Discovery of McbB, an Enzyme Catalyzing the β‐Carboline Skeleton Construction in the Marinacarboline Biosynthetic Pathway

Chen, Qi; Ji, Changtao; Song, Yongxiang; Huang, Hongbo; Ma, Junying; Tian, Xinpeng; Ju, Jianhua

doi:10.1002/ange.201303449

Cited by 24 publications

(16 citation statements)

References 27 publications

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“…Recent studies have revealed that the microbial bC alkaloids are produced by totally different enzyme families from those of plants (Aroonsri et al, 2012;Chen et al, 2013). Among them, the recently reported McbB from Marinactinospora thermotolerans is a novel enzyme in marinacarboline biosynthesis.…”

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confidence: 99%

“…Among them, the recently reported McbB from Marinactinospora thermotolerans is a novel enzyme in marinacarboline biosynthesis. Based on heterologous gene-expression experiments, McbB was proposed to be a multifunctional enzyme that catalyzes the PS reaction of L-tryptophan and oxaloacetaldehyde, as well as decarboxylation and oxidation reactions, to produce 1-acetyl-3-carboxy-b-carboline (1) and 1-acetyl-b-carboline (2) ( Figure 1) (Chen et al, 2013). However, in some cases the decarboxylation and oxidation of tetrahydro-bC are catalyzed by other proteins such as oxygenase and aromatase (Santos et al, 1998), or even non-enzymatically (Nemet and Varga-Defterdarovic, 2008), to generate partially or fully aromatized bC scaffold.…”

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confidence: 99%

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Structural Basis for β-Carboline Alkaloid Production by the Microbial Homodimeric Enzyme McbB

Mori

Hoshino

Sahashi

et al. 2015

Chemistry & Biology

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The β-carboline (βC) alkaloids occur throughout nature and exhibit diverse biological activities. In contrast to βC alkaloid synthesis in plants, the biosynthesis in microorganisms remains poorly understood. The recently reported McbB from Marinactinospora thermotolerans is a novel enzyme proposed to catalyze the Pictet-Spengler (PS) reaction of L-tryptophan and oxaloacetaldehyde to produce the βC scaffold of marinacarbolines. In this study, we solved the crystal structure of McbB complexed with L-tryptophan at 2.48 Å resolution, which revealed the novel protein folding of McbB and the totally different structure from those of other PS condensation catalyzing enzymes, such as strictosidine synthase and norcoclaurine synthase from plants. Structural analysis and site-directed mutagenesis confirmed that the previously proposed catalytic Glu97 at the active-site center functions as an acid and base catalyst. Remarkably, the structure-based mutants R72A and H87A, with expanded active-site cavities, newly accepted bulky phenylglyoxal as the aldehyde substrate, to produce 1-benzoyl-3-carboxy-β-carboline.

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confidence: 99%

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confidence: 99%

Structural Basis for β-Carboline Alkaloid Production by the Microbial Homodimeric Enzyme McbB

Mori

Hoshino

Sahashi

et al. 2015

Chemistry & Biology

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“…2A and SI Appendix, Table S1). We also inactivated mtdB (encoding a peptidase S15), mtdC (encoding an unknown protein), mtdD (encoding an unknown protein), the orf1 (encoding an inositol-phosphate phosphatase), and orf2 genes using our established λ-RED-mediated PCR-targeting mutagenesis strategy in M. thermotolerans SCSIO 00652 by replacing them with an apramycin resistance gene cassette (33,40). Genotypes of the mutants were confirmed by PCR (SI Appendix).…”

Section: Resultsmentioning

confidence: 99%

Deciphering the sugar biosynthetic pathway and tailoring steps of nucleoside antibiotic A201A unveils a GDP- l -galactose mutase

Zhu

Chen

Song

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Galactose, a monosaccharide capable of assuming two possible configurational isomers (D-/L-), can exist as a six-membered ring, galactopyranose (Galp), or as a five-membered ring, galactofuranose (Galf). UDP-galactopyranose mutase (UGM) mediates the conversion of pyranose to furanose thereby providing a precursor for D-Galf. Moreover, UGM is critical to the virulence of numerous eukaryotic and prokaryotic human pathogens and thus represents an excellent antimicrobial drug target. However, the biosynthetic mechanism and relevant enzymes that drive L-Galf production have not yet been characterized. Herein we report that efforts to decipher the sugar biosynthetic pathway and tailoring steps en route to nucleoside antibiotic A201A led to the discovery of a GDP-L-galactose mutase, MtdL. Systematic inactivation of 18 of the 33 biosynthetic genes in the A201A cluster and elucidation of 10 congeners, coupled with feeding and in vitro biochemical experiments, enabled us to: (i) decipher the unique enzyme, GDP-Lgalactose mutase associated with production of two unique D-mannose-derived sugars, and (ii) assign two glycosyltransferases, four methyltransferases, and one desaturase that regiospecifically tailor the A201A scaffold and display relaxed substrate specificities. Taken together, these data provide important insight into the origin of L-Galf-containing natural product biosynthetic pathways with likely ramifications in other organisms and possible antimicrobial drug targeting strategies.nucleoside antibiotic A201A | biosynthesis | GDP-L-galactose mutase | methyltransferase | desaturase

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“…The type strain was isolated from marine sediment collected from the northern South China Sea (Tian et al 2009). During the last 5 years, Prof. Ju's group have discovered many bioactive metabolites from this taxon (Huang et al 2011;Ma et al 2012;Zhu et al 2012;Zhou et al 2012;Chen et al 2013). The current study reports the isolation of a new species of this genus obtained from a specimen of Salsola ferganica Drob, a traditional Chinese medicinal plant.…”

Section: Introductionmentioning

confidence: 96%

Marinactinospora endophytica sp. nov., isolated from a medicinal plant

Liu

Khieu

Gao

et al. 2015

Antonie van Leeuwenhoek

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A novel endophytic actinobacterium, designated strain YIM 690053(T), was isolated from healthy stems of Salsola ferganica Drob, and was characterized using a polyphasic taxonomic approach. The strain formed well-branched substrate mycelia with no fragmentation, and abundant aerial mycelia that differentiated into long spores. The strain was found to grow at 10-55 °C, pH 6.0-10.0 and in the presence of 0-7 % (w/v) NaCl. The genomic DNA G+C content was determined to be 70.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 690053(T) belongs to the genus Marinactinospora, and it shared the highest 16S rRNA gene sequence similarity with Marinactinospora thermotolerans SCSIO 00652(T) (98.96 %). However, the DNA-DNA relatedness values between the two strains was 37.85 ± 2.77 %. Based on the differences in the molecular and biochemical characteristics from its closest relative, strain YIM 690053(T) is concluded to represent a novel species of the genus Marinactinospora, for which the name Marinactinospora endophytica sp. nov. is proposed. The type strain is YIM 690053(T)(=KCTC 29664(T) = DSM 46799(T)).

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Discovery of McbB, an Enzyme Catalyzing the β‐Carboline Skeleton Construction in the Marinacarboline Biosynthetic Pathway

Cited by 24 publications

References 27 publications

Structural Basis for β-Carboline Alkaloid Production by the Microbial Homodimeric Enzyme McbB

Structural Basis for β-Carboline Alkaloid Production by the Microbial Homodimeric Enzyme McbB

Deciphering the sugar biosynthetic pathway and tailoring steps of nucleoside antibiotic A201A unveils a GDP- l -galactose mutase

Marinactinospora endophytica sp. nov., isolated from a medicinal plant

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