Wartmann T, Kunze I, Duc B M, Manteuffel R, Kunze G
Institute of Plant Genetics and Crop Plant Research, Gatersleben, Germany.
Microbiol Res. 1995 May;150(2):113-20. doi: 10.1016/S0944-5013(11)80045-5.
Seven Arxula adeninivorans strains originating from The Netherlands (CSIR 1136, CSIR 1138 and CBS 8244T), South Africa (CSIR 1147, CSIR 1148 and CSIR 1149) and Siberia (Ls3) were compared concerning their secretory glucoamylase. Within the spectrum of secretory polypeptides obtained by SDS-polyacrylamide gel electrophoresis, the glucoamylase corresponding polypeptide, could be identified by means of specific antibodies directed against the glucoamylase of strain Ls3. The molecular masses of the glucoamylases vary from 84 to 95 kDa. After endoglycosidase F treatment of the secretory proteins, the N-linked carbohydrate content for each glucoamylase could be quantified at about 25%. Glucoamylase activity staining of secretory proteins separated under nondenaturing conditions revealed one glucoamylase active protein for the Dutch strains and two active forms for the strains originating from South Africa and Siberia. Highest activities of glucoamylase can be measured at the end of the exponential growth phase for each strain. The Dutch strain CSIR 1138 secretes the highest activity. Optimum values for temperature and pH were determined and compared. By means of pulsed field gel electrophoresis and Southern analysis, the glucoamylase corresponding gene could be localized on the second of the four chromosomes of each yeast strain.
对来自荷兰（CSIR 1136、CSIR 1138和CBS 8244T）、南非（CSIR 1147、CSIR 1148和CSIR 1149）和西伯利亚（Ls3）的7株嗜腺嘌呤阿氏酵母菌株的分泌型糖化酶进行了比较。在通过SDS-聚丙烯酰胺凝胶电泳获得的分泌多肽谱中，可通过针对Ls3菌株糖化酶的特异性抗体鉴定出与糖化酶相对应的多肽。糖化酶的分子量在84至95 kDa之间。对分泌蛋白进行内切糖苷酶F处理后，每种糖化酶的N-连接碳水化合物含量可定量为约25%。在非变性条件下分离的分泌蛋白的糖化酶活性染色显示，荷兰菌株有1种糖化酶活性蛋白，而来自南非和西伯利亚的菌株有2种活性形式。每种菌株在指数生长期结束时可测得最高的糖化酶活性。荷兰菌株CSIR 1138分泌的活性最高。测定并比较了温度和pH的最佳值。通过脉冲场凝胶电泳和Southern分析，可将糖化酶对应基因定位在每个酵母菌株四条染色体中的第二条上。
