解淀粉芽孢杆菌M1摇瓶发酵条件优化及脱氮性能评价

doi:10.13560/j.cnki.biotech.bull.1985.2018-0920

摘要/Abstract

摘要： 旨在提高解淀粉芽孢杆菌M1液体发酵产芽孢量,并考察其对实际废水的反硝化脱氮效果。首先采用单因子实验研究了碳源、氮源、初始pH值、温度、转速和装液量等因子对M1液体发酵产芽孢量的影响。然后对其中显著性因子：氮源浓度、接种量、装液量、温度4个因素进行正交试验,进一步优化发酵条件。结果显示,优化后的培养基成分为：5 g/L碳源（可溶性淀粉）、10 g/L 氮源（酵母粉∶蛋白胨=2∶1）、1 g/L NaCl;最佳培养条件为：初始pH6.5、发酵温度34℃、摇床转速180 r/min、培养瓶装液量30%、接种量7%。在此条件下,芽孢杆菌M1芽孢数可达到 6.8×10⁸ CFU/mL,高于优化前的2.08×10⁸ CFU/mL。将芽孢杆菌M1投加于反应器中,与不加菌种相比,反硝化脱氮效果提升15%-34%。

关键词: 解淀粉芽孢杆菌, 液体发酵, 产芽孢量, 正交试验, 反硝化脱氮

Abstract: This work aims to increase the total spores of Bacillus amyloliquefaciens M1 in fermentation,and investigate the anaerobic denitrification effect in wastewater. First,single factor experiments were used to investigate the effects of species and concentrations of various carbon and nitrogen sources,initial pH,temperature,shaking speed,medium volume etc. on the spore yield of M1.Then orthogonal experiments were carried out to optimize 4 significant factors of nitrogen source concentration,inoculation amount,liquid loading amount and temperature. Results showed that the optimal composition of the medium were as such：5 g/L carbon source（soluble starch）,10 g/L nitrogen source[yeast powder and tryptone mix（2∶1,V/V）],1 g/L NaCl. The optimal cultural condition was as：initial pH6.5,temperature 34℃,shaking speed 180 r/min,medium capacity 30%（75 mL/250mL）,and inoculation rate 7%. Under the optimal fermentation condition,the total spores yield of B. amyloliquefaciens M1 was up to 6.8×10⁸ CFU/mL,2.3 times higher than that cultured in LB medium（2.08×10⁸ CFU/mL）. Compared with the control test,test with M1 addition showed 15%-34% higher denitrification.

Key words: Bacillus amyloliquefaciens, liquid fermentation, sporulation rate, orthogonal experiment, nitrogen removal by anaerobic denitrification·

李丽, 严月根, 吴华明. 解淀粉芽孢杆菌M1摇瓶发酵条件优化及脱氮性能评价[J]. 生物技术通报, 2019, 35(5): 102-108.

LI Li, YAN Yue-gen, WU Hua-ming. Optimization of Shake Flask-fermentation Condition for Bacillus amyloliquefaciens M1 Strain and Its Denitrification Performance[J]. Biotechnology Bulletin, 2019, 35(5): 102-108.

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