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中国生物工程杂志

China Biotechnology
China Biotechnology
China Biotechnology  2011, Vol. 31 Issue (03): 55-60    DOI:
    
The Effects of NH4+ on L-tryptophan Fermentation
HUANG Jing1, SHI Jian-ming1, HUO Wen-ting1, XU Qing-yang1, XIE Xi-xian1, WEN Ting-yi2, CHEN Ning1
1. College of Biological Engineering, Tianjin University of Science & Technology, Key Laboratory of Industrial Microbiology, Tianjin 300457, China;
2. Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China
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Abstract  

Objective:To study the effects of NH4+ on biosynthesis of L-tryptophan. Methods:Fed-batch fermentation of E. coli TRTH with addition of ammonium sulfate carried out in 30-Liter fermentor and biomass, yield of L-tryptophan, consumption rate of glucose, plasmid stability were measured. The concentration of acetate, NH4+, lactate and pyruvate were investigated by BioProfile 300A Nova and high performance liquid chromatography (HPLC). The metabolic flux balance model of L-tryptophan synthesis by E. coli was established. Based on this model, the practical metabolic flux distribution of E. coli TRTH were determined with the linear program planted in MATLAB software. Results:During the fed-batch culture, E. coli TRTH was able to maintain higher growth rate at exponential phase, finally, the biomass and the yield of L-tryptophan were increased by 51.07% and 46.54% respectively by controlling the concentration of NH4+ less than 120 mmol/L. As the increase of the concentration of NH4+, the concentration of pyruvate, lactate and acetate were slightly increased, but cell plasmid stability was decreased. Data indicated that EMP pathway and TCA cycle were decreased by 7.31% and 22.04% respectively, PP pathway was increased by 7.14% compared with addition 5g/L of ammonium sulfate at 14h during the fed-batch culture. Conclusion:High concentrations of NH4+ terminated the cell growth, decreased the consumption rate of glucose and inhibited the L-tryptophan production. Controlled the concentration of NH4+ less than 120 mmol/L, the biomass and the yield of L-tryptophan were significantly increased, which accessed high cell density cultivation.

Key words： E. coli      NH4+      L-Tryptophan      High cell density cultivation     
Received: 24 November 2010      Published: 01 April 2011
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HUANG Jing
SHI Jian-Meng
SUO Wen-Ting
XU Qiang-Yang
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YUN Ting-Yi
CHEN Ning
Cite this article:

HUANG Jing, SHI Jian-ming, HUO Wen-ting, XU Qing-yang, XIE Xi-xian, WEN Ting-yi, CHEN Ning. The Effects of NH4+ on L-tryptophan Fermentation. China Biotechnology, 2011, 31(03): 55-60.

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https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2011/V31/I03/55

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