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使用Analytab API ZYM系统测定荧光假单胞菌的胞外和细胞相关水解酶谱。

Determination of the extracellular and cell-associated hydrolase profiles of Pseudomonas fluorescens sp. using the Analytab API ZYM system.

作者信息

McKellar R C

出版信息

J Dairy Sci. 1986 Mar;69(3):658-64. doi: 10.3168/jds.S0022-0302(86)80453-2.

Abstract

The extracellular and cell-associated hydrolase profiles of a number of Pseudomonas fluorescens strains were examined with the Analytab API ZYM system. Esterase/lipase was the only strong extracellular enzyme activity detected (mean 3.33): weak esterase, lipase, and leucine aminopeptidase activities were found with some strains (mean activities of 1.08, 1.53, and 1.40, respectively). Very strong leucine aminopeptidase activity (4.5) was associated with the cells. Cell-associated trypsin, esterase/lipase, acid phosphatase, and phosphoamidase were also found. Neither extracellular nor cell-associated hydrolase profiles changed significantly when cells were grown in skim milk or mineral salts medium at either 5 or 20 degrees C. Similarly, added calcium did not seem required for synthesis of any of the enzymes. The extracellular enzyme profiles differed considerably from those of the cell-associated enzymes for all strains tested. An extracellular proteinase-deficient mutant of strain 32A (RM14) failed to produce significant quantities of extracellular esterase/lipase activity. Production of cell-associated enzymes was unaffected by the mutation. These results suggest that the Analytab API ZYM system may be useful in identifying psychrotrophs isolated from milk.

摘要

利用Analytab API ZYM系统检测了多种荧光假单胞菌菌株的胞外和细胞相关水解酶谱。酯酶/脂肪酶是唯一检测到的强胞外酶活性(平均为3.33):在一些菌株中发现了弱酯酶、脂肪酶和亮氨酸氨肽酶活性(平均活性分别为1.08、1.53和1.40)。非常强的亮氨酸氨肽酶活性(4.5)与细胞相关。还发现了与细胞相关的胰蛋白酶、酯酶/脂肪酶、酸性磷酸酶和磷酸酰胺酶。当细胞在脱脂牛奶或矿物盐培养基中于5℃或20℃培养时,胞外和细胞相关水解酶谱均未发生显著变化。同样,似乎合成任何一种酶都不需要添加钙。对于所有测试菌株,胞外酶谱与细胞相关酶谱有很大差异。菌株32A(RM14)的胞外蛋白酶缺陷突变体未能产生大量胞外酯酶/脂肪酶活性。该突变不影响细胞相关酶的产生。这些结果表明,Analytab API ZYM系统可能有助于鉴定从牛奶中分离出的嗜冷菌。

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