The heterologous production of terpenes by the thermophile Parageobacillus thermoglucosidasius in a consolidated bioprocess using waste bread.

The heterologous production of terpenes by the thermophile Parageobacillus thermoglucosidasius in a consolidated bioprocess using waste bread.
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嗜热副杆菌热葡糖苷酶在使用废面包的综合生物过程中异源生产萜烯。

DOI:
10.1016/j.ymben.2020.11.005
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发表时间:
2021
影响因子:
8.4
通讯作者:
Styles MQ
Styles MQ
中科院分区:
工程技术1区
文献类型:
--
作者:
Styles MQ

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热糖sidasiparageobacillus thermoglucosidasiusa是一种基因上易受控制的嗜热菌,在高温下生长迅速,在65℃时翻倍时间与大肠杆菌的最短翻倍时间相当。它能够使用各种各样的底物,包括碳水化合物低聚物,并且已经开发用于乙醇的工业生产。在这项研究中,P。thermoglucosidasiusNCIMB11955通过引入异源甲基戊酸途径产生倍半萜τ-muurolol,该途径是由几种嗜热古菌的基因和最近表征的耐热性τ-muurolol合成酶构建的。热葡萄糖苷自然使用甲基赤藓糖醇磷酸途径生产萜烯前体,焦磷酸异戊烯基,而古细菌使用甲戊酸途径的一种版本。通过引入正交古菌途径,可以增加倍半萜生物合成的通量。构建如此大的代谢途径存在遗传载体导入和稳定性方面的问题,因此通过偶联方式引入重组质粒,并开发了一种耐热丝氨酸整合酶系统,用于将大途径整合到染色体上。最后,通过使异源途径麦芽糖诱导,我们证明了新菌株能够在一个巩固的生物过程中直接使用废面包作为自诱导碳源来生产萜烯。
Parageobacillus thermoglucosidasiusis a genetically tractable thermophile that grows rapidly at elevated temperatures, with a doubling time at 65 °C comparable to the shortest doubling times ofEscherichia coli. It is capable of using a wide variety of substrates, including carbohydrate oligomers, and has been developed for the industrial production of ethanol. In this study,P. thermoglucosidasiusNCIMB11955 has been engineered to produce the sesquiterpene τ-muurolol by introduction of a heterologous mevalonate pathway constructed using genes from several thermophilic archaea together with a recently characterised thermostable τ-muurolol synthase.P. thermoglucosidasiusnaturally uses the methylerythritol phosphate pathway for production of the terpene precursor, isopentenyl pyrophosphate, while archaea use a version of the mevalonate pathway. By introducing the orthogonal archaeal pathway it was possible to increase the flux through to sesquiterpene biosynthesis. Construction of such a large metabolic pathway created problems with genetic vector introduction and stability, so recombinant plasmids were introduced by conjugation, and a thermostable serine integrase system was developed for integration of large pathways onto the chromosome. Finally, by making the heterologous pathway maltose-inducible we demonstrate that the new strain is capable of using waste bread directly as an autoinduction carbon source for the production of terpenes in a consolidated bioprocess.
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