title: Purification and characterization of a novel β-agarase of Paenibacillus sp. SSG-1 isolated from soil.
abstract: Agar is a polysaccharide polymer material, generally extracted from seaweed. Most agar degradation strains were isolated from seawater. In order to find new species resources and novel agarase from soil, an agar-degrading bacterium Paenibacillus sp. SSG-1 was isolated from soil. Agarase SSG-1a was purified to homogeneity by 30.2 fold with a yield of 4.8% through ammonium sulfate precipitation, DEAE FF chromatography and native-PAGE separation. The tandem mass spectrometry (MS/MS) results indicated that purified SSG-1a should be a novel β-agarase. The molecular mass of SSG-1a was estimated to be 77 kDa. The optimal temperature and pH for SSG-1a were 50°C and pH 6.0, respectively. Moreover, SSG-1a was stable in pH range of 4.0-10.0 and at temperature up to 40°C. It could hydrolyze the β-1,4 linkage of agarose to produce neoagarohexaose (95 mol%) and neoagarooctaose (5 mol%). Metal ion Mn(2+) and reducing reagents (β-Me and DTT) could increase its activity by 150% and 60%, respectively.
authors.affiliation: Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu 610065, PR China.
authors.forename: Tao
authors.initials: T
authors.lastname: Song
authors.affiliation: Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu 610065, PR China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu 610065, PR China.
authors.forename: Hui
authors.initials: H
authors.lastname: Xu
authors.affiliation: Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu 610065, PR China.
authors.forename: Weijia
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu 610065, PR China; Chengdu Ronsen Pharmaceuticals Co. Ltd., Chengdu 610061, PR China.
authors.forename: Baojin
authors.initials: B
authors.lastname: Fei
authors.affiliation: Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu 610065, PR China.
authors.forename: Dairong
authors.initials: D
authors.lastname: Qiao
authors.affiliation: Microbiology and Metabolic Engineering of Key Laboratory of Sichuan Province, College of Life Science, Sichuan University, Chengdu 610065, PR China. Electronic address: geneium@scu.edu.cn.
authors.forename: Yi
authors.initials: Y
authors.lastname: Cao
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002852:Chromatography, Ion Exchange; D004795:Enzyme Stability; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000302:isolation & purification; D006868:Hydrolysis; D007700:Kinetics; D058889:Molecular Typing; D008970:Molecular Weight; D056507:Paenibacillus / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012685:Sepharose / Q000737:chemistry; D012988:Soil Microbiology; D053719:Tandem Mass Spectrometry
chemical_list: D001426:Bacterial Proteins; D012336:RNA, Ribosomal, 16S; D012685:Sepharose; D006026:Glycoside Hydrolases; C019624:agarase
keywords: Agarase; Native-PAGE separation; Neoagarohexaose; Paenibacillus; Protein purification