Ei-lchiro Ochiai Principles in Bioinorganic Chemistry
The University of British Columbia
Vancouver, B.C., Canada
Basic inorganic exercises
The biochemical roles played by essential inorganic elements and
compounds are 1) structural, 2) carrying and transporting electrons
and oxygen, 3) catalytic roles in oxidation-reduction reactions, and
4) catalytic roles in acid-base and other reactions.
Many inorganic elements and their compounds are now
known to be essential to organisms (J). Organic compounds
are of course essential, because they provide organisms with
such essentia! compounds as proteins, nucleotides, carbohy-
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drates, vitamins, and so forth. Inorganic compounds, partic-
ularly metallic ions and complexes, are essential cofactors in
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a variety of enzymes and proteins. They conceivably provide
essential services which cannot be or can only poorly be ren-
dered by organic compounds. The roles played by essential
inorganic elements and compounds are (1) structural (2)
carrying and transporting electrons and oxygen, (3) catalytic
roles in oxidation-reduction (including oxygenation) reactions
and (4) catalytic roles in acid-base and other reactions. The
type of question we are interested in here is why a certain in-
organic element (compound) is specifically required for a
certain function in biological systems. For example, why is
cobalt uniquely required for vitamin B12 and the enzymatic
reactions dependent on Bi2? Why not iron or copper? We
think that questions of this type may be answered at least
partially in terms of basic inorganic chemistry. This is what
this article attempts to show.
First let us lay out some basic principles regarding the above
question. They are
1) Rule of abundance
2) Rule of efficiency
3) Rule of basic fitness
4) Evolutionary improvement of efficiency and specificity
We will illustrate these principles by a few examples.
Rule of Abundance
The rule of abundance may be stated as: When a function
can be accomplished by two or more entities, organisms would
utilize the more abundant, readily available one. Examples
of this rule are numerous. As a general rule, the lighter ele-
ments are more abundant. The elements essential to organ-
isms are mostly light elements, lighter than atomic number
34 (selenium), with exceptions of iodine and molybdenum.
Molybdenum, as Mo042_, is present in a rather high concen-
tration, as high as iron in sea water. The concentration of zinc
in sea water is also on the same order of magnitude as that of
iron; the copper concentration is about half as high. These four
elements are the most frequently found in the catalytic sites
of enzymes. The use of the most abundant alkali metals, so-
dium and potassium, in controlling ion balance and enzyme
activities is also in accord with this rule.
Most organisms utilize calcium compounds such as car-
bonate and phosphate as protective and skeletal material.
Undoubtedly this is due to the insolubility of calcium car-
bonate and phosphate. However, the corresponding strontium
compounds are equally insoluble and could substitute calcium
compounds. It is obvious that calcium is much more abundant
than strontium (1, 2). An interesting exception is a group of
marine protist, radioralian; their outer skeletons are made of
either strontium sulfate or silica. Somehow they could not
develop a mechanism to utilize more abundant calcium or
their specific need may not be satisfied by calcium com-
pounds.
Zinc(II) in zinc-enzymes can be in most cases replaced by
cobalt(II) in vitro without losing catalytic activity (3). If or-
ganisms are grown in cobalt-rich media, they can produce
enzymes in which Zn(II) is replaced by Co(II). For example,
when Escherichia coli is grown in the presence of relatively
high concentration of 60Co(II), a catalytically active 60Co(II}-
alkaline phosphatase is obtained (4). Thus, zinc and cobalt
seem to be interchangeable, but organisms selected zinc be-
cause zinc is much more abundant both in sea and earth’s
upper crust.
A general discussion of the relationship between the
abundance of elements and their essentiality or toxicity was
made elsewhere (1, 2).
Rule of Efficiency
The rule of efficiency asserts that organisms would choose
the more efficient entity as long as it is readily available.
Flavodoxins (5) and ferredoxins function as electron-carriers
in very similar ways, being interchangeable in most cases.
However, their compositions are entirely different. Flavodoxin
contains flavin mononucleotide (FMN) as the prosthetic
group, whereas the functional units in ferredoxins are iron-
sulfur complexes. Flavodoxin in general is less efficient than
ferredoxin, and the synthesis of flavodoxin occurs only during
growth in iron-poor media in a number of microorganisms. For
example, in the case of Peptostreptococcus elsdenii, iron-rich
cells were found not to contain flavodoxin and iron-deficien-
cies brought about its de novo synthesis (5).
Rule of Basic Fitness
Primitive (in historical sense) organisms tried to improve
their survival chances by tapping whatever resources were
available to them. For example, they might have explored
many transition metal complexes in order to effect the oxi-
dation-reduction reactions. As the results of this endeavor,
they selected iron, copper, and molybdenum and their com-
plexes to effect their desired oxidation-reduction reactions.
This third basic principle might be stated as: an inorganic
element (generally a metal) to be selected should have a basic
ability or potential to carry out the desired function. That is,
a certain element (or elements) would inherently fit to a
particular function.
As mentioned earlier, most organisms utilize calcium
compounds as protective material. An important group of
organisms, diatom, use silica as their outer cover, despite the
fact that silicon is much less abundant than calcium and
strontium in sea water. Diatom is a kind of algae. There are
a vast number of different kinds of algae and all algae other
than diatom do not have any cover of inorganic material,
though there are a few green and red algae, mostly extinct,
which have slightly calcified walls. Somehow the predecessor
of diatom happened to try a coat of silica and found it very
useful for its survival. Perhaps some other algae might have
Volume 55, Number 10, October 1978 / 631

Figure 1. The reduction potentials of some important metal ions and other redox
systems. O represents the reduction potential of a metal ion in an aqueous
medium either at pH = 0 or at pH = 14, or the reduction potential of a simple
metal complex. See the text for a more detailed explanation.
tried limy coats but did not find them useful. Probably it is
due to the fact that silica forms transparent material whereas
limy coats are often much less transparent. Algae, of course,
have to absorb sunlight to support their photosynthesis.
An element may have ranges of capacities modulated by
external factors such as coordinating ligands. However, these
ranges are not limitless, and are more or less confined within
certain values, particularly when the kinds of external factors
are limited. Some of the important restrictions in the case of
ordinary biological systems are: (1) the medium is water, (2)
the range of temperature is rather narrow, and (3) the possible
coordinating ligands are limited; they are proteins, carbohy-
drates, nucleotides, lipids, and a few other specific ligands such
as porphyrins.
The catalytic effects of a metal ion can be characterized by
many factors (6). They are: (1) continuous parameters such
as reduction potential and Lewis acidity and (2) discontinuous
(discrete) parameters such as number of valence electrons and
favorable coordination number (structure). The parameters
in category (1) are modulated by ligands and other external
factors such as pH of the medium, but those in category (2)
are much less influenced by ligands and others.
Let us take reduction potential as an example and illustrate
how the rule of basic fitness may work. Figure 1 shows the
reduction potentials for several important metal ions and
other redox systems. The dotted lines represent the oxidation
and reduction of water and the pH-dependency of the re-
duction potentials. The points shown by O are the reduction
potentials of the aquo species at pH 0 and those of the cor-
=
responding hydroxo complexes at pH 14, The two points at
=
pH 0 and 14 are joined by a straight line which does not nec-
essarily represent the pH dependence of the reduction po-
tential. The reduction potentials for a few ordinary complexes
are shown on the extended broken lines. The reduction po-
tentials (at pH 7) of the metalloenzymes and metalloproteins
containing copper and iron are shown by • or A on the re-
spective straight lines or the broken lines in Figure 2.
The following items are some of the points that we can infer
from Figures 1 and 2.
(1) Since the species whose reduction potentials lie outside of the
decomposition lines of water (dotted lines) would (in thermo-
632 / Journal of Chemical Education
V
+ 1.2-
+08-
+0.4-
0-
-04-
-0.8-
Figure 2. The reduction potentials of iron- and copper-enzymes and proteins
at pH 7. The point represents merely the reduction potential value; the position
on the pH axis has no meaning. See Figure 1 for the explanation of the dotted
lines and the straight solid or broken lines.
dynamic sense) either oxidize or reduce water, they may not be
suitable for a catalytic entity working in aqueous media. This
consideration alone would probably exclude Co(III)/Co(II),
Sn(IV)/Sn(II), and Cr(III)/Cr(II) systems as candidates for
redox catalysts (enzymes).
2) The entire range of the reduction potentials of a variety of
iron-enzymes and proteins is confined in the potential range of
the iron-aquo system. The potentials of iron-sulfur proteins are
closer to that of the Fe2UIS3/2FenS system. These facts may
indicate that the prebiotic catalysts for those reactions carried
out today by iron-containing enzymes and proteins might have
been Fe(H20)6-n (OH)„ and FeS/Fe2S3 or their simple com-
plexes. These simple compounds were later incorporated into
porphyrins and proteins, leading to the formation of more ef-
ficient and selective catalysts; the potentials, however, did not
change very widely. In other words, these simple compounds
that might have been readily available to precursors of organ-
isms or primitive organisms are basically fit to the specified
requirements (chemical reactions). It should be noted, however,
that iron porphyrins themselves might have been the first cat-
alysts, since porphyrins are known to be one of the most ancient
compounds synthesized abiotically (7).
3) The situation of copper species is slightly different from that
of iron. Cuprous aquo species are not stable and dispropor-
tionate into Cu(0) and Cu(II). This would preclude the use of
the simple aquo Cu(II)/Cu(I) system as an oxidation-reduction
catalyst. Instead, such simple complexes as Cu^CL/CuCh-,
CuII(pyridine)2CuI(pyridine)2, or similar complexes could have
provided the basis for prebiotic or primitive catalyst. Cu(II)-Cl~
species could indeed be predominant in a marine environment.
The reduction potentials of these compounds are basically fit
to those of present day copper enzyme and proteins.
4) The reduction potentials of 02_/H202 and 02/02_ are +0.96 V
and —0.45 V (at pH 7), respectively. Suppose that the mecha-
nism of superoxide dismutase reaction is simply as follows
02 + M"+ + 2H+ —
H202 + M'l+1+
M"tl+ + 02“ -*
M"+ + 02
Then it would be inferred that a redox system whose potential
lies somewhere in the middle of the range +0.96 V to -0.45 V
would function as a catalyst for the superoxide dismutase re-
action. Figure 1 indicates that such systems are Fe(III)/Fe(II),
Cu(II)/Cu(I), and Mn(IlI)/Mn(II). In fact, superoxide dismu-
tases from different sources contain copper, iron, or manganese
 (8). A superoxide dismutase containing copper was found to
have a reduction potential of +0.42 V (9). As far as the reduction
potential is concerned, the aquo V(IV)/V(III) system is also fit
for superoxide dimutase function. One of the functions of
hemovanadin, which is a vanadium-containing protein found
in a marine invertebrate, ascidian, might well be a superoxide
dismutase.
Let us turn to another example, oxygen-carrying proteins
(10). This function can be translated into the reversibility of
the reaction to bind oxygen; that is
M"+ + 02^ [Mn+1+02-]: AGa
2Mn+ + 02 [Mfl+1+022-M"+1+]: AGb
—
efficient in the senses of kinetics and specificity. The substrate
specificity and the efficiency seem to be controlled mainly by
the protein portions of metalloproteins or metalloenzymes.
Thus, the improvement of these functions must have taken
place in early stages of evolution, involving the mutations in
the genetic codes. We know little about this, but we may be
able to rationalize some of the results from an inorganic
standpoint. What we are concerned with here is the efficiency
of many enzymes. Their rates are usually very much higher
than those of simple model compounds. Vallee and Williams
(13) proposed that the structure of the active site of an en-
zyme, particularly a metalloenzyme, is rather distorted or
strained and that it is responsible for its high catalytic activity.
They called it “Entatic Effect.” Let us now translate this into

In order for these reactions to be reversible, AGa or A Gb chemical language.

should be roughly —2 to —12 kcal/mole (9). If AG„ (or A Gb) Very generally speaking, the rate is most dependent on the
< —2 kcal/mole, the forward reaction occurs to only a small activation energy. The activation energy can be defined in
extent. On the other hand, if AGa (AGb) < —13 kcal/mole, the terms of absolute rate theory as the potential energy difference
reaction goes essentially to completion and the reverse reac- between the initial and the transition state, that is, the acti-
tion occurs to a negligible extent. A rough estimate for AGa vation energy AG*, for the process ML + S ^ (MLS)*, where
and AGb was made from reduction potentials and is given in ML is a metal complex and S is a substrate including electron.
the table (10). This table indicates: (1) the reversible oxy- A significant portion of AG* comes from the energy necessary
to reorganize the structure of ML so that ML take the struc-
Rough Estimates of AGa (kcal/mole) and AGb (kcal/mole) ture in (MLS)*. Therefore, the closer the structure of the
M"*
initial state ML is to that of the transition state, the lesser AG*
AGa AGb
becomes. A few examples will suffice to illustrate this
Ti(ll)aq + 1.7 -29.2 point.
Ti(lll) (in 5 F H3P04) + 6.9 -19.0 Blue copper proteins (14) such as plastocyanin and azurin
V(lliaq +4.4 -24.0 function as electron-carriers, in which copper oscillates be-
Cr(ll)aq + 0.9 -31.0 tween Cu(II) and Cu(I). The favorable coordination structure
Mrx(ll) (in 8 FH2S04) +47.7 +62.6
+ 5.2 -22.4
of Cu(II) is square planar and that of Cu(I) is tetrahedral. The
Mn(ll)/CN~
Fe(1l) (pH
<
2) +28.0 +23.2 reduction of a square planar Cu(II), therefore, would require
+ 10.0 -11.8 a significant rearrangement in the structure about copper ion.
Fe(ll) (pH ~
6)
Fe(OH)2 -2.6 -38.0 One way to reduce this cost is to start with a compound whose
Fe(il)/CN- + 16.6 -4.4 structure is between the regular square planar and the regular
Co(ll)aq +52.7 +72.6 tetrahedral one. There is now convincing evidence that the
Co(ll)NH3 + 12.6 -7.6 coordination structure about copper atom in the blue proteins
Co{ll)/CN~ -9.0 -50.8 is indeed distorted-tetrahedral (see (15) for review).
Cu(l)/aq + 13.8 -4,8
Other examples are zinc enzymes (16). The structural
Cu(I(/NH3 + 10.1 -12.6
studies by X-ray crystallography and by spectroscopy of the
cobalt (Il)-substituted enzymes have established that the
genation of type (a) is very probable with some Fe(II) and coordination structures about zinc atom in carbonic anhy-
Co(II) complexes and possibly some Cr(II) complexes, and (2)
the reversible oxygenation of type (b) may be carried out by drase, carboxypeptidase A, and alkaline phosphatase are
distorted tetrahedra, being between a regular tetrahedron and
some Fe{II), Co(II), and Cu(I) complexes. This conclusion fits
a five-coordinate trigonal bipyramid (3). The function of these
the known facts (9).
An example of the effect of discrete parameters seems to enzymes is to hydrate the substrates. Not only the substrate
but also the water molecule would have to be activated. The
be vitamin Bi2, cobalamin. The mechanisms of Bi2 coen-
distorted tetrahedral structures of the enzymes usually have
zyme-dependent enzymatic reactions are still under intensive one water molecule and three amino acid residues coordinated
study and are far from settled (11,12). However, the impor- about the zinc ion. If the coordination structure about the zinc
tant requirements for the candidates for the job appear to be
atom is distorted from tetrahedron in such a way that the
rather well defined. They are: (I) it should readily take three
substrate can readily approach the zinc ion (forming a five-
consecutive oxidation states, for example, I, II, and III in
coordinate bipyramidal type of structure), the whole reaction
aqueous media, (2) the lowest oxidation state should be highly would be facilitated. In a regular tetrahedron, this binding of
nucleophilic, and (3) the middle oxidation state should per- an additional ligand would not be very easy.
haps have one unpaired electron (11). The requirement (2)
implies that the lowest oxidation state of the catalytic metal Literature Cited
ion should have ds or configuration. This condition may (1) Ochiai, E. -I., “Bioinorganic Chemistry: An Introduction," Aliya and Bacon. Inc.,
be satisfied by Fe(O), Co(I), Ni(O), and Cu(I). Fe(O), and 1977.
(2) Ochiai, E. -I., J. CHEM. EDUC., 51, 235 (1974); Chap. 1 and 17 in ref. (J),
Ni(O) are not readily attainable, particularly in aqueous (3) Ochiai, E. -I., Chap. 13 in ref, {/).
media. This leaves us Co(I) and Cu(I). Cu(I) is not particularly (4) Harris, M. I., and Coleman, J. E., J. Biol. Chem., 244, 709 (1964).
(5) Mayhew, S. G., and Ludwig, M. L., in “The Enzymes, 3rd Ed,, XII,” (Editor: Boyer,
nucleophilic and Cu(III) is not readily obtained. Only a P. D.) Academic Press, 1975, p, 57.
Co(III)/Co(II)/Co(I) system satisfies all these requirements. (6) Ochiai, E. -I., Coordin. Chem. Rev., 3, 49 (1968).
This explains why cobalt (cobalamin) uniquely fits the job (7) e.g., Miller, S. L., and Orgel, L. E., “The Origins of Life on Earth," Prentice-Hall, Inc.,
1974.
(11). (8) Fee, J. A., and di Corleto, P. Q-, Biochemistry, 12, 4893 (1973).
Other applications of the rule of basic fitness to more spe- (9) Fridovich, I., in “Free Radicals in Biology,” Vol. 1, (Editor: Pryor, W. A.), Academic
Press, 1976, p. 239.
cific cases will be discussed elsewhere. (10) Ochiai, E. -I., J. Inorg. Nucl. Chem., 35, 3375 (1973); Chap. 10 in ref. (7).
(11) Ochiai, E. -1 Inorg. Nucl. Chem., 37, 351 (1975); Chap. 12 in ref. (7).
Evolutionary Improvement of Efficiency and Specificity (12) Abeles, R. H., in “Biological Aspects of Inorganic Chemistry,” (Editors: Addison, A.
W., et al.), Wiley-Interscience, 1977, p. 245.
The rule of basic fitness dictates which element(s) is suit- (13) Vallee, B. L., and Williams, R. J. P., Proc. Nat. Acad. Sci. [US\, 59,498 (1968).
able for a specific enzymatic reaction only in terms of ther- (14) Fee, J. A., Struct, and Bondg., 23, 1 (1975).
(15) Yokoi, H., and Addison, A. W., Inorg. Chem., 16, 1341 (1977).
modynamics. The element or its simpler compounds selected (16) Chlebowski, J. F., and Coleman, J. E., in “Metal Ions in Biological Systems,” Vol. 6,
may be able to do the required function but may not be very [Editor: Sigel, H.), Marcel Dekker, 1976, p. 1,

Volume 55, Number 10, October 1978 l 633