Identification of the Grincamycin Gene Cluster Unveils Divergent Roles for GcnQ in Different Hosts, Tailoring the <scp>l</scp>-Rhodinose Moiety

Zhang, Yun; Huang, Hongbo; Chen, Qi; Luo, Min; Sun, Aijun; Song, Yongxiang; Ma, Junying; Ju, Jianhua

doi:10.1021/ol401253p

Cited by 49 publications

(85 citation statements)

References 18 publications

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“…The cosmid pCSG2404 (Table S2†), obtained in our previous work in screening the xiamycin gene cluster, 19 was found to contain the intact PTM gene cluster. However, introduction of the cosmid pCSG2801 (Table S2†), a derivative of pCSG2404 in which the neo gene (encoding kanamycin resistance) was replaced by a gene cassette containing aac(3)IV (encoding apramycin resistance), oriT (for conjugation), and Int φ C31 (for specific integration into Streptomyces species) from pSET152A′B 25 (Table S2†) into Streptomyces lividans TK64, resulted in no observed production of PTM-like compounds (Fig. 2B, trace ii), similar to the control strain S. lividans TK64/pSET152 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Activation and characterization of a cryptic gene cluster reveals a cyclization cascade for polycyclic tetramate macrolactams

Saha

Zhang

et al. 2017

Chem. Sci.

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Section: Resultsmentioning

confidence: 99%

Activation and characterization of a cryptic gene cluster reveals a cyclization cascade for polycyclic tetramate macrolactams

Saha

Zhang

et al. 2017

Chem. Sci.

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“…The putative cluster also encodes a dedicated phosphopantetheinyl transferase (SqnCC), eight deoxysugar biosynthesis enzymes (SqnS1–SqnS8), three GTs (SqnG1–G3) and a sugar-modifying dehydrogenase (SqnQ) homologous to AknOx and GcnQ (Table S1). 14,16 To probe the role of the identified cluster in the biosynthesis of saquayamycins 1 – 7 , the sqnCC gene encoding a phosphopantetheinyl transferase (PPTase) was replaced with an apramycin resistance marker through homologous recombination (Figure S1). The resultant mutant, ΔCC does not produce saquayamycins either on solid media (no brown color characteristic of WT was observed, Figure S1) or liquid fermentation (Figures S2).…”

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confidence: 99%

Two Cooperative Glycosyltransferases Are Responsible for the Sugar Diversity of Saquayamycins Isolated from Streptomyces sp. KY 40-1

2017

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Glycosyltransferases are key enzymes involved in the biosynthesis of valuable natural products providing an excellent drug-tailoring tool. Herein, we report the identification of two cooperative glycosyltransferases from the sqn gene cluster directing the biosynthesis of saquayamycins in Streptomyces sp. KY40-1: SqnG1 and SqnG2. Gene inactivation of sqnG1 leads to 50 folds decrease in saquayamycin production, while inactivation of sqnG2 leads to complete production loss suggesting that SqnG2 acts as dual O- and C-glycosyltransferase. Gene inactivation of a third putative glycosyltransferase-encoding gene, sqnG3, does not majorly affect saquayamycin production suggesting that SqnG3 has no or supportive role in glycosylation. The data indicate that SqnG1 and SqnG2 are solely and possibly cooperatively responsible for the sugar diversity observed in saquayamycins 1–7. This is the first evidence of a proposed glycosyltransferase complex with dual O- and C-glycosyltransferase activity, utilizing NDP-activated d-olivose, l-rhodinose as well as an unusual amino sugar, presumably 3,6-dideoxy-l-idosamine.

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“…We subsequently selected cosmid 142H harboring the genes from mtdC to orf3, integrated elements for conjugation from the pSET152AB vector (41), and transferred them into S. lividans TK64 through conjugation. The recombinant S. lividans TK64/ 142H was fermented using the same medium as had been the case for the wild-type producer.…”

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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Identification of the Grincamycin Gene Cluster Unveils Divergent Roles for GcnQ in Different Hosts, Tailoring the l-Rhodinose Moiety

Cited by 49 publications

References 18 publications

Activation and characterization of a cryptic gene cluster reveals a cyclization cascade for polycyclic tetramate macrolactams

Activation and characterization of a cryptic gene cluster reveals a cyclization cascade for polycyclic tetramate macrolactams

Two Cooperative Glycosyltransferases Are Responsible for the Sugar Diversity of Saquayamycins Isolated from Streptomyces sp. KY 40-1

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

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