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J Bacteriol. 1997 July; 179(13): 4129–4137.
PMCID: PMC179231

6-phospho-alpha-D-glucosidase from Fusobacterium mortiferum: cloning, expression, and assignment to family 4 of the glycosylhydrolases.


The Fusobacterium mortiferum malH gene, encoding 6-phospho-alpha-glucosidase (maltose 6-phosphate hydrolase; EC, has been isolated, characterized, and expressed in Escherichia coli. The relative molecular weight of the polypeptide encoded by malH (441 residues; Mr of 49,718) was in agreement with the estimated value (approximately 49,000) obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the enzyme purified from F. mortiferum. The N-terminal sequence of the MalH protein obtained by Edman degradation corresponded to the first 32 amino acids deduced from the malH sequence. The enzyme produced by the strain carrying the cloned malH gene cleaved [U-14C]maltose 6-phosphate to glucose 6-phosphate (Glc6P) and glucose. The substrate analogs p-nitrophenyl-alpha-D-glucopyranoside 6-phosphate (pNP alphaGlc6P) and 4-methylumbelliferyl-alpha-D-glucopyranoside 6-phosphate (4MU alphaGlc6P) were hydrolyzed to yield Glc6P and the yellow p-nitrophenolate and fluorescent 4-methylumbelliferyl aglycons, respectively. The 6-phospho-alpha-glucosidase expressed in E. coli (like the enzyme purified from F. mortiferum) required Fe2+, Mn2+, Co2+, or Ni2+ for activity and was inhibited in air. Synthesis of maltose 6-phosphate hydrolase from the cloned malH gene in E. coli was modulated by addition of various sugars to the growth medium. Computer-based analyses of MalH and its homologs revealed that the phospho-alpha-glucosidase from F. mortiferum belongs to the seven-member family 4 of the glycosylhydrolase superfamily. The cloned 2.2-kb Sau3AI DNA fragment from F. mortiferum contained a second partial open reading frame of 83 residues (designated malB) that was located immediately upstream of malH. The high degree of sequence identity of MalB with IIB(Glc)-like proteins of the phosphoenol pyruvate dependent:sugar phosphotransferase system suggests participation of MalB in translocation of maltose and related alpha-glucosides in F. mortiferum.

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  • Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol. 1990 Oct 5;215(3):403–410. [PubMed]
  • Bakken V, Högh BT, Jensen HB. Utilization of amino acids and peptides by Fusobacterium nucleatum. Scand J Dent Res. 1989 Feb;97(1):43–53. [PubMed]
  • Bennett KW, Eley A. Fusobacteria: new taxonomy and related diseases. J Med Microbiol. 1993 Oct;39(4):246–254. [PubMed]
  • Bolstad AI, Jensen HB, Bakken V. Taxonomy, biology, and periodontal aspects of Fusobacterium nucleatum. Clin Microbiol Rev. 1996 Jan;9(1):55–71. [PMC free article] [PubMed]
  • Brook I, Walker RI. The relationship between Fusobacterium species and other flora in mixed infection. J Med Microbiol. 1986 Mar;21(2):93–100. [PubMed]
  • Burland V, Plunkett G, 3rd, Daniels DL, Blattner FR. DNA sequence and analysis of 136 kilobases of the Escherichia coli genome: organizational symmetry around the origin of replication. Genomics. 1993 Jun;16(3):551–561. [PubMed]
  • Claesson R, Edlund MB, Persson S, Carlsson J. Production of volatile sulfur compounds by various Fusobacterium species. Oral Microbiol Immunol. 1990 Jun;5(3):137–142. [PubMed]
  • Dzink JL, Socransky SS. Amino acid utilization by Fusobacterium nucleatum grown in a chemically defined medium. Oral Microbiol Immunol. 1990 Jun;5(3):172–174. [PubMed]
  • Embree JE, Williams T, Law BJ. Hepatic abscesses in a child caused by Fusobacterium necrophorum. Pediatr Infect Dis J. 1988 May;7(5):359–360. [PubMed]
  • Feng DF, Doolittle RF. Progressive alignment and phylogenetic tree construction of protein sequences. Methods Enzymol. 1990;183:375–387. [PubMed]
  • George WL, Kirby BD, Sutter VL, Citron DM, Finegold SM. Gram-negative anaerobic bacilli: Their role in infection and patterns of susceptibility to antimicrobial agents. II. Little-known Fusobacterium species and miscellaneous genera. Rev Infect Dis. 1981 May-Jun;3(3):599–626. [PubMed]
  • Hengstenberg W, Kohlbrecher D, Witt E, Kruse R, Christiansen I, Peters D, Pogge von Strandmann R, Städtler P, Koch B, Kalbitzer HR. Structure and function of proteins of the phosphotransferase system and of 6-phospho-beta-glycosidases in gram-positive bacteria. FEMS Microbiol Rev. 1993 Sep;12(1-3):149–163. [PubMed]
  • Henrissat B, Bairoch A. New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J. 1993 Aug 1;293(Pt 3):781–788. [PubMed]
  • Henrissat B, Callebaut I, Fabrega S, Lehn P, Mornon JP, Davies G. Conserved catalytic machinery and the prediction of a common fold for several families of glycosyl hydrolases. Proc Natl Acad Sci U S A. 1995 Jul 18;92(15):7090–7094. [PubMed]
  • Henry S, DeMaria A, Jr, McCabe WR. Bacteremia due to Fusobacterium species. Am J Med. 1983 Aug;75(2):225–231. [PubMed]
  • Hofstad T. Pathogenicity of anaerobic gram-negative rods: possible mechanisms. Rev Infect Dis. 1984 Mar-Apr;6(2):189–199. [PubMed]
  • Jones TH, Bergvall V, Bradshaw JP. Carotid artery stenoses and thrombosis secondary to cavernous sinus thromboses in Fusobacterium necrophorum meningitis. Postgrad Med J. 1990 Sep;66(779):747–750. [PMC free article] [PubMed]
  • Langworth BF. Fusobacterium necrophorum: its characteristics and role as an animal pathogen. Bacteriol Rev. 1977 Jun;41(2):373–390. [PMC free article] [PubMed]
  • Liljeström PL, Liljeström P. Nucleotide sequence of the melA gene, coding for alpha-galactosidase in Escherichia coli K-12. Nucleic Acids Res. 1987 Mar 11;15(5):2213–2220. [PMC free article] [PubMed]
  • Meadow ND, Fox DK, Roseman S. The bacterial phosphoenolpyruvate: glycose phosphotransferase system. Annu Rev Biochem. 1990;59:497–542. [PubMed]
  • Meins M, Jenö P, Müller D, Richter WJ, Rosenbusch JP, Erni B. Cysteine phosphorylation of the glucose transporter of Escherichia coli. J Biol Chem. 1993 Jun 5;268(16):11604–11609. [PubMed]
  • Moore WE, Holdeman LV, Smibert RM, Good IJ, Burmeister JA, Palcanis KG, Ranney RR. Bacteriology of experimental gingivitis in young adult humans. Infect Immun. 1982 Nov;38(2):651–667. [PMC free article] [PubMed]
  • Parker LL, Hall BG. Characterization and nucleotide sequence of the cryptic cel operon of Escherichia coli K12. Genetics. 1990 Mar;124(3):455–471. [PubMed]
  • Pearson WR, Lipman DJ. Improved tools for biological sequence comparison. Proc Natl Acad Sci U S A. 1988 Apr;85(8):2444–2448. [PubMed]
  • Postma PW, Lengeler JW, Jacobson GR. Phosphoenolpyruvate:carbohydrate phosphotransferase systems of bacteria. Microbiol Rev. 1993 Sep;57(3):543–594. [PMC free article] [PubMed]
  • Reizer J, Michotey V, Reizer A, Saier MH., Jr Novel phosphotransferase system genes revealed by bacterial genome analysis: unique, putative fructose- and glucoside-specific systems. Protein Sci. 1994 Mar;3(3):440–450. [PubMed]
  • Reizer J, Paulsen IT, Reizer A, Titgemeyer F, Saier MH., Jr Novel phosphotransferase system genes revealed by bacterial genome analysis: the complete complement of pts genes in mycoplasma genitalium. Microb Comp Genomics. 1996;1(3):151–164. [PubMed]
  • Reizer A, Reizer J. Progressive multiple alignment of protein sequences and the construction of phylogenetic trees. Methods Mol Biol. 1994;25:319–325. [PubMed]
  • Reizer J, Saier MH, Jr, Deutscher J, Grenier F, Thompson J, Hengstenberg W. The phosphoenolpyruvate:sugar phosphotransferase system in gram-positive bacteria: properties, mechanism, and regulation. Crit Rev Microbiol. 1988;15(4):297–338. [PubMed]
  • Robrish SA, Fales HM, Gentry-Weeks C, Thompson J. Phosphoenolpyruvate-dependent maltose:phosphotransferase activity in Fusobacterium mortiferum ATCC 25557: specificity, inducibility, and product analysis. J Bacteriol. 1994 Jun;176(11):3250–3256. [PMC free article] [PubMed]
  • Robrish SA, Oliver C, Thompson J. Sugar metabolism by fusobacteria: regulation of transport, phosphorylation, and polymer formation by Fusobacterium mortiferum ATCC 25557. Infect Immun. 1991 Dec;59(12):4547–4554. [PMC free article] [PubMed]
  • Robrish SA, Thompson J. Regulation of fructose metabolism and polymer synthesis by Fusobacterium nucleatum ATCC 10953. J Bacteriol. 1990 Oct;172(10):5714–5723. [PMC free article] [PubMed]
  • Roseman S. Sialic acid, serendipity, and sugar transport: discovery of the bacterial phosphotransferase system. FEMS Microbiol Rev. 1989 Jun;5(1-2):3–11. [PubMed]
  • Saier MH, Jr, Reizer J. Proposed uniform nomenclature for the proteins and protein domains of the bacterial phosphoenolpyruvate: sugar phosphotransferase system. J Bacteriol. 1992 Mar;174(5):1433–1438. [PMC free article] [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Slots J, Potts TV, Mashimo PA. Fusobacterium periodonticum, a new species from the human oral cavity. J Dent Res. 1983 Sep;62(9):960–963. [PubMed]
  • Smith DW. A complete, yet flexible, system for DNA/protein sequence analysis using VAX/VMS computers. Comput Appl Biosci. 1988 Mar;4(1):212–212. [PubMed]
  • Socransky SS, Haffajee AD. The bacterial etiology of destructive periodontal disease: current concepts. J Periodontol. 1992 Apr;63(4 Suppl):322–331. [PubMed]
  • Thompson J, Gentry-Weeks CR, Nguyen NY, Folk JE, Robrish SA. Purification from Fusobacterium mortiferum ATCC 25557 of a 6-phosphoryl-O-alpha-D-glucopyranosyl:6-phosphoglucohydrolase that hydrolyzes maltose 6-phosphate and related phospho-alpha-D-glucosides. J Bacteriol. 1995 May;177(9):2505–2512. [PMC free article] [PubMed]
  • Thompson J, Nguyen NY, Robrish SA. Sucrose fermentation by Fusobacterium mortiferum ATCC 25557: transport, catabolism, and products. J Bacteriol. 1992 May;174(10):3227–3235. [PMC free article] [PubMed]
  • Thompson J, Robrish SA, Bouma CL, Freedberg DI, Folk JE. Phospho-beta-glucosidase from Fusobacterium mortiferum: purification, cloning, and inactivation by 6-phosphoglucono-delta-lactone. J Bacteriol. 1997 Mar;179(5):1636–1645. [PMC free article] [PubMed]
  • Würsch P, Koellreutter B. Maltotriitol inhibition of maltose metabolism in Streptococcus mutans via maltose transport, amylomaltase and phospho-alpha-glucosidase activities. Caries Res. 1985;19(5):439–449. [PubMed]
  • Xavier KB, Martins LO, Peist R, Kossmann M, Boos W, Santos H. High-affinity maltose/trehalose transport system in the hyperthermophilic archaeon Thermococcus litoralis. J Bacteriol. 1996 Aug;178(16):4773–4777. [PMC free article] [PubMed]
  • Yamamoto H, Uchiyama S, Fajar AN, Ogasawara N, Sekiguchi J. Determination of a 12 kb nucleotide sequence around the 76 degrees region of the Bacillus subtilis chromosome. Microbiology. 1996 Jun;142(Pt 6):1417–1421. [PubMed]

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