Yao Kaiya, Gao Shuhong, Wu Yanjie, Zhao Zhen, Wang Wen, Mao Quangui
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China.
Topfond Pharmaceutical Co., Ltd, Zhumadian, Henan, 463000, China.
Folia Microbiol (Praha). 2018 Jan;63(1):105-113. doi: 10.1007/s12223-017-0544-5. Epub 2017 Aug 19.
Spiramycin is a 16-membered macrolide antibiotic produced by Streptomyces ambofaciens and used in human medicine for the treatment of various respiratory tract and genital infections. Several impurities were detected in spiramycin-fermentation broth, especially impurities D and F, which decreased the separation-extraction yield and increased production cost. Dextrins, as the main carbon source, influence the accumulation of spiramycin and impurities. In this work, two types of dextrin from vendor Y and Z were compared to study their influences on spiramycin production. Our results showed that final spiramycin production with dextrin Z was enhanced twofold as compared with dextrin Y; however, the content of impurities F and D were higher with dextrin Z relative to dextrin Y. Several parameters (adenosine triphosphate, total sugar, reducing sugar, and reducing sugar to total sugar) were analyzed to reveal differences in the fermentation process. In vitro dextrin hydrolysis by amylase revealed structural differences in the two types of dextrin, and real-time quantitative polymerase chain reaction analyses showed that the transcription of srm7 and srm21 (involved in forosaminyl methylation) was enhanced and potentially related to the reduced formation of impurity F with dextrin Y. Furthermore, the srm20/srm33 ratio, representing flux balance of forosaminyl and mycarosyl, was ~ 1, implying that forosaminyl and mycarosyl biosynthesis were well balanced, resulting in reduced production of impurity D with dextrin Y.
螺旋霉素是一种由产二素链霉菌产生的16元大环内酯类抗生素,在人类医学中用于治疗各种呼吸道和生殖器感染。在螺旋霉素发酵液中检测到了几种杂质,尤其是杂质D和F,它们降低了分离提取产率并增加了生产成本。糊精作为主要碳源,会影响螺旋霉素和杂质的积累。在这项工作中,对供应商Y和Z的两种糊精进行了比较,以研究它们对螺旋霉素生产的影响。我们的结果表明,与糊精Y相比,使用糊精Z时螺旋霉素的最终产量提高了两倍;然而,相对于糊精Y,糊精Z的杂质F和D含量更高。分析了几个参数(三磷酸腺苷、总糖、还原糖以及还原糖与总糖的比例)以揭示发酵过程中的差异。淀粉酶对糊精的体外水解揭示了两种糊精的结构差异,实时定量聚合酶链反应分析表明,srm7和srm21(参与福洛糖胺基甲基化)的转录增强,并且可能与糊精Y杂质F形成减少有关。此外,代表福洛糖胺基和碳霉糖基通量平衡的srm20/srm33比率约为1,这意味着福洛糖胺基和碳霉糖基的生物合成平衡良好,导致糊精Y杂质D的产量降低。