The yeast peroxisome: A dynamic storage depot and subcellular factory for squalene overproduction
The yeast peroxisome: A dynamic storage depot and subcellular factory for squalene overproduction
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DOI：
10.1016/j.ymben.2019.11.001
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发表时间：
2020-01-01
影响因子：
8.4
通讯作者：
Wei, Dong-Zhi
中科院分区：
文献类型：
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作者：
Liu, Guo-Song;Li, Tian;Wei, Dong-Zhi
工程微生物从可再生原料中生产萜烯是传统生产方法的一个有前途的替代方案。通常，萜烯不易由微生物细胞分泌，它们在细胞内的分布通常是模糊的，并且由于储存的限制，往往是萜烯过量生产的限制因素。在这里，我们确定在酿酒酵母细胞质中过量产生的角鲨烯以类似油滴的形式分布。有趣的是，这些可疑的油滴被证实是膨胀的过氧化物酶体，它们随着角鲨烯的产生而膨胀，这表明酿酒酵母中的过氧化物酶体是储存角鲨烯的动态仓库。鉴于此，利用过氧化物酶体作为亚细胞区室进行角鲨烯合成，相对于亲本菌株，角鲨烯滴度提高了138倍（1312.82 mg/L），表明葡萄球菌过氧化物酶体是合成萜烯的高效亚细胞工厂。通过细胞质工程和过氧化物酶体工程的双重调控，进一步将角鲨烯滴度提高到1698.02 mg/L。优化两段补料分批发酵法后，角鲨烯滴度达到11.00 g/L，为已有报道的最高滴度。这为角鲨烯在过氧化物酶体中的合成和储存提供了新的见解，并揭示了利用过氧化物酶体通过双细胞质-过氧化物酶体工程过量生产萜烯的潜力。
Engineering microbes to produce terpenes from renewable feedstock is a promising alternative to traditional production approaches. Generally, terpenes are not readily secreted by microbial cells, and their distribution within cells is usually obscure and often a restricting factor for the overproduction of terpenes due to the storage limitation. Here, we determined that squalene overproduced in the cytoplasm of Saccharomyces cerevisiae was distributed in a form similar to oil droplets. Interestingly, these suspected oil droplets were confirmed to be inflated peroxisomes that were swollen along with the production of squalene, indicating that peroxisomes in S. cerevisiae are dynamic depots for the storage of squalene. In view of this, harnessing peroxisomes as subcellular compartments for squalene synthesis was performed, achieving a 138-fold improvement in squalene titer (1312.82 mg/L) relative to the parent strain, suggesting that the peroxisome of S. cerevisiae is an efficient subcellular factory for the synthesis of terpenes. By dual modulation of cytoplasmic and peroxisomal engineering, the squalene titer was further improved to 1698.02 mg/L. After optimizing a two-stage fed-batch fermentation method, the squalene titer reached 11.00 g/L, the highest ever reported. This provides new insight into the synthesis and storage of squalene in peroxisomes and reveals the potential of harnessing peroxisomes to overproduce terpenes in S. cerevisiae through dual cytoplasmic-peroxisomal engineering.