制癌链霉菌ATCC15944发酵液中抗肿瘤组分的制备和初步鉴定

Preparation and Initial Characterization of Anti-Tumor Component from Streptomyces carcino-staticus ATCC15944

新制癌菌素是1965年发现的烯二炔类抗肿瘤抗生素,聚(苯乙烯-马来酸)-NCS偶联物在日本主要用于临床的肝癌介入治疗,目前该药物在国内还没有生产。为了制备新制癌菌素。新制癌菌素产生菌Streptomyces carcino-staticus ATCC15944被在28℃、250 r/min条件下液体发酵,振荡培养72 h。发酵液经预处理后,离心后取上清液经硫酸铵分级沉淀、纯水膜透析分离纯化得到组分SIPI-ZX-1。HPLC分析表明产物纯度大于90%。SDS-PAGE和质谱数据显示,该蛋白质相对分子质量约为11 ku。该产物的抗人肝癌细胞的体外活性被测定。该结果与新制癌菌素的分子质量和抗肿瘤活性数值基本一致。本制备方法简捷,成本较低,本工作为进一步研究新制癌菌素的制备工艺奠定了基础。

Neocarzinostatin, an enediyne antitumor antibiotic, was found in 1965. The styrene maleic acid-based polymer of neocarzi- nostatin was mainly used for the interventional therapy of liver cancer in Japan. This drug still wasn＇t produced in our country. For the preparation of enediyne antibiotics neocarzinostatin,Streptomyces carcino-staticus ATCC15944, neocarzinostatin producing strain, was fermented in the liquid culture media. The flasks were shaken in an incubator at 28 ℃ and 250 r/min for 72 h. After pre- treatment of the broth, one component was obtained from the supernatant, and named SIPI-ZX-1 by ammonium sulfate fractionation salting-out and dialysis. The HPLC analysis showed that the purity of SIPI-ZX-1 was greater than 90% and SDS-PAGE as well as mass spectrometry data showed that the protein molecular weight was about 11 ku. The activity of SIPI-ZX-1 against human liver tumor cell in vitro was determined. The results here are consistent with the molecular weight of the neocarcinostatin and antitumor activity. The methods described here are convenient and low-cost. Our work provides the ground for the further study of the preparation of the neocarcinostatin in large scale.