title: High-level expression of a sika deer (Cervus nippon) Cu/Zn superoxide dismutase in Pichia pastoris and its characterization.
abstract: Production of a sika deer Cu/Zn-SOD was achieved in Pichia pastoris after the reconstituted expression vector pPIC9K was transformed into the strain GS115. By employing Saccharomyces cerevisiae secretion signal peptide (α-factor) under the regulation of the methanol-inducible promoter of the gene of alcohol oxidase 1 (AOX1), sika deer Cu/Zn-SOD with a molecular mass of 16kDa was expressed while recombinant sika deer Cu/Zn-SOD with an activity of 3500U/mL was obtained from a 5L bioreactor. After two successive steps of chromatography on DEAE-650C and Superdex75, recombinant sika deer Cu/Zn-SOD was obtained with 13.8% yield, 14.5-fold purification, and a specific activity of 3447U/mg. Its optimum temperature and optimum pH were 40°C and 7.0, respectively.
authors: College of Biological Science and Technology, Fuzhou University, PR China.
authors: Ren-Kuan
authors: RK
authors: Li
authors: Cai-Li
authors: CL
authors: Fu
authors: Ping
authors: P
authors: Chen
authors: Tzi Bun
authors: TB
authors: Ng
authors: Xiu-Yun
authors: XY
authors: Ye
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D019149:Bioreactors; D003001:Cloning, Molecular; D003670:Deer / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism*; D013696:Temperature
chemical_list: D011994:Recombinant Proteins; D013482:Superoxide Dismutase