Cloning,characterization and functional expression of a new beta-D-fructofuranosidase (Os beta fruct2) cDNA from Oryza sativa |
| |
Authors: | Fu Ru-Huei Wang Yuan-Liang Sung Hsien-Yi |
| |
Affiliation: | (1) Biochemistry Laboratory, Department of Agricultural Chemistry, National Taiwan University, No. 1, Sec. 4, Roosevelt Road, Taipei, 106, Taiwan |
| |
Abstract: | A new cDNA (Osfruct2) encoding an acid –d-fructofuranosidase from rice has been cloned, sequenced and expressed in Pichia pastoris. The full-length cDNA is 2453 base pairs long and encodes a pre-pro-protein of 682 amino acids. The cDNA fragment coding for mature enzyme was sub-cloned into vector pPICZA for extracellular expression in the methylotrophic yeast. The recombinant product was purified by Ni2+-nitrilotriacetic acid agarose column and biochemically characterized. The enzyme could hydrolyze sucrose and raffinose. Molecular mass is 66 kDa. The activity optimum was at pH 4.8 and 40 °C. |
| |
Keywords: | cloning /content/t265j1j056447nr6/xxlarge946.gif" alt=" beta" align=" MIDDLE" BORDER=" 0" >-d-fructofuranosidase functional expression Pichia pastoris rice |
本文献已被 PubMed SpringerLink 等数据库收录! |