title: A novel cold-adapted β-galactosidase isolated from Halomonas sp. S62: gene cloning, purification and enzymatic characterization.
abstract: A novel 1,170 bp β-galactosidase gene sequence from Halomonas sp. S62 (BGalH) was identified through whole genome sequencing and was submitted to GenBank (Accession No. JQ337961). The BGalH gene was heterologously expressed in Escherichia coli BL21(DE3) cells, and the enzymatic properties of recombinant BGalH were studied. According to the polyacrylamide gel electrophoresis results and the sequence alignment analysis, BGalH is a dimeric protein and cannot be classified into one of the known β-galactosidase families (GH1, GH2, GH35, GH42). The optimal pH and temperature were determined to be 7.0 and 45 °C, respectively; the K m and K cat were 2.9 mM and 390.3 s(-1), respectively, for the reaction with the substrate ortho-nitrophenyl-β-D-galactopyranoside. At 0-20 °C, BGalH exhibited 50-70 % activity relative to its activity under the optimal conditions. BGalH was stable over a wide range of pHs (6.0-8.5) after a 1 h incubation (>93 % relative activity) and was thermostable at 50 °C and below (>60 % relative activity). The enzyme hydrolyzes lactose completely in milk over 24 h at 7 °C. The characteristics of this novel β-galactosidase suggest that BGalH may be a good candidate for medical researches and food industry applications.
authors.affiliation: Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai 200433, China.
authors.forename: Guo-Xiang
authors.initials: GX
authors.lastname: Wang
authors.forename: Yun
authors.initials: Y
authors.lastname: Gao
authors.forename: Bo
authors.initials: B
authors.lastname: Hu
authors.forename: Xiao-Ling
authors.initials: XL
authors.lastname: Lu
authors.forename: Xiao-Yu
authors.initials: XY
authors.lastname: Liu
authors.forename: Bing-Hua
authors.initials: BH
authors.lastname: Jiao
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D003001:Cloning, Molecular; D003080:Cold Temperature; D004795:Enzyme Stability; D020632:Halomonas / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D012623:Seawater / Q000382:microbiology*; D013379:Substrate Specificity; D001616:beta-Galactosidase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D001616:beta-Galactosidase