Hashimoto H, Kaneko Y, Iwaasa T, Wokotsuka T
Appl Microbiol. 1973 Apr;25(4):584-8. doi: 10.1128/am.25.4.584-588.1973.
A thermophilic fungus, Penicillium duponti K1014, produced an acid protease in a submerged culture. Maximum enzyme production, in a 30-liter fermentor at a preselected optimum growth temperature of 50 C, occurred after 3 days. The productivity of the enzyme was associated with changes of viscosity and pH of culture broth during the growth. The acid protease was isolated from the culture filtrates and was purified ninefold by alcohol precipitation, column chromatography on O-(diethylaminoethyl)cellulose, batchwise treatment with O-(carboxymethyl)cellulose, and by gel filtration. The purified enzyme was homogeneous on disk electrophoresis and sedimentation analysis. The sedimentation constant s(20.w), calculated at a concentration of 0.64%, was 3.98S.
嗜热真菌杜邦青霉K1014在深层培养中产生一种酸性蛋白酶。在30升发酵罐中，于预先选定的50℃最佳生长温度下，培养3天后酶产量达到最大值。酶的生产率与生长过程中培养液的粘度和pH值变化有关。从培养滤液中分离出酸性蛋白酶，并通过乙醇沉淀、O-（二乙氨基乙基）纤维素柱色谱、O-（羧甲基）纤维素分批处理和凝胶过滤将其纯化了9倍。纯化后的酶在圆盘电泳和沉降分析中呈均一性。在浓度为0.64%时计算出的沉降常数s(20.w)为3.98S。
