Pseudomonas aeruginosa produces two distinct ADP-ribosyl transferases, exotoxin A and exoenzyme S, which differ in a number of properties including substrate specificity. Exoenzyme S was purified from culture supernatants of P. aeruginosa DG1. The procedure for purification consists of four major steps: ammonium sulfate precipitation, anion-exchange chromatography on DEAE-Sephacel, acetone precipitation in the presence of 1 M NaCl, and G-100 Superfine gel filtration chromatography. Exoenzyme S was monitored during purification by an assay for ADP-ribosyl transferase activity, mouse toxicity, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The purified material exhibited ADP-ribosyl transferase activity, reacted with monoclonal antibodies to exoenzyme S, and was toxic to mice and a variety of tissue culture cell lines.
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机译：铜绿假单胞菌产生两种不同的ADP-核糖基转移酶，外毒素A和外酶S，它们在包括底物特异性在内的许多特性上都不同。从铜绿假单胞菌DG1的培养上清液中纯化外切酶S。纯化过程包括四个主要步骤：硫酸铵沉淀，DEAE-Sephacel上的阴离子交换色谱，在1 M NaCl存在下的丙酮沉淀和G-100超细凝胶过滤色谱。通过测定ADP-核糖基转移酶活性，小鼠毒性和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳的方法监测纯化过程中的外切酶S。纯化的物质表现出ADP-核糖基转移酶活性，与抗外切酶S的单克隆抗体反应，并且对小鼠和多种组织培养细胞系具有毒性。
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