Abstract
In this research, single-factor and response surface experiments were conducted in the fed-batch fermentation process to improve the yield of iturin A. The effect of adding various concentrations of precursor amino acids l-asparagine (Asn), l-aspartic acid (Asp), l-glutamic acid (Glu), l-glutamine (Gln), l-Serine (Ser) and l-proline (Pro) at different adding times (3 and 12 h) on iturin A production and cell growth was studied. The respective addition of amino acids (Asp 0.28 g/L; Asn 0.36 g/L; Glu 0.20, 0.28 and 0.360 g/L; Gln 0.20, 0.28 and 0.36 g/L; Pro 0.12, 0.20, 0.28 and 0.36 g/L) at 3 h was shown to improve cell growth but did not affect the yield of iturin A. Meanwhile, the individual addition of the same amino acids at 12 h improved cell growth and increased the yield of iturin A. Excellent correlation was obtained between the predicted and measured values, suggesting that the regression model was accurate and reliable; highly significant (P < 0.0001), and the determination coefficient (R2 = 0.975). When 0.0752 g/L Asn; 0.1992 g/L Gln and 0.1464 g/L Pro were added at 12 h, the yield of iturin A reached 0.85 g/L, which is 32.81%-fold higher than that of the initial process. Therefore, this study obtained optimal parameters for iturin A production by the experimental method, and process validation gave high iturin A yields (0.85 g/L) during a 60 h fermentation. These findings could guide an up-scaling of the fermentation process.
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source was soybean protein (total nitrogen concentration 4.0 g/L). The fermentation conditions were as follows: initial pH 8.0, temperature 30 °C, rotating speed 150 rpm, fermentation time 60 h. The samples were taken at 0, 3, 6, 9, 12, 15, 18, 21, 24, 28, 32, 36, 42, 48, 54 and 60 h, and stored at −20 °C
source was soybean protein (total nitrogen concentration 4.0 g/L). The fermentation conditions were as follows: initial pH 8.0, temperature 30℃, rotating speed 150 rpm, fermentation time 60 h. The samples were taken at 0, 3, 6, 9, 12, 15, 18, 21, 24, 28, 32, 36, 42, 48, 54 and 60 h, and stored at −20 °C
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Acknowledgements
The authors thank the Chinese Academy of Sciences (CAS) Light of West China Program (Grant No. 2017XBZG_XBQNXZ_B_006), the Key Laboratory of Environmental and Applied Microbiology of the Chengdu Institute of Biology CAS (grant No. KLCAS-2018-3) and Innovation Academy for Seed Design CAS for financial support of this work.
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HY carried out the experiments. HY and JZ established the feeding method. HY, JY, DL and JZ participated in the Separation and extraction of iturin A. HY, ZL, DS and HW participated in the statistical analysis. HT conceived of the study, and participated in the design and coordination and helped to draft the manuscript. All authors read and approved the final manuscript.
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Yue, H., Zhong, J., Li, Z. et al. Optimization of iturin A production from Bacillus subtilis ZK-H2 in submerge fermentation by response surface methodology. 3 Biotech 11, 36 (2021). https://doi.org/10.1007/s13205-020-02540-7
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DOI: https://doi.org/10.1007/s13205-020-02540-7