Coenzyme Aの定量の改良法について

概要

論文の詳細を見る
A method for the assya of coenzyme A has been established, in which yeast enzyme preparation is used instead of pigeon liver enzymes for the acetylation of sulfanilamide in the method of KAPLAN and LIPMANN. The enzyme solution from yeast adequate for co-enzyme A assay can be prepared as follows : cells of Hansenula anomala grown on acetate-glucose medium is acetone-dried.One g of acetone-dried yeast is mixed with 1 g of glass powder and 1.5 ml of M/50 KHCO_3 solution, and after grinding in morter for ca. 15 min., the viscous mass is extracted with additional 3.5 ml of M/50 KHCO_3 solution. Repetition of freezing and thawing is effective in removing inactive protein from the extract. The extract can be stored in frozen state for months without appreciable loss of activity. Thawed extract is centrifuged at 10,000 r.p.m. for 30 min, and resulting supernatant is incubated at 30℃ for hrs to decompose inherent coenxyme A. This procedure is summarized sche-matically in Fig. 1(p. 386).The enzyme solution thus obtained catalyzes the acetylation of sulfanylamide in the presence of acetate, ATP and CoA. The standard curve depicted by plotting the amount of sulfanylamide acetylated against that of coenzyme A present in a reaction mixture with the above yeast enzyme solution is shown in Fig. 3(p. 387), which is essentially similar to that with pigeon liver enzymes shown in Fig. 2. Yeast cells for the source of enzymes in our assay method should be harvested from acetate-glucose medium as the activity of enzyme extract is very low from cells grown on glucose-medium without added acetate. Examples are shown in Table 1 (p. 388). The coenzyme A contents in cells of various microbes and in mold brans estimated by our method are recorded in Table 2(p. 388), in which extraction of CoA is effected by boiling aqueous suspension of a sample for 5 min.
社団法人日本生物工学会の論文
1958-09-15

Coenzyme Aの定量の改良法について

スポンサーリンク

概要

著者

関連論文

スポンサーリンク