Bacillus sp.WH4是一株锰氧化能力强的锰氧化细菌,在锰污染的水源净化方面具有较好的应用前景.为提高发酵液中Bacillus sp.WH4的活菌数,应用响应面法优化其发酵培养基.Plackett-Burman设计获得对活菌数有显著影响的4个因素:豆粕粉、KH2PO4、MgSO4、FeSO4;最陡爬坡试验确定中心组合实验的最大响应区域;中心组合设计和响应面分析优化出4个显著因素的最佳浓度.优化后的最适培养基(w/%)组成为:蔗糖2、豆粕粉0.96、K2HPO4 0.8、KH2PO4 0.19、MgSO4.7H2O 0.11、CaCl2 0.02、NaCl 0.5、FeSO4.7H2O 0.02.发酵试验表明,最优培养基的活菌数可达4.2×109 CFU mL-1,比初始发酵培养基提高近3倍.
Bacillus sp.WH4 is a manganese-oxidizing bacterium possessing the strong activity to oxidize soluble manganese ion into insoluble manganese dioxide.It would be a promising application candidate for the cleaning of water resources polluted by manganese.To maximize the viable cell of Bacillus sp.WH4 in a fermentation broth,its fermentation medium was optimized using response surface method(RSM).Firstly,four significant factors,soybean meal,KH2PO4,MgSO4 and FeSO4,were screened out by Plackett-Burman(PB) design.Secondly,the maximum response region for the central composite design(CCD) was determined using the method of steepest ascent.Finally,the optimal concentration for each significant factor was determined using CCD and RSM analysis.The optimized fermentation medium contained sucrose 2,soybean meal 0.96,K2HPO4 0.8,KH2PO4 0.19,MgSO4.7H2O 0.11,CaCl2 0.02,NaCl 0.5 and FeSO4.7H2O 0.02(w/%).The results of the fermentation test confirmed that the viable count of Bacillus sp.WH4 in the optimized fermentation medium was 4.2×109 CFU mL-1,which is nearly four times higher than that in the initial fermentation medium.