GC-MS Method for the Quantitative Analysis of Limonene from Genetically Engineered Saccharomyces cerevisiae

摘要

In this study, a gas chromatography-mass spectrometry (GC-MS) analysis method was developed and validated for the quantitative analysis of limonene that was attempted to be industrially mass-produced from genetically engineered Saccharomyces cerevisiae (S. cerevisiae). Owing to the high potential economic value of limonene, (e.g., for biofuels, fragrant additives, pharmaceuticals, or organic solvents), it is important to develop and validate a readily available quantitative analysis method for limonene produced from genetically engineered yeast or microorganisms. The engineered S. cerevisiae was cultured using two-phase extractive fermentation with n-dodecane, which is known to alleviate monoterpene toxicity, used as an extractive solvent. The quantitative GC-MS analysis of the n-dodecane overlay that contained limonene produced from the engineered S. cerevisiae was validated to be in accord with the international standards set forth by the Food and Drug Administration.