Abstract：
:Actinomycetes produce a large variety of pharmaceutically active compounds, yet production titers often require to be improved for discovery, development and large-scale manufacturing. Here, we describe a new technique, multiplexed site-specific genome engineering (MSGE) via the 'one integrase-multiple attB sites' concept, for the stable integration of secondary metabolite biosynthetic gene clusters (BGCs). Using MSGE, we achieved five-copy chromosomal integration of the pristinamycin II (PII) BGC in Streptomyces pristinaespiralis, resulting in the highest reported PII titers in flask and batch fermentations (2.2 and 2g/L, respectively). Furthermore, MSGE was successfully extended to develop a panel of powerful Streptomyces coelicolor heterologous hosts, in which up to four copies of the BGCs for chloramphenicol or anti-tumour compound YM-216391 were efficiently integrated in a single step, leading to significantly elevated productivity (2-23 times). Our multiplexed approach holds great potential for robust genome engineering of industrial actinomycetes and novel drug discovery by genome mining.
journal_name
Metab Engjournal_title
Metabolic engineeringauthors
Li L,Zheng G,Chen J,Ge M,Jiang W,Lu Ydoi
10.1016/j.ymben.2017.01.004subject
Has Abstractpub_date
2017-03-01 00:00:00pages
80-92eissn
1096-7176issn
1096-7184pii
S1096-7176(16)30220-8journal_volume
40pub_type
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