Data_Sheet_1_One-Step Purification of Microbially Produced Hydrophobic Terpenes via Process Chromatography.xlsx

Novel and existing terpenes are already being produced by genetically modified microorganisms, leading to new process challenges for the isolation and purification of these terpenes. Here, eight different chromatographic resins were characterized for the packed bed adsorption of the model terpene β-caryophyllene, showing their applicability on an Escherichia coli fermentation mixture. The polystyrenic Rensa® RP (Ø 50 μm) shows the highest affinity, with a maximum capacity of >100 g L−1 and the best efficiency, with a height equivalent of a theoretical plate (HETP) of 0.022 cm. With this material, an optimized adsorption-based purification of β-caryophyllene from a fermentation mixture was developed, with the green solvent ethanol for desorption. A final yield of >80% and a purity of >99% were reached after only one process step with a total productivity of 0.83 g h−1 L−1. The product solution was loaded with a volume ratio (feed to column) of >500 and the adapted gradient elution yielded a 40 times higher concentration of β-caryophyllene. The adsorption-based chromatography represents therefore a serious alternative to the liquid-liquid extraction and achieves desired purities without the utilization of hazardous solvents.

目前已有研究通过基因工程微生物生产新型及现有萜类(terpenes)，这为该类萜类的分离纯化工艺带来了全新挑战。本研究中，针对模型萜类β-石竹烯(β-caryophyllene)的填充床吸附(packed bed adsorption)过程，我们对八种不同色谱树脂(chromatographic resins)开展了表征，证实其可适用于大肠杆菌(Escherichia coli)发酵混合体系。其中，聚苯乙烯基Rensa® RP色谱树脂（粒径Ø50μm）展现出最高的吸附亲和力，最大吸附容量超过100 g·L⁻¹，且吸附效率最优，理论塔板当量高度(HETP)仅为0.022 cm。基于该树脂材料，本研究开发了从发酵混合液中纯化β-石竹烯的优化吸附工艺，以绿色溶剂乙醇作为解吸剂。仅通过单步工艺即可实现超过80%的最终收率与超过99%的产品纯度，总生产能力达0.83 g·h⁻¹·L⁻¹。该工艺可实现进料液与柱体积比超过500的上样量，经适配优化的梯度洗脱可使β-石竹烯的浓度提升40倍。综上，基于吸附的色谱法可作为液液萃取(liquid-liquid extraction)的可靠替代方案，无需使用有害溶剂即可达到预期的产品纯度。