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Production of Tagatose by Whole-cell Bioconversion from Fructose Using Corynebacterium glutamicum

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Abstracts

Tagatose is a rarely occurring sugar found in food and sweet fruits. It is a potential sweetener with low calories but a sweetness similar to that of sucrose. In this study, we developed a whole-cell biocatalyst using Corynebacterium glutamicum for direct bioconversion of fructose to tagatose. First, we constructed a biological conversion pathway for the conversion of fructose to tagatose by expressing tagatose 4-epimerase (TN) from Thermotoga neapolitana in C. glutamicum. Next, to increase the expression level of the enzyme, we engineered copy number of plasmid. Plasmid library was constructed by randomizing the cgrI antisense RNA region in the plasmid, and the plasmid with high-copy number was isolated using fluorescence-activated single cell sorting (FACS)-based high-throughput screening. The isolated plasmid, pHCP7, had 2-fold higher copy numbers than the original plasmid. A higher expression level and conversion yield could be achieved using pHCP7. Finally, we examined a novel tagatose 4-epimerase TN(KNF4E) isolated from Kosmotoga olearia. The gene expression level was further increased (33.9% of total fraction) through codon optimization and expression in pHCP7, and a conversion yield as high as 21.7% was achieved.

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Acknowledgements

This work was supported by research grants from CJ CheilJedang Institute of Biotechnology, and by the National Research Foundation of Korea (NRF) grant funded by the Ministry of Science and ICT (MSIT) [No. NRF-2020R1A2C2012537], Republic of Korea.

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering (BK Plus Program), Korea Advanced Institute of Science and Technology (KAIST), Daejeon, 34141, Korea

    Eun Jung Jeon & Ki Jun Jeong

  2. Synthetic Biology Research Center, Synthetic Biology and Bioengineering Research Institute, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon, 34141, Korea

    Eun Jung Jeon

  3. CJ Cheiljedang Food Research Institute, Suwon, 16495, Korea

    Young-Mi Lee, Eun Jung Choi & Seong-Bo Kim

  4. Global Leaders College, Yonsei University, Seoul, 03722, Korea

    Seong-Bo Kim

  5. Institute for the BioCentury, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, 34141, Korea

    Ki Jun Jeong

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Contributions

EJJ designed the study, performed the experiments, analyzed the data and drafted the manuscript. EJC, YML, SBK participated in the execution of the experiments and data interpretation. KJJ initiated and supervised the study, and revised manuscript. All authors read and approved the final manuscript.

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Correspondence to Ki Jun Jeong.

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EJJ, SBK, and KJJ declare no direct and indirect conflict of interest that relate to this manuscript. YMJ and EJC are full-time employees of CJ Cheiljedang, however the funding sponsors had no role in the performance of the experiments and in the collection, analysis and interpretation of the data. Neither ethical approval required nor informed consent was required for this study.

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Jeon, E.J., Lee, YM., Choi, E.J. et al. Production of Tagatose by Whole-cell Bioconversion from Fructose Using Corynebacterium glutamicum. Biotechnol Bioproc E 28, 419–427 (2023). https://doi.org/10.1007/s12257-022-0304-5

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