title: Purification and Characterization of a Fucoidanase (FNase S) from a Marine Bacterium Sphingomonas paucimobilis PF-1.
abstract: The Search for enzyme activities that efficiently degrade marine polysaccharides is becoming an increasingly important area for both structural analysis and production of lower-molecular weight oligosaccharides. In this study, an endo-acting fucoidanase that degrades Miyeokgui fucoidan (MF), a sulfated galactofucan isolated from the sporophyll (called Miyeokgui in Korean) of Undaria pinnatifida, into smaller-sized galactofuco-oligosaccharides (1000-4000 Da) was purified from a marine bacterium, Sphingomonas paucimobilis PF-1, by ammonium sulfate precipitation, diethylaminoethyl (DEAE)-Sepharose column chromatography, and chromatofocusing. The specific activity of this enzyme was approximately 112-fold higher than that of the crude enzyme, and its molecular weight was approximately 130 kDa (FNase S), as determined by native gel electrophoresis and 130 (S1), 70 (S2) and 60 (S3) kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The optimum pH and temperature of FNase S were pH 6.0-7.0 and 40-45 °C, respectively. FNase S activity was enhanced by Mn2+ and Na+ (115.7% and 131.2%), but it was inhibited by Ca2+, K+, Ba2+, Cu2+ (96%, 83.7%, 84.3%, and 89.3%, respectively), each at 1 mM. The Km, Vmax and Kcat values of FNase S on MF were 1.7 mM, 0.62 mg·min-1, and 0.38·S-1, respectively. This enzyme could be a valuable tool for the structural analysis of fucoidans and production of bioactive fuco-oligosaccharides.
authors.affiliation: Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea, Bucheon, Gyeonggi-do 420-743, Korea. wj0504@gstep.re.kr.|Biocenter, Gyeonggi Institute of Science and Technology Promotion (GSTEP), Suwon, Gyeonggi-do 443-270, Korea. wj0504@gstep.re.kr.
authors.forename: Woo Jung
authors.initials: WJ
authors.lastname: Kim
authors.affiliation: Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea, Bucheon, Gyeonggi-do 420-743, Korea. pjwnbd@hanmail.net.
authors.forename: Joo Woong
authors.initials: JW
authors.lastname: Park
authors.affiliation: Department of Pharmaceutical Science, University of Gachon, Yeonsu-dong, Yeonsu-gu, InCheon 406-799, Korea. jkpark@gachon.ac.kr.
authors.forename: Jae Kweon
authors.initials: JK
authors.lastname: Park
authors.affiliation: Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea, Bucheon, Gyeonggi-do 420-743, Korea. myenwls@hanmail.net.
authors.forename: Doo Jin
authors.initials: DJ
authors.lastname: Choi
authors.affiliation: Department of Biotechnology and The Biomaterial Engineering Research Center, The Catholic University of Korea, Bucheon, Gyeonggi-do 420-743, Korea. yongil382@catholic.ac.kr.
authors.forename: Yong Il
authors.initials: YI
authors.lastname: Park
mesh_terms: D059001:Aquatic Organisms / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases / Q000302:isolation & purification*; D008970:Molecular Weight; D011134:Polysaccharides / Q000378:metabolism; D020578:Sphingomonas / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D011134:Polysaccharides; C007789:fucoidan; D006026:Glycoside Hydrolases
keywords: Sphingomonas sp.; fucoidan; fucoidanase; galactofuco-oligosaccharides