본문 바로가기
HOME> 논문 > 논문 검색상세

논문 상세정보

Biotechnology and bioprocess engineering v.14 no.5, 2009년, pp.651 - 661   SCI SCIE
본 등재정보는 저널의 등재정보를 참고하여 보여주는 베타서비스로 정확한 논문의 등재여부는 등재기관에 확인하시기 바랍니다.

Novel Synthesis of Mannosamine Conjugated Magnetic Nanoparticles for Purification and Stabilization of Human Lysosomal $\beta$-mannosidase

Samra, Zahoor Qadir    (Institute of Biochemistry and Biotechnology, Quaid-i-Azam Campus, University of Punjab   ); Athar, Muhammad Amin    (Institute of Biochemistry and Biotechnology, Quaid-i-Azam Campus, University of Punjab  );
  • 초록

    Human $\beta$ -mannosidase (MANB) was purified to homogeneity directly from lysosomes by using mannosamine conjugated magnetic ( $Fe_3O_4$ ) nanoparticles, DE-52 cellulose, and sephadex G-200 chromatography. $Fe_3O_4$ nanoparticles were synthesized and utilized ammonia to attach the amino group on the nanoparticles. The particles were covalently attached with Dmannosamine by cross linker glutaraldehyde and confirmed by FTIR spectroscopy. In FTIR analysis, the peaks appeared at 2,356.6 $cm^{-1}$ for -N = CH linkage and at 3,378.4 $cm^{-1}$ , 3,664.9 $cm^{-1}$ for -OH groups confirmed the conjugation of D-mannosamine with $Fe_3O_4$ nanoparticles. Results showed a single band of 97 kDa of purified MANB in SDS-PAGE. The isoelectric point was 4.5 and the $K_m$ and Vmax values were 2.51 mM and $0.315\;{\mu}M/min/mg$ , respectively. The purification fold was 329 with 68% yield. The optimal activity was at pH 5.0 and 75% activity was stable in 20% glycerol at $4^{\circ}C$ . The enzyme activity was inhibited by $Ni^{2+}$ , $Zn^{2+}$ , $Cd^{2+}$ , $Cu^{2+}$ , $Mo^{2+}$ , $Ag^{+1}$ , iodoacetate, SDS, DMF, DMSO, ethanol, and acetone; slightly reduced by $Pb^{2+}$ , $Co^{2+}$ , EDTA, DTT, and $\beta$ -mercaptoethanol. The activity was not affected by $Mg^{2+}$ , $Mn^{2+}$ , $Sn^{2+}$ , $Ca^{2+}$ , $Fe^{3+}$ , PMSF, Triton X-100, D-mannosamine, D-mannose, D-mannitol, D-glucose, and D-fructose. The homogeneity of MANB enzyme was further confirmed by 2D-PAGE and immunoblot. This is the first novel report of conjugation of D-mannosamine with $Fe_3O_4$ nanoparticles for purification of human MANB enzyme.

  • 주제어

    Mannosamine .   magnetic nanoparticles .   FTIR .   western blot .   2D-PAGE.  

  • 참고문헌 (45)

    1. Ademark, P., J. Lundqvist, P. Hagglund, M. Tenkanen, N. Torto, F. Tjerneld, and H. Stalbrand (1999) Hydrolytic properties of a $\beta$-mannosidase purified from Aspergillus niger. J Biotechnol. 75: 281-289 
    2. Jones, M. Z., E. J. Rathke, D. A. Gage, C. E. Costello, K. Murakami, M. Ohta, and F. Matsuura (1992) Oligosaccharides accumulated in the bovine beta-mannosidosis kidney. J. Inherit. Metab. Dis. 15: 57-67 
    3. Sopher, B. L., C. E. Traviss, K. T. Cavanagh, M. Z. Jones, and K. H. Friderici (1993) Bovine kidney beta-mannosidase: purification and characterization. Biochem. J. 15: 343-347 
    4. Mo, B. and J. D. Bewley (2002) Beta-mannosidase (EC activity during and following germination of tomato (Lycopersicon esculentum Mill.) seeds. Purification, cloning, and characterization. Planta. 215: 141-152 
    5. Van, P. J., C. H. Hokke, L. Dorland, M. Duran, J. P. Kamerling, and J. F. Vliegenthart (1990) Accumulation of mannosyl-beta(1----4)-N-acetylglucosamine in fibroblasts and leukocytes of patients with a deficiency of beta-mannosidase. Clin. Chim. Acta. 187: 55-60 
    6. Alkhayat, A. H., S. A. Kraemer, J. R. Leipprandt, M. Macek, W. J. Kleijer, and K. H. Friderici (1998) Human beta-mannosidase cDNA characterization and first identification of a mutation associated with human betamannosidosis. Hum. Mol. Genet. 7: 75-83 
    7. Poenaru, L., S. Akli, F. Rocchiccioli, P. Eydoux, and P. Zamet (1992) Human beta-mannosidosis: a 3-year-old boy with speech impairment and emotional instability. Clin. Genet. 41: 331-340 
    8. Cooper, A., J. E. Wraith, W. J. Savage, M. Thornley, and M. J. Noronha (1991) Beta-mannosidase deficiency in a female infant with epileptic encephalopathy. J Inherit. Metab. Dis. 14: 18-22 
    9. Rodriguez-Serna, M., R. Botella-Estrada, A. Chabas, M. J. Coll, V. Oliver, M. I. Febrer, and A. Aliaga (1996) Angiokeratoma corporis diffusum associated with beta-mannodeficiency. Arch. Dermatol. 132: 1219-1222 
    10. Kleijer, W. J., G. C. Geilen, D. O. P. Van, R. Wevers, and F. J. Los (1992) Prenatal analyses in a pregnancy at risk for beta-mannosidosis. Prenat. Diagn. 12: 841-843 
    11. Huang, S. H., M. H. Liao, and D. H. Chen (2003) Direct binding and characterization of lipase onto magnetic nanoparticles. Biotechnol. Prog. 19: 1095 -1100 
    12. Bradford, M. M. (1976) A dye binding assay for protein. Ann. Biochem. 72: 248-254 
    13. Lovell, K. L., R. J. Kranich, and K. T. Cavanagh (1994) Biochemical and histochemical analysis of lysosomal enzyme activities in caprine beta-mannosidosis. Mol. Chem. Neuropathol. 21: 61-74 
    14. Van, P. J., L. Dorland, M. Duran, C. H. Hokke, J. P. Kamerling, and J. F. Vliegenthart (1990) Sialyl-alpha 2-6-mannosyl-beta 1-4-N-acetylglucosamine, a novel compound occurring in urine of patients with $\beta$-mannosidosis. J. Biol. Chem. 265: 19685-19689 
    15. Bauer, M. W., E. J. Bylina, R. V. Swanson, and R. M. Kelly (1996) Comparison of a beta-glucosidase and a beta-mannosidase from the hyperthermophilic archaeon Pyrococcus furiosus. Purification, characterization, gene cloning, and sequence analysis. J. Biol. Chem. 271: 23749-23755 
    16. Sopher, B. L., C. E. Traviss, K. T. Cavanagh, M. Z. Jones, and K. H. Friderici (1993) Bovine kidney beta-mannosidase: purification and characterization. Biochem. J. 289: 343-347 
    17. Iwasaki, Y., A. Tsuji, K. Omura, and Y. Suzuki (1989) Purification and characterization of beta-mannosidase from human placenta. J Biochem (Tokyo) 106: 331-335 
    18. Tartaj, P., M. D. Morales, S. V. Verdaguer, T. G. Carre, and C. J. Serna (2003) The preparation of magnetic nanoparticles for applications in biomedicine. J. Phys. D. Appl. Phys. 36: 182-197 
    19. Uchino, Y., T. Fukushige, S. Yotsumoto, T. Hashiguchi, H. Taguchi, N. Suzuki, I. Konohana, and T. Kanzak (2003) Morphological and biochemical studies of human beta-mannosidosis: identification of a novel beta-mannosidase gene mutation. Br. J. Dermatol. 149: 23-29 
    20. Safarik, I. and M. Safarikova (2004) Magnetic techniques for the isolation and purification of proteins and peptides. Bio.Magn. Res. Technol. 2: 1-17 
    21. Samra, Z. Q. and M. A. Athar (2008) Bioinfocomparative studies of beta-mannosidases. Pak. J. Zool. (submitted /accepted) 
    22. Kurakake, M. and T. Komaki (2001) Production of betamannanase and beta-mannosidase from Aspergillus awamori K4 and their properties. Curr. Microbiol. 42: 377-380 
    23. Towbin, H. K., T. Staehelin, and J. Gordon (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose membrane. Proc. Natl. Acad. Sci. 76: 4350-4359 
    24. Guadalupi, R., M. Bernard, A. Orlacchio, M. J. Foglietti, and C. Emiliani (1996) Purification and properties of human urinary beta-D-mannosidase. Biochim. Biophys. Acta. 1293: 9-16 
    25. Samra, Z. Q., S. N. Mirza, N. Dar, Z. Mahmood, and M. A. Athar (2006) Characterization of Lactate dehydrogenase enzyme immobilized to magnetic nano particles. Pak. J. Zool. 38: 327-332 
    26. Abbitt, B., M. Z. Jones, T. R. Kasari, R. W. Storts, J. W. Templeton, P. S. Holland, and P. E. Castenson (1991) Beta-mannosidosis in twelve Salers calves. J. Am. Vet. Med. Assoc. 198: 109-113 
    27. Liao, M. H. and D. H. Chen (2002) Preparation and characterization of a novel magnetic nanoadsorbant. J. Mater. Chem. 12: 3654-3659 
    28. Levade, T., D. Graber, V. Flurin, M. B. Delisle, M. T. Pieraggi, M. F. Testut, J. P. Carriere, and R. Salvayre (1994) Human beta-mannosidase deficiency associated with peripheral neuropathy. Ann. Neurol. 35: 116-119 
    29. Walker, J. (1994) Methods in molecular biology: Basic protein and peptide protocol. Humana Press Inc, Totowa, NJ, USA 
    30. Bedilu, R., K. A. Nummy, A. Cooper, R. Wevers, J. Smeitink, W. J. Kleijer, and K. H. Friderici (2002) Variable clinical presentation of lysosomal beta-mannosidosis in patients with null mutations. Mol. Genet. Metab. 77: 282-290 
    31. Plummer, D. T. (1987) An introduction to practical biochemistry. 3rd ed., McGraw Hill book company, UK 
    32. Kouassi, K. G., I. Joseph, J. Irudayaraj, and M. Gregory (2005) Examination of cholesterol oxidase attachment to magnetic nanoparticles. J. Nanobiotechnol. 3: 1-9 
    33. Bernard, M., M. Sioud, F. Percheron, and M. J. Foglietti (1986) Beta-mannosidase in human serum and urine. A comparative study. Int. J. Biochem. 18: 1065-1068 
    34. Percheron, F., M. J. Foglietti, M. Bernard, and B. Ricard (1992) Mammalian beta-D-mannosidase and beta-mannosidosis. Biochimie. 74: 5-11 
    35. Liao, M. H. and D. H. Chen (2001) Immobilization of yeast alcohol dehydrogenase on magnetic nanoparticles for improving its stability. Biotechnol. Lett. 23: 1723-1727 
    36. Kleijer, W. J., P. Hu, R. Thoomes, M. Boer, J. G. Huijmans, W. Blom, O. P. Van Diggelen, E. Seemanova, and M. Macek (1990) Beta-mannosidase deficiency: heterogeneous manifestation in the first female patient and her brother. J Inherit Metab. Dis. 13: 867-872 
    37. Gort, L., J. Duque, J. M. Fabeiro, A. Zulaica, M. J. Coll, and A. Chabas (2006) Molecular analysis in two betamannosidosis patients: Description of a new adult case. Mol. Genet. Metab. 89: 398-400 
    38. Cooper, A., C. Hatton, M. Thornley, and I. B. Sardharwalla (1988) Human beta-mannosidase deficiency: biochemical findings in plasma, fibroblasts, white cells, and urine. J. Inherit. Metab. Dis. 11: 17-29 
    39. Duffaud, G. D., C. M. McCutchen, P. Leduc, K. N. Parker, and R. M. Kelly (1997) Purification and characterization of extremely thermostable beta-mannanase, beta-mannosidase, and alpha-galactosidase from the hyperthermophilic eubacterium Thermotoga neapolitana 5068. Appl. Environ. Microbiol. 63: 169-177 
    40. Bryan, L., S. Schmutz, S. D. Hodges, and F. F. Snyder (1990) Bovine beta-mannosidase deficiency. Biochem. Biophys. Res. Commun. 173: 491-495 
    41. Jones, M. Z., J. G. Cunningham, A. W. Dade, D. M. Alessi, U. V. Mostosky, J. R. Vorro, J. T. Benitez, and K. L. Lovell (1983) Caprine beta-mannosidosis: clinical and pathological features. J. Neuropathol. Exp. Neurol. 42: 268-285 
    42. Kawaguchi, H. (2000) Functional polymer microspheres. Prog. Polym. Sci. 25: 1171-1210 
    43. Render, J. A., K. L. Lovell, and M. Z. Jones (1988) Otic pathology of caprine beta-mannosidosis. Vet. Pathol. 25: 437-442 
    44. Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685 
    45. Pearce, R. D., J. W. Callahan, P. B. Little, L. R. Klunder, and J. T. Clarke (1990) Caprine beta-D-mannosidosis: characterization of a model lysosomal storage disorder. Can. J. Vet. Res. 54: 22-29 

 활용도 분석

  • 상세보기

    amChart 영역
  • 원문보기

    amChart 영역


  • 원문이 없습니다.

유료 다운로드의 경우 해당 사이트의 정책에 따라 신규 회원가입, 로그인, 유료 구매 등이 필요할 수 있습니다. 해당 사이트에서 발생하는 귀하의 모든 정보활동은 NDSL의 서비스 정책과 무관합니다.

원문복사신청을 하시면, 일부 해외 인쇄학술지의 경우 외국학술지지원센터(FRIC)에서
무료 원문복사 서비스를 제공합니다.

NDSL에서는 해당 원문을 복사서비스하고 있습니다. 위의 원문복사신청 또는 장바구니 담기를 통하여 원문복사서비스 이용이 가능합니다.

이 논문과 함께 출판된 논문 + 더보기