Characterization of a novel alkaline arylsulfatase from Marinomonas sp. FW-1 and its application in the desulfation of red seaweed agar.
A bacterial strain capable of hydrolyzing sulfate ester bonds of p-nitrophenyl sulfate (pNPS) and agar was isolated from the coast area of Qingdao, China. It was identified as Marinomonas based on its 16S rRNA gene sequence and named as Marinomonas sp. FW-1. An arylsulfatase with a recovery of 13 % and a fold of 12 was purified to a homogeneity using ion exchange and gel filtration chromatographies. The enzyme was composed of a single polypeptide chain with the molecular mass of 33 kDa estimated using SDS-PAGE. The optimal pH and temperature of arylsulfatase were pH 9.0 and 45, respectively. Arylsulfatase was stable over pH 8-11 and at temperature below 55 °C. The K m and V max of this enzyme for the hydrolysis of pNPS were determined to be 13.73 and 270.27 μM/min, respectively. The desulfation ratio against agar from red seaweed Gelidium amansii and Gracilaria lemaneiformis were 86.11 and 89.61 %, respectively. There was no difference between the DNA electrophoresis spectrum on the gel of the arylsulfatase-treated G. amansii agar and that of the commercial agarose. Therefore, this novel alkaline arylsulfatase might have a great potential for application in enzymatic conversion of agar to agarose.
College of Food Science and Engineering, Ocean University of China, No. 5 Yushan Road, Qingdao, 266003, China.
Xueyan
X
Wang
Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China.
Delin
D
Duan
College of Food Science and Engineering, Ocean University of China, No. 5 Yushan Road, Qingdao, 266003, China.
Jiachao
J
Xu
College of Food Science and Engineering, Ocean University of China, No. 5 Yushan Road, Qingdao, 266003, China.
Xin
X
Gao
College of Food Science and Engineering, Ocean University of China, No. 5 Yushan Road, Qingdao, 266003, China. xiaotingfu@ouc.edu.cn.
Xiaoting
X
Fu
D000362:Agar / Q000737:chemistry* / Q000378:metabolism*; D001192:Arylsulfatases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D002681:China; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D053544:Marinomonas / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D008970:Molecular Weight; D009578:Nitrobenzenes / Q000378:metabolism; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012624:Seaweed / Q000737:chemistry*; D012685:Sepharose / Q000737:chemistry / Q000378:metabolism; D013696:Temperature
D009578:Nitrobenzenes; D012336:RNA, Ribosomal, 16S; C023379:4-nitrophenyl sulfate; D000362:Agar; D012685:Sepharose; D001192:Arylsulfatases
Agarose; Arylsulfatase; Characterization; Desulfation; Purification