The Subunit Structure of Lactate Dehydrogenase from <i>Streptococcus cremoris</i> US3

Dynon, M K; Jago, G. R.; Davidson, Barrie E.; Gething, Mary‐Jane

doi:10.1111/j.1432-1033.1972.tb02104.x

Cited by 18 publications

(10 citation statements)

References 25 publications

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“…The molecular weight estimate of 132000-135000 with a subunit molecular weight estimate of 34000 show close similarity to the values obtained from S. cremoris [37], P. cerevisiue [38], L. acidophilus [39], L.plantarurn [40] and seems to conform to the general pattern for the tetrameric arrangement of L-lactate dehydrogenases. However, this general pattern appears not to exist outside this bacterial family, since an estimate of 100000 was reported for fructose-l,6bisphosphate-activated L( + )-lactate dehydrogenase from Actinonzyces viscosus.…”

Section: Immunological Relationships Of Lactate Dehydrogenuse From Lasupporting

confidence: 81%

Purification, Properties and Immunological Relationship of l(+)‐Lactate Dehydrogenase from Lactobacillus casei

Gordon

Doelle

1976

European Journal of Biochemistry

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The fructose-1,6-bisphosphate-activated L-lactate dehydrogenase (EC 1.1 .I .27) from Lactobacillus casei ATCC 393 has been purified to homogenity by including affinity chromatography (cibacronblue -Sephadex-G-200) and preparative polyacrylamide gel electrophoresis into the purification procedures. The enzyme has an M , of 132000-135000 with a subunit M , of 34000. The pH optimum was found to be 5.4 in sodium acetate buffer. Tris/maleate and citrate/phosphate buffers inhibited enzyme activity at this pH. The enzyme was completely inactivated by a temperature increase from 60 "C to 70 "C. Pyruvate saturation curves were sigmoidal in the absence of fructose 1,6-bisphosphate. In the presence of 20 pM fructose 1,6-bisphosphate a K, of 1 .OmM for pyruvate was obtained, whereas fructose 1,6-bisphosphate had no effect on the K,,, of 0.01 mM for NADH. The use of pyruvate analogues revealed two types of pyruvate binding sites, a catalytic and an effector site. The enzyme from L. casei appears to be subject to strict metabolic control, since ADP, ATP, dihydroxyacetone phosphate and 6-phosphogluconate are strong inhibitors.Immunodiffusion experiments with a rabbit antiserum to L. casei lactate dehydrogenase revealed that L. casei ATCC 393 L( +)-lactate dehydrogenase is probably not immunologically related to group D and group N streptococci. Of 24 lactic acid bacterial strains tested only 5 strains did cross-react :Fructose 1,6-bisphosphate activation of lactate dehydrogenase is found widely in several streptococcal groups [I -61. This phenomenon also exists among bifidobacteria [7], fermentative mycoplasmas [S, 91, certain coagulase-negative and mannitol-negative staphylococci [lo], and an actinomycete [l 11. However, Lactobacillus casei is the only member of its genus which possesses a fructose-l,6-bisphosphate-activated lactate dehydrogenase [12,32], with the enzyme being specific for L(+)-lactate [12]. This lactate isomer is also the major end-product of glucose fermentation [13]. Immunological and hence evolutionary links have been established between the isofunctional malic enzyme from Streptococcus fuecalis and L. cusei [14, 151. Further evidence of a close relationship between these two genera was obtained [16] using an antiserum prepared from purified S. faecalis fructose-l,6-bisphosphate aldolase. Difficulties in assaying and purifying L. casei lactate dehydrogenase [17] precluded this lactic acid bacterium from an immunological study of the relationships among lactate dehydrogenase from lactobacilli and leuconostocs [18].The presented paper reports the purification of L( +)-lactate dehydrogenase from L. casei to a homogenous preparation and establishes the kinetic characteristics of this enzyme. It also investigates the effects of various divalent metal ions, reaction products, metabolic intermediates and adenylates in an effort to elucidate the metabolic control of glucose utilization as well as the immunological relationship to the lactate dehydrogenases of other lactic acid bacteria.

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Section: Immunological Relationships Of Lactate Dehydrogenuse From Lasupporting

confidence: 81%

Purification, Properties and Immunological Relationship of l(+)‐Lactate Dehydrogenase from Lactobacillus casei

Gordon

Doelle

1976

European Journal of Biochemistry

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“…There is general agreement that the L(+)-nLDH's have molecular weights of about 140,000 (6,20,39,40,41,43,45,52,111), but the molecular weight of L. acidophilus nLDH was only 120,000 (35). The nLDH molecule is a tetramer (20,39,40,43,45). It may be split into subunits in different ways; the LDH of S. cremoris is irreversibly denatured into units of 72,000 daltons at pH 9.0 (52).…”

Section: Purification and Properties Of Bacterial Ldhsmentioning

confidence: 99%

“…Mammalian nLDH has a high cysteine content, whereas bacterial nLDH cysteine content is low. The significance of these differences is discussed elsewhere for the lactobacilli (45) and streptococci (20). These differences are reflected in different responses to a number of inhibitors ( Tables 5 and 6).…”

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confidence: 95%

Bacterial lactate dehydrogenases.

Garvie¹

1980

Microbiological Reviews

338

201

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“…Molecular size and arrangement of both lactate dehydrogenases from P. cerevisiae are of interest since they conform to the general pattern that is gradually becoming apparent for all lactic acid bacteria. The dimeric arrangement for D-LDH from P. cerevisiae with a molecular weight of about 70,000 shows a close similarity to the enzymes from Leuconostoc lactis (11,16) and Lactobacillus leichmannii (11), whereas the tetrameric arrangement for L-+ LDH from P. cerevisiae with a molecular weight of about 140,000 shows a close similarity to the enzyme from Streptococcus cremoris (7). Native molecular weights of both of these enzymes from P. cerevisiae are comparable to values reported for many other lactic acid bacteria (5,8,10).…”

Section: Resultsmentioning

confidence: 81%

Production of racemic lactic acid in Pediococcus cerevisiae cultures by two lactate dehydrogenases

Gordon¹,

Doelle²

1975

J Bacteriol

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Nicotinamide adenine dinucleotide (NAD)-dependent D(-)-and L(+)-lactate dehydrogenases have been partially purified 89and 70-fold simultaneously from cell-free extracts of Pediococcus cerevisiae. Native molecular weights, as estimated from molecular sieve chromatography and electrophoresis in nondenaturing polyacrylamide gels, are 71,000 to 73,000 for D(-)-lactate dehydrogenase and 136,000 to 139,000 for L(+)-lactate dehydrogenase. Electrophoresis in sodium dodecyl sulfate-containing gels reveals subunits with approximate molecular weights of 37,000 to 39,000 for both enzymes. By lowering the pyruvate concentration from 5.0 to 0.5 mM, the pH optimum for pyruvate reduction by D(-)-lactate dehydrogenase decreases from pH 8.0 to 3.6. However, L(+)-lactate dehydrogenase displays an optimum for pyruvate reduction between pH 4.5 and 6.0 regardless of the pyruvate concentration. The enzymes obey Michaelis-Menten kinetics for both pyruvate and reduced NAD at pH 5.4 and 7.4, with increased affinity for both substrates at the acid pH. a-Ketobutyrate can be used as a reducible substrate, whereas oxamate has no inhibitory effect on lactate oxidation by either enzyme. Adenosine triphosphate causes inhibition of both enzymes by competition with reduced NAD. Adenosine diphosphate is also inhibitory under the same conditions, whereas NAD acts as a product inhibitor. These results are discussed with relation to the lactate isomer production during the growth cycle of P. cerevistae.

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The Subunit Structure of Lactate Dehydrogenase from Streptococcus cremoris US3

Cited by 18 publications

References 25 publications

Purification, Properties and Immunological Relationship of l(+)‐Lactate Dehydrogenase from Lactobacillus casei

Purification, Properties and Immunological Relationship of l(+)‐Lactate Dehydrogenase from Lactobacillus casei

Bacterial lactate dehydrogenases.

Production of racemic lactic acid in Pediococcus cerevisiae cultures by two lactate dehydrogenases

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