title: High-level soluble expression of recombinant human manganese superoxide dismutase in Escherichia coli, and its effects on proliferation of the leukemia cell.
abstract: Manganese superoxide dismutase (Mn-SOD) is one of the major enzymes responsible for the defense against oxidative damage due to reactive oxygen species (ROS) in the mitochondria. The present study aimed to produce and evaluate the genetically engineered manganese superoxide dismutase protein. A recombinant plasmid containing DNA segment coding Mn-SOD protein was transformed into Escherichia coli (E. coli) Rosetta-gami strain, for expression. After induction with IPTG, an expected molecular mass of 25 kDa was detected by SDS-PAGE. After Ni-NTA affinity chromatography purification, the purity rate came up to 95%. UV spectroscopy data for our preparations indicated that a peak at 275 nm existed in the spectrum. SOD activity assay showed that the activity of the rhMn-SOD was 1890.9 U/mg. The ORAC level of rhMn-SOD was 151492.2 uM Trolox equiv/mg. Furthermore, in vitro bioactivity assay indicated that the rhMn-SOD protein can inhibit the proliferation of the leukemia K562 cells.
authors.affiliation: College of Pharmacy, Jinan University, Guangzhou 510632, China. novelfunction@163.com
authors.forename: Wang
authors.initials: W
authors.lastname: Feng
authors.forename: Shao
authors.initials: S
authors.lastname: Mei
authors.forename: Yang
authors.initials: Y
authors.lastname: Wenjie
authors.forename: Huang
authors.initials: H
authors.lastname: Luyuan
mesh_terms: D000595:Amino Acid Sequence; D048708:Cell Growth Processes / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D020014:K562 Cells; D004915:Leukemia, Erythroblastic, Acute / Q000188:drug therapy* / Q000473:pathology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D016415:Sequence Alignment; D012995:Solubility; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013482:Superoxide Dismutase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*
chemical_list: D011994:Recombinant Proteins; D013482:Superoxide Dismutase