Abstract
A marine bacterial strain, Fucobacter marina, produced extracellular sulfated fucoglucuronomannan (SFGM) lyase when cultivated in the presence of crude SFGM obtained from fucoidan of Kjellmaniella crassifolia (brown algae) by cetyl pyridinium chloride fractionation. For the SFGM lyase assay, SFGM fraction separated from K. crassifolia fucoidan by anion exchange column chromatography was used as the substrate. The extracellular SFGM lyase was purified to homogeneity on an electrophoresis gel with 4240-fold purity at 13.8% yield. The enzyme proved to be a monomer, since gel filtration and sodium dodecyl sulfate polyacrylamide gel electrophoresis gave the same relative molecular mass of 67,000. The enzyme specifically digested SFGM but did not digest any other uronic-acid-containing polysaccharides tested. The optimum conditions for the enzyme reaction were around pH 7.5, 43°C, and 0.4 M NaCl concentration. The enzyme was strongly inhibited by CuCl2 and ZnCl2, and also by some sulfhydryl reagents.
Similar content being viewed by others
References
T. Bitter H.M. Muir (1962) ArticleTitleA modified uronic acid carbazole reaction. Anal Biochem 4 330–334 Occurrence Handle1:CAS:528:DyaF3sXnsVyrtw%3D%3D
M. Dubois K.A. Gilles J.K. Hamilton P.A. Rebers F. Smith (1956) ArticleTitleColorimetric method for determination of sugars and related substances. Anal Chem 28 350–356 Occurrence Handle1:CAS:528:DyaG28XjvFynsg%3D%3D
T. Sakai H. Kimura I. Kato (2002) ArticleTitleA marine strain of Flavobacteriaceae utilizes brown seaweed fucoidan. Mar Biotechnol 4 399–405 Occurrence Handle1:CAS:528:DC%2BD38XosVGrurY%3D
T. Sakai H. Kimura K. Kojima K. Shimanaka K. Ikai I. Kato (2003) ArticleTitleMarine bacterial sulfated fucoglucuronomannan (SFGM) lyase digests brown algal SFGM into trisaccharides. Mar Biotechnol 5 70–78 Occurrence Handle10.1007/s10126-002-0056-3 Occurrence Handle1:CAS:528:DC%2BD3sXit1Sgs7w%3D
N. Seno K. Anno K. Kondo S. Nagase S. Saito (1970) ArticleTitleImproved method for electrophoretic separation and rapid quantitation of isomeric chondroitin sulfates on cellulose acetate strips. Anal Biochem 37 197–202 Occurrence Handle1:CAS:528:DyaE3cXltFOjsL8%3D Occurrence Handle5506556
T. Takeuchi K. Murata I. Kusakabe (1994) ArticleTitleA method for depolymerization of alginate using the enzyme system of Flavobacterium multivolum. Nippon Shokuhin Kogyo Gakkaishi 41 505–511 Occurrence Handle1:CAS:528:DyaK2cXlslOrs7o%3D
T. Yamagata H. Saito O. Habuchi S. Suzuki (1968) ArticleTitlePurification and properties of bacterial chondroitinases and chondrosulfatases. J Biol Chem 243 1523–1535 Occurrence Handle1:CAS:528:DyaF1cXosVGisw%3D%3D Occurrence Handle5647268
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Sakai, T., Kimura, H. & Kato, I. Purification of Sulfated Fucoglucuronomannan Lyase from Bacterial Strain of Fucobacter marina and Study of Appropriate Conditions for Its Enzyme Digestion . Mar. Biotechnol. 5, 380–387 (2003). https://doi.org/10.1007/s10126-002-0083-0
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10126-002-0083-0