4-羟基环孢素高产菌株诱变筛选及培养基优化

doi:10.7523/j.issn.2095-6134.2017.05.005

中国科学院大学学报 ›› 2017, Vol. 34 ›› Issue (5): 558-566.DOI: 10.7523/j.issn.2095-6134.2017.05.005

4-羟基环孢素高产菌株诱变筛选及培养基优化

金丽娜, 闻建平

天津大学化工学院, 天津 300072

收稿日期:2016-09-29 修回日期:2016-11-28 发布日期:2017-09-15
通讯作者: 闻建平,E-mail:jpwen@tju.edu.cn
基金资助:
国家973项目（2013CB733600）、国家自然科学基金（21376171）、国家自然科学基金重点项目（21236005）资助

Mutagenesis, screening, and medium optimization of high[(4'-OH)MeLeu]⁴-CsA producing strain-Nonomuraea dietziae

JIN Lina, WEN Jianping

School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China

Received:2016-09-29 Revised:2016-11-28 Published:2017-09-15

摘要/Abstract

摘要： 以4-羟基环孢素转化菌Nonomuraea dietziae为出发菌，采用紫外-氯化锂复合诱变的方法，筛选得到一株产量达208 mg/L的稳定遗传菌J263，较出发菌164 mg/L的产量提高26.8%。在此基础上，采取单因素法和响应面法针对诱变菌的发酵培养基碳氮源进行系统优化，并对金属离子的添加进行考察。最终获得最佳的培养基配方（甘油26.5 g/L，糊精11.6 g/L，蛋白胨9.7 g/L，酵母浸粉3.5 g/L，大豆饼粉15.0 g/L，大豆油2.0 g/L，硫酸镁0.40 g/L，硫酸锰0.010 g/L，钼酸铵0.010 g/L）。在最优化条件下，4-羟基环孢素产量可达324 mg/L，较出发菌提高97.6%。

关键词: Nonomuraea dietziae, 4-羟基环孢素, 紫外-氯化锂诱变, 响应面优化

Abstract: An ultraviolet-lithium chloride mutagenesis strategy was carried out on the[(4'-OH)MeLeu]⁴-CsA producing strain, Nonomuraea dietziae.The production of mutant strain reached 208mg/L and increased by 26.8%. Then the fermentation medium was optimized by single-factor and response surface methods. The optimal medium was determined as follows (materials/concentrations in g/L):glycerol/26.5, dextrin/11.6, peptone/9.7, yeast extract powder/3.5, soybean meal/15.0, soybean oil/2.0, MgSO₄·7H₂O/0.40, MnSO₄·H₂O/0.010, and (NH₄)₂MoO₄/0.010. The final production reached 324mg/L and increased by 97.6% compared with the initial value.

Key words: Nonomuraea dietziae, [(4'-OH)MeLeu]⁴-CsA, ultraviolet lithium chloride mutagenesis, response surface optimization

中图分类号:

Q939.9

金丽娜, 闻建平. 4-羟基环孢素高产菌株诱变筛选及培养基优化[J]. 中国科学院大学学报, 2017, 34(5): 558-566.

JIN Lina, WEN Jianping. Mutagenesis, screening, and medium optimization of high[(4'-OH)MeLeu]⁴-CsA producing strain-Nonomuraea dietziae[J]. , 2017, 34(5): 558-566.

参考文献

[1] Lawen A, Traber R, Geyl D, et al. Cell-free biosynthesis of new cyclosporins[J]. The Journal of Antibiotics, 1989, 42(8):1283-1289.
[2] Borel J F. Comparative study of in vitro and in vivo drug effects on cell-mediated cytotoxicity[J]. Immunology, 1976, 31(4):631-641.
[3] Scribner A, Houck D, Huang Z, et al. Synthesis and biological evaluation of ⁸ cyclosporin A analogs as potential anti-HCV agents[J]. Bioorganic & Medicinal Chemistry Letters, 2010, 20(22):6542-6546.
[4] Peel M, Scribner A. Cyclophilin inhibitors as antiviral agents[J]. Bioorganic & Medicinal Chemistry Letters, 2013, 23(16):4485-4492.
[5] Evers M, Barrière J C, Bashiardes G, et al. Synthesis of non-immunosuppressive cyclophilin-Binding cyclosporin A derivatives as potential anti-HIV-1 drugs[J]. Bioorganic & Medicinal Chemistry Letters, 2003, 13(24):4415-4419.
[6] Kim S-N, Ahn H-J, Lee C-W, et al. Use of nonimmunosuppressive cyclosporin derivatives for treating hair loss:US, US20040161399. (2004-08-19). http://www.freepatentsonline.com/y2004/0161399.html.
[7] Kim C D, Lee M H, Sohn K C, et al. Induction of synapse associated protein 102 expression in cyclosporin A-stimulated hair growth[J]. Experimental Dermatology, 2008, 17(8):693-699.
[8] Lee M J, Kim H B, Yoon Y J, et al. Identification of a cyclosporine-specific P450 hydroxylase gene through targeted cytochrome P450 complement (CYPome) disruption in Sebekia benihana[J]. Applied and environmental microbiology, 2013, 79(7):2253-2262.
[9] Takahashi T, Kamimura A. Cyclosporin A promotes hair epithelial cell proliferation and modulates protein kinase C expression and translocation in hair epithelial cells[J]. Journal of Investigative Dermatology, 2001, 117(3):605-611.
[10] Bistolas N, Wollenberger U, Jung C, et al. Cytochrome P450 biosensors:a review[J]. Biosensors & Bioelectronics, 2005, 20(12):2408-2423.
[11] Kuhnt M, Bitsch F, France J, et al. Microbial biotransformation products of cyclosporin A[J]. The Journal of Antibiotics, 1996, 49(8):781-787.
[12] Lee M J, Han K, Kim E S. Targeted gene disruption and functional complementation of cytochrome P450 hydroyxlase involved in cyclosporin A hydroxylation in Sebekia benihana [J]. J Microbiol Biotechnol, 2011, 21(1):14-19.
[13] Park N S, Myeong J S, Park H J, et al. Characterization and culture optimization of regiospecific cyclosporin hydroxylation in rare actinomycetes species[J]. Journal of Microbiology & Biotechnology, 2005, 15(1):188-191.
[14] 章丽, 戴梦, 郑桂珍, 等. 等离子体-紫外复合诱变选育四羟基环孢菌素衍生物高产菌株[J]. 微生物学杂志, 2014, 34(1):68-71.
[15] 刘静, 戴梦, 章丽, 等. 4-羟基环孢菌素A衍生物-CyA生产菌的接合转移[J]. 化学与生物工程, 2013, 30(4):44-46.
[16] Stackebrandt E, Wink J, Steiner U, et al. Nonomuraea dietzii sp. nov[J]. International Journal of Systematic and Evolutionary Microbiology, 2001, 51(4):1437-1441.
[17] Sebek O K, Dolak L A. Microbial hydroxylation of novobiocin and related compounds[J]. The Journal of Antibiotics, 1984, 37(2):136-142.
[18] 黄乐平, 汤锋, 刘启阳, 等. 紫外-氯化锂复合诱变选育聚半乳糖醛酸酶高产菌株[J]. 安徽农业科学, 2011, 39(22):13275-13276.
[19] 袁琳. 采用氯化锂和紫外线复合诱变方法筛选四环素高产菌株[J]. 宁夏医学杂志, 2000, 22(6):340-341.
[20] 虞龙, 张宇, 龚文静, 等. 氯化锂-紫外-离子束复合诱变红霉素高产菌株研究[J]. 原子能科学技术, 2011, 45(7):780-784.
[21] Wang P, Hongbing Y U, Wang L, et al. Production of levulinic acid from sucrose catalyzed by ferric chloride supported on activated carbon[J]. Chemical Industry & Engineering Progress, 2009, 28(5):817-821.
[22] Ahmadi M, Vahabzadeh F, Bonakdarpour B, et al. Application of the central composite design and response surface methodology to the advanced treatment of olive oil processing wastewater using Fenton's peroxidation[J]. Journal of Hazardous Materials, 2005, 123(1):187-195.
[23] 高宏伟, 张萍, 石彦鹏, 等. 紫外-氯化锂复合诱变选育泰妙菌素高产菌株[J]. 中国兽药杂志, 2011, 45(3):18-19.
[24] 邱雁临, 梁亮, 汪亮. 紫外线与氯化锂复合诱变选育L-组氨酸产生菌[J]. 现代食品科技, 2008, 24(3):217-219.
[25] 章丽, 戴梦, 郑桂珍, 等. 四羟基环孢菌素衍生物转化菌株发酵条件优化[J]. 中国抗生素杂志, 2014, 39(4):249-252.
[26] Gunnarsson N, Mortensen U H, Sosio M, et al. Identification of the Entner-Doudoroff pathway in an antibiotic-producing actinomycete species[J]. Molecular Microbiology, 2004, 52(3):895-902.
[27] Conway T. The Entner-Doudoroff pathway:history, physiology and molecular biology[J]. FEMS Microbiology Reviews, 1992, 9(1):1-27.
[28] Hamedi J, Malekzadeh F,Niknam V. Improved production of erythromycin by Saccharopolyspora erythraea by various plant oils[J]. Biotechnology Letters, 2002, 24(9):697-700.
[29] Qi H, Zhao S, Fu H, et al. Enhancement of ascomycin production in Streptomyces hygroscopicus var. ascomyceticus by combining resin HP20 addition and metabolic profiling analysis[J]. Journal of Industrial Microbiology and Biotechnology, 2014, 41(9):1365-1374.
[30] Zhao S, Huang D, Qi H, et al. Comparative metabolic profiling-based improvement of rapamycin production by Streptomyces hygroscopicus[J]. Applied Microbiology and Biotechnology, 2013, 97(12):5329-5341.
[31] Xia M, Huang D, Li S, et al. Enhanced FK506 production in Streptomyces tsukubaensis by rational feeding strategies based on comparative metabolic profiling analysis[J]. Biotechnology and Bioengineering, 2013, 110(10):2717-2730.

4-羟基环孢素高产菌株诱变筛选及培养基优化

Mutagenesis, screening, and medium optimization of high[(4'-OH)MeLeu]⁴-CsA producing strain-Nonomuraea dietziae

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

编辑推荐

Metrics

本文评价

访问统计

联系我们

4-羟基环孢素高产菌株诱变筛选及培养基优化

Mutagenesis, screening, and medium optimization of high[(4'-OH)MeLeu]4-CsA producing strain-Nonomuraea dietziae

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 0

编辑推荐

Metrics

本文评价

访问统计

联系我们

Mutagenesis, screening, and medium optimization of high[(4'-OH)MeLeu]⁴-CsA producing strain-Nonomuraea dietziae