Department of Biotechnology, School of Life Sciences, Pondicherry University, Pondicherry, India.
Prep Biochem Biotechnol. 2011;41(1):40-52. doi: 10.1080/10826068.2010.534221.
Statistics-based experimental designs were used to develop a cost-effective medium for enhanced production of viable cells and bacteriocin by probiotic Enterococcus faecium MC13. Carbon, nitrogen, and mineral sources were first screened by one-variable-at-a-time (OVAT) methods. In order to increase yield production, the selected variables were further statistically optimized using response-surface methodology (RSM) with central composite design (CCD). The maximum and minimum levels of the selected variables were determined and a set of 34 experimental runs was performed. The optimum concentrations of the tested variables for production of viable cells (12.24 log CFU mL(-1)) and bacteriocin activity (25,600 AU mL(-1)) were tryptone (10.0 g/L), peptone (6.0 g/L), maltose (3.0 g/L), glucose (9.0 g/L), NaCl (15.0 g/L), sodium citrate (2.5 g/L), sodium acetate (1.0 g/L), and dipotassium PO(4) (0.1 g/L). Threefold increased yield of bacteriocin was achieved in optimized medium compared to the unoptimized counterpart, and this was two times less cost than commercial MRS medium.
采用基于统计学的实验设计,开发出一种经济有效的培养基,用于提高益生菌屎肠球菌 MC13 活细胞和细菌素的产量。采用单变量法(OVAT)对碳源、氮源和矿物质源进行初步筛选。为了提高产量,进一步采用响应面法(RSM)和中心复合设计(CCD)对选定的变量进行统计学优化。确定选定变量的最大和最小水平,并进行了一组 34 次实验运行。测试变量的最佳浓度为生产活细胞(12.24 log CFU mL(-1)) 和细菌素活性(25600 AU mL(-1)) 为胰蛋白胨(10.0 g/L)、蛋白胨(6.0 g/L)、麦芽糖(3.0 g/L)、葡萄糖(9.0 g/L)、NaCl(15.0 g/L)、柠檬酸钠(2.5 g/L)、乙酸钠(1.0 g/L)和磷酸二氢钾(0.1 g/L)。与未优化的培养基相比,优化后的培养基中细菌素的产量增加了两倍,成本比商业 MRS 培养基低两倍。