title: Purification of a fucoidan from kelp polysaccharide and its inhibitory kinetics for tyrosinase.
abstract: High-speed countercurrent chromatography (HSCCC) was used to separate kelp polysaccharide. HSCCC was performed using an aqueous two-phase solvent system composed of PEG1000-K₂HPO₄-KH₂PO₄-H₂O (0.5:1.25:1.25:7.0, w/w) by eluting a lower aqueous phase at 2.0 mL/min at 600 rpm, yielding two separate fractions, KPS-1 and KPS-2. The KPS-2 fraction was further purified by DEAE-Sepharose fast flow anion-exchange column chromatography to provide 3 fractions, KPS-2-1, KPS-2-2 and KPS-2-3. GPC-HPLC analysis indicated that KPS-2-1 fraction was a purified fucoidan. FT-IR analysis showed that KPS-2-1 was a sulphated polysaccharide. An analysis of enzymatic kinetics showed that the purified fucoidan was a competitive inhibitor of tyrosinase toward l-tyrosine, and the inhibitory constant Ki obtained from double-reciprocal plots was 0.9907 mg/mL.
authors.affiliation: College of Food Science and Biotechnology, Zhejiang Gongshang University, 149 Jiaogong Road, Hangzhou 310035, Zhejiang Province, People's Republic of China. Electronic address: yup9202@gmail.com.
authors.forename: Ping
authors.initials: P
authors.lastname: Yu
authors.forename: Haisen
authors.initials: H
authors.lastname: Sun
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003377:Countercurrent Distribution; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000302:isolation & purification; D044423:Kelp / Q000737:chemistry*; D007700:Kinetics; D014442:Monophenol Monooxygenase / Q000037:antagonists & inhibitors* / Q000737:chemistry; D010710:Phosphates / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry / Q000302:isolation & purification; D017680:Potassium Compounds / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D014443:Tyrosine / Q000737:chemistry*
chemical_list: D004791:Enzyme Inhibitors; D010710:Phosphates; D011134:Polysaccharides; D017680:Potassium Compounds; D012997:Solvents; D011092:Polyethylene Glycols; D014443:Tyrosine; C007789:fucoidan; C013216:potassium phosphate; D014442:Monophenol Monooxygenase
keywords: Fucoidan; High-speed countercurrent chromatography; Kelp polysaccharide; Purification; Tyrosinase