Cloning, characterization and functional expression of a new beta-D-fructofuranosidase (Os beta fruct2) cDNA from Oryza sativa.
A new cDNA (Os beta fruct2) encoding an acid beta-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 pPICZalphaA 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 degrees C.
Fu
Ru-Huei
RH
Biochemistry Laboratory, Department of Agricultural Chemistry, National Taiwan University, No. 1, Sec. 4, Roosevelt Road, Taipei 106, Taiwan. f85623601@ms86.ntu.edu.tw
Wang
Yuan-Liang
YL
Sung
Hsien-Yi
HY
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D006026:Glycoside Hydrolases / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D008969:Molecular Sequence Data; D012275:Oryza / Q000201:enzymology / Q000235:genetics; D010843:Pichia / Q000737:chemistry* / Q000201:enzymology* / Q000254:growth & development*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D043324:beta-Fructofuranosidase
D018076:DNA, Complementary; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase