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. 1991 May;57(5):1418–1422. doi: 10.1128/aem.57.5.1418-1422.1991

Cloning, sequencing, and expression of the N-acyl-D-mannosamine dehydrogenase gene from Flavobacterium sp. strain 141-8 in Escherichia coli.

H Yamamoto-Otake 1, Y Koyama 1, T Horiuchi 1, E Nakano 1
PMCID: PMC182964  PMID: 1854199

Abstract

The gene coding for N-acyl-D-mannosamine dehydrogenase (NAM-DH) from Flavobacterium sp. strain 141-8 was cloned and expressed under the control of a lac promoter in Escherichia coli JM109. The DNA sequence of the gene was determined, and an open reading frame encoding a polypeptide composed of 272 amino acid residues (Mr, 27,473) was identified. The E. coli transformants which showed over 200-fold higher NAM-DH activity than did the Flavobacterium strain produced the enzyme as a protein fused with beta-galactosidase. Despite being a fusion, NAM-DH produced by E. coli transformants appeared unchanged in pH optimum, Km, and substrate specificity from Flavobacterium sp. strain 141-8. This newly recombinant enzyme may be applicable to the quantitative determination of sialic acid in serum.

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Selected References

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  1. Borck K., Beggs J. D., Brammar W. J., Hopkins A. S., Murray N. E. The construction in vitro of transducing derivatives of phage lambda. Mol Gen Genet. 1976 Jul 23;146(2):199–207. doi: 10.1007/BF00268089. [DOI] [PubMed] [Google Scholar]
  2. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  3. GROSS E., WITKOP B. Nonenzymatic cleavage of peptide bonds: the methionine residues in bovine pancreatic ribonuclease. J Biol Chem. 1962 Jun;237:1856–1860. [PubMed] [Google Scholar]
  4. Horiuchi T., Kurokawa T. Purification and properties of N-acyl-D-mannosamine dehydrogenase from Flavobacterium sp. 141-8. J Biochem. 1988 Sep;104(3):466–471. doi: 10.1093/oxfordjournals.jbchem.a122491. [DOI] [PubMed] [Google Scholar]
  5. Koyama Y., Kitao S., Yamamoto-Otake H., Suzuki M., Nakano E. Cloning and expression of the creatinase gene from Flavobacterium sp. U-188 in Escherichia coli. Agric Biol Chem. 1990 Jun;54(6):1453–1457. [PubMed] [Google Scholar]
  6. Koyama Y., Nakano E. Cloning of the glpK gene of Escherichia coli K-12 and its overexpression using a 'sleeper' bacteriophage vector. Agric Biol Chem. 1990 May;54(5):1315–1316. [PubMed] [Google Scholar]
  7. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  8. Silver H. K., Karim K. A., Archibald E. L., Salinas F. A. Serum sialic acid and sialyltransferase as monitors of tumor burden in malignant melanoma patients. Cancer Res. 1979 Dec;39(12):5036–5042. [PubMed] [Google Scholar]
  9. Sugahara K., Sugimoto K., Nomura O., Usui T. Enzymatic assay of serum sialic acid. Clin Chim Acta. 1980 Dec 22;108(3):493–498. doi: 10.1016/0009-8981(80)90360-5. [DOI] [PubMed] [Google Scholar]
  10. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  11. Young R. A., Davis R. W. Yeast RNA polymerase II genes: isolation with antibody probes. Science. 1983 Nov 18;222(4625):778–782. doi: 10.1126/science.6356359. [DOI] [PubMed] [Google Scholar]

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