Activation and enhancement of caerulomycin A biosynthesis in marine-derived Actinoalloteichus sp. AHMU CJ021 by combinat
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Microbial Cell Factories Open Access
RESEARCH
Activation and enhancement of caerulomycin A biosynthesis in marine‑derived Actinoalloteichus sp. AHMU CJ021 by combinatorial genome mining strategies Yunchang Xie1, Jiawen Chen1, Bo Wang3,4, Tai Chen3,4, Junyu Chen1, Yuan Zhang2, Xiaoying Liu2* and Qi Chen2*
Abstract Background: Activation of silent biosynthetic gene clusters (BGCs) in marine-derived actinomycete strains is a feasible strategy to discover bioactive natural products. Actinoalloteichus sp. AHMU CJ021, isolated from the seashore, was shown to contain an intact but silent caerulomycin A (CRM A) BGC-cam in its genome. Thus, a genome mining work was preformed to activate the strain’s production of CRM A, an immunosuppressive drug lead with diverse bioactivities. Results: To well activate the expression of cam, ribosome engineering was adopted to treat the wild type Actinoalloteichus sp. AHMU CJ021. The initial mutant strain XC-11G with gentamycin resistance and CRM A production titer of 42.51 ± 4.22 mg/L was selected from all generated mutant strains by gene expression comparison of the essential biosynthetic gene-camE. The titer of CRM A production was then improved by two strain breeding methods via UV mutagenesis and cofactor engineering-directed increase of intracellular riboflavin, which finally generated the optimal mutant strain XC-11GUR with a CRM A production titer of 113.91 ± 7.58 mg/L. Subsequently, this titer of strain XC-11GUR was improved to 618.61 ± 16.29 mg/L through medium optimization together with further adjustment derived from response surface methodology. In terms of this 14.6 folds increase in the titer of CRM A compared to the initial value, strain XC-GUR could be a well alternative strain for CRM A development. Conclusions: Our results had constructed an ideal CRM A producer. More importantly, our efforts also had demonstrated the effectiveness of abovementioned combinatorial strategies, which is applicable to the genome mining of bioactive natural products from abundant actinomycetes strains. Keywords: Caerulomycin A, Genome mining, Combinatorial strategies, Ribosome engineering, Strain improvement, Marine-derived Actinoalloteichus
*Correspondence: [email protected]; [email protected] 2 School of Life Sciences, Anhui Medical University, Hefei 230032, China Full list of author information is available at the end of the article
Background Actinomycetes, the acknowledged producers of bioactive secondary metabolites, are historically important bioresources in the discovery and development of pharmaceutical molecules [1–5]. In particular, marine actinomycetes
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