β-榄香烯是抗肿瘤药物榄香烯的主要活性成分,来源于温郁金成熟块茎的倍半萜,广泛应用于临床抗肿瘤辅助治疗。然而,从植物中提取β-榄香烯的方法有很多弊端,如产量低、纯化困难和环境污染等问题。该研究通过在解脂耶氏酵母中表达莴苣来源的吉玛烯A合成酶,重组菌株β-榄香烯产量为23.19 mg/L。采用模块化工程分步表达参与β-榄香烯合成的基因ERG10、ERG13、HMGR、ERG12、ERG8、IDI、ERG19、ERG20-LTC2,以研究其对β-榄香烯产量的影响。通过增加前体模块,β-榄香烯产量提高至(881.33±19.27)mg/L。进一步增加ERG20-LTC2基因拷贝数并优化培养基β-榄香烯的产量达到(1 668.13±41.01)mg/L,是初始菌株产量的71倍。该研究为β-榄香烯的工业化生产提供了一定的参考,也为解脂耶氏酵母萜类天然产物的生物合成奠定了基础。
β-elemene, the primary active ingredient of the antitumor drug elemene, is a type of sesquiterpene extracted from the mature tubers of Curcuma wenyujin.It has been widely used in clinical settings as an adjuvant therapy for treating tumors.However, the extraction method of β-elemene from plants is plagued by a low yield, difficulties in purification, and potential environmental pollution issues.This study constructed a recombinant strain in Yarrowia lipolytica.By expressing germacrene A synthase from Lactuca sativa, an initial β-elemene yield of 23.19 mg/L was obtained.Modular engineering was utilized to sequentially express genes associated with β-elemene synthesis, including ERG10, ERG13, HMGR, ERG12, ERG8, IDI, ERG19, and ERG20-LTC2, with an (881.33±19.27) mg/L production.Furthermore, by increasing the copy number of ERG20-LTC2 and medium optimization, the β-elemene production reached (1 668.13±41.01) mg/L, which was 71-fold higher than that of the initial strain.This study provides practical guidance for the industrial production of β-elemene and establishes a solid foundation for the biosynthesis of terpene natural products in Y.lipolytica.
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