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J4  2011, Vol. 45 Issue (10): 1868-1876    DOI: 10.3785/j.issn.1008-973X.2011.10.028
化学与生物工程、环境工程     
新型抗真菌代谢产物发酵条件的响应面优化
陈正杰1,2, 朱婉萍1,2, 张书衍2, 吴绵斌1,
贺娟1, 陈华2, 孟强2
1.浙江大学 化学工程与生物工程学系,浙江 杭州310027;2.浙江省抗真菌药物研究重点实验室,浙江 台州 318000
Optimization of fermentation conditions for production of a novel
antifungal metabolite by response surface methodology
CHEN Zheng-jie1,2, ZHU Wan-ping1,2, ZHANG Shu-yan2,
WU Mian-bin1, HE Juan1,CHEN Hua2,MENG Qiang2
1. Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China;
2. Zhejiang Key Laboratory of Antifungal Drugs, Taizhou 318000, China
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摘要:

从浙江东海土壤中筛选获得1株菌株(ZS09-33)能产生抑制白色念珠菌的代谢产物,经16S rRNA序列测定和Genbank数据库搜索,确定其属于Bacillus amyloliquefaciens. PlackettBurman实验发现,影响抗真菌效价的主要因素为:黄豆饼粉、可溶性淀粉以及(NH4)2SO4.最陡爬坡实验和Box-Behnken实验得到响应面(RSM)优化的最佳培养基质量浓度为:黄豆饼粉2.72 g/L,可溶性淀粉 26.67 g/L,(NH4)2SO4 3.95 g/L,K2HPO4 1.8 g/L,NaCl 2 g/L,MgSO4 0.5 g/L,CaCO3 0.72 g/L,CuSO4 0.02 g/L,FeSO4 0.02 g/L,MnSO4 0.02 g/L,初始pH 6.5.在此条件下发酵液抗真菌效价达16 806.48 U/mL,与模型值相符.与单因素优化结果相比,效价提高了239%.产物经ESI/MS、1H-NMR及13C-NMR确定为相对分子质量为402的大环内脂类物质.

Abstract:

A strain ZS09-33, antagonising human pathogenic fungi—Candida albicans, was screened from soil samples of East Sea, Zhejiang Province, China. The strain was identified as Bacillus amyloliquefaciens based on sequencing 16S rRNA gene and blasting in Genbank. Then a two-level Plackett-Burman design was established to determine which factors in the fermentation medium influence the potency activity of metabolites. The concentrations of soybean bran, starch soluble and (NH4)2SO4 were optimized using the response surface methodology (RSM). The optimum composition of medium was as follows (per liter): soybean flour 2.72, soluble starch 26.67, (NH4)2SO4 3.95, K2HPO4 1.8, NaCl 2, MgSO4 0.5, CaCO3 0.72, CuSO4 0.02, FeSO4 0.02, MnSO4 0.02, initial pH 6.5. Under the RSM-optimized conditions, the potency activity of culture broth was 16 806.48 U/mL, which accorded well with the model-predicted value and resulted in a 2.39fold increase in antifungal activity compared to the original culture conditions obtained by singlefactor methodology. After analyzed by ESI/MS, 1H-NMR and 13C-NMR, the substance was determined as a macrolides antibiotic with relative molecular weight 402.

出版日期: 2011-10-01
:  Q 814.2  
基金资助:

浙江科技厅计划资助项目(2009C33019); 台州市浙江大学科技合作专项资金资助项目(07ZJU001,08ZJU007).

通讯作者: 吴绵斌,男,副教授.     E-mail: wumb@zju.edu.cn
作者简介: 陈正杰(1971—),男,高级工程师,从事微生物药物的研究与开发.E-mail: zjchen@hisunpharm.com
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引用本文:

陈正杰, 朱婉萍, 张书衍, 吴绵斌, 贺娟, 陈华, 孟强. 新型抗真菌代谢产物发酵条件的响应面优化[J]. J4, 2011, 45(10): 1868-1876.

CHEN Zheng-jie,ZHU Wan-ping,ZHANG Shu-yan,WU Mian-bin,HE Juan,CHEN Hua. Optimization of fermentation conditions for production of a novel
antifungal metabolite by response surface methodology. J4, 2011, 45(10): 1868-1876.

链接本文:

https://www.zjujournals.com/eng/CN/10.3785/j.issn.1008-973X.2011.10.028        https://www.zjujournals.com/eng/CN/Y2011/V45/I10/1868

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