采用均匀设计法设计和二次多项式逐步回归分析,对一株高效杀塔玛亚历山大藻微泡菌BS03(Microbulbifer sp.)产杀藻活性物质的发酵培养条件进行优化.通过单因素实验筛选出碳源、氮源、pH、培养时间和接种量为显著影响因子,并对5个显著影响因子采用U15(155)水平对培养基进行优化.结果表明BS03最适发酵培养条件为:蔗糖8 g/L,蛋白胨10.50 g/L,初始pH值7.5,培养时间32 h,接种量3.00%.验证试验结果显示,在此条件下该菌发酵液的干重为4.725 g/L,较优化前增加了31.35%,LD50为0.768%,较优化前降低了25.14%.研究结果为杀藻活性物质以及杀藻机理的研究奠定了理论基础.
The optimal parameters of fermentation conditions of marine bacterium Microbulbifer sp.BS03 producing algicidal substances against Alexandrium tamarense were obtained by using Uniform Design and quadric polynomial regression methods.Five main factors were achieved through single factor experiments,that is,carbon source,nitrogen source,pH,incubation time and inoculum size.The U15(155) design result showed the optimum conditions of experiment were as follows: sugar 8 g/L,peptone 10.5 g/L,initial pH 7.5,incubation time 32 h,and inoculum size of 3.00%.According to the verification experiment,under the optimal conditions,the dry biomass was 4.725 g/L which was increased by 31.35% and LD50was 0.768% which was decreased by 25.14% compared with those of the basic fermentation conditions.This outcome will help us to separate active substances from complex components in medium and thus to reveal the mechanism of algicidal activity in the future.