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Combining Genome Shuffling with Streptomycin Resistance to Improve Poly-γ-L-diaminobutanoic Acid Production in Bacillus pumilus

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Abstract

A breeding approach combining genome shuffling with streptomycin resistance was developed in this research to improve the poly-γ-L-diaminobutanoic acid (γ-PAB) production in Bacillus pumilus LS-1. By this unique strategy, recombinants from the third round of genome shuffling could tolerate 18 µg/L of streptomycin and exhibited higher γ-PAB yield as 152.2 mg/L in shake-flask fermentation, 3-fold over the parent. In batch fermentation, B. pumilus GS3-9 could produce γ-PAB as high as 1284.7 mg/L in two days, 2.4-fold higher than the control, which was the highest productivity ever reported. In addition, the optimal pH in B. pumilus for γ-PAB synthesis was changed after atmospheric and room temperature plasma (ARTP) mutagenesis and protoplast fusion, because lower pH environment is favorable for accumulation of intracellular ATP. Some key enzymes in GS3-9 showed higher activities than those in parent, suggesting a greater flux to TCA circle and DAP pathway, which was a reason for enhanced γ-PAB production.

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Acknowledgments

This work was financially supported by Shandong Natural Science Foundation (ZR2019BC044), and open project from the Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University (KLIB-KF202007).

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Authors and Affiliations

  1. Marine College, Shandong University, Weihai, 264209, China

    Shu Li & Wei Cong

  2. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China

    Liang Wang

  3. Weihai Food and Drug Administration, Testing Center, Weihai, 264210, China

    Nan Wang

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  1. Shu Li
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  4. Wei Cong
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Correspondence to Shu Li.

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Li, S., Wang, L., Wang, N. et al. Combining Genome Shuffling with Streptomycin Resistance to Improve Poly-γ-L-diaminobutanoic Acid Production in Bacillus pumilus. Biotechnol Bioproc E 26, 630–640 (2021). https://doi.org/10.1007/s12257-020-0320-2

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