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Efficient brazzein production in yeast (Kluyveromyces lactis) using a chemically defined medium

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Abstract

The sweet-tasting protein brazzein offers considerable potential as a functional sweetener with antioxidant, anti-inflammatory, and anti-allergic properties. Here, we optimized a chemically defined medium to produce secretory recombinant brazzein in Kluyveromyces lactis, with applications in mass production. Compositions of defined media were investigated for two phases of fermentation: the first phase for cell growth, and the second for maximum brazzein secretory production. Secretory brazzein expressed in the optimized defined medium exhibited higher purity than in the complex medium; purification was by ultrafiltration using a molecular weight cutoff, yielding approximately 107 mg L−1. Moreover, the total media cost in this defined medium system was approximately 11% of that in the optimized complex medium to generate equal amounts of brazzein. Therefore, the K. lactis expression system is useful for mass-producing recombinant brazzein with high purity and yield at low production cost and indicates a promising potential for applications in the food industry.

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Abbreviations

BCA:

Bicinchoninic acid

BSA:

Bovine serum albumin

DMSO:

Dimethyl sulfoxide

GRAS:

Generally recognized as safe

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

SIAM:

Single-interval adjustment matrix

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Acknowledgements

This research was supported by a National Research Foundation of Korea Grant funded by the Korean Government (no. 2018R1D1A1B07043467) and the Chung-Ang University Graduate Research Scholarship Grants in 2018.

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Author notes

  1. Se-Woong Park and Byung-Ha Kang contributed equally to this work.

Authors and Affiliations

  1. Department of Chemistry, College of Natural Sciences, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul, 06974, Republic of Korea

    Se-Woong Park, Byung-Ha Kang, Hyeong-Min Lee, Sung-Jun Lee, Han-Seul Kim, Hye-Won Choi, Tae Jung Park & Kwang-Hoon Kong

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  1. Se-Woong Park
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Park, SW., Kang, BH., Lee, HM. et al. Efficient brazzein production in yeast (Kluyveromyces lactis) using a chemically defined medium. Bioprocess Biosyst Eng 44, 913–925 (2021). https://doi.org/10.1007/s00449-020-02499-y

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