본문 바로가기
HOME> 논문 > 논문 검색상세

논문 상세정보

Food science and biotechnology v.14 no.3 = no.63, 2005년, pp.317 - 322   피인용횟수: 2

Affinity Immobilization of Dextransucrase on Dextran-based Support and the Production of Leucrose

Han, Nam-Soo    (Department of Food Science and Technology, Research Center for Bioresource and Health, Chungbuk National University   ); Kang, Seung-Yeon    (Department of Food Science and Technology, Research Center for Bioresource and Health, Chungbuk National University   ); Lee, Soo-Bok    (Department of Food Nutrition, Yonsei University   ); Robyt, John F.    (Department of Biochemistry and Biophysics, Iowa State University  );
  • 초록

    A simple and convenient method of immobilizing dextransucrase via an affinity interaction is described, along with the use of this system to synthesize leucrose. Dextransucrase was produced in sucrose-free medium by fermenting a constitutive mutant of Leuconostoc mesenteroides NRRL B-512F and was separated using an ultrafiltration membrane. The purified enzyme was free of dextran polymer, which previously was always found with the sucrose-induced enzyme. Therefore, it was possible to immobilize the enzyme on dextran-based resins using an affinity interaction. Sephadex G-200 was the best resin for immobilizing the dextransucrase and gave a fast flow rate through the packed column. The immobilized dextransucrase retained more than 80% of its specific activity after immobilization ( $K_m\;=\;18.1\;mM$ and $k_{cat}\;=\;450\;sec^{-1}$ vs. 13.1 mM and $640\;sec^{-1}$ , respectively, for the free enzyme). The immobilized dextransucrase showed improved stability over a pH range of 4.0 to 6.5 and at moderately high temperatures over $40^{\circ}C$ . When immobilized dextransucrase was used to synthesize leucrose via the transfer reaction with sucrose and fructose, about 74% of the sucrose was converted into leucrose after one day, and the half-life of the enzyme activity was 15 days. Regeneration of the resin by supplementation with dextransucrase enabled the recovery of the initial activity of the system, but both the reaction and the flow rate were lower, probably owing to the accumulation of dextran inside the resin.


  • 주제어

    dextransucrase .   Leuconostoc mesenteroides .   immobilization .   leucrose.  

  • 참고문헌 (34)

    1. Dextrans;Industrial Gums , De Belder, A.N.;Whistler, R.L.(ed.);BeMiller, J.N.(ed.) , / v.,pp.399-425,
    2. The mechanism of acceptor reactions of Leuconostoc mesenteroides B-512F dextransucrase , Robyt, J.F.;Walseth, T.E. , Carbohydr. Res. / v.61,pp.433-445,
    3. Relative, quantitative effects of acceptors in the reaction of Leuconostoc mesenteroides B-512F dextransucrase , Robyt, J.F.;Eklund, S.H. , Carbohydr. Res. / v.121,pp.279-286,
    4. Leucrose, a ketodisaccharide of industrial design;Carbohydrates as Organic Raw Materials , Schwengers, D.;Lichtenthaler, F.W.(ed.) , / v.,pp.183-195,
    5. Immobilization and properties of Leuconostoc mesenteroides dextransucrase , Kaboli, H.;Reilly, P.J. , Biotech. Bioeng. / v.22,pp.1055-1069,
    6. On the production of dextran by free and immobilized dextransucrase , Monsan, P.;Lopez, A. , Biotech. Bioeng. / v.23,pp.2027-2037,
    7. Investigation of production of dextran and dextransucrase by Leuconostoc mesenteroides immobilized within porous stainless steel , El-Sayed, A.M.M.;Abdul-Wahid, K.;Coughlin, R.W. , Biotech. Bioeng. / v.40,pp.617-624,
    8. Unconventional immobilization of dextransucrase with alginate , Reischwitz, A.;Reh, K.D.;Buchholz, K. , Enzyme Microb. Technol. / v.17,pp.457-461,
    9. Productivity of immobilized dextransucrase for leucrose formation , Reh, K.D.;Noll-Borchers, M.;Buchholz, K. , Enzyme Microb. Technol. / v.19,pp.518-524,
    10. Hemoglobin encapsulation in chitosan/calcium alginate beads , Huguet, M.L.;Groboillot, A.;Neufeld, R.J.;Poncelet, D.;Dellacherie, E. , J. Appl. Polymer Sci. / v.51,pp.1427-1432,
    11. Calcium-alginate beads coated with polycationic polymers: comparison of chitosan and DEAE-dextran , Huguet, M.L.;Neufeld, R.I.;Dellacherie, E. , Process Biochem. / v.31,pp.347-353,
    12. Cloning and sequencing of a gene coding for a novel dextransucrase from Leuconostoc mesenteroides NRRL B-1299 synthesizing only ${\alpha}$(1-6) and ${\alpha}$(1-3) linkages , Monchois, B.;Willemot, R.M.;Remaud-Simeon, M.;Croux, C.;Monsan, P. , Gene / v.182,pp.23-32,
    13. Aggregated form of dextransucrases from Leuconostoc mesenteroides NRRL B-512F and its constitutive mutant , Funane, K.;Yamada, M.;Shiraiwa, M.;Takahara, H.;Yamamoto, N.;Ichishima, E.;Kobayashi, M. , Biosci. Biotech. Biochem. / v.53,pp.776-780,
    14. Large-scale preparation of highly purified dextransucrase from a high-producing constitutive mutant of Leuconostoc mesenteroides B-512FMC , Kitaoka, M.;Robyt, J.F. , Enzyme Microb. Technol. / v.23,pp.386-391,
    15. Production and selection of mutants of Leuconostoc mesenteroides constitutive for glucansucrases , Kim, D.;Robyt, J.F. , Enzyme Microb. Technol. / v.16,pp.659-664,
    16. Mechanism of dextran activation of dextransucrase , Robyt, J.F.;Kim, D.;Yu, L. , Carbohydr. Res. / v.266,pp.293-299,
    17. Properties of Leuconostoc mesenteroides B-512FMC constitutive dextransucrase , Kim, D.;Robyt, J.F. , Enzyme Microb. Technol. / v.16,pp.1010-1015,
    18. Milligram to gram scale purification and characterization of dextransucrase from Leuconostoc mesenteroides NRRL B-512F , Miller, A.W.;Eklund, S.H.;Robyt, J.F. , Carbohydr. Res. / v.147,pp.119-133,
    19. Characterization of the multiple forms and main component of dextransucrase from Leuconostoc mesenteroides NRRL B-512F , Kobayashi, M.;Matsuda, K. , Biochem. Biophys. Acta / v.614,pp.46-62,
    20. Immobilization of thermostable maltogenic amylase from Bacillus stearothermophilus for continuous production of branched oligosaccharides , Kang, G.J.;Kim, M.J.;Kim, J.W.;Park, K.H. , J. Agric. Food Chem. / v.45,pp.4168-4172,
    21. Covalent immobilization of ${\alpha}$-amylase onto PHEMA microspheres: preparation and application to fixed bed reactor , Agric, M.Y.;Hasirci, V.;Alaeddinoglu, N.G. , Biomaterials / v.13,pp.704-709,
    22. Entrapment of urease in glycol containing polymeric matrices and estimation of effective diffusion coefficients of urea , Dermircioglu, H.;Beyenal, H.;Tanyolac, A.;Hasirci, N. , Polymer / v.36,pp.4091-4096,
    23. Mechanisms in the glucansucrase synthesis of polysaccharides and oligosaccharides from sucrose , Robyt, J.F. , Adv. Carbo. Chem. Biochem. / v.51,pp.133-168,
    24. The preparation, properties and structure of the disaccharide leucrose , Stodola, F.H.;Sharpe, E.S.;Koepsell, H.J. , J. Am. Chem. Soc. / v.78,pp.2514-2518,
    25. Dextran synthesis using immobilized Leuconostoc mesenteroides dextransucrase , Monsan, P.;Paul, F.;Auriol, D.;Lopez, A. , Methods Enzymol. / v.136,pp.239-254,
    26. In vitro synthesis of oligosaccharides by acceptor reaction of dextransucrase from Leuconostoc mesenteroides , Pereira, A.M.;Costa, F.A.;Rodrigues, M.I.;Maugeri, F. , Biotechnol. Lett. / v.20,pp.397-401,
    27. Bioaffinity based immobilization of enzymes , Saleemuddin, M. , Adv. Biochem. Eng. Biotechnol. / v.64,pp.203-226,
    28. Use of peptide libraries to map the substrate specificity of a peptide-modifying enzyme: a 13 residue consensus peptide specifies biotinylation in Escherichia coli , Schatz, P.J. , Biotechnology / v.11,pp.1138-1143,
    29. Calmodulin as a versatile tag for antibody fragments , Neri, D.;De Lalla, C.;Petrul, H.;Neri, P.;Winter, G. , Biotechnology / v.13,pp.373-377,
    30. Solid-phase refolding of cyclodextrin glycosyltransferase adsorbed on cationic-exchange resin , Kweon, D.H.;Lee, D.H.;Han, N.S.;Seo, J.H. , Biotech. Prog. / v.20,pp.277-283,
    31. Improved adsorption to starch of a beta-galactosidase fusion protein containing the starch-binding domain from Aspergillus glucoamylase , Chen, L.J.;Ford, C.;Kusnadi, A.;Nikolov, Z.L. , Biotechnol. Prog. / v.7,pp.225-229,
    32. Separation of MBP fusion proteins through affinity membranes , Cattoli, F.;Sarti, G.C. , Biotechnol. Prog. / v.18,pp.94-100,
    33. Cellulose-binding domains: biotechnological applications , Levy, I.;Shoseyov, O. , Biotechnol. Adv. / v.20,pp.191-213,
    34. Conserved repeat motifs and glucan binding by glucansucrases of oral streptococci and Leuconostoc mesenteroides , Shah, D.S.;Joucla, G.;Remaud-Simeon, M.;Russell, R.R. , J. Bacteriol. / v.186,pp.8301-8308,
  • 이 논문을 인용한 문헌 (2)

    1. 2005. "" Food science and biotechnology, 14(6): 818~822     
    2. 2009. "" Food science and biotechnology, 18(5): 1083~1090     

 저자의 다른 논문

  • Han, Nam-Soo (38)

    1. 2000 "Refolding of Bacillus macerans Cyclodextrin Glucanotransferase Expressed as Inclusion Bodies in Recombinant Escherichia coli" Journal of microbiology and biotechnology 10 (5): 632~637    
    2. 2002 "김치로부터 고활성 dextransucrase를 생성하는 저온성 Leuconostoc mesenteroides 균주선발" 한국식품과학회지 = Korean journal of food science and technology 34 (6): 1085~1090    
    3. 2002 "구리흡착 단백질 유전자를 함유하는 재조합 효모의 중금속 흡착" 한국생물공학회지 = Korean journal of biotechnology and bioengineering 17 (1): 38~43    
    4. 2002 "Display of Bacillus macerans Cyclodextrin Glucanotransferase on Cell Surface of Saccharomyces cerevisiae" Journal of microbiology and biotechnology 12 (3): 411~416    
    5. 2002 "Glycomics Reference" 생물산업 : 한국미생물·생명공학회 소식지 = Bioindustry news 15 (1): 33~36    
    6. 2002 "Quantitative Analysis of Leuconostoc mesenteroides and Lactobacillus plantarum Populations by a Competitive Polymerase Chain Reaction" Journal of microbiology and biotechnology 12 (5): 801~806    
    7. 2002 "Simultaneous Biocatalytic Synthesis of Panose During Lactate Fermentation in Kimchi" Journal of microbiology and biotechnology 12 (1): 46~52    
    8. 2003 "신규 기능성당 L-아라비노스: 생리활성, 이용, 생산방법" 한국식품과학회지 = Korean journal of food science and technology 35 (5): 757~763    
    9. 2004 "Statistical Optimization of Medium Composition for Growth of Leuconostoc citreum" Biotechnology and bioprocess engineering 9 (4): 278~284    
    10. 2005 "Comparison of D- and L-Lactic Acid Contents in Commercial Kimchi and Sauerkraut" Food science and biotechnology 14 (1): 64~67    
  • Lee, Soo-Bok (3)

 활용도 분석

  • 상세보기

    amChart 영역
  • 원문보기

    amChart 영역

원문보기

무료다운로드
  • NDSL :
유료다운로드

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