Purification and Properties of Lactate Racemase from Lactobacillus sake

TETSUO HIYAMA; SAKUZO FUKUI; KAKUO KITAHARA

Purification and Properties of Lactate Racemase from Lactobacillus sake

TETSUO HIYAMA, SAKUZO FUKUI, KAKUO KITAHARA

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1968 Volume 64 Issue 1 Pages 99-107

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Published: July 25, 1968 Received: February 12, 1968 Available on J-STAGE: June 30, 2008 Accepted: - Advance online publication: - Revised: -
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Date of correction: June 30, 2008 Reason for correction: - Correction: ABSTRACT Details: Wrong : A lactate racemase [EC 5. 1. 2. 1] was isolated and purified about 190 fold in specific activity from the sonic extract of Lactobacillus sake. The purified preparation was almost homogeneous by ultracentrifugal and electrophoretical analyses. Characteristic properties of the enzyme were as follows: (a) absorption spectrum showed a single peak at 274 mμ, (b) molecular weight was 25, 000, (c) the enzyme did not require any additional cofactors and showed no activity of lactate dehydrogenase, (d) K_m values were 1.7×10^-2M and 8.0×10^-2M for D- and L-lactate, respectively, (e) optimal pH was 5.8-6.2, (f) an equilibrium point was at a molar ratio of 1/1 (L-isomer/D-isomer), (g) the enzyme activity was inhibited by atebrin, adenosine monosulfate, oxamate and some of Fechelating agents, (h) pyruvate and acrylate were not incorporated into lactate during the reaction, and (i) exchange reaction of hydrogen between lactate and water did not occur during the reaction.
Right : A lactate racemase [EC 5. 1. 2. 1] was isolated and purified about 190 fold in specific activity from the sonic extract of Lactobacillus sake. The purified preparation was almost homogeneous by ultracentrifugal and electrophoretical analyses. Characteristic properties of the enzyme were as follows: (a) absorption spectrum showed a single peak at 274 mμ, (b) molecular weight was 25, 000, (c) the enzyme did not require any additional cofactors and showed no activity of lactate dehydrogenase, (d) K_m values were 1.7×10^-2M and 8.0×10^-2M for D- and L-lactate, respectively, (e) optimal pH was 5.8-6.2, (f) an equilibrium point was at a molar ratio of 1/1 (L-isomer/D-isomer), (g) the enzyme activity was inhibited by atebrin, adenosine monosulfate, oxamate and some of Fechelating agents, (h) pyruvate and acrylate were not incorporated into lactate during the reaction, and (i) exchange reaction of hydrogen between lactate and water did not occur during the reaction.

Date of correction: June 30, 2008 Reason for correction: - Correction: CITATION Details: Wrong : (1) H. Katagiri and K. Kitahara, J. Agr. Chem. Soc. Japan, 12, 844 (1936)
(2) W. A. Wood and I. C. Gunsalus, J. Biol. Chem., 190, 403 (1951)
(3) G. D. Shockman and G. Toennies, Arch. Biochem. Biophys., 50, 1, 9 (1954)
(4) M. Tanaka, Y. Kato, and S. Kinoshita, Biochem. Biophys. Res. Commun., 4, 114 (1961)
(5) T. C. Stadtman and P. Elliott, J. Biol. Chem., 228, 983 (1958)
(6) A. Ichihara, S. Furiya, and M. Suda, J. Biochem., 48, 277 (1960)
(7) H. Amos, J. Am. Chem. Soc., 76, 3858 (1954)
(8) C. F. Gunsalus, R. Y. Stanier, and I. C. Gunsalus, J. Bacterial., 66, 548 (1953)
(9) H. Katagiri, H. Sugimori, and K. Imai, Agr. Biol. Chem., 25, 281 (1961)
(10) D. Dennis and N. O. Kaplan, Biochem. Z., 338, 485 (1963)
(11) T. Hiyama, S. Mizushima, and K. Kitahara, J. Gen. Appl. Microbiol., 11, 51 (1965)
(12) T. Hiyama, Y. Koga, and S. Fukui, J. Gen. Appl. Microbiol., 13, 121 (1967)
(13) H. D. Harn and F. H. Bruns, Biochim, Biophys. Acta, 21, 378 (1956)
(14) L. Ornstein and B. J. Davis, Ann. N. N. Acad. Sci., 121, Art 2, 321, 404 (1964)
(15) P. Andrews, Biochem. J, 91, 222 (1964)
(16) S. Mizushima, T. Hiyama, and K. Kitahara, J. Gen. Appl. Microbiol., 10, 33 (1964)
(17) S. Mizushima and K. Kitahara, J Gen. Appl. Microbial., 8, 130 (1962)
(18) F. Egami and K. Yagi, J. Biochem., 43, 153 (1956)
(19) D. Dennis and N. O. Kaplan, J. Biol. Chem., 235, 810 (1960)
(20) H. Takahashi, T. Hattori, and B. Maruo, Anal. Chem., 35, 1982 (1963)
(21) S. S. Shapiro and D. Dennis, Biochemistry, 4, 2283 (1965)
(22) K. Kitahara, A. Obayashi, and S. Fukui, Proc. Int. Symp. Enz. Chem., Tokyo and Kyoto, 1957, p. 460 (1958)

Right : (1) H. Katagiri and K. Kitahara, J. Agr. Chem. Soc. Japan, 12, 844 (1936)
(2) W. A. Wood and I. C. Gunsalus, J. Biol. Chem., 190, 403 (1951)
(3) G. D. Shockman and G. Toennies, Arch. Biochem. Biophys., 50, 1, 9 (1954)
(4) M. Tanaka, Y. Kato, and S. Kinoshita, Biochem. Biophys. Res. Commun., 4, 114 (1961)
(5) T. C. Stadtman and P. Elliott, J. Biol. Chem., 228, 983 (1958)
(6) A. Ichihara, S. Furiya, and M. Suda, J. Biochem., 48, 277 (1960)
(7) H. Amos, J. Am. Chem. Soc., 76, 3858 (1954)
(8) C. F. Gunsalus, R. Y. Stanier, and I. C. Gunsalus, J. Bacteriol., 66, 548 (1953)
(9) H. Katagiri, H. Sugimori, and K. Imai, Agr. Biol. Chem., 25, 281 (1961)
(10) D. Dennis and N. O. Kaplan, Biochem. Z., 338, 485 (1963)
(11) T. Hiyama, S. Mizushima, and K. Kitahara, J. Gen. Appl. Microbiol., 11, 51 (1965)
(12) T. Hiyama, Y. Koga, and S. Fukui, J. Gen. Appl. Microbiol., 13, 121 (1967)
(13) H. D. Harn and F. H. Bruns, Biochim, Biophys. Acta, 21, 378 (1956)
(14) L. Ornstein and B. J. Davis, Ann. N. N. Acad. Sci., 121, Art 2, 321, 404 (1964)
(15) P. Andrews, Biochem. J, 91, 222 (1964)
(16) S. Mizushima, T. Hiyama, and K. Kitahara, J. Gen. Appl. Microbiol., 10, 33 (1964)
(17) S. Mizushima and K. Kitahara, J. Gen. Appl. Microbiol., 8, 130 (1962)
(18) F. Egami and K. Yagi, J. Biochem., 43, 153 (1956)
(19) D. Dennis and N. O. Kaplan, J. Biol. Chem., 235, 810 (1960)
(20) H. Takahashi, T. Hattori, and B. Maruo, Anal. Chem., 35, 1982 (1963)
(21) S. S. Shapiro and D. Dennis, Biochemistry, 4, 2283 (1965)
(22) K. Kitahara, A. Obayashi, and S. Fukui, Proc. Int. Symp. Enz. Chem., Tokyo and Kyoto, 1957, p. 460 (1958)

Date of correction: June 30, 2008 Reason for correction: - Correction: PDF FILE Details: -

Abstract

A lactate racemase [EC 5. 1. 2. 1] was isolated and purified about 190 fold in specific activity from the sonic extract of Lactobacillus sake. The purified preparation was almost homogeneous by ultracentrifugal and electrophoretical analyses. Characteristic properties of the enzyme were as follows: (a) absorption spectrum showed a single peak at 274 mμ, (b) molecular weight was 25, 000, (c) the enzyme did not require any additional cofactors and showed no activity of lactate dehydrogenase, (d) K_m values were 1.7×10^-2M and 8.0×10^-2M for D- and L-lactate, respectively, (e) optimal pH was 5.8-6.2, (f) an equilibrium point was at a molar ratio of 1/1 (L-isomer/D-isomer), (g) the enzyme activity was inhibited by atebrin, adenosine monosulfate, oxamate and some of Fechelating agents, (h) pyruvate and acrylate were not incorporated into lactate during the reaction, and (i) exchange reaction of hydrogen between lactate and water did not occur during the reaction.

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