Laboratoire de Protection et Amélioration des Plantes (LPAP) Team of Biopesticides, Centre of Biotechnology of Sfax, Sfax University, P.O. Box 1177, 3018, Sfax, Tunisia.
Bioprocess Biosyst Eng. 2012 Nov;35(9):1505-13. doi: 10.1007/s00449-012-0740-2. Epub 2012 May 5.
Optimization of a fermentation process for bioinsecticides production by Photorhabdus temperata strain K122 was investigated into fully controlled 3-L fermenter using an optimized medium (OM). Development of large-scale inocula showed that the composition of the growth medium greatly influenced the physiological state of P. temperata cells. The effect of pH, agitation and dissolved oxygen concentration (DO) on the growth, culturability and oral toxicity of P. temperata cells were also investigated. Indeed, maintaining the pH at 7 and controlling DO concentration at 50 % saturation throughout the fermentation process, improved biomass production, CFU counts and oral toxicity by 41.1, 35 and 32.1 %, respectively, as compared to cultures carried out in 500 mL shake flasks. At such conditions, 8 g/L glucose fed-batch fermentation, enhanced cell lysis and variants small colony (Vsm) polymorphism appearance. To overcome such limitations, glucose concentration should be maintained at 4 g/L. In this case, P. temperata cells were produced at high cell density and culturability reaching 4.5 and 1.2 × 10(9) cells/mL, respectively. In addition, the stability of the primary form was maintained for a long period in the stationary growth phase and Vsm polymorphism was completely avoided that can be crucial for scale-up the bioprocess of P. temperata bioinsecticide.
优化了 Photorhabdus temperata K122 菌株生产生物杀虫剂的发酵过程,在 3-L 发酵罐中使用优化的培养基(OM)进行了全控制发酵。开发大规模接种物表明,生长培养基的组成极大地影响了 P. temperata 细胞的生理状态。还研究了 pH、搅拌和溶解氧浓度(DO)对 P. temperata 细胞生长、可培养性和口服毒性的影响。事实上,与在 500 mL 摇瓶中进行的培养相比,通过将 pH 维持在 7 并在整个发酵过程中控制 DO 浓度为 50%饱和度,可以将生物量、CFU 计数和口服毒性分别提高 41.1%、35%和 32.1%。在这些条件下,8 g/L 葡萄糖补料分批发酵增强了细胞裂解和变体小菌落(Vsm)多态性的出现。为了克服这些限制,应将葡萄糖浓度维持在 4 g/L。在这种情况下,P. temperata 细胞以高细胞密度和可培养性生产,分别达到 4.5 和 1.2×10(9)细胞/mL。此外,在静止生长阶段,原始形态的稳定性得以维持很长时间,并且完全避免了 Vsm 多态性,这对于扩大 P. temperata 生物杀虫剂的生物过程至关重要。