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. 1977 Apr;16(1):163–173. doi: 10.1128/iai.16.1.163-173.1977

Polyol Metabolism by a Caries-Conducive Streptococcus: Purification and Properties of a Nicotinamide Adenine Dinucleotide-Dependent Mannitol-1-Phosphate Dehydrogenase

Albert T Brown 1, Robert D Bowles 1
PMCID: PMC421504  PMID: 873604

Abstract

The mannitol-1-phosphate dehydrogenase (M1PDH) (EC 1.1.1.17) from Streptococcus mutans strain FA-1 was purified to approximately a 425-fold increase in specific activity with a 29% recovery of total enzyme units, using a combination of (i) streptomycin sulfate and ammonium sulfate precipitation and (ii) diethyl-aminoethyl-cellulose (DE-52), agarose A 0.5M, and agarose-nicotinamide adenine dinucleotide (NAD) affinity column chromatography. Polyacrylamide gel electrophoresis of the purified enzyme preparation showed a single protein component that coincided with a band of M1PDH activity. The enzyme had a molecular weight of approximately 45,000 and was stable for long periods of time when stored at −80°C in the presence of β-mercaptoethanol. Its activity was not affected by mono- or divalent cations, and high concentrations of ethylenedia-minetetraacetic acid were not inhibitory. The M1PDH catalyzed both the NAD-dependent oxidation of mannitol-1-phosphate and the reduced NAD (NADH)-dependent reduction of fructose-6-phosphate. The forward reaction was highly specific for mannitol-1-phosphate and NAD, whereas the reverse reaction was highly specific for NADH and fructose-6-phosphate. The Km values for mannitol-1-phosphate and NAD were 0.15 and 0.066 mM, respectively, and the Km values for fructose-6-phosphate and NADH were 1.66 and 0.016 mM, respectively. The forward and reverse reactions catalyzed by the M1PDH from S. mutans appeared to be under cellular control. Both adenosine 5′-triphosphate and fructose-6-phosphate were negative effectors of the forward reaction, whereas adenosine 5′-diphosphate served as a negative effector of the reverse reaction catalyzed by the enzyme.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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