Fujishiro K, Sanada Y, Tanaka H, Katunuma N
J Biochem. 1980 May;87(5):1321-6. doi: 10.1093/oxfordjournals.jbchem.a132870.
Protease B [EC 3.4.22.9] was purified from baker's yeast by plasmolysis of yeast, acid activation, acid precipitation, and column chromatographies on QAE-Sephadex, SP-Sephadex, D-tryptophan methyl ester-Sepharose 4B and Sephadex G-100. The purified enzyme was inhibited by phenylmethylsulfonyl fluoride and sulfhydryl-blocking reagents. Chymostatin and antipain at extremely low concentrations (1 micro M) inhibited the protease B. The effects of the enzyme on various yeast enzymes were examined by measuring their inactivation. The enzyme inactivated 6-phosphogluconate dehydrogenase [EC 1.1.1.44] and uricase [EC 1.7.3.3], but not malate dehydrogenase [EC 1.1.1.37], alcohol dehydrogenase [EC 1.1.1.1], glutamate dehydrogenase [EC 1.4.1.3], glucose-6-phosphate dehydrogenase [EC 1.1.1.49] or hexokinase [EC 2.7.1.1].
蛋白酶B [EC 3.4.22.9] 是通过对面包酵母进行质壁分离、酸激活、酸沉淀以及在QAE-葡聚糖凝胶、SP-葡聚糖凝胶、D-色氨酸甲酯-琼脂糖凝胶4B和葡聚糖凝胶G-100上进行柱色谱法从面包酵母中纯化得到的。纯化后的酶受到苯甲基磺酰氟和巯基封闭试剂的抑制。极低浓度（1微摩尔）的抑肽酶和抗蛋白酶能抑制蛋白酶B。通过测量各种酵母酶的失活情况来检测该酶对它们的影响。该酶能使6-磷酸葡萄糖酸脱氢酶 [EC 1.1.1.44] 和尿酸酶 [EC 1.7.3.3] 失活，但不能使苹果酸脱氢酶 [EC 1.1.1.37]、乙醇脱氢酶 [EC 1.1.1.1]、谷氨酸脱氢酶 [EC 1.4.1.3]、葡萄糖-6-磷酸脱氢酶 [EC 1.1.1.49] 或己糖激酶 [EC 2.7.1.1] 失活。
