ARTP 技术选育吩嗪-1-甲酰胺高产菌株及发酵优化

摘要

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.%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2093 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2810 mg/L，是目前国际上通过诱变育种获得的较高 PCN 产量。