Purification and Characterization of Extracellular β-Galactosidase Secreted by Supension Cultured Rice (Oryza sativaL.) Cells

Abstract

A b-galactosidase was puriˆed 1300-fold by lactosylSepharose 4B and Sephacryl S-200 column chromatographies from the cultured medium of a rice-cell suspension. The puriˆed enzyme appeared as 47 kD and 40 kD polypeptides on SDS-PAGE and had a speciˆc activity of 65.1 units W mg. Optimum activity was observed at pH 3.5 and 609 C. The enzyme released galactose from galactoxyloglucan and pectic galactans.

Biochemical characterization of a strictly specific beta‐galactosidase from the digestive juice of the palm weevil Rhynchophorus palmarum larvae

“…The enzyme characterized in this study showed a pH optimum at 5.0, similar to the b-galactosidases from Sterigmatomyces elviae (Onishi & Tanaka 1995) and Helix aspersa liver (Barnett 1971) but different to the beta-galactosidases from chicken seminal plasma (3.4-4.0; Droba & Broba 1987) and suspension-cultured rice (Oryza sativa) cells (3.5; Kaneko & Kobayashi 2003). The similarity in the molecular weights determined by denaturing SDS-PAGE and native gel filtration suggests that b-galactosidase is likely to be monomeric, as found in human liver (Mutoh et al 1988) and Thermus sp.IB-21 (Kang et al 2005) beta-galactosidases.…”

“…Entomological Science (2007) 10, [343][344][345][346][347][348][349][350][351][352] for the beta-galactosidases from cold-adapted bacterium (26°C) (Fernandes et al 2002) and Aspergillus nidulans (30°C) (Diaz et al 1996), but it is lower when compared to those reported for beta-galactosidases from Kluyveromyces lactis (60°C; Ramirez & Rivas 2003) and Sterigmatomyces elviae (85°C; Onishi & Tanaka 1995). The enzyme characterized in this study showed a pH optimum at 5.0, similar to the b-galactosidases from Sterigmatomyces elviae (Onishi & Tanaka 1995) and Helix aspersa liver (Barnett 1971) but different to the beta-galactosidases from chicken seminal plasma (3.4-4.0;Droba & Broba 1987) and suspension-cultured rice (Oryza sativa) cells (3.5; Kaneko & Kobayashi 2003). The similarity in the molecular weights determined by denaturing SDS-PAGE and native gel filtration suggests that b-galactosidase is likely to be monomeric, as found in human liver (Mutoh et al 1988) and Thermus sp.IB-21 (Kang et al 2005) beta-galactosidases.…”

“…IB-21 (Kang et al 2005). However, it might seem low when compared with those reported for beta-galactosidases from the suspension-cultured rice (Oryza sativa) cells (Kaneko & Kobayashi 2003) and Alternaria tenius (Letunova et al 1981).…”

AtBGAL10 Is the Main Xyloglucan β-Galactosidase in Arabidopsis, and Its Absence Results in Unusual Xyloglucan Subunits and Growth Defects

“…1, B and G) could be due to one or more of the other 17 members of family 35 of glycosyl hydrolases (Ahn et al, 2007). A rice ortholog of AtBGAL8 had activity against both xyloglucan and galactans (Kaneko and Kobayashi, 2003). Similarly, a pea ortholog of AtBGAL16 could remove Gal from both xyloglucan and pectic polysaccharides (Dwevedi and Kayastha, 2009).…”

Genomic and expression analysis of glycosyl hydrolase family 35 genes from rice (Oryza sativa L.)

“…Konno and Tsumuki [ 36 ] identified both soluble and cell-wall-bound Bgal, of which a 42 kDa soluble Bgal was purified and shown to release galactose from larch wood and rice cell wall arabinogalactans. Likewise, Kaneko and Kobayashi [ 37 ] isolated a Bgal with 40 and 47 kDa subunits from the medium of rice suspension cells. Recently, Chantarangsee et al [ 38 ] characterized two recombinant Bgal isozymes, including the 90 kDa OsBgal1 and 72 kDa OsBgal2, which had different expression patterns, though both are found throughout plant growth.…”