Wissing F
J Bacteriol. 1975 Feb;121(2):695-9. doi: 10.1128/jb.121.2.695-699.1975.
A cell-free preparation with cyanide-producing activity was obtained from a bacterium, strain C, of the genus Pseudomonas. To preserve activity, an oxidizing agent, e.g., phenazine methosulphage (PMS), had to be added to the cell suspension before disruption by sonic treatment. By the procedure described, a total homogenate made from a 15% (wet weight) bacterial suspension in tris(hydroxymethyl)aminomethane-hydrochloride buffer (0.05 M, pH 8.2) and with PMS (0.4mM) exhibited about 8% of the activity obtained from a suspension of untreated bacteria. In the presence of flavine-adenine dinucleotide (0.3 mM) and PMS (0.4mM), the activity was augmented to about 16% of that of the intact cells. By gradient centrifugation the homogenate was separated into three fractions. The main enzyme activity was associated with those fractions which by electron microscopy were found to consist of membranous structures.
从假单胞菌属的一种细菌(菌株C)中获得了具有产氰活性的无细胞制剂。为了保持活性,在通过超声处理破坏细胞之前,必须向细胞悬液中添加一种氧化剂,例如吩嗪硫酸甲酯(PMS)。按照所述方法,由在三(羟甲基)氨基甲烷 - 盐酸缓冲液(0.05M,pH 8.2)中含PMS(0.4mM)的15%(湿重)细菌悬体制成的总匀浆表现出约8%从未经处理的细菌悬液中获得的活性。在存在黄素腺嘌呤二核苷酸(0.3mM)和PMS(0.4mM)的情况下,活性增加到完整细胞活性的约16%。通过梯度离心将匀浆分离成三个部分。主要酶活性与通过电子显微镜发现由膜结构组成的那些部分相关。