Journal of Biology ›› 2022, Vol. 39 ›› Issue (2): 110-.doi: 10.3969/j.issn.2095-1736.2022.02.110

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Establishment of determination method and optimization of fermentation conditions of erucamide in Bacillus megaterium L2

  

  1. 1.Key laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region (Ministry of
    Education), Collaborative Innovation Center for Mountain Ecology & Agro-Bioengineering (CICMEAB),
    College of Life Sciences/Institute of Agro-bioengineering, Guizhou University, Guiyang 550025, China;
    2.Institute of Products Detection & Determination of Guizhou Province, Guiyang 550003, China
  • Online:2022-04-18 Published:2022-04-18

Abstract: A method for the determination of erucamide in B. megaterium L2 was established by gas chromatography-mass spectrometry (GC-MS), and then single factor experiments and response surface methods were used to study the fermentation medium and fermentation conditions. Design-expert 8.06 was used to design a three-factor response surface experiment for peptone addition, temperature, and pH. The response value was erucamide yield, and finally response surface analysis was performed. Under GC-MS chromatographic conditions, the erucamide content and peak area showed a good linear relationship (R2=0.9974), the relative standard deviations (RSD) of precision, repeatability and stability were all lower than 10%, and the recovery rate was 82.78%-98.14%, and the content of erucamide in the fermentation broth of B. megaterium was determined to be 0.940 mg/L. The optimal fermentation conditions were as follows: glucose concentration of 5 g/L, peptone concentration of 16.40 g/L, NaCl concentration of 3 g/L, inoculation 2.0%, culture temperature 34.8 ℃, shaking speed 150 r/min, initial pH 5.80. Under these conditions, the yield of erucamide was 1.325 mg/L, which was 39.03% higher than that before optimization. It can provide a reference for the determination of erucamide in B. megaterium. The fermentation medium and conditions of B. megaterium L2 significantly increased erucamide production, they provided reliable support for the subsequent research of erucamide.

Key words: erucamide, Bacillus megaterium, gas chromatography-mass spectrometry (GC-MS), response surface method, optimization fermentation

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