摘要
目的 优化海洋来源的蓝灰异壁放线菌OUCMDZ-413突变株CRM05的发酵条件,提高浅蓝霉素H的发酵产量并测定其对海洋环境病原细菌的抑菌活性。方法 探索培养基组成、接种量、种龄、发酵天数、盐度、pH、碳源、氮源、前体以及大孔吸附树脂添加量,再采用单因素实验和正交实验确定最优发酵条件。结果 优化后浅蓝霉素H的最佳发酵条件为:摇瓶发酵11 d(28℃、180 r/min)、装液量150/500 mL(体积分数)、优化培养基(20 g可溶性淀粉、10 g蛋白胨、10 g酵母浸膏、4 g NaCl、4 g CaCO_(3)、0.1 g 2-吡啶甲酸、40 g XAD-16大孔吸附树脂、1 L陈海水、灭菌前用1 mol/L盐酸调节pH 6.5)、2%接种量、5 d种龄(即菌落的A_(600)2.72)。结论 以优化条件发酵菌株CRM05,浅蓝霉素H的分离产量达到434 mg/L,是优化前液体发酵产量的10倍和优化后固体发酵产量的1.6倍。浅蓝霉素H可显著抑制3种海洋弧菌—溶藻弧菌、创伤弧菌和副溶血弧菌,其最小抑菌浓度分别为8、16和16μg/mL,活性强于环丙沙星(相应的最小抑菌浓度分别为64、64和32μg/mL)。
Objective To optimize the fermentation conditions for the caerulomycin H-producing strain CRM05 mutant from the marine-derived Actinoalloteichus cyanogriseus OUCMDZ-413 that we previously numbered as WH1-2216-6,and then assay the antibacterial activity against the pathogenic bacteria in the marine environment.Methods The optimal composition of the culture medium,inoculum size,inoculum age,cultivation time,salinity,pH,carbon sources,nitrogen sources,precursor,and additive amount of XAD-16 macroreticular resin were investigated by means of the single factor experiment,and the orthogonal experiment.Results The highest production of caerulomycin H was obtained through the liquid fermentation at 28℃and shaking 11 d(180 r/min)in a 500 mL conical flask containing 150 mL of an optimized medium and 2%inoculation volume with 5d-old strain CRM05(A_(600)2.72).The optimized medium was prepared by dissolving soluble starch 20 g,peptone 10 g,yeast extract 10 g,NaCl 4 g,CaCO_(3)4 g,and 2-pyridinic acid 0.1 g in 1 L of natural seawater with pH 6.5(adjusted by 1 mol/L HCl),and then adding 40 g of XAD-16 resin.Conclusion The isolation yield of caerulomycin H reached 434 mg/L after optimization,which is respectively 10-and 1.6-folds increase in the yields of the previous unoptimized liquid fermentation and the optimized solid fermentation.In addition,caerulomycin H showed stronger inhibitory activity than ciprofloxacin(a positive control)against three marine vibrio strains,Vibrio alginolyticus,V.vulnificus and V.parahaemolyticus with the minimum inhibition concentrations of 8,16 and 16μg/mL for caerulomycin H and 64,64 and 32μg/mL for ciprofloxacin,respectively.
作者
高谕康
刘培培
李燕楠
邱培菊
朱伟明
Gao Yukang;Liu Peipei;Li Yan nan;Qiu Peiju;Zhu Weiming(Key Laboratory of Marine Drugs,Ministry Education of China,School of Medicine and Pharmacy,Ocean University of China,Qingdao 266003)
出处
《中国抗生素杂志》
CAS
CSCD
北大核心
2023年第11期1251-1260,共10页
Chinese Journal of Antibiotics
基金
国家自然科学基金委员会-山东省人民政府联合基金(No.U1906213)
国家自然科学基金(No.41876172)。
关键词
发酵优化
浅蓝霉素H
抑菌活性
海洋环境病原细菌
突变株CRM05
Fermentation optimization
Caerulomycin H
Antibacterial activity
Marine pathogenic bacteria
Mutant strain CRM05
