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Pasteur Institute, Paris, France: BY Monod
Pasteur Institute, Paris, France: BY Monod
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THE
GROWTH OF
BACTERIAL
CULTURES
BY JACQUES MONOD
PasteurInstitute, Paris, France
INTRODUCTION
The study of the growth of bacterial cultures does not constitute a specialized subject or branch of research: it is the basic
method of Microbiology. It would be a foolish enterprise,
and
doomed to failure,
to attempt reviewing briefly a "subject"
which covers actually our whole discipline. Unless, of course, we
considered the formal laws of growth for their own sake, an approach which has repeatedly proved sterile.
In the present review
we shall consider bacterial growth as a method for the study of
bacterial physiology and biochemistry. More precisely, we shall
concern ourselves with the quantitative
aspects of the method,
with the interpretation of quantitative data referring to bacterial
growth. Furthermore, we shall consider z exclusively the positive
phases of growth, since the study of bacterial "death," i.e., of the
negative phases of growth, involves distinct problems and methods. The discussion will be limited to populations considered
genetically homogeneous. The problems of mutation and selection
in growing cultures have been excellently dealt with in recent
review articles by Delbriick (1) and Luria (2).
No attempt is made at reviewing the literature
on a subject
which, as we have just seen, is not really a subject at all. The
papers and results quoted have been selected as illustrations of the
points discussed.
DEFINITION
OF GROWTH PHASES
AND GROWTH CONSTANTS
Division
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per unit
x = xx2;
after n divisions it will be
n.
x -- xt.2
If r is the number of divisions per unit time, we have at time t~:
log2x2 - log2xl
...............
[11
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6
TIME
FzG.1.--Phases o[ growth.Lowercurve: log bacterial density. Uppercurve:
variations of growthrate. Vertical dottedlines markthe limits of phases.Figures
refer to phasesas definedin text (see p. 373).
GROWTH CONSTANTS
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~
I /
/
~
/I
I
I
i
I
FIG. 2.--Lag time and growth lag. ~lid line~ob~rved growth. Dotted
line ~ "ideal growth" (without lag). T~=lag time. L=growthlag. (~
p. 375.)
very limited. Centrifugal techniques have been found of value
(9).
The most widely used methods, by far, are based on determinations of transmitted or scattered light. (Actually, the introduction
around 1935 of instruments fitted with photoelectric
cells has
contributed to a very large degree to the development of quantitative studies of bacterial
growth.) We cannot go here into the
physical aspects of this problem [for a discussion of these see
(10)]. What should be noted is that in spite of the widespread use
of the optical techniques, not enough efforts have been made to
check them against direct estimations of cell concentrations or
bacterial densities. Furthermore a variety of instruments, based
on different principles, are in use. The readings of these instruments are often quoted without reference to direct estimations as
arbitrary units of turbidity, the word being used in an undefined
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GROWTH
OF BACTERIALCULTURES
377
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metabolic activity
OF THE
of bacterial
cells
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15o $ 200
[/~. I~IANNffOL
PER
cm
FIG. 3.--Total growthof E. oli in synthetic mediumwith organic source
(mannltol)as limiting factor. Ordinates:arbitrary units. Oneunit is equivalent
to 0.8/~g. dry weightper ml. (11).
50
measure
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growth of purple
baeterla
with acetate
as
0.5
1.0
2.0
3.0
Total growth
(mg/ml.)
0.18
0.36
0.70
1.12
0.36
0.36
0.35
0.37
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~1.0
Fx6. 4.~Growth
rate of E. col in synthetic medium
as a function of glucose
concentration.Solidline is drawnto equation(2) withRK= 1.35 divisionsper hour,
-4 (11). Temperature
C.
and Ct =0.22MX10
be determined by a single master reaction. But such a situation
could hardly be assumed to prevail, in any one case, without direct
experimental evidence. Somerecent attempts at making use of the
master reaction concept in the interpretation
of bacterial growth
rates are quite unconvincing in that respect (19).
Rate-concentration relations.--Notwithstanding
these difficulties, relatively simple empirical laws are found to express conveniently the relation between exponential growth rate and concentration of an essential nutrient. Examples are provided in Figs. 4 and
5. Several mathematically different formulations could be made to
fit the data. But it is both convenient and logical to adopt a
hyperbolic equation:
C
R = RK........................
Ct+C
[21
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max~mumo
o.o~o
~0.o2s
-~ 0.o~o
o.o|o
!
O.1
t
I
O~.
O.~
GLUCOSE(Mx)
~--~/~5 (20~.
organic sources are compared under otherwise identical conditions.
There is no doubt that it is related to the activity of the specific
enzyme systems involved in the breakdown of the different compounds, and it can be used with advantage for the detection of
specific changes (e.g., hereditary variation) affecting one or another of these systems (30).
The value of C~ should similarly be expected to bear some
more or less distant relation to the apparent dissociation constant
of the enzyme involved in the first step of the breakdown of a
given compound. Furthermore, since a change of conditions affecting primarily the velocity of only one rate-determlning step will,
in general (but not necessarily), be only partially reflected in the
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knowledge, this has been clearly observed only once (25), actually
during the residual growth of a streptomycin-requiring B. subtilis
in a mediumcontaining no streptomycin (Fig. 6). The interpretation is obvious, albeit surprising. Growth must be limited by one
enzyme or system of enzymes, the activity of which is constant. In
other words, in the absence of streptomycin, one rate-determining
enzyme ceases to be formed, so that by being outgrown by the
, f
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HOURS
FIG. 7.--Growth of E. toll in synthetic medium with glucose (circles)
and
xylose (squares) as organic source. Culture previously maintained on arabinose
medium, temperature 37 C. Growth on glucose proceeds without any lag. Lag
time (T;) on xylose is approximatlvely 2.5 hours (46).
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~05
~4
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~6o
~.,~o
0
~o
2o
Io
Hours
FI6. 9.--Diauxie. Growthof E. coli in synthetic mediumwith glucose+sorbitol as carbon source.
The figures betweenarrows indicate total growth correspondingto each cycle.
(a) Glucose50/zg. per ml.; sorbitol 150 #g. per ml.
(b) Glucose100/~g. per ml.; sorbitol 100/~g. per ml.
(c) Glucose 150 #g. per ml.; sorbitol 50 #g. per ml.
Total growthcorrespondingto first cycle is proportional to glucose concentration. Total growthof secondcycle is proportional to sorbitol concentration(1 I).
of interactions between closely related compoundsin the formation of specific enzymesand has proved valuable in the study
of certain aspects of enzymatic adaptations (11, 30, 35). It may
perhaps be susceptible of certain technical applications, e.g., for
the quantitative analysis of certain mixtures of carbohydrates.
CONCLUDING REMARKS
The time-honored methodof looking at a tube, shaking it, and
looking again before writing downa + or a 0 in the lab-book
has led to many a great discovery. Its gradual replacement by
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LITERATURE CITED
1. DELBR~CK,
M., Ann. Missouri Botan. Garden, 32, 223-33 (1945)
2. LURIA,S. E., Bact. Revs., 11, 1-40 (1947)
3. HENRICI,
A. T., MorphologiVariation and the Rate of Growth"of Bacteria, 194
pp. (C. C Thomas,Springfield, Ill., 1928)
4. WILSON,
G. S., .d. Bact., 7, 405 (1922)
5. I~LLy, C. D., ANDRAHN,
O., ar. Bact., 23, 147 (1932)
6. LODGE,
R. M., ANDHINSHELWOOn,
C. N., J. Chem.Sot., 213-219 (1943)
7. FISHER,K. C., ANDARMSTRONG,
F. H., ar. Gen. Physiol., 30, 263 (1947)
8. MCILWAIN,
H., Biochem.f., 38, 97-105 (1944)
9. VAN
NIEL,C. B., Bact. Revs., 8~ 1-118 (1944)
10. DOGNOI~,
A., in Techni,~ues de laboratoire, 197-210 (Masson& Cie, Paris,
1947)
11. MONOD,
J., Recherchessur la croissanee des cultures bact~riennes,211 pp. (Hermann& Cie, Paris, 1942)
12. DtlBOS,R. J., ar. Exptl. Med., 65, 873-83(1937)
13. MONOD,
J., Ann. inst. Pasteur, 68, 444 (1942)
14. MoNoi),J., Ann. inst. Pasteur (In press)
15. JORI)AI~,R. C., Am)JACOBS,
S. E., d. Bact., 48, 579 (1944)
16. LwovI~,A., VANNIEL,C. B., RYAN,
F. J., AN~)TATI~M,
E. L., Cold Spring
HarborSymposiaQuant. Biol., 11, 302-3 (1946)
17. Bt/RTOI~I,A. C., J. Cellular Comp.Physiol., 9~ 1 (1936)
18. HINSHELWOOn,
C. N., The Chemical Kinetics of the Bacterial Cell, 284 pp.
(Clarendor/Press, Oxford, 1946)
19. JOHNSON,
F. H., A~r~LEWIN,I., dr. Cellular Comp.Physiol., 28, 47 (1946)
20. SC~Am*ER,
W., Ann. inst. Pasteur, 74, 458-63 (1948)
21. MCILWAIN,
H., Biol. Keys., 19, 135 (1944)
22. McILwAII%
H., Advances in Enzymol., 7~ 409-60 (1947)
23. WYss,0., Proc. Soc. Exptl. Biol. Med., 48, 122 (1941)
24. KOHN,
H. I., ANDHARRIS,
J. S., J. Pharmacol.Exptl. Therap., 73, 343 (1941)
25. SCHAE~VER,
P., Compt.rend., 228, 277-79 (1949)
26. LwoF~,A., ANDMONOn,
J., Ann. inst. Pasteur, 73, 323 (1947)
27. MCILWAm,
H., FrLDES,P., GL~t~SxOI~m,
G. P., Am)K~IG~T,B. C. J. G., Biochem.at., 33, 223(1939)
28. LODGE,
R. M., ANDHINSHELWOOD,
C. N., ar. Chem.Soc., 1692-97 (1939)
29. MOREL~
M., Ann. inst. Pasteur, 67, 449 (1941)
30. MONOD,
J., Growth, 11,223-89 (1947)
31. MoNon,J., Ann. inst. Pasteur, 69, 179 (1943)
32. POLLOCK,
M. R., ANDWAINWRIGHT,
S. D., Brit. J. Exptl. Path., 29, 223-40
(1948)
33. STANIER,
R. Y., ar. Bact., ~4, 339 (1947)
34. COHEN,
S. S., ar. Biol. Chem.,177, 607-19(1949)
35. MoNon,J., Ann. inst. Pasteur, 71, 37 (1945)
36. SPIEGELMAN,
S., Am)DUNN,
R., Y. Gen. Physiol., 31, 153-73 (1947)
37. SPIE6ELM.~.~,
S., Cold Spring HarborSymposiaQuant. Biol., 11, 256-77 (1946)
38. WINSLOW,
C. E., ANDWALK~R,
H. H., Bact. Revs., 3, 147-86 (1939)
39. HERSHI~Y,
A. D., ar. Bact., 37~290 (1939)
Annual Reviews
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394
HERSHEY,
A. D., Proc. Soc. Exptl. Biol. Med., 38, 127-28 (1938)
LWOF~,
A., Cold Spring HarborSymposia Quant. Biol., 11~ 139-55 (1946)
LEMON,
C. G., or. Hyg., 33, 495 (1937)
TO~LEY,
W.W.C., ANDWX~.SON,
G. S., Principles of Bacteriology and Immunity, 3rd Ed., 2054pp. (Williams &Wilkins, 1946)
44. LWoF~,A., QUERmO,
A., ~,N~ LATASTE,
C., Compt.rend. soc. biol., 130~ 1580
(1939)
45. MONOD~
J. (Unpublished data)
46. MoNo~,J. (Unpublished data)
Annu. Rev. Microbiol. 1949.3:371-394. Downloaded from arjournals.annualreviews.org
by California Institute of Technology on 09/17/07. For personal use only.
40.
41.
42.
43.
MONOD