Makhmud S A, Machur V A, Zl'-khadad M E, Zl'-gamal S E
Prikl Biokhim Mikrobiol. 1978 May-Jun;14(3):466-70.
Isolation and purification of glucoamylase from Aspergillus foetidus ATCC 14916 were examined. Ethanol and ammonium sulphate proved to be the best precipitants. With their use the yield of glucoamylase was 99.0 and 92.3%, respectively. Treatment of the culture liquid with 0.5% CaCl2 and activated charcoal clarified the filtrate noticeably and left glucoamylase activity unchanged. During submerged cultivation the gungus synthesized no glucosyl transferase. A method to inactivate alpha-amylase through acidification of the reaction mixture to bring pH to 2.5 and incubation for 15 min at 60 degrees C was developed. Glyoamylase was partially purified from ballast proteins by means of (NH4)2SO4 at 19% of total saturation. Glucoamylase was obtained at 79% saturation. The infrared spectrum was recorded in the fraction obtained by (NH4)2SO4 precipitation at 39, 53, 66, 72 and 79% saturation. In all the cases the spectra were identical. This is another proof of purity of the resultant preparation.
对来自臭曲霉ATCC 14916的糖化酶进行了分离和纯化研究。结果表明乙醇和硫酸铵是最佳沉淀剂。使用它们时,糖化酶的产率分别为99.0%和92.3%。用0.5%氯化钙和活性炭处理培养液能显著澄清滤液,且糖化酶活性不变。在深层培养过程中,该真菌不合成葡萄糖基转移酶。开发了一种通过将反应混合物酸化至pH 2.5并在60℃孵育15分钟来使α-淀粉酶失活的方法。通过硫酸铵在总饱和度19%的条件下从杂质蛋白中部分纯化糖化酶。在79%饱和度下获得糖化酶。记录了在39%、53%、66%、72%和79%饱和度下通过硫酸铵沉淀获得的馏分的红外光谱。在所有情况下,光谱都是相同的。这是所得制剂纯度的又一证据。