Improvement of Aspergillus nidulans penicillin production by targeting AcvA to peroxisomes
Improvement of Aspergillus nidulans penicillin production by targeting AcvA to peroxisomes
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DOI：
10.1016/j.ymben.2014.07.002
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发表时间：
2014-09-01
影响因子：
8.4
通讯作者：
Fischer, Reinhard
中科院分区：
文献类型：
--
作者：
Herr, Andreas;Fischer, Reinhard
构巢曲霉能够合成青霉素，并可作为研究其生物合成调控的模型。形成β内酰胺环三肽只需要三种酶，该三肽由t-半胱氨酸、t-缬氨酸和t-氨基己二酸组成。而两种酶。 AcvA 和 lpnA 定位于细胞质，AatA 位于过氧化物酶体中。在这里，我们测试了一种提高青霉素产量的新策略，即改变参与生物合成的酶的驻留。我们测试了将 AcvA 或 lpnA（或两者）靶向过氧化物酶体是否会增加青霉素产量。事实上，AcvA 过氧化物酶体靶向导致了 3.2 倍的增加。相比之下，将 IpnA 靶向过氧化物酶体会导致青霉素生产完全丧失。与野生型相比，acvA、ipnA 或 aatA 的过表达分别导致青霉素多 1.4、2.8 和 3.1 倍。同时过度表达所有三种酶甚至会导致青霉素增加 6 倍。 acvA 过氧化物酶体靶向和该基因的过度表达相结合，导致青霉素滴度增加了 5 倍。最后，通过pexIC A菌株的过表达增加了过氧化物酶体的数量，过氧化物酶体数量增加一倍，产生的青霉素量增加了2.3倍。这些结果表明，青霉素的产生可以在多个调节水平上触发，其中之一是酶的亚细胞定位。 (C) 2014 年国际代谢工程学会。由爱思唯尔公司出版。保留所有权利
Aspergillus niduluns is able to synthesize penicillin and serves as a model to study the regulation of its biosynthesis. Only three enzymes are required to form the beta lactam ring tripeptide, which is comprised of t-cysteine, t-valine and t-aminoadipic acid. Whereas two enzymes. AcvA and lpnA localize to the cytoplasm, AatA resides in peroxisomes. Here, we tested a novel strategy to improve penicillin production, namely the change of the residence of the enzymes involved in the biosynthesis. We tested if targeting of AcvA or lpnA (or both) to peroxisomes would increase the penicillin yield. Indeed, AcvA peroxisomal targeting led to a 3.2-fold increase. In contrast, targeting IpnA to peroxisomes caused a complete loss of penicillin production. Overexpression of acvA, ipnA or aatA resulted in 1.4, 2.8 and 3.1-fold more penicillin, respectively in comparison to wildtype. Simultaneous overexpression of all three enzymes resulted even in 6-fold more penicillin. Combination of acvA peroxisomal targeting and overexpression of the gene led to 5-fold increase of the penicillin titer. At last, the number of peroxisomes was increased through overexpression of pexIC A strain with the double number of peroxisomes produced 2.3 times more penicillin. These results show that penicillin production can be triggered at several levels of regulation, one of which is the subcellular localization of the enzymes. (C) 2014 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved