Department of Chemistry, College of Natural Sciences, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul, 06974, Republic of Korea.
Bioprocess Biosyst Eng. 2021 Apr;44(4):913-925. doi: 10.1007/s00449-020-02499-y. Epub 2021 Jan 27.
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. 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.
甜蛋白 Brazzein 具有抗氧化、抗炎和抗过敏等特性,有望成为一种有功能的甜味剂。在这里,我们优化了一种化学成分确定的培养基,以在乳酸克鲁维酵母中生产分泌型重组 Brazzein,可应用于大规模生产。我们研究了确定培养基的组成,以适应发酵的两个阶段:第一阶段用于细胞生长,第二阶段用于 Brazzein 的最大分泌生产。在优化的确定培养基中表达的分泌型 Brazzein 比在复杂培养基中表现出更高的纯度;通过超滤使用分子量截止值进行纯化,可得到约 107mg/L 的产量。此外,在该确定培养基系统中,总培养基成本约为优化复杂培养基的 11%,可产生等量的 Brazzein。因此,该 K. lactis 表达系统可用于大规模生产高纯度和高产量的重组 Brazzein,生产成本低,在食品工业中有广阔的应用前景。