In order to improve the fermentation process of natamycin by Streptomyces lydicus A02, the main parameters for the fed batch fermentation are optimized. The range of suitable pH values is screened by an assisted optimization using a shaker with the function of automatic feeding to regulate pH values. Then the experiments in 30 L fermentor are carried out by controlling pH values within the suitable range as the main regulation point. The relationship between the natamycin yields and the fermentation parameters is obtained through the software Biostar 5.0. It is shown that the optimized parameter combination for the fed batch fermentation of natamycin by S. lydicus A02 is as follows: The culture temperature is 30℃, the pH value is 6.25-6.29, the DO is 20%-30%, the OUR is 25-15 mmol/(L·h), the CER is 20-12 mmol/(L·h), the amino-N is 0.20-0.23 g/L, and the reducing sugar remains at about 1.5% in 88 h then drops gradually to 0.32% at 112 h, the end of fermentation, when the natamycin yield is up to 2.02 g/L in the supernatant and 6.98 g/L in the total fermented broth. It can be concluded that the fermentation level of natamycin by S. lydicus A02 could be increased effectively by the optimization of the main fermentation parameters.
[1] 隋勤, 刘伟成, 裘季燕, 等. 利迪链霉菌A02的抑菌谱及其抑菌活性的 稳定性[J]. 植物保护, 2007, 33(5): 67-71. Sui Qin, Liu Weicheng, Qiu Jiyan, et al. Determination on the antimicrobial spectrum and the stability of the antagonistic activity of Streptomyces lydicus A02[J]. Plant Protection, 2007, 33(5): 67-71.
[2] 刘霆, 陶万强, 王合, 等. 拮抗菌A02代谢产物对几种果树病害的活性[J]. 北方园艺, 2011(17): 13-16. Liu Ting, Tao Wanqiang, Wang He, et al. The antifungal activity of the metabolite of biocontrol strain A02 against some fruit tree diseases[J]. Northern Horticulture, 2011(17): 13-16.
[3] 隋勤, 刘伟成, 卢彩鸽, 等. 利迪链霉菌A02抗真菌活性产物的分离和 结构鉴定[J]. 生物工程学报, 2009, 25(6): 840-846. Sui Qin, Liu Weicheng, Lu Caige, et al. Extraction and structural identification of the antifungal metabolite of Streptomyces lydicus A02[J]. Chinese Journal of Biotechnology, 2009, 25(6): 840-846.
[4] Gill J A, Martin J F. Biotechnology of antibiotics[M]. 2nd ed. New York: Marcel Dekker, 1997: 551-576.
[5] 卢彩鸽, 刘伟成, 杨剑芳, 等. 利迪链霉菌A01活性产物对甘蓝枯萎病 菌麦角甾醇含量的影响[J]. 植物病理学报, 2011, 41(S1): 43-49. Lu Caige, Liu Weicheng, Yang Jianfang, et al. Effect of bioactive metabolite of Streptomyces lydicus A01 on the content of ergosterol of Fusarium oxysporum f. sp. conglutinans[J]. Acta Phytopathologica Sinica, 2011, 41(S1): 43-49.
[6] Oostendorp J G. Natamycin[J]. Antonie van Leeuwenhoek, 1981, 47 (2): 170-171.
[7] 魏宝东, 孟宪军. 天然生物性食品防腐剂纳他霉素的特性及其应用[J]. 辽宁农业科学, 2004(2): 24-26. Wei Baodong, Meng Xianjun. The characteristics and applications of natamycin-the natural food preservative[J]. Liaoning Agricultural Sciences, 2004(2): 24-26.
[8] 孙远功, 乎玉侠, 冯晰. 纳他霉素在柑桔防腐保鲜中的应用[J]. 食品 研究与开发, 2006, 27(7): 190-192. Sun Yuangong, Hu Yuxia, Feng Xi. The application of natamycin in the antiseptic and freshness of orange[J]. Food Research and Development, 2006, 27(7): 190-192.
[9] 张鹏, 姜兴印, 房锋, 等. 纳他霉素对灰葡萄孢不同生育阶段菌体的 毒力及其生物学性状的影响[J]. 农药学学报, 2008, 10(2): 205-210. Zhang Peng, Jiang Xingyin, Fang Feng, et al. Toxicity of natamycin to Botrytis cinerea at different growth stages and its effect on biological characteristics[J]. Chinese Journal of Pesticide Science, 2008, 10(2): 205-210.
[10] 花玉鹏, 文才艺, 刘伟成, 等. 响应曲面法优化利迪链霉菌A02发酵 培养基[J]. 中国生物防治学报, 2011, 27(4): 520-527. Hua Yupeng, Wen Caiyi, Liu Weicheng, et al. Optimization of medium components for production of natamycin by Streptomyces lydicus A02 with response surface methodology[J]. Chinese Journal of Biological Control, 2011, 27(4): 520-527.
[11] 张龙翔, 张婷芳, 李令媛. 生化实验方法和技术[M]. 第2版. 北京: 高 等教育出版社, 1997: 1-3. Zhang Longxiang, Zhang Tingfang, Li Lingyuan. Methods and techniques of biochemistry experiments[M]. 2nd ed. Beijing: Higher Education Press, 1997: 1-3.
[12] 李巧枝, 程绎南. 生物化学实验技术[M]. 北京: 中国轻工业出版社, 2011: 83-85. Li Qiaozhi, Cheng Yinan. Biochemistry experiment techniques[M]. Beijing: China Light Industry Press, 2011: 83-85.
[13] 熊智强. 还原型谷胱甘肽高产菌的胁迫生理特性与高密度发酵过程 优化技术[D]. 上海: 华东理工大学, 2010: 27. Xiong Zhiqiang. Cell physiology and process optimization in high-celldensity cultivation of GSH high-yield strain[D]. Shanghai: East China University of Science and Technology, 2010: 27.
[14] 魏宝东, 郑凤娥, 孟宪军, 等. 发酵液中纳他霉素含量快速检测方法 研究[J]. 食品研究与开发, 2007, 28(2): 132-136. Wei Baodong, Zheng Fenge, Meng Xianjun, et al. The research of rapid determination methods of natamycin in fermentation solution[J]. Food Research and Development, 2007, 28(2): 132-136.
[15] 胡海洋, 乔春明, 葛菁萍, 等. 纳他霉素的特性和生产研究状况[J]. 中 国现代药物应用, 2009, 3(2): 200-201. Hu Haiyang, Qiao Chunming, Ge Jingping, et al. The characteristic and production research condition of a new polyene macrolide antimycotics-Natamycin[J]. Chinese Journal of Modern Drug Application, 2009, 3(2): 200-201.
[16] 李卫宁, 李宁, 魏昭, 等. 褐黄孢链霉菌WZ-18产纳他霉素发酵条件的优化[J]. 食品科技, 2010, 35(8): 18-22. Li Weining, Li Ning, Wei Zhao, et al. Optimization of fermentation conditions of natamycin from Streptomyces gilvosporeus WZ-18[J]. Food Science and Technology, 2010, 35(8): 18-22.
[17] 骆健美, 金志华, 岑沛霖. 褐黄孢链霉菌纳他霉素发酵条件优化[J]. 高校化学工程学报, 2006, 20(1): 68-73. Luo Jianmei, Jin Zhihua, Cen Peilin. Optimizing the fermentation conditions for producing natamycin by Streptomyces gilvosporeus[J]. Journal of Chemical Engineering of Chinese Universities, 2006, 20(1): 68-73.
[18] 魏宝东, 林雨舒. 纳他霉素高产菌株发酵罐发酵条件研究[J]. 食品与 生物技术学报, 2010, 29(3): 448-452. Wei Baodong, Lin Yushu. Study of natamycin high-yield strains fermenter fermentation condition[J]. Journal of Food Science and Biotechnology, 2010, 29(3): 448-452.
[19] 郝晓兵, 卢英华, 凌雪萍. 褐黄孢链霉菌生产纳他霉素工艺条件研究[J]. 厦门大学学报: 自然科学版, 2009, 48(6): 866-870. Hao Xiaobing, Lu Yinghua, Ling Xueping, et al. Study of cultivation conditions for natamycin production by Streptomyces gilvosporeus[J]. Journal of Xiamen University: Natural Science, 2009, 48(6): 866-870.
