|Budget Amount *help
¥1,700,000 (Direct Cost : ¥1,700,000)
Fiscal Year 1991 : ¥400,000 (Direct Cost : ¥400,000)
Fiscal Year 1990 : ¥1,300,000 (Direct Cost : ¥1,300,000)
Aspergillus niger alpha-glucosidase catalyzed the slow hydroysis of beta-Dglucopyranosyl fluoride to form alpha-D-glucose. Maltotriose competitively inhibited the hydrolysis, and beta-D-glucosyl fluoride in turn competitively inhibited the hydrolysis of p-nitrophenyl alpha-D-glucopyranoside, indicating that beta-D-glucosyl fluoride is bound at the same site as known substrates for the alpha-glucosidase.
Also alpha-glucosidases from sugar beet and rice catalyzed the hydrolysis of beta-D-glucosyl fluoride to form alpha-D-glucose. The reaction was slow, with V/K 1-2% of that for PNP alpha-D-glucoside hydrolysis, but was not due to any impurity in the substrate or to contaminating beta-glucosidase or glucoamylase. Maltopentaose was a linear competitive inhibitor of PNP alpha-glucoside hydrolysis in each case. Almond beta-glucosidase, promoted alpha-D-glucosyl fluoride hydrolysis to form beta-glucose at an exceedingly low rate. This weak reaction did not stem from any impurity in substance,
or to contamination with alpha-glucosidase or glucoamylase, but it was partly(ca. 20%)attributable to a trace of accompanying trehalase.
That the disfavored D-glucosyl fluoride in each case was converted to a product of the same configuration as from enitols or favored D-glucosyl substrates provides ndw evidence for a separate stage of catalysis in which the steric outcome of reactions of a glucosidase is strictly conserved by protein structure regardless of substrate configuration.
Coffee bean alpha-galactosidase was found to catalyze the hydration of D-galactal and D-galacto-octenitol, each a known substrate for beta-galactosidase. The enzyme protonated each substrate from beneath the plane of the ring, as assumed for alpha-D-galactosides. These results provide an unequivocal assignment of the orientation of an acidic catalytic group to the alpha-galactosidase reaction center. The findings are discussed in light of the concept that catalysis by glycosidases involves a "plastic" protonation phase and a "conserved" product configuration phase.
5.テンサイおよびコメ起源αーグルコシダ-ゼについてβーglucosyl fluoride作用能を調べ,これら酵素もβー型基質のCーF切断を解媒することを確認した。これら事実から,一般にグリコシラ-ゼは基質のアノマ-を厳格には選択しないと言う結論に達することができた。 Less