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Establishing a Mathematical Equations and Improving the Production of l-tert-Leucine by Uniform Design and Regression Analysis

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Abstract

l-tert-Leucine (l-Tle) and its derivatives are extensively used as crucial building blocks for chiral auxiliaries, pharmaceutically active ingredients, and ligands. Combining with formate dehydrogenase (FDH) for regenerating the expensive coenzyme NADH, leucine dehydrogenase (LeuDH) is continually used for synthesizing l-Tle from α-keto acid. A multilevel factorial experimental design was executed for research of this system. In this work, an efficient optimization method for improving the productivity of l-Tle was developed. And the mathematical model between different fermentation conditions and l-Tle yield was also determined in the form of the equation by using uniform design and regression analysis. The multivariate regression equation was conveniently implemented in water, with a space time yield of 505.9 g L−1 day−1 and an enantiomeric excess value of >99 %. These results demonstrated that this method might become an ideal protocol for industrial production of chiral compounds and unnatural amino acids such as chiral drug intermediates.

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Acknowledgments

This work was supported by the State Key Program of the National Natural Science Foundation of China (No. 21336009), the National Natural Science Foundation of China (Nos. 41176111 and 41306124), the Foundation of South Oceanographic Research Center of China in Xiamen (No. 14GYY011NF11), and the Public Science and Technology Research Funds Projects of Ocean (No. 201505032-6).

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Authors and Affiliations

  1. Department of Chemical and Biochemical Engineering, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, 361005, China

    Wei Jiang, Chao-zhen Xu, Si-zhi Jiang, Tang-duo Zhang, Shi-zhen Wang & Bai-Shan Fang

  2. The Key Lab for Synthetic Biotechnology of Xiamen City, Xiamen University, Xiamen, 361005, China

    Wei Jiang, Chao-zhen Xu & Bai-Shan Fang

  3. The Key Laboratory for Chemical Biology of Fujian Province, Xiamen University, Xiamen, Fujian, 361005, China

    Bai-Shan Fang

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  1. Wei Jiang
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Correspondence to Bai-Shan Fang.

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The authors declare that they have no conflict of interest.

Authors’ Contributions

WJ and BSF designed the study. WJ carried out the experiments and wrote the paper. CZX carried out the optimizing of fermentation conditions. SZJ, TDZ, and SZW carried out the uniform design logic. BSF supervised the work and finalized the manuscript. All authors have read and approved the final manuscript.

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Highlights

• A multilevel factorial experimental design was performed for l-Tle production.

• A mathematical model relating fermentation conditions and conversion rate was established.

• The multivariate regression equation was verified and implemented in l-Tle synthesis.

• The space time yield, 505.9 g L−1 day−1, of the l-Tle was obtained in this work.

• The l-Tle was synthesized in the coenzyme regeneration system with an enantiomeric excess of >99 %.

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Jiang, W., Xu, Cz., Jiang, Sz. et al. Establishing a Mathematical Equations and Improving the Production of l-tert-Leucine by Uniform Design and Regression Analysis. Appl Biochem Biotechnol 181, 1454–1464 (2017). https://doi.org/10.1007/s12010-016-2295-1

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