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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
Optimization of iturin A production from Bacillus subtilis ZK-H2 in submerge fermentation by response surface
Hua Yue1,2, Juan Zhong1,2, Zhemin Li1,2
1CAS Key Laboratory of Environmental and Applied Microbiology, Environmental Microbiology Key Laboratory of Sichuan Province Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu, 610041 People's Republic of China.
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.
Electronic Supplementary Material:
The online version of this article (10.1007/s13205-020-02540-7) contains supplementary material, which is available to authorized users.
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