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1 ( 1 ) : 3 - - 8 , 1959
~T]~PA.N K U B : [ N
Souhrn
Summary
The author has studied the relation of the catalase activity in thermal
blue-green algae to temperature. Experimental data were taken from the
first phase of the reaction, so that oxidation of the enzyme b y the substrate
was eliminated. The relation of the activity of a homogenate of thermal blue-
green algae to temperature gives a curve similar to that obtained b y SIZER
(1944) for pure enzyme preparations isolated from beef livers. The resistance
of the enzyme to high temperatures is the same in thermal blue-green algae
as in mesopholic seaweeds. The catalase of thermal blue-green algae does
not exhibit any special characteristics differing from the catalase of mesophiles.
Introduction
The thermophily of some organisms still remains incompletely explained,
although we know b y now for certain that their metabolism is essentially
the same as that of mesophiles, that no natural division with regard to temper-
ature exists between thermophiles and mesophfles and that thermophily is
a characteristic which organisms of the same species, can gain and lose (Sporo-
vibrio desul/uricans ).
4 ~. KUBIN
On the basis of her experiments ALLEN (1950) has come to the conclusion
that the nature of thermophily is not to be sought in the specific resistance
of plasmatic proteins to high temperatures. In her opinion the more intense
metabolism at high temperatures is responsible for the higher resistance of
thermophilic organisms, the synthesis of their enzymes and other plasmatic
components taking place more rapidly than their denaturation. This assump-
tion has been based on the direct relationship between consumption of nutri-
ents from the environment and rising temperature. With insufficient nutrients
thermophilic species or strains (Bacteria) perish at high temperatures as
quickly as mesophilic species.
Blue-green algae are characterised by the wide temperature limits within
which they can exist and the thermophily of many species can be regarded
rather as facultative than strict. In connection with the study of the tempera-
ture dependence of photosynthesis and respiration in blue-green algae the
author also observed the dependence of catalase activity on temperature.
- - dC/dt ~ k E
- - dC/dt = kC
P. Vm
h = 2h 1 + - - ,
Vg
p V = (P + h ) . V g (1)
pV = (P + 2h~). ( V g + Vm) (2)
Vg -[- V m P . Vm
h = 2h 1 -- -~ - -
Vg Vg
Since
Vg-~- V m ~ V g ,
then
Vg + V m
Vgo --
a n d therefore this expression can be ignored in the formula. W i t h i n t h e pressure range of the
W a r b u r g m a n o m e t e r it is possible in m o s t cases to ignore the error (less t h a n 30/o) so produced.
F o r example, if Vg = 16,850 1., t t h l ~ 300 ram. a n d V m ~ 800/tl. (the v o l u m e of gas correspond-
ing to 1 ram. of m a n o m e t r i c liquid m u s t be k n o w n ; in o u r case it is 2.67 td.) Calculating h
according to the original f o r m u l a the r e s u l t a n t value is 1105 ram., w h e n using the simplified
f o r m u l a h is 1075 ram.
W i t h i n the pressure range of the W a r b u r g m a n o m e t e r the relation b e t w e e n h a n d h 1 is also
practically linear, w h i c h can be usefully e m p l o y e d in the graphic evaluation of the results.
Time intervals corresponding to a change in pressure of 10 ram. of m a n o m e t r i c liquid were
registered photographically. A spring m a c h i n e was used in which a c o n s t a n t speed was ensured
b y a centrifugal regulator. To the machine there was a t t a c h e d a d r u m w i t h grooves for the in-
sertion of polarographic p a p e r a n d w i t h a signalling a p p a r a t u s to a n n o u n c e the beginning a n d
the end of the record paper. The speed of r o t a t i o n w a s one t u r n in t w o a n d a h a l f m i n u t e s (r
= 8 cm.). Opposite the d r u m there w a s a slit w i t h t w o signal electric b u l b s one above the other.
The c o n t a c t s of m e t r o n o m e were connected to the circuit of one b u l b recording the interval of
one second. The circuit of the other b u l b was connected b y the pressing of a switch at the m o m e n t
I ! I !
300 ' i
I
/-
I
/
250 0,8
200
,.; 0,6
-,t,
0///
/0
o
:L
100
q2
5C
G I | I | I
0 20 40 60 80 sec. 0 20 40 6O
~
Fig. 2. R a t e of liberation of o x y g e n f r o m Fig. 3. Relation of the catalytic activity of
0.1 M solution of h y d r o g e n peroxide a t va- the Oscillatoria h o m o g e n a t e to t e m p e r a t u r e .
rious t e m p e r a t u r e s . The reaction w a s catal- The m e a s u r e of activity of the h o m o g e n a t e
ysed b y a h o m o g e n a t e of the alga Oscilla. is the original rate of decomposition of h y d r o -
toria at pI-I = 7. gen peroxide expressed in micromols of I~O~
distributed in seconds.
Results
sumed exponential curve cannot be fully explained since the work was not
carried out with the pure enzyme but with a suspension of homogenised mate-
rial. Therefore, not even the logarithmic expression of the constant in relation
to 1/T gives a straight line (fig. 4).
Activation energies for individual temperature intervals were calculated from
Arrhenius's equation ~ 2-303 9 R logI/T1
k 2 -__
- l ol/T2
g k~ (where I~ is the gas constant
expressed as 1.987 cal./mol.). They
are set out in the following table: o.5
0 to 50 ~ C 4100 cal.
0 to 10 ~ C 5500 cal. t ~
10 to 40 ~ C 2900 cal. O0
40 to 50 ~ C 6400 cal.
50 to 60 ~ C 59,000 cal.
References
FRv~o~, J. S., Si~O~DS, S.: General Biochemistry. (pp. 235--260). New York, 1954.
POTTER, V. 1~., ELVEH~EYl, C. A.: (cit. after Kleinzeller, A., MAlek, J., Vrba, R.: Manometrick6
motherly a jejich pou~iti v biologii a biochemii. Praha, 1954): Manometric methods and their
application in biology and biochemistry. - - J. biol. Chem. 114 : 495--504, 1936.
SIZER, I. W.: Temperature activation and inactivation of the crystalline catalase-hydrogen
peroxide system. - - J. biol. Chem. 154 : 461--473, 1944.
TAKAGI, M.: On the stability against heat of the catalase in marine algae. - - Bull. Jap. Soc. Sci.
Fish. 19 (8) : 886--888, 1953.
Address: ~t~pAn Kubin, prom. biol., Botanical Garden of the Slovak Academy of Sciences,
Komcnsk~ho 67, Ko~ice, Czechoslovakia.
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