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Enhancing L-malate production of Aspergillus oryzae by nitrogen regulation strategy

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Abstract

Regulating morphology engineering and fermentation of Aspergillus oryzae makes it possible to increase the titer of L-malate. However, the existing L-malate-producing strain has limited L-malate production capacity and the fermentation process is insufficiently mature, which cannot meet the needs of industrial L-malate production. To further increase the L-malate production capacity of A. oryzae, we screened out a mutant strain (FMME-S-38) that produced 79.8 g/L L-malate in 250-mL shake flasks, using a newly developed screening system based on colony morphology on the plate. We further compared the extracellular nitrogen (N1) and intracellular nitrogen (N2) contents of the control and mutant strain (FMME-S-38) to determine the relationship between the curve of nitrogen content (N1 and N2) and the L-malate titer. This correlation was then used to optimize the conditions for developing a novel nitrogen supply strategy (initial tryptone concentration of 6.5 g/L and feeding with 3 g/L tryptone at 24 h). Fermentation in a 7.5-L fermentor under the optimized conditions further increased the titer and productivity of L-malate to 143.3 g/L and 1.19 g/L/h, respectively, corresponding to 164.9 g/L and 1.14 g/L/h in a 30-L fermentor. This nitrogen regulation-based strategy cannot only enhance industrial-scale L-malate production but also has generalizability and the potential to increase the production of similar metabolites.

Key Points

• Construction of a new screening system based on colony morphology on the plate.

• A novel nitrogen regulation strategy used to regulate the production of L-malate.

• A nitrogen supply strategy used to maximize the production of L-malate.

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Data availability

The data that support the figures within this paper and other findings of this study are available from the corresponding author upon reasonable request.

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Funding

This work was financially supported by the Key Program of the National Natural Science Foundation of China (22038005), the National Key R & D Program of China (2018YFA0901400, 2020YFA0908500), the General Program of National Natural Science Foundation of China (21978113), and the national first-class discipline program of Light Industry Technology and Engineering (LITE2018-08).

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Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China

    Lihao Ji, Qiuling Luo, Qiang Ding, Wenxiu Tang, Xiulai Chen & Liming Liu

  2. International Joint Laboratory on Food Safety, Jiangnan University, Wuxi, 214122, China

    Lihao Ji, Qiuling Luo, Qiang Ding, Wenxiu Tang, Xiulai Chen & Liming Liu

  3. College of Food Engineering, Anhui Science and Technology University, Chuzhou, 233100, Anhui, China

    Ju Wang

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  1. Lihao Ji
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  2. Ju Wang
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  3. Qiuling Luo
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  4. Qiang Ding
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  5. Wenxiu Tang
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  6. Xiulai Chen
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  7. Liming Liu
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Contributions

Ji L and Luo Q conceived and designed research. Ji L and Wang J conducted experiments. Ji L analyzed the data and wrote the manuscript. Wang J, Ding Q, and Tang W contributed with scientific discussions and commented on the manuscript. Chen X and Liu L supervised the work and revised the manuscript. All authors read and approved the manuscript.

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Correspondence to Liming Liu.

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Ji, L., Wang, J., Luo, Q. et al. Enhancing L-malate production of Aspergillus oryzae by nitrogen regulation strategy. Appl Microbiol Biotechnol 105, 3101–3113 (2021). https://doi.org/10.1007/s00253-021-11149-6

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