Subtilisin-Involved Morphology Engineering for Improved Antibiotic Production in Actinomycetes
Subtilisin-Involved Morphology Engineering for Improved Antibiotic Production in Actinomycetes
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DOI：
10.3390/biom10060851
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发表时间：
2020-06-01
期刊：
影响因子：
5.5
通讯作者：
Bai, Linquan
中科院分区：
文献类型：
--
作者：
Wu, Yuanting;Kang, Qianjin;Bai, Linquan
In the submerged cultivation of filamentous microbes, including actinomycetes, complex morphology is one of the critical process features for the production of secondary metabolites. Ansamitocin P-3 (AP-3), an antitumor agent, is a secondary metabolite produced byActinosynnema pretiosumATCC 31280. An excessive mycelial fragmentation ofA. pretiosumATCC 31280 was observed during the early stage of fermentation. Through comparative transcriptomic analysis, a subtilisin-like serine peptidase encoded geneAPASM_4178was identified to be responsible for the mycelial fragmentation. Mutant WYT-5 with theAPASM_4178deletion showed increased biomass and improved AP-3 yield by 43.65%. We also found that the expression ofAPASM_4178is specifically regulated by an AdpA-like protein APASM_1021. Moreover, the mycelial fragmentation was alternatively alleviated by the overexpression of subtilisin inhibitor encoded genes, which also led to a 46.50 +/- 0.79% yield increase of AP-3. Furthermore,APASM_4178was overexpressed in salinomycin-producingStreptomyces albusBK 3-25 and validamycin-producingS. hygroscopicusTL01, which resulted in not only dispersed mycelia in both strains, but also a 33.80% yield improvement of salinomycin to 24.07 g/L and a 14.94% yield improvement of validamycin to 21.46 g/L. In conclusion, our work elucidates the involvement of a novel subtilisin-like serine peptidase in morphological differentiation, and modulation of its expression could be an effective strategy for morphology engineering and antibiotic yield improvement in actinomycetes.
