Open AccessExpression of Rice (Oryza sativaL. var. Nipponbare) α-Galactosidase Genes inEscherichia coliand Characterization
Author(s) -
Suhong Li,
Wook-Dong Kim,
Satoshi Kaneko,
Prukutty A. PREMA,
Mitsutoshi Nakajima,
Hideyuki Kobayashi
Publication year - 2007
Publication title -
bioscience biotechnology and biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.509
H-Index - 116
eISSN - 1347-6947
pISSN - 0916-8451
DOI - 10.1271/bbb.60554
Subject(s) - raffinose , galactosidases , oryza sativa , biochemistry , stachyose , escherichia coli , melibiose , glycoside hydrolase , enzyme , biology , alpha galactosidase , molecular mass , affinity chromatography , chemistry , gene , beta galactosidase , fabry disease , disease , pathology , sucrose , medicine
Two putative alpha-galactosidase genes from rice (Oryza sativa L. var. Nipponbare) belonging to glycoside hydrolase family 27 were cloned and expressed in Escherichia coli. These enzymes showed alpha-galactosidase activity and were purified by Ni Sepharose column chromatography. Two purified recombinant alpha-galactosidases (alpha-galactosidase II and III; alpha-Gal II and III) showed a single protein band on SDS-PAGE with molecular mass of 42 kDa. These two enzymes cleaved not only alpha-D-galactosyl residues from the non-reducing end of substrates such as melibiose, raffinose, and stachyose, but also liberated the galactosyl residues attached to the O-6 position of the mannosyl residue at the reducing-ends of mannobiose and mannotriose. In addition, these enzymes clipped the galactosyl residues attached to the inner-mannosyl residues of mannopentaose. Thus, alpha-Gal II catalyzes efficient degalactosylation of galactomannans, such as guar gum and locust bean gum.
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