ARTP技术选育吩嗪-1-甲酰胺高产菌株及发酵优化

doi:10.13560/j.cnki.biotech.bull.1985.2016.01.029

生物技术通报 ›› 2016, Vol. 32 ›› Issue (1): 174-179.doi: 10.13560/j.cnki.biotech.bull.1985.2016.01.029

ARTP技术选育吩嗪-1-甲酰胺高产菌株及发酵优化

谭剑, 熊欣, 梁万利, 彭华松, 张雪洪

上海交通大学微生物代谢国家重点实验室, 上海 200240

收稿日期:2015-05-08 出版日期:2016-01-09 发布日期:2016-01-22
作者简介:谭剑, 男, 硕士, 研究方向：天然产物的生物合成与调控;E-mail：kevintan_2014@163.com
基金资助:
国家自然科学基金项目(31270084), 国家高技术研究发展计划(“863”计划)(2012AA022107), 国家重点基础研究发展计划 (“973”计划) (2012CB721005)

Breeding of a Phenazine-1-carboxamid-producing Strain by ARTP Mutation and Its Optimization of Fermentation

TAN Jian, XIONG Xin, LIANG Wan-li, PENG Hua-song, ZHANG Xue-hong

State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240

Received:2015-05-08 Published:2016-01-09 Online:2016-01-22

摘要/Abstract

摘要： 从一株高产吩嗪-1-甲酰胺(PCN)的绿针假单胞菌P3株出发, 利用常压室温等离子体诱变技术进行诱变育种, 从初筛的20株突变株中获得了一株PCN产量达到2 093 mg/L的突变株P3-9, 为出发菌株的125%。随后通过单因素实验考察了各种营养因子对该高产菌株合成PCN的影响, 结果表明发酵培养基的最佳碳源、氮源分别为甘油和蛋白胨, 外源添加Fe³⁺或Fe²⁺对于积累PCN有显著促进作用, 而添加苯丙氨酸、色氨酸和酪氨酸对PCN产量无明显影响。优化后, 该突变株的PCN产量高达2 810 mg/L, 是目前国际上通过诱变育种获得的较高PCN产量。

关键词: 吩嗪-1-甲酰胺, 常压室温等离子体, 培养基优化

Abstract: Using an atmospheric and room temperature plasma(ARTP)jet to mutagenize a phenazine-1-carboxamide-producting strain Pseudomonas Chlororaphis P3, the mutant strain P3-9 with the highest phenazine-1-carboxamide(PCN)yield of more than 2 093 mg/L was obtained from the primary-screened 20 mutant ones, and it was 125% that by the original strain. Furthermore, single factor experiment was used to investigate the effects of varied nutrient factors on the PCN synthesis of P3-9. The results showed that the best carbon source and nitrogen source were glycerol and tryptone respectively. Adding Fe³⁺ or Fe²⁺ had a significant effect on PCN production, and no measurable effect while adding aromatic amino acids of phenylalanine, tryptophan and tyrosine. After the optimization, the PCN yield of strain P3-9 reached 2 810 mg/L, which was the highest yield of PCN by mutation breeding in the world so far.

Key words: phenazine-1-carboxamide, atmospheric and room temperature plasma, medium optimization

谭剑, 熊欣, 梁万利, 彭华松, 张雪洪. ARTP技术选育吩嗪-1-甲酰胺高产菌株及发酵优化[J]. 生物技术通报, 2016, 32(1): 174-179.

TAN Jian, XIONG Xin, LIANG Wan-li, PENG Hua-song, ZHANG Xue-hong. Breeding of a Phenazine-1-carboxamid-producing Strain by ARTP Mutation and Its Optimization of Fermentation[J]. Biotechnology Bulletin, 2016, 32(1): 174-179.

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ARTP技术选育吩嗪-1-甲酰胺高产菌株及发酵优化

Breeding of a Phenazine-1-carboxamid-producing Strain by ARTP Mutation and Its Optimization of Fermentation

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