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(高效抗肿瘤抗生素calicheamicin的发酵和分离纯化研究 陈华1,2*，郑玲辉1，姜璐琰1，王继栋1，王玲萍1，白骅1 (1. 浙江海正药业股份有限公司中研院微生物研究所, 浙江台州 318000; 2. 浙江省抗真菌药物研究重点实验室, 浙江台州 318000) 摘要：目的 从一株产抗肿瘤抗生素calicheamicin (CLM)原始菌种出发，旨在提高发酵单位和建立分离纯化CLM的工艺。方法 通过微波、NTG等物理化学方法选育CLM高产突变菌种，利用有机溶剂萃取、硅胶分离、制备HPLC精制等方法分离纯化CLM。结果 从原始菌种Micromonospora echinospora ssp. Calichensis出发选育出CLM高产菌种，发酵单位达到50 μg?mL-1，并分离出目标活性组分CLM γ1I。结论 通过提高发酵单位和建立分离提取工艺路线，为该产品的产业化开发打好了基础。 关键词：Calicheamicin; 高产菌种; 分离纯化 中图分类号： 文献标识码：A 文章编号： Fermentation and Purification of a High Active Antitumor Antibiotic Calicheamicin CHEN Hua1,2*, ZHENG Lin-hui1, JIANG Lu-yan1, WANG Ji-dong1, WANG Lin-ping1, BAI Hua1 (1. The RD center, Zhejiang Hisun Pharmaceutical Co. Ltd., Taizhou 318000; 2. Zhejiang Key Laboratory of Antifungal Drugs, Taizhou 318000, China) Abstract: OBJECTIVE The aims of this study were to improve the production of calicheamicin (CLM) and to purify the antitumor antibiotic. Methods The microwave and NTG were applied as mutagenic source for breeding high-yielding mutant strain. The isolation procedures of organic solvent extraction, silica gel chromatography, and preparative HPLC were used to purify CLM γ1I. RESULTS A high-yielding mutant strain with CLM production of 50 μg?mL-1 was screening out. The purified CLM γ1I was obtained after the isolation steps. CONCLUSION The production of CLM γ1I was improved and the purification steps were established. The results of this study were useful for further industrialization of this product. Key words: Calicheamicin; high-yielding strain; purification 恶性肿瘤是威胁人类健康的最严重疾病。长期以来，肿瘤治疗一直都是医学界研究的一大课题，寻找治疗肿瘤的新药更是药物研究人员的首要任务。很多抗肿瘤药物都是由微生物代谢产生的，如doxorubicin, geldanamycin, epothilone B, bleomycin等。其中烯二炔类抗肿瘤抗生素具有特殊的分子结构和作用机制，对多种肿瘤细胞均有强烈的杀伤作用，是迄今发现的活性最强的一类抗肿瘤抗生素[1]。───单克隆抗体的“弹头”进行开发[2-3]。其中以CLM作为“弹头”的靶向治疗药物GO (Gemtuzumab ozogamicin)已被FDA批准用于治疗[4]。 然而，CLM由于发酵单位过低，文献报道不到10 μg?mL-1，而且存在多种同系物[5]，对其分离纯化和进一步开发带来很大难度。本文从一株产CLM原始菌种出发，从提高发酵单位和建立分离提取工艺进行了研究，得出较好的结果。 1 材料 1.1 菌种 实验所用原始菌种为棘孢小单孢菌Calichensis。高产菌种91