Laboratory of Biopesticides, Centre of Biotechnology of Sfax, Sfax University, P.O. Box '1177', 3018, Sfax, Tunisia.
Laboratory of Enzyme Engineering and Microbiology, National School of Engineering of Sfax (ENIS), Sfax University, BP 1173, 3038, Sfax, Tunisia.
Sci Rep. 2021 Jan 12;11(1):779. doi: 10.1038/s41598-020-80773-5.
This study investigates the optimization of the culture conditions for enhancing Photorhabdus temperata biopesticide production using wastewater (WS4) as a raw material. Box-Behnken design (BBD) was used to evaluate the effects of carbon to nitrogen ratio (C/N), sodium chloride concentration and inoculum size on P. temperata biomass production and insecticidal activity. For an enhanced biopesticide production, the optimum operating conditions were as follows: inoculum size = 4%; C/N ratio = 12.5 and [NaCl] = 4 g/L for two responses. 1.95 and 2.75 fold improvements in oral toxicity and biomass production were respectively obtained in the cost-effective medium developed in this study (WS4 I) using the three variables at their optimal values. Under the optimized conditions, WS4 I-grown cells exhibited higher membrane integrity according to flow cytometry analysis since dead cells presented only 9.2% compared to 29.2% in WS4. From batch fermentations carried out in WS4 I and WS4, P. temperata kinetic parameters in terms of biomass production and substrate consumption rates were modeled. The obtained results showed that the maximum specific growth rate in WS4 I was of 0.43 h while that obtained in WS4 was of 0.14 h. In addition, the efficiency of P. temperata to metabolize organic carbon was enhanced by optimizing the culture conditions. It reached 72.66% instead of 46.18% in the control fermentation after 10 h of incubation. Under the optimized conditions, P. temperata cells showed the highest specific consumption rate resulting in a toxin synthesis improvement.
本研究旨在通过优化培养条件,利用废水(WS4)作为原料来提高 Photorhabdus temperata 生物农药的产量。采用 Box-Behnken 设计(BBD)评估了碳氮比(C/N)、氯化钠浓度和接种量对 P. temperata 生物量和杀虫活性的影响。为了提高生物农药的产量,最佳操作条件如下:接种量=4%;C/N 比=12.5,[NaCl]=4 g/L,这两个响应的优化值。在本研究开发的经济型培养基(WS4 I)中,使用这三个变量的最佳值,可分别使口服毒性和生物量提高 1.95 倍和 2.75 倍。根据流式细胞术分析,在优化条件下,WS4 I 培养的细胞表现出更高的膜完整性,因为死细胞仅占 9.2%,而 WS4 中死细胞占 29.2%。在 WS4 I 和 WS4 中进行分批发酵,对 P. temperata 生物量生产和基质消耗速率的动力学参数进行了建模。结果表明,在 WS4 I 中的最大比生长速率为 0.43 h,而在 WS4 中的最大比生长速率为 0.14 h。此外,通过优化培养条件,提高了 P. temperata 代谢有机碳的效率。在 10 小时的孵育后,它从对照发酵中的 46.18%提高到 72.66%。在优化条件下,P. temperata 细胞表现出最高的比消耗率,从而提高了毒素的合成。