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中国农学通报 ›› 2013, Vol. 29 ›› Issue (15): 68-72.doi: 10.11924/j.issn.1000-6850.2012-2633

所属专题: 生物技术

• 农学 农业基础科学 • 上一篇    下一篇

解磷巨大芽孢杆菌液体发酵培养条件的优化

王金玲 刘晓平 赵凤艳 吕长山 高照亮   

  • 收稿日期:2012-07-27 修回日期:2012-08-29 出版日期:2013-05-25 发布日期:2013-05-25
  • 基金资助:
    哈尔滨市科技局重点攻关项目

Optimization of Culture Conditions for Phosphate-solubilizing Bacteria Bacillus megaterium in Submerged Fermentation

  • Received:2012-07-27 Revised:2012-08-29 Online:2013-05-25 Published:2013-05-25

摘要: 为了获得高密度培养的解磷细菌菌剂,以一株解磷巨大芽孢杆菌为研究对象,以OD600为依据测定活菌数量,优化最佳液体发酵培养条件。首先采用单因素试验对其培养条件如接种量、装液量、发酵温度、起始pH、发酵时间、摇床转速等进行筛选,获得单因素试验最佳值;在此基础上,利用L9(34)正交试验对其培养条件进行优化及验证,并制作菌株生长曲线。研究结果表明:该株巨大芽孢杆菌的最佳培养条件为:发酵温度30℃、起始pH 8.0、装液量20 mL/250 mL三角瓶、接种量3%、摇床转速250 r/min、培养时间22 h。在此最优条件下培养,以平板涂布法计数,发酵液最终活菌数达到3.2×109 cfu/mL以上。培养2~8 h,菌体生长处于对数期,8 h后菌体生长进入稳定期,22 h后进入衰亡期。试验结果为工业生产解磷巨大芽孢杆菌菌剂提供了基础数据。

关键词: 谷氨酰胺, 谷氨酰胺

Abstract: To obtain high density bacterial fertilizer of phosphate-solubilizing bacteria, culture conditions for a strain of phosphate-solubilizing bacteria Bacillus megaterium isolated from soil were optimized, and OD600 of the broth was used as a standard to judge the number of viable cells. Firstly, single factor experiments were used to research the effects of different factors of inoculation amount, medium volume, culture temperature, initial pH, incubation time, and rotate speed on submerged fermentation and the best values of single factors were obtained. Then orthogonal experiment of L9(34) was used to optimize the optimal culture conditions. After that, growth curve of the strain was described. The results showed that the optimal culture conditions were: temperature of 30℃, initial pH of 8.0, medium volume of 20 mL/250 mL flask, inoculation amount of 3%, rotate speed of 250 r/min, and incubation time of 22 h; under the optimal culture conditions, the number of viable cells of Bacillus megaterium reached above 3.2×109 cfu/mL. The period between 2 hours to 8 hours after inoculation was logarithmic phase for cell growth, then cell growth was in the stable stage, and after 22 hours, decline phase started. The results could provide basic data for production of phosphate-solubilizing bacterial fertilizer for agricultural application.

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