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. 1971 Jul;107(1):217–222. doi: 10.1128/jb.107.1.217-222.1971

Partial Purification and Properties of a Highly Specific Trehalose Phosphate Phosphatase from Mycobacterium smegmatis

Mike Matula 1, Mike Mitchell 1, Alan D Elbein 1
PMCID: PMC246907  PMID: 4327508

Abstract

A specific trehalose phosphate phosphatase was purified approximately 50-fold from Mycobacterium smegmatis. The enzyme had a pH optimum of about 7.0 and was stimulated by Mg2+. The optimum concentration of Mg2+ was about 1.5 × 10−3m. Of other divalent cations tested, only Co2+ showed some activity. The Km for trehalose phosphate was found to be about 1.5 × 10−3m. The enzyme showed slight activity toward mannose-6-P and fructose-6-P but was inactive on a large number of other phosphorylated compounds. Citrate was a competitive inhibitor of the enzyme both with respect to trehalose phosphate concentration and Mg2+ concentration. This inhibition appears to be due to chelation of Mg2+ by this compound. Ethylenediaminetetraacetic acid and NaF were also inhibitors of the enzyme, but these inhibitions were noncompetitive.

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Selected References

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

  1. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  2. BIRCH G. G. TREHALOSES. Adv Carbohydr Chem. 1963;18:201–225. doi: 10.1016/s0096-5332(08)60243-x. [DOI] [PubMed] [Google Scholar]
  3. CABIB E., LELOIR L. F. The biosynthesis of trehalose phosphate. J Biol Chem. 1958 Mar;231(1):259–275. [PubMed] [Google Scholar]
  4. CANDY D. J., KILBY B. A. Site and mode of trehalose biosynthesis in the locust. Nature. 1959 Jun 6;183(4675):1594–1595. doi: 10.1038/1831594a0. [DOI] [PubMed] [Google Scholar]
  5. Elbein A. D. Carbohydrate metabolism in streptomycetes. II. Isolation and enzymatic synthesis of trehalose. J Bacteriol. 1967 Nov;94(5):1520–1524. doi: 10.1128/jb.94.5.1520-1524.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Elbein A. D. Trehalose phosphate synthesis in Streptomyces hygroscopicus: purification of guanosine diphosphate D-glucose: D-glucose-6-phosphate 1-glucosyl-transferase. J Bacteriol. 1968 Nov;96(5):1623–1631. doi: 10.1128/jb.96.5.1623-1631.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. FRIEDMAN S. Occurrence of trehalose-6-phosphatase in Phormia regina Meig. Arch Biochem Biophys. 1960 Jun;88:339–343. doi: 10.1016/0003-9861(60)90246-0. [DOI] [PubMed] [Google Scholar]
  8. GOLDMAN D. S., LORNITZO F. A. Enzyme systems in the mycobacteria. XII. The inhibition of the transglycosidase-catalyzed formation of trehalose 6-phosphate. J Biol Chem. 1962 Nov;237:3332–3338. [PubMed] [Google Scholar]
  9. Hey A. E., Elbein A. D. Partial prufication and properties of a trehalase from Streptomyces hygroscopicus. J Bacteriol. 1968 Jul;96(1):105–110. doi: 10.1128/jb.96.1.105-110.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Liu C., Patterson B. W., Lapp D., Elbein A. D. Trehalose phosphate synthesis from uridine diphosphate glucose or guanosine diphosphate glucose. Activation of uridine diphosphate-glucose: trehalose phosphate synthetase by polynucleotides. J Biol Chem. 1969 Jul 10;244(13):3728–3731. [PubMed] [Google Scholar]
  11. MACDONALD D. L., WONG R. Y. A CHEMICAL SYNTHESIS OF TREHALOSE 6-PHOSPHATE. Biochim Biophys Acta. 1964 May 11;86:390–392. doi: 10.1016/0304-4165(64)90066-2. [DOI] [PubMed] [Google Scholar]
  12. MENDICINO J. Sucrose phosphate synthesis in wheat germ and green leaves. J Biol Chem. 1960 Dec;235:3347–3352. [PubMed] [Google Scholar]
  13. MURPHY T. A., WYATT G. R. THE ENZYMES OF GLYCOGEN AND TREHALOSE SYNTHESIS IN SILK MOTH FAT BODY. J Biol Chem. 1965 Apr;240:1500–1508. [PubMed] [Google Scholar]
  14. Roth R., Sussman M. Trehalose synthesis in the cellular slime mold Dictyostelium discoideum. Biochim Biophys Acta. 1966 Aug 10;122(2):225–231. doi: 10.1016/0926-6593(66)90064-6. [DOI] [PubMed] [Google Scholar]
  15. WINDER F., BRENNAN P. THE ACCUMULATION OF FREE TREHALOSE BY MYCOBACTERIA EXPOSED TO ISONIAZID. Biochim Biophys Acta. 1964 Aug 19;90:442–444. doi: 10.1016/0304-4165(64)90222-3. [DOI] [PubMed] [Google Scholar]
  16. WYATT G. R., KALE G. F. The chemistry of insect hemolymph. II. Trehalose and other carbohydrates. J Gen Physiol. 1957 Jul 20;40(6):833–847. doi: 10.1085/jgp.40.6.833. [DOI] [PMC free article] [PubMed] [Google Scholar]

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