Vitamin B6 Antagonists and Growth of Microorganisms. II. S-Desoxypyridoxal and Related Compounds
Vitamin B6 Antagonists and Growth of Microorganisms. II. S-Desoxypyridoxal and Related Compounds
Vitamin B6 Antagonists and Growth of Microorganisms. II. S-Desoxypyridoxal and Related Compounds
TABLE I
Inhibitory Analogs of Vitamin Be
a In some earlier literature (9, 14, 15) this compound has been referred to as
“ethylpyridoxine”.
TABLE II
The Comparative Inhibitory Potencies of Vitamin Be Antagonists for
Saccharomyces carlsbergensis
Reversing agent”
Inhibitor Pyridoxal Pyridoxamine Pyridoxine
Inhibition ratio*
5-Desoxypyridoxal 440 190 1,499
5-Desoxypyridoxamine 12,099 l-,800 35,999
5-Desoxypyridoxine 430 120 6,509
4-Desoxypyridoxinec 27 ‘28 135
w-Methylpyridoxinec 16 16 43
a Fifty millimicrograms of pyridoxamine dihydrochloride or equimolar
amounts of pyridoxal hydrochloride or pyridoxine hydrochloride per 6 ml. were
added unless otherwise indicated.
* The molar ratio of inhibitor to reversing agent at which growth was de-
pressed to 5Oye of that obtained in the absence of added inhibitor. The inhibition
ratio between 5desoxypyridoxal and pyridoxamine in the presence of 100 and
1009 nmg. of the vitamin was 150 and 60, respectively.
c Determined in presence of 109 mFg. of reversing agent.
TABLE III
The Comparative Inhibitory Potencies of Vitamin Be Antagonists for
Streptococcus faecalis
Reversing agent
Pyridoxamine
Inhibitor Pyridoxal P&doxamine phosphate
Inhibition ratio”
5-Desoxypyridoxal 2,700 48,000 4,40@
5-Desoxypyridoxamine 400 5,000 7ob
5-Desoxypyridoxine >200,000 >200,000 >800,000
4-Desoxypyridoxine - >500,000 >80,ooo
w-Methylpyridoxine - >200,000 >75,000
a Defined as in Table II and determined in the presence of 5 mpg. of pyridox-
amine dihydrochloride, 10 mpg. of pyridoxal hydrochloride, or 15 wg. of calcium
pyridoxamine phosphate.
b These represent the average of the values determined in the presence of 10,
20,50, and 100 mpg. of calcium pyridoxamine phosphate. With Sidesoxypyridoxal
as the inhibitor, the individual values were 6700,3400,3100, and 4400, respectively.
With 5-desoxypyridoxamine as the inhibitor the corresponding values were 91,
38, 91, and 60.
promoting growth in the absence of inhibitors (5), but is far less active
than pyridoxamine in reversing the inhibitory effects of these analogs.
Under the conditions of growth used here, synthesis of n-alanine
represents the sole reaction for which an external source of vitamin Be
is required by S. jaecalis, and when this amino acid is supplied preformed,
growth occurs in the absence of this vitamin (6, 7). When the sole
function for which vitamin Ba is required is by-passed in this way,
5desoxypyridoxal and 5-desoxypyridoxamine no longer inhibit growth
of 8. jaecalis.
The effectiveness of these inhibitors for various lactic acid bacteria
VITAMIN Bg ANTAGONISTS. II 411
TABLE IV
The Comparative Inhibitory Potencies of Vitamin Be Antagonists for
Lactobacillus helveticus
Inhibitor Inhibition ratid
5-Desoxypyridoxal ‘19,000
5-Desoxypyridoxsmine >91o,ooo
5-Desoxypyridoxine >910,000
0 Determined in the presence of 10 mg. of calcium pyridoxamine phosphate/
10 ml. Pyridoxal and pyridoxamine are essentially inactive as growth factors
for this organism (8).
4 -DESOXYP(fUCClXIN~ HC I 5- DESOXYPYRIDOXAL
MICROGRAMS PER 6 MILLILITERS
FIG. 1. Inhibition of S. carlsbergensis by 4-desoxypyridoxine, 5-desoxypyri-
doxal, and mixtures of the two. All tubes contained 69 wg. of pyridoxamine di-
hydrochloride and 10 pg. of thiamine chloride in addition to the usual
constituents of the medium (10).
1: Response to 4-desoxypyridoxine.
I: As in 1 plus 2 I.cg.of 5-desoxypyridoxal/b ml.
3: Response to 5-desoxypyridoxsl.
.J: As in 3 plus 0.6 pg. of 4-desoxypyridoxine/b ml.
c,yMETHYLPYRlDOXlNE
MICROGRAMS PER 6 ML.
FIG. 2. The inhibition of S. carlsbergensis by w9ethylpyridoxine in the ab-
sence and presence of other inhibitors. Each tube contains thiamine and pyri-
doxamine as in Fig. 1.
f: w-Methylpyridoxine alone.
8: w-Methylpyridoxine plus 0.6 pg. of 4-desoxypyridoxine/6 ml.
3: w-Methylpyridoxine plus 2.0 pg. of 5-desoxypyridoxal/6 ml.
DISCUSSION
SUMMARY
REFERENCES
1. RABINOWITZ, J. C., AND SNELL, E. E., Arch. Biochem. and Biophye. 43, 399
(1953).
2. HEYL. D., HARRIS, S. A., AND FOLKERS, K., J. Am. Chem. Sot. in press.
VITAMIN B,y ANTAGONISTS. II 415
3. RABINOWITZ, J. C., AND SNELL, E. E., J. Biol. Chem. 169, 631 (1947).
4. SNELL, E. E., AND RANNEFELD, A. N., J. Biol. Chem. 167, 475 (1945).
5. RABINOWITZ, J. C., AND SNELL, E. E., J. Biol. Chem. 189, 643 (1947).
6. HOLDEN, J. T., AND SNELL, E. E., J. Biol. Chem. 178, 799 (1949).
7. SNELL, E. E., Symposium sur le Metabolism Microbien, p. 47, IIe Congr&s
International de Biochimie, Paris, 1952.
8. MCNUTT, W. S., AND SNELL, E. E., J. Biol. Chem. 173,801 (1948).
9. WOOLLEY, D. W., A Study of Antimetabolites. John Wiley and Sons, New
York, 1952.
10. RABINOWITZ, J. C., AND SNELL, E. E., Anal. Chem. 19, 277 (1947).
11. UMBREIT, W. W., AND WADDELL, J. W., Proc. Sot. Exptl. Biol. Med. ‘IO, 293
(1949).
12. SNELL, E. E., AND SHIVE, W., J. Biol. Chem. 168,551 (1945).
13. SHIVE, W., AND SNELL, E. E,, Science 162, 401 (1945).
14. ROBBINS, W. J., Am. J. Botany 29, 241 (1942).
15. ROBBINS, W. J., AND MA, R., Bull. Torrey Botan. Club 69, 342 (1942).