Chemical Engineering and Process Development Division, National Chemical Laboratory, Pune, 411008, India.
J Ind Microbiol Biotechnol. 2010 Mar;37(3):297-306. doi: 10.1007/s10295-009-0673-1. Epub 2009 Dec 5.
Mutants of Gibberella fujikuroi with different colony characteristics, morphology and pigmentation were generated by exposure to UV radiation. A mutant, Mor-189, was selected based on its short filament length, relatively high gibberellin A(4) (GA(4)) and gibberellin A(3) (GA(3)) production, as well as its lack of pigmentation. Production of GA(4) by Mor-189 was studied using different inorganic and organic nitrogen sources, carbon sources and by maintaining the pH of the fermentation medium using calcium carbonate. Analysis of GA(4) and GA(3) was done by reversed-phase high-performance liquid chromatography and LC-MS. The mutants of G. fujikuroi produced more GA(4) when the pH of the medium was maintained above 5. During shake flask studies, the mutant Mor-189 produced 210 mg l(-1) GA(4) in media containing wheat gluten as the nitrogen source and glucose as the carbon source. Fed-batch fermentation in a 14 l agitated fermenter was performed to evaluate the applicability of the mutant Mor-189 for the production of GA(4). In 7-day fed-batch fermentation, 600 mg l(-1) GA(4) were obtained in the culture filtrate. The concentration of GA(4) and GA(3) combined was 713 mg l(-1), of which GA(4) accounted for 84% of the total gibberellin. These values are substantially higher than those published previously. The present study indicated that, along with maintenance of pH and controlled glucose feeding, the use of wheat gluten as the sole nitrogen source considerably enhances GA(4) production by the mutant Mor-189.
通过紫外线照射,产生了具有不同菌落特征、形态和色素的藤仓赤霉菌突变体。根据其短菌丝长度、相对较高的赤霉素 A(4)(GA(4))和赤霉素 A(3)(GA(3))产量以及缺乏色素的特点,选择了一个突变体 Mor-189。使用不同的无机和有机氮源、碳源,并使用碳酸钙维持发酵培养基的 pH 值,研究了 Mor-189 产生 GA(4)的情况。通过反相高效液相色谱法和 LC-MS 分析了 GA(4)和 GA(3)。当培养基的 pH 值保持在 5 以上时,藤仓赤霉菌突变体产生更多的 GA(4)。在摇瓶研究中,突变体 Mor-189 在含有面筋作为氮源和葡萄糖作为碳源的培养基中产生 210mg/L 的 GA(4)。在 14 升搅拌发酵罐中进行分批补料发酵,以评估突变体 Mor-189 用于生产 GA(4)的适用性。在 7 天的分批补料发酵中,在培养滤液中获得了 600mg/L 的 GA(4)。GA(4)和 GA(3)的总浓度为 713mg/L,其中 GA(4)占总赤霉素的 84%。这些值明显高于以前发表的值。本研究表明,通过维持 pH 值和控制葡萄糖的添加,使用面筋作为唯一的氮源,可以显著提高突变体 Mor-189 产生 GA(4)的能力。