Facile Production of (+)-Aristolochene and (+)-Bicyclogermacrene in Escherichia coli Using Newly Discovered Sesquiterpene Synthases from Penicillium expansum,Journal of Agricultural and Food Chemistry

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Facile Production of (+)-Aristolochene and (+)-Bicyclogermacrene in Escherichia coli Using Newly Discovered Sesquiterpene Synthases from Penicillium expansum
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2022-05-04 , DOI: 10.1021/acs.jafc.2c01885
Zheng-Yu Huang ₁ , Qing-Yang Wu ₁ , Chun-Xiu Li ₁ , Hui-Lei Yu ₁ , Jian-He Xu ₁

Affiliation

Penicillium expansum, producer of a wide array of secondary metabolites, has the potential to be a source of new terpene synthases. In this work, a platform was constructed with Escherichia coli BL21(DE3) by enhancing its endogenous 2-methyl-d-erythritol-4-phosphate pathway to supply sufficient terpenoid precursors. Using this precursor-supplying platform, we discovered two sesquiterpene synthases from P. expansum: PeTS1, a new (+)-aristolochene synthase, and PeTS4, the first microbial (+)-bicyclogermacrene synthase. To enhance the sesquiterpene production by PeTS1, we employed a MBP fusion tag to improve the heterologous protein expression, resulting in the increase of aristolochene production up to 50 mg/L in a 72 h flask culture, which is the highest production reported to date. We also realized the first biosynthesis of (+)-bicyclogermacrene, achieving 188 mg/L in 72 h. This work highlights the great potential of this microbial platform for the discovery of new terpene synthases and opens new ways for the bioproduction of other valuable terpenoids.

中文翻译：

使用新发现的来自扩展青霉的倍半萜合酶在大肠杆菌中轻松生产 (+)-Aristolochene 和 (+)-Bicyclogermacrene

Penicillium expansum是多种次级代谢物的生产者，有可能成为新的萜烯合酶的来源。在这项工作中，通过增强其内源性 2-甲基-d-赤藓糖醇-4-磷酸途径以提供足够的萜类前体，构建了一个与大肠杆菌 BL21(DE3)的平台。使用这个前体供应平台，我们从P. expansum中发现了两种倍半萜合酶：Pe TS1，一种新的 (+)-马兜铃烯合酶，和Pe TS4，第一个微生物 (+)-双环锗烯合酶。提高Pe的倍半萜生产TS1，我们采用 MBP 融合标签来改善异源蛋白表达，导致在 72 小时烧瓶培养中马兜铃烯的产量增加高达 50 mg/L，这是迄今为止报道的最高产量。我们还实现了 (+)-双环锗烯的首次生物合成，在 72 小时内达到 188 mg/L。这项工作突出了该微生物平台在发现新萜合酶方面的巨大潜力，并为生物生产其他有价值的萜类化合物开辟了新途径。

更新日期：2022-05-04

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