α‐Galactosidase from the yeast Torulaspora delbrueckii IFO 1255

Abstract

Y. ODA AND K. TONOMURA. 1996. T h e yeast Torulaspora delbrueckii IF0 1255 was selected as the strain fermenting melibiose from 35 strains of Torulaspora species. T h e strain IF0 1255 produced extracellular and cell-associated forms of a-galactosidase when grown on either melibiose or galactose as the sole carbon source. Most of the enzyme was located outside of the cell membrane : the periplasmic space, or cell walls, or both. a-Galactosidase was purified to homogeneity from the cell-free extract of the strain IF0 1255 by acid treatment and column chromatography on DEAE-Toyopearl650M and ButylToyopearl 650M. T h e molecular weight of the purified enzyme was estimated to be 88 000 by SDS-polyacrylamide gel electrophoresis and 530 000 by gel filtration. T h e enzyme contained 50% of its molecular weight as carbohydrate. Optimum pH and temperature were 4-5-55 and 55"C, respectively. T h e enzyme was inhibited strongly by Ag2+, Hgz+ and Cu2+ each at 1 mmol 1-'. T h e K,,, (mmol 1-I) for p-, o-, mnitrophenyl a-D-galactopyranoside, melibiose, raffinose and stachyose were 2.8, 1.3, 2.8, 4.2, 170 and 230, respectively, and V,,, (pmol min-I mg protein-') for those substrates were 310, 140, 21, 22, 30 and 44, respectively. T h e properties of agalactosidase from T. delbrueckii IF0 1255 were similar to those from the related species, Saccharomyces cerevisiae.

Response surface methodology-based optimization of production media and purification of α-galactosidase in solid-state fermentation by Fusarium moniliforme NCIM 1099

“…Activity of enzyme was completely inhibited by Hg 2+ , similar reports were observed in case of α-galactosidase from Cucurbita pepo plant and various fungal sources like Torulaspora delbrueckii IFO, G. lucidum, and white rot fungus Pleurotus florida (Gaudreault and Webb 1983, Oda and Tonomura 1996, Ramlingum et al 2007). It was observed that EDTA has no negative effect on enzyme activity, while on the other hand, it showed inhibitory effect on Cucurbita pepo (Gaudreault and Webb 1983).…”

Purification and Characterization of Aspergillus terreus α-Galactosidases and Their Use for Hydrolysis of Soymilk Oligosaccharides

“…terreus GR α-galactosidase presented optimum temperature at 65°C and stability for 40 min at this temperature [2]. α-Galactosidase from Aspergillus fumigatus [30], Monascus pilosus [31], and Toluraspora delbrueckii IFO 1255 [32] showed higher activity at 55°C. Differently, α-galactosidases from Penicillium griseoroseum and Penicillium sp.…”

Influence of cultivating conditions on the alpha-galactosidase biosynthesis from a novel strain of Penicillium sp. in solid-state fermentation

“…There are extensive applications of a-galactosidase in food, feed and pharmaceutical industries (Cruz and Park 1982;Dey et al 1993 2 ; Gdala et al 1997). Various micro-organisms produce a-galactosidase, such as fungi (Dey et al 1993;Kotwal et al 1998 3 ), yeasts (Oda and Tonomura 1996) and bacteria (Garro et al 1994). It has been documented that several a-galactosidases could be synthesized by Penicillium ochrochloron and P. purpurogenum (Park et al 1991), P. simplicissimum (Luonteri et al 1998) and P. brevicompactum (Mckay 1991), but they were produced in submerged culture and the enzyme activity was low.…”