Flavoenzyme CrmK-mediated substrate recycling in caerulomycin biosynthesis

Zhu, Yiguang; Picard, Marie-Ève; Zhang, Qingbo; Barma, Julie; Després, Xavier Murphy; Mei, Xiangui; Zhang, Liping; Duvignaud, Jean-Baptiste; Couture, Manon; Zhu, Weiming; Shi, Rong

doi:10.1039/c6sc00771f

Cited by 17 publications

(12 citation statements)

References 51 publications

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“…2). The cam contains 20 ORFs including essential dipyridine ring biosynthetic genes and backbone post-modification genes [22,25,[27][28][29] (Fig. 2 and Table S7), which indicates that Actinoalloteichus sp.…”

Section: Genomic Analysis Of the Crm A Biosynthetic Potential In Actimentioning

confidence: 99%

“…Then, the 2, 2'-dipyridine ring is finally converted into the CRM A by a series of post-modification reactions, including aminohydrolysis, oxime formation and methylation [22,[26][27][28][29]. CRM A can induce the generation of regulatory T cells to avoid T cell responses and change the function of B cells, which significantly inhibits the Mixed Lymphocyte Reaction (MLR) [30,31].…”

Section: Introductionmentioning

confidence: 99%

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Activation and enhancement of Caerulomycin A biosynthesis in marine-derived Actinoalloteichus sp. AHMU CJ021 by combinatorial genome mining strategies

Xie

Chen

Wang

et al. 2020

Preprint

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Backgrounds: 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 bioproduction of CRM A, an immunosuppressive drug lead with diverse bioactivities.Results: To well activate the expression of cam, ribosomal engineering was adopted to treat the wild type Actinoalloteichus sp. AHMU CJ021. The initial mutant strain XC-11G with gentamycin resistance and CRM A bioproduction 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 bioproduction was then improved by two strain breeding methods via UV mutagenesis and cofactor engineering-directed increasing of intracellular riboflavin, which finally generated the optimal mutant strain XC-11GUR with a CRM A bioproduction 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 14.7 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 have constructed an ideal CRM A producer. More importantly, our efforts also have demonstrated the effectiveness of abovementioned combinatorial strategies, which is applicable to the genome mining of bioactive natural products from abundant actinomycetes strains.

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Section: Genomic Analysis Of the Crm A Biosynthetic Potential In Actimentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Activation and enhancement of Caerulomycin A biosynthesis in marine-derived Actinoalloteichus sp. AHMU CJ021 by combinatorial genome mining strategies

Xie

Chen

Wang

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…2). The cam contains 20 ORFs including essential dipyridine ring biosynthetic genes and backbone post-modi cation genes [23,26,[28][29][30] (Fig. 2 and Table S4), which indicates that Actinoalloteichus sp.…”

Section: Genomic Analysis Of the Crm A Biosynthetic Potential In Actimentioning

confidence: 99%

“…This new metabolic engineering strategy named as cofactor engineering has been well applied to microbial second metabolites development [39][40][41]. Previous study had revealed that the biosynthesis of CRM A required several essential avoenzymes, including CamD, which completed the formation of the picolinic acid precursor; CamH, which catalyzed the formation of the oxime group, and CamK, which maintained the substrate recycling process [23,28,29]. Thus, enhancing the intracellular ribo avin supplement could facilitate the formation of these avoenzymes, which may accelerate CRM A production in turn.…”

Section: Enhancement Of Crm a Production By Uv Mutagenesismentioning

confidence: 99%

“…The biosynthesis of this marine drug lead is initiated by the formation of a core 2, 2'-dipyridine skeleton, which is catalyzed by a unique hybrid polyketide-nonribosomal peptide (PKS-NRPS) assembly line [22][23][24][25][26]. Then, the 2, 2'-dipyridine skeleton is nally converted into the CRM A by a series of post-modi cation reactions, including aminohydrolysis, oxime formation and methylation [23,[27][28][29][30]. CRM A can induce the generation of regulatory T cells to avoid T cell responses and change the function of B cells, which signi cantly inhibits the Mixed Lymphocyte Reaction (MLR) [31,32].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Activation and enhancement of Caerulomycin A biosynthesis in marine-derived Actinoalloteichus sp. AHMU CJ021 by combinatorial genome mining strategies

Xie

Chen

Wang

et al. 2020

Preprint

View full text Add to dashboard Cite

Backgrounds: 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 have constructed an ideal CRM A producer. More importantly, our efforts also have demonstrated the effectiveness of abovementioned combinatorial strategies, which is applicable to the genome mining of bioactive natural products from abundant actinomycetes strains.

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Catalysis of Extracellular Deamination by a FAD‐Linked Oxidoreductase after Prodrug Maturation in the Biosynthesis of Saframycin A

Tang

Song²,

Zhang³

et al. 2017

Angewandte Chemie

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The biosynthesis of antibiotics in bacteria is usually believed to be an intracellular process,atthe end of which the matured compounds are exported outside the cells.T he biosynthesis of saframycin A( SFM-A), an antitumor antibiotic,r equires ac ryptic fatty acyl chain to guide the construction of ap entacyclic tetrahydroisoquinoline scaffold; however,t he follow-up deacylation and deamination steps remain unknown. Herein we demonstrate that SfmE, am embrane-bound peptidase,h ydrolyzes the fatty acyl chain to release the amino group;and SfmCy2, asecreted oxidoreductase covalently associated with FAD, subsequently performs an oxidative deamination extracellularly.T hese results not only fill in the missing steps of SFM-A biosynthesis,but also reveal that aF AD-binding oxidoreductase catalyzesa nu nexpected deamination reaction through an unconventional extracellular pathway in Streptmyces bacteria.

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Flavoenzyme CrmK-mediated substrate recycling in caerulomycin biosynthesis

Abstract: Biochemical and structural investigations into the flavoenzyme CrmK reveal a substrate recycling/salvaging mechanism in caerulomycin biosynthesis.

Cited by 17 publications

References 51 publications

Activation and enhancement of Caerulomycin A biosynthesis in marine-derived Actinoalloteichus sp. AHMU CJ021 by combinatorial genome mining strategies

Activation and enhancement of Caerulomycin A biosynthesis in marine-derived Actinoalloteichus sp. AHMU CJ021 by combinatorial genome mining strategies

Activation and enhancement of Caerulomycin A biosynthesis in marine-derived Actinoalloteichus sp. AHMU CJ021 by combinatorial genome mining strategies

Catalysis of Extracellular Deamination by a FAD‐Linked Oxidoreductase after Prodrug Maturation in the Biosynthesis of Saframycin A

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