Research
DRAWDE INAMINE (52), Developmental Laboratories, Eli Lilly and Company,
Microbiology Department, Merck Indianapolis, Indiana
Sharp and Dohme Research Labora- YRNEH SULUAP (44), Department o/Bio-
,seirot Rahway, New Jersey logical Chemistry, Harvard Medical
KENNETH JOHNSON (53f), Biochemistry School, Boston, Massachusetts
Laboratory, National Research Council D. NAMLREP (60, 61), School ]o Phar-
]o Canada, Ottawa, Canada macy, The University o/ Wisconsin,
KCIREDERF HGANAVAK (4), 132 Blue Madison, Wisconsin
Ridge Road, Indianapolis, Indiana RETEP REDNEAFP (41), Institut r~] Bio-
L. A. KOMINEK (35), Fermentation Re- logische Chemie und Erntihrungswis-
search and Development, The Upjohn senscha/t, Universitat Hohenheim,
Company, Kalamazoo, Michigan Hohenheim, Germany
SHINICHI ODNOK (11, 12), Institute ]o NOTRUB M. LLEGOP (36), Department o/
Microbial Chemistry, Shinagawa-ku, ,ygoloiborciM St. Louis University
Tokyo, Japan School ]o Medicine, St. Louis, Mis-
ZOFIA AKSWoRoB-o~_~.,RUK (42), The iruos
Rocke]eller University, New York, JOHN N. RETROP (1), Department ]o
New York Microbiology, Lederle Laboratories,
NERCS G. DNALAL (43), Department ]o Pearl River, New York
Biochemistry, University ]o Oslo, ,olsO M. H. DNOMHCIR (6, 53C, 53d), Depart-
Norway ment o/ Bacteriology, The Medical
GNUS G. EEL (45), The Rocke]eller Uni- School, University o/ Bristol, Bristol,
versity, New York, New York England
YELBOR J. LIGHT (39), Department ]o RETEPSNAH REDEIR (41), Institut r~i] Bio-
Chemistry, Florida Stale University, logische Chemie und Erntihrungswis-
Tallahasee, ad~TolF senscha]t, Universittit Hohenheim,
FRITZ LIPMANN (45), The Rocke]elIer Hohenheim, Germany
University, New York, New York JOHN H. NOSTREBOR (9), Fermentation
F. LYNEN (38), Max-Planck-Institut r~] Research and Development, The -pU
Biochemie, Munich, Germany john Company, Kalamazoo, Michigan
YRAG G. INOCRAM (13), Lilly Research XAM RSHR (31), Institut r~] Biochem-
Laboratories, Eli Lilly and Company, ische Technologie und MikrobioIogie,
Indianapolis, Indiana Technische Hochschule Wien, Wien,
It. F. MEYER (35), Fermentation Re- Austria
search and Development, The Upjohn NODROG W. Ross (5, 53e), Glaxo Re-
Company, Kalamazoo, Michigan search Limited, Green]ord, Middlesex,
PHmIP A. MILLER (46, 47), Department England
]o Microbiology, Ho]]mann-La Roche .W A. EGDIVAS (54a, 54b), Biochemical
CONTRIBUTORS TO VOLUME XLIII xi
Services Unit, Beecham Pharmaceuti- of Edinburgh, Edinburgh, Scotland
cals, Betchworth, Surrey, England KivosHi Tswi (9, 15), Control Analyti-
W. V. SHAW (57), Department of Bio- lac Research and Development, The
chemistry, University of Leicester, Upjohn Company, Kalamazoo, Michi-
Leicester, England gan
DRAWOH NAMBEGEIS (18), Princeton Ap- .W UEMATSU (59), Department of Bio-
plied Research Corporation, Princeton, logical Organic Chemistry, Albert Ein-
New Jersey stein Medical Center, Philadelphia,
RIVAHAM M. SIMLOT (41), Agricultural Pennsylvania
Experimental Station, University of OAMAH UMEZAWA (11, 12), Institute of
Udaipur, Udaipur (Rajasthan), India Microbial Chemistry, Shinagawa-ku,
EGROEG SLOMP (19), Fermentation Re- Tokyo, Japan
search and Development, The Upjohn TREBUH EHGEAHREDNAV (54a, 54c), Rega
Company, Kalamazoo, Michigan Institute, University of Leuven,
JOHN SOGN (16), The Rockefeller Uni- Leuven, Belgium
versity, New York, New York L. C. VINING (56), Department of Biol-
ERODOEHT S. IKSOLOKOS (17), College of ogy, Dalhonsie University, Halifax,
Pharmacy, The Ohio State University, Nova Scotia, Canada
Columbus, Ohio BETNTiiG VOGEL (38, 39), Max-PIanck-
MARILYN K. SPEEDIE (34), School of Institute iir Biologie, Tubingen,
Pharmacy, Oregon State University, Germany
Corvallis, Oregon JAMES B. WALKER (21, 22, 23, 24, 25, 26,
BRIAN RECNEPS (32), Department of Bio- 27, 28, 49, 50, 51), Department of Bio-
chemistry, University of Dublin, Trin- chemistry, Rice University, Houston,
ity College, Dublin, Ireland Texas
R. J. KINLODAHUS (37, 59), Department TEBA6RAM S. WALKER (50, 51), Depart-
of Biological Organic Chemistry, Al- ment of Biochemistry, Rice Univer-
bert Einstein Medical Center, Phila- sity, Houston, Texas
delphia, Pennsylvania TRINE-LIsE ZIMMER (43), Department of
DIVAD ~. REHCTAHT (53a, 53b), Depart- Biochemistry, University of Oslo, OsIo,
ment of Molecular Biology, University Norway
METHODS IN ENZYMOLOGY
EDITED BY
Sidney P. Colowick and Nathan O. Kaplan
VANDERBILT UNIVERSITY DEPARTMENT OF CHEMISTRY
SCHOOL OF MEDICINE UNIVERSITY OF CALIFORNIA
NASHVILLE, TENNESSEE AT SAN DIEGO
LA JOLLA, CALIFORNIA
I. Preparation and Assay fo Enzymes
II. Preparation and Assay fo Enzymes
III. Preparation and Assay fo Substrates
.VI Special Techniques for the Enzymologist
.V Preparation and Assay fo Enzymes
VI. Preparation and Assay fo Enzymes )deunitnoC(
Preparation and Assay fo Substrates
Special Techniques
VII. Cumulative Subject Index
XV
METHODS IN ENZYMOLOGY
EDITORS-IN-CHIEF
Sidney P. Colowick Nathan 0. Kaplan
EMULOV VIII. Complex Carbohydrates
Edited by HTEBAZILE F. NEUFELD DNA ROTCIV GRUBSNIG
EMULOV IX. Carbohydrate Metabolism
Edited by WILLIS A. DOOW
EMULOV X. Oxidation and Phosphorylation
Edited by DLANOR W. KOORBATSE DNA DRANYAM E. NAMLLUP
EMULOV XI. Enzyme Structure
Edited by C. H. W. SRIH
EMULOV XII. Nucleic Acids (Parts A and B)
Edited by ECNERWAL NAMSSORG DNA KIVIE EVADLOM
EMULOV XIII. Citric Acid Cycle
Edited by J. M. NIETSNEWOL
EMULOV XIV. Lipids
Edited by J. M. NIETSNEWOL
EMULOV XV. Steroids and Terpenoids
Edited by DNOMYAR B. NOTYALC
EMULOV XVI. Fast Reactions
Edited by HTENNEK NITSUK
EMULOV XVII. Metabolism of Amino Acids and Amines (Parts A and B)
Edited by TREBREH ROBAT DNA AILEC WHITE ROBAT
EMULOV XVIII. Vitamins and Coenzymes (Parts A, B, and C)
Edited by DLANOD B. KCIMROCCM DNA LEUMEL D. THGIRW
EMULOV XIX. Protcolytic Enzymes
Edited by EDURTREG E. NNAMLREP DNA OLZSAL DNAROL
EMULOV XX. Nucleic Acids and Protein Synthesis (Part C)
Edited by KIVIE EVADLOM DNA ECNERWAL NAMSSORG
iivx
xviii METHODS IN ENZYMOLOGY
EMULOV XXI. Nucleic Acids (Part D)
Edited by ECNERWAL NAMSSORG DNA KIVIE EVADLOM
EMULOV XXII. Enzyme Purification and Related Techniques
Edited by MAILLIW B. YBOKAJ
EMULOV XXIII. Photosynthesis (Part A)
Edited by YNOHTNA NAS ORTEIP
EMULOV XXlV. Photosynthesis and Nitrogen Fixation (Part B)
Edited by YNOHTNA NAS ORTEIP
EMULOV XXV. Enzyme Structure (Part B)
Edited by C. H. W. HIRS DNA EGRES N. FFEHSAMIT
EMULOV XXVI. Enzyme Structure (Part C)
Edited by C. H. W. HIRS DNA EGRES N. FFEHSAMIT
EMULOV XXVII. Enzyme Structure (Part D)
Edited by C. H. W. HIRS DNA EGRES N. FFEHSAMIT
EMULOV XXVIII. Complex Carbohydrates (Part B)
Edited by ROTCIV GRUBSNIG
EMULOV XXIX. Nucleic Acids and Protein Synthesis (Part E)
Edited by ECNERWAL NAMSSORG DNA KIVlE EVADLOM
EMULOV
XXX. Nucleic Acids and Protein Synthesis (Part F)
Edited by KIVlE EVADLOM DNA ECNERWAL NAMSSORG
EMULOV XXXI, Biomembranes (Part A)
Edited by YENDIS REHCSIELF DNA RETSEL REKCAP
EMULOV XXXII. Biomembranes (Part B)
Edited by YENDIS REHCSIELF DNA RETSEL REKCAP
EMULOV XXXIII. Cumulative Subject Index Volumes I-XXX
Edited by AHTRAM G. DENNIS DNA DRAWDE A. SINNED
EMULOV XXXIV. Affinity Techniques (Enzyme Purification: Part B)
Edited by MAILLIW B. YBOKAJ DNA MEIR KEHCLIW
METHODS IN ENZYMOLOGY xix
EMULOV XXXV. Lipids (Part B)
Edited by JOHN M. NIETSNEWOL
EMULOV XXXVI. Hormone Action (Part A: Steroid Hormones)
Edited by TREB W. YELLAM'O DNA LEOJ G. NAMDRAH
EMULOV XXXVII. Hormone Action (Part B: Peptide Hormones)
Edited by TREB W. YELLAM'O DNA LEOJ G. NAMDRAH
EMULOV XXXVIII. Hormone Action (Part C: Cyclic Nucleotides)
Edited by LEOJ G. NAMDRAH DNA TREB W. YELLAM'O
EMULOV XXXIX. Hormone Action (Part D: Isolated Cells, Tissues, and
Organ Systems)
Edited by LEOJ G. NAMDRAH DNA TREB W. YELLAM'O
EMULOV XL. Hormone Action (Part E: Nuclear Structure and Function)
Edited by TREB W. YELLAM'O DNA LEOJ G. NAMDRAH
EMULOV XLI. Carbohydrate Metabolism (Part B)
Edited by W. A. DOOW
EMULOV XLII. Carbohydrate Metabolism (Part C)
Edited by W. A. WooD
EMULOV XLIII. Antibiotics
Edited by JOHN H. HSAH
1 ANTIBIOTIC-PRODUCING MICROORGANISMS 3
1 Cultural Conditions for Antibiotic-Producing
Microorganisms
By JOHN N. PORTER
I. Introduction . . . . . . . . . . . . . . . . . . 3
II. Culture Maintenance . . . . . . . . . . . . . . . . 3
III. Culture Preservation . . . . . . . . . . . . . . . . . 8
IV. Antibiotic Production in Liquid Culture . . . . . . . . . . II
V. Selected Antibiotic Fermentations by Fungi . . . . . . . . . 41
A. Cephalosporin C . . . . . . . . . . . . . . . . 14
B. Griseofulvin . . . . . . . . . . . . . . . . . 51
C. Penicillin . . . . . . . : . . . . . . . . . . 61
VI. Selected Antibiotic Fermentations by Eubacteria . . . . . . . 61
A. Bacitracin . . . . . . . . . . . . . . . . . . 61
B. Polymyxins . . . . . . . . . . . . . . . . . 71
VII. Selected Antibiotic Fermentations by Actinomycetes . . . . . . 18
A. Chloralnphenicol . . . . . . . . . . . . . . . . 81
B. Chlortetracycline . . . . . . . . . . . . . . . . 19
C. Erythromycin . . . . . . . . . . . . . . . . . 20
D. Gentamicin . . . . . . . . . . . . . . . . . 12
E. Neomycin . . . . . . . . . . . . . . . . . . 22
F. Streptomycin . . . . . . . . . . . . . . . . . 23
I. Introduction
Of the approximately 3000 known antibiotics about 70% are derived
from actinomycetes, especially species of Streptomyces, 20% from fungi,
and 10% from eubacteria. Most of the producing organisms, but by no
means all, were isolated from soils in programs designed to discover new
antibiotics. Some of the more prominent culture collections from which
many of these organisms are obtainable are the following: ATCC--
American Type Culture Collection, Rockville, Maryland; CBS--Cen-
traalbureau voor Schimmelcultures, Baarn, The Netherlands ; CMI--Com-
monwealth Mycological Institute, Kew, England; IFO--Institute for
Fermentation, Osaka, Japan; NCIB--National Collection of Industrial
Bacteria, Aberdeen, Scotland ; NRRL--Northern Utilization Research and
Development Division, U.S. Department of Agriculture, Peoria, Illinois.
II. Culture Maintenance
The growth requirements of microorganisms cover a very wide range
of conditions. However, few antibiotic producers are particularly fastidi-
ous, and most can be readily cultivated by the proper selection of media
and other environmental factors.