Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.
Enzyme Microb Technol. 2011 Jan 5;48(1):33-8. doi: 10.1016/j.enzmictec.2010.09.002. Epub 2010 Sep 15.
Statistical experimental design was used to optimize the concentration of trace elements for production of antifungal compound, 2,4-diacetylphloroglucinol (DAPG), from fluorescent pseudomonad R62 in shake-flask cultivation. The selection of the trace metal ions, influencing DAPG production, was done using Plackett-Burman design (PBD). Only Zn(2+), Mn(2+) and MoO(4)(2-) were the most significant components (p<0.05). A quadratic model was used to fit the response. Application of response surface methodology (RSM) revealed that the optimum values of the salts of the trace elements Zn(2+) (ZnSO(4)·7H(2)O), Mn(2+) (MnCl(2)·4H(2)O), and MoO(4)(2-) (Na(2)MoO(4)·2H(2)O) were 83, 42 and 135μM, respectively, to achieve 125 mg/L of DAPG, which was nearly 13-fold more compared to its production in basal synthetic medium in shake flask. The studies in 14L bioreactor resulted in 135 mg/L of DAPG at the end of 36 h of cultivation. The culture broth containing 125 mg/L of DAPG was found to be sufficient for keeping the bio-inoculant viable in non-sterile talcum powder-based formulations (which contained 25μg DAPG/g carrier) when stored at 28°C for 6 months. The structure of the purified DAPG was confirmed using (1)H NMR and mass spectrometry.
采用统计实验设计优化荧光假单胞菌 R62 摇瓶发酵生产 2,4-二乙酰基间苯三酚（DAPG）的微量元素浓度。采用 Plackett-Burman 设计（PBD）选择影响 DAPG 产量的痕量金属离子。只有 Zn(2+)、Mn(2+) 和 MoO(4)(2-)是最重要的成分（p<0.05）。使用二次模型拟合响应。响应面法（RSM）的应用表明，微量元素盐的最佳值为 Zn(2+)（ZnSO(4)·7H(2)O）、Mn(2+)（MnCl(2)·4H(2)O）和 MoO(4)(2-)（Na(2)MoO(4)·2H(2)O）分别为 83、42 和 135μM，以实现 125mg/L 的 DAPG，这几乎是摇瓶基础合成培养基中产量的 13 倍。在 14L 生物反应器中的研究在 36 小时的培养结束时产生了 135mg/L 的 DAPG。含有 125mg/L DAPG 的培养液在 28°C 下储存 6 个月时，在不含无菌滑石粉基配方（其中含有 25μg DAPG/g 载体）中足以保持生物接种剂的活力。使用（1）H NMR 和质谱法确认了纯化 DAPG 的结构。
