摘要
为提升巨大芽胞杆菌(Bacillus megaterium)ZS-3有效菌体数量,降低ZS-3发酵成本,通过单因素、正交与响应面相结合的方法探究ZS-3产活菌数最高的碳源、氮源、无机盐、转速、装液量、pH、温度和接菌量等培养条件.单因素与正交实验筛选得ZS-3最适培养基与比例为碳源1.0%(质量分数)、氮源0.5%、磷酸氢二铵0.5%和十二水合硫酸铝钾0.5%;响应面优化后ZS-3培养条件为装液量40%、转速200 r/min、pH值6.67、培养温度30.37℃、接种量1.31%(体积分数)时生物量达到最高,优化后培养的菌体数量可达到1.21×109 cfu/mL.经过优化后ZS-3活菌数为原始培养条件产生的活菌数的3.9倍.降低了发酵成本,为未来可能的工业化扩大发酵生产提供参考与数据支持.
Abstract
In order to increase the number of effective bacteria of Bacillus megaterium ZS-3 and reduce the fermenta-tion cost of ZS-3,combination of single factor,orthogonal,and response surface method were used to investigate the fermentation conditions to produce the highest number of viable ZS-3,including carbon source,nitrogen source,inor-ganic salt,rotation speed,liquid filling volume,pH,temperature,inoculation volume,and other culture conditions.The results showed that strain ZS-3 optimal carbon,nitrogen source and inorganic salt were soluble starch 1.0%(mass fraction),corn milk dry powder 0.5%,diammonium hydrogen phosphate 0.5%,aluminum potassium sulfate dodecahydrate 0.5%;optimal cultivation conditions were rotation speed of 200 r/min,liquid filling volume 40%,pH of 6.67,30.37 ℃,and inoculation volume 1.31% (v/v),the biomass reached the highest.After optimization,the number of cultured cells reached 1.21 × 109 cfu/mL,the number of viable cells in ZS-3 is 3.9 times those of the vi-able cells produced under the original cultivation conditions.The fermentation cost reduced and provided theoretical basis and data support for the possible industrial expansion of fermentation production in the future.