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Published on: September 20, 2016
Engineering Yarrowia lipolytica for Enhanced Gastrodin Production via High-Throughput Screening and
Mengchen Hu1, Yijin He1, Yifan Ma1
1School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, China.
Abstract:
Gastrodin, the principal bioactive component of the renowned herb Gastrodia elata, is extensively utilized in medicinal drugs and nutraceuticals. This study seeks to enhance microbial production of gastrodin through high-throughput screening (HTS) and transcriptomics-guided optimization. Initially, atmospheric pressure and room temperature plasma (ARTP)-mediated mutagenesis were employed to develop a library of mutant strains. Furthermore, a transcription factor-based biosensor and a high-throughput solid-phase extraction mass spectrometry (HP-SPE-MS) were evaluated to establish an HTS method for gastrodin. Consequently, mutant strain MT8 was isolated, producing 9.8 g/L of gastrodin in YPD medium, which represents a 55.6% increase compared to the control strain. Next, key genes identified via transcriptomics were overexpressed in strain MT8, with the overexpression of gene YALI2E01737g, a gene involved in the synthesis of aromatic amino acids, significantly enhancing gastrodin production to reach 10.1 g/L. In addition, fermentation process optimization further improved gastrodin titer up to 13.1 g/L in shaking flasks. This study demonstrated the utility of HTS techniques to enhance gastrodin production and paved the way for its future industrial application.
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