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Application of a dissolved oxygen control strategy to increase the expression of Streptococcus suis glutamate dehydrogenase in Escherichia coli

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Abstract

The accumulation of acetate in Escherichia coli inhibits cell growth and desired protein synthesis, and cell density and protein expression are increased by reduction of acetate excretion. Dissolved oxygen (DO) is an important parameter for acetate synthesis, and the accumulation of acetate is inversely correlated to DO level. In this study, the effect of DO levels on glutamate dehydrogenase (GDH) expression was investigated, and then different DO control strategies were tested for effects on GDH expression. DO control strategy IV (50% 0–9 h, 30% 9–18 h) provided the highest cell density (15.43 g/L) and GDH concentration (3.42 g/L), values 1.59- and 1.99-times higher than those achieved at 10% DO. The accumulation of acetate was 2.24 g/L with DO control strategy IV, a decrease of 40.74% relative to that achieved for growth at 10% DO. Additionally, under DO control strategy IV, there was lower expression of PoxB, a key enzyme for acetate synthesis, at both the transcriptional and translational level. At the same time, higher transcription and protein expression levels were observed for a glyoxylate shunt gene (aceA), an acetate uptake gene (acs), gluconeogensis and anaplerotic pathways genes (pckA, ppsA, ppc, and sfcA), and a TCA cycle gene (gltA). The flux of acetate with DO strategy IV was 8.4%, a decrease of 62.33% compared with the flux at 10% DO. This decrease represents both lower flux for acetate synthesis and increased flux of reused acetate.

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Acknowledgements

This study was funded by the Major Science and Technology Innovation Project of Shandong (2019JZZY020606), the Development of Science and Technology Plan Program of Binzhou (2015ZC0107), the Technological Innovation Project of Shandong (201741916018), the Key Specialized Research and Development Breakthrough program in Henan Province (202102110101, 182102110347), Young Talent Lifting Project in Henan Province (2020HYTP041,2021HYTP038) , the National Natural Science Foundation of China (31702263) and the Youth Backbone Teacher Project of Colleges and Universities of Henan Province (2020GGJS163).

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Author notes

  1. Likun Cheng and Chunguang Zhao are co-first authors.

Authors and Affiliations

  1. Shandong Research Center of High Cell Density Fermentation and Efficient Expression Technology, Shandong Lvdu Bio-Science and Technology Co., Ltd., Shandong Binzhou Animal Science and Veterinary Medicine Academy, Binzhou, 256600, China

    Likun Cheng, Xiuyan Yang, Chuwen Lin, Xiubao Zhao, Jinliang Wang & Zhiqiang Shen

  2. Research and Development Center, Ningxia Eppen Biotech Co., Ltd., Yinchuan, 750000, China

    Chunguang Zhao

  3. Department of Clinical Pharmacy, Affiliated Hospital of Binzhou Medical University, Binzhou, 256600, China

    Xiuyan Yang, Zheng Song & Jing Wang

  4. College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, 453003, China

    Lei Wang & Xiaojing Xia

Authors
  1. Likun Cheng
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  2. Chunguang Zhao
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  3. Xiuyan Yang
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  4. Zheng Song
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  9. Lei Wang
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Contributions

LC, XX and ZS designed the research. LC and XX performed the experiments. LC, CZ, ZS, and CL acquired and analyzed the data. XZ, JW, JW, and LW contributed to writing of the manuscript. Finally, all authors have reviewed and approved the final submitted manuscript.

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Correspondence to Likun Cheng.

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Cheng, L., Zhao, C., Yang, X. et al. Application of a dissolved oxygen control strategy to increase the expression of Streptococcus suis glutamate dehydrogenase in Escherichia coli. World J Microbiol Biotechnol 37, 60 (2021). https://doi.org/10.1007/s11274-021-03025-2

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